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生物英语短篇作文高中作文

生物英语短篇作文高中作文Biological Diversity。

Biological diversity, or biodiversity, refers to the variety of life on earth, including the variety of species, ecosystems, and genetic diversity within species. It is essential for the functioning of ecosystems and the services they provide, such as food, clean water, and air, and for human well-being.Biodiversity is threatened by human activities such as habitat destruction, pollution, overexploitation of resources, and climate change. These activities have led to the extinction of many species and the degradation of ecosystems, which in turn threatens human well-being.To protect biodiversity, we need to take actions at different levels. At the individual level, we can reduce our ecological footprint by consuming less, using renewable energy, and supporting conservation efforts. At thecommunity level, we can promote sustainable practices, such as organic farming and eco-tourism, and protect natural habitats. At the national and international level, we need policies and regulations that promote sustainable development, protect endangered species and habitats, and address the root causes of biodiversity loss.Conserving biodiversity is not only a moral and ethical duty, but also a practical necessity for our survival and well-being. We depend on biodiversity for our food, medicines, and other resources, and for the services that ecosystems provide. By protecting biodiversity, we are also protecting our own future.In conclusion, biodiversity is a crucial aspect of our planet's health and well-being. It is threatened by human activities, but we can take action to protect it. By conserving biodiversity, we are not only protecting the natural world, but also our own future.。

生物给人类的科技贡献英文作文

生物给人类的科技贡献英文作文English:Biology has made significant contributions to technology in numerous ways. One of the most important ways is through the field of medicine, where biological discoveries have led to the development of vaccines, antibiotics, and various medical technologies that have saved countless lives. Biotechnology, a branch of biology, has also revolutionized many industries with advancements in genetic engineering, pharmaceuticals, agriculture, and environmental protection. The study of genetics has paved the way for personalized medicine and gene therapy, providing solutions to genetic disorders and improving the overall quality of healthcare. Furthermore, biomimicry, which involves learning from nature to design innovative technologies, has led to the creation of new materials, structures, and systems that are more efficient, sustainable, and resilient. Overall, biology continues to play a crucial role in driving technological advancements that benefit society as a whole.中文翻译:生物学在许多方面为技术的发展做出了重要贡献。

生物医学工程医疗仪器论文中英文资料外文翻译文献

生物医学工程医疗仪器论文中英文资料外文翻译文献Present status and problems of domestic medical instrument engineering. Biomedical EngineehngIn recent years considerable progress has been achieved in domestic medical instrument engineering. Many plants and scientific-research organizations of machine-making and the defense industry have changed their profile toward production of medical equipment.However, medical equipment often meets a reluctant market because of funding cuts in health services. Medical organizations often cannot satisfy even their basic requirements for diagnostic and therapeutic devices.Also, health service organizations tend to buy foreign rather than domestic medical equipment because the former are easily available on the domestic market and prices for both are comparable because of inflation.The transition to a market economy in Russia has had substantial impact on the relations between domestic manufactur- ers and consumers of medical devices. The spectrum and quality of available items has been significantly extended in recentyears. It should be also noted that available models of medical devices are continuously updated, which makes them sufficiently competitive.Representative information on the updating dynamics of domestic medical equipment is summarized in Table 1. The data were provided by the VNIIMP-VITA Joint-Stock Company, which compiles a data bank of such information.Generally, new items account for 37% of total production of medical devices. Routinely produced devices (duration of production, 2-5 years) account for 28%. Medical devices of long-term production (5-10 years) account for 17% and obsolete nomenclature (devices produced for more than 10 years) accounts for 18%.It is seen from Table 1 that in recent years there has been considerable progress in the updating of production of medical equipment. For example, according to the VNIIMP-VITA Joint-Stock Company, the share of the items that have been produced for no longer than 5 years in 1988 did not exceed 35%, while now it is 65%. For the items that have been produced for more than 10 years such shares are 40 and 18%, respectively.Updating of produced medical devices was encouraged by the increase in the number of designers and manufacturers, particularly those of former defense industry facilities. In collaboration with foreign partners they set up joint ventures for producing medical equipment.Analysis of the updating of the various groups of medical equipment is of substantialinterest.It is seen from Table 1 that detoxication devices contribute dominantly to the group of items that have been updated within the standard period of up to 5 years (100% of production, including modern devices for hemodialysis and hemosorption).Comparatively high updating indices are observed for devices for functional diagnosis: 72% of these devices have been produced for no longer than 5 years, and obsolete devices account for only 9% of total production. However, it should be noted that although production of some obsolete devices has been terminated, equipment of similar functional capacity is still urgently needed.Relatively low updating indices are observed among the devices for intensive care and resuscitation: 16% of new items and comparatively many obsolete devices (26%). Among new models apparatuses for artificial lung ventilation are worth mention. However, some apparatuses, which have been developed long ago are still on the market because they have good performance, are quite reliable, and still are in demand. This reduces the updating index of the group as a whole.All-Russian Scientific-Research Institute for Medical Instrument Engineering, Rusaian Academy of Medical Sciences (VNIIMP-VITA Joint-Stock Company), Moscow. Translated from Meditsinskaya Tekhnika, No. 1, pp. 4-9, January-February, 1996. Original article submitted August 23, 1995.0006-3398/96/3001-0001515.00 y Plenum Publishing CorporationTABLE 1. Updating of Basic Groups of Medical Devices and Apparatuses (% of total nomenclature)这里有个表The lowest updating indices are observed for devices for examining a patient's body structures. These are: ophthalmological, otolaryngological, and anthropometric devices, endoscopes, etc. The share of obsolete devices is high (44%), while the devices which have been produced for no more than 5 years account for only 20% of total production.It should be noted that these results on medical equipment updating are important general estimates, although they do not take into consideration specific achievements and shortcomings in the production of individual items. Therefore, some corresponding amendments are required.Our survey of available information, including the VNIIMP-VITA Joint-Stock Company data bank, materials presented at various exhibitions, and recent literature, shows that domestic medical industry has developed a number of original medical devices and apparatuses which were designed to replace similar obsolete models. However, many types of important and necessary medical devices still do not meet contemporary requirements, and some types of devices are not produced at all.For example, in recent years production of some sophisticated medical devices (apparatuses for intensive care, resuscitation, and anesthesiology; devices for artificial lung ventilation, respiratory narcosis devices, extracorporeal circulation) significantly rose, particularly at the former defense industry facilities, and their quality has been significantly improved. The functional performance of the devices is generally on par with foreign analogs.Perfusion units have also been improved and their production has expanded. This allowed the demand of the health service organizations for such equipment to be satisfied completely. Modern domestic hemodialysis devices (Renart-10, Renan- 10RT, etc.) have beendeveloped and brought into wide clinical practice.The development and production of diagnostic magnetic resonance imaging systems (Obraz-3, TOROS) are considerable breakthroughs in domestic medical industry. This substantially extends diagnostic capacities of many health service organizations and provides them with topical diagnosis previously unavailable domestically, although it is quite common in developed foreign countries.Domestic medical industry has begun production of pulse oximeters; these are of particular use in surgery and resuscita-tion. This bridged a substantial gap in the spectrum of available domestic medical devices.The Bilitest bilirubin meter, which has been recently developed and produced in Russia, fully meets the requirements of maternity and children's hospitals in devices for diagnosing jaundice.A high-standard radioimmunochemical laboratory was opened at the VNIIMP-VITA Joint-Stock Company to supply customers with necessary radioimmunochemical assay kits.A number of high-quality medical devices and instruments have been developed at the electronic industry plants andinstitutes. The following devices are particularly worthy of mention:- artificial cardiac valves of the Emitron Plant, which are on par with the best foreign analogs;- pH meters (Istok State Scientific-Manufacturing Association);- Ikar long-term (up to 24 h) cardiomonitors with electronic memory (Kometa Central Scientific-Manufacturing Association);- radiothermographs and racliothermoscopes for detecting deeply located thermal fields in the human body (Oktyabr' Manufacturing Association and Design Bureau for Ecological and Medical Equipment);- original thermal imaging system (Institute of Radioelectronics and Automatics, Russian Academy of Sciences; OPTROS, Ltd.);- original computer-assisted system Cardiac Rhythms for monitoring oatient condition and pulsimetry (Institute of Chemical Physics, Russian Academy of Sciences; Ekos, Ltd.);- video system for endoscopic imaging (Zenit Scientific-Manufacturing A~sociation; Elektron Scientific-Research and Manufacturing Association);- streamlined technology for producing disposable and reusable syringes, injection needles, and surgical threads.A number of other problems of domestic medical instrument-making industry have been successfully solved in recent years.For example, the number and quality of therapeutic devices, particularly for laser therapy, is quite sufficient. Research studies are carried out by many organizations including former defense industry facilities. Technologies which have been developed for other purposes give fruitful results in medical industry.According to our data, more than 150 models of such medical devices have been developed over the last 5 years. Some 100 of them are commercially available. Although domestic medical devices are often superior ot foreign analogs in terms of working performance and they are definitely less expensive, many of them are not in short demand and are virtually not used.However, this activity in many other areas of medical instrument engineering cannot beconsidered as sufficiently successful and rational.It should be noted that many newly developed models of domestic medical devices compare unfavorably with foreign analogs. This is particularly the case for X-ray and ultrasonic devices, electrocardiographic monitors, laboratory equipment, etc. Nevertheless, according to the VNIIMP-VITA Joint-Stock Company databank, certain positive trends have been observed in recent years even in these areas. However, most problems still remain unsettled and the conditions required to solve them have not yet been established.It is important to note that the serially produced X-ray apparatus RUM-20 (Mosrentgen Joint-Stock Company) has been significantly updated. The updated model RUM-20M-SG312 is commercially available in combination with the Sapfir domestic image intensifier or an image intensifier of a French manufacturer. The Kruiz fiat image intensifier has been developed at theAll-Russian Scientific-Research Institute for Medical Instrument Engineering in collaboration with MELZ Manufacturing Association and Mosrentgen Joint-Stock Company. This device is designed to replace existing fluorescent screens in the X-ray diagnostic apparatuses RUM-10, RUM-20, RUM-20M, and others. The use of the Kruiz image intensifier significantly increases image information content and allows threefold decrease in the radiation load on patients and medical personnel.The G 202-5 system for lit-par-lit raster imaging of patients in lying position has been developed at the Mosrentgen Joint-Stock Company. This device is commercially available with the PURS power source. It allows both manual and automatic X-ray photography and organ-oriented X-ray examination.The RTS-61 mobile X-ray video diagnostic apparatus has been developed at the Elektron Scientific-Research and Manufacturing Association. This device is designed to be used in surgery, orthopedics, and traumatology.Among the defense industry facilities which have reoriented their production to medical market the Scientific-Research Institute for Electromechanics (Istra) is worth mention. In collaboration with Phillips (Germany) and borrowing their technology and circuitry, the Institute for Electromechanics developed the Mammodiagnost mammographic scanner, which meets international standards of operating performance.The Rentgen-48 X-ray tomographic diagnostic systems with a rotary support table and the Rentgen-60 X-ray diagnostic systems with a remote control support table have been developed at the Sevkavrentgen Plant and received positive recognition by practicing physicians.The models of X-ray diagnostic devices listed above are examples of achievements of domestic medical industry.However, many important and significant problems of the development of domestic medical X-ray equipment remain unsettled, and it is unreasonable to expect that they will be solved in the foreseeable future unless special measures are taken.For example, the most common RUM-20 X-ray apparatuses with the Sapfir image intensifier are equipped with the obsolete X-ray image converter REP-1. To replace the REP-1 image converter, the Moscow Plant for Electronic Tubes has developed the Buer image converter of improved design. This device offers better image contrast, reduced clark background noise, and has an output fiberoptic window of improved design. However, the Buer image converter is not yet commercially available.Digital X-ray diagnostic devices are not yet commercially available from domesticmanufacturers either.The Design Bureau for Medical Engineering in collaboration with Medtekh, Ltd. (Novosibirsk) have developed the Diaskan X-ray digital scanner. Serial production of this device is in progress at the Design Bureau for Medical Engineering.However, devices of sufficient quality are not yet commercially available.Domestic medical industry does not produce X-ray tomographs. Their production in Chelyabinsk has been suspended.Electrocardiographic monitors are very important devices for functional diagnosis. However, domestic medical industry fails substantially behind leading foreign manufacturers and there is a disproportion in the development and production of necessary devices and apparatuses. Many automatic systems for ECG processing, including syndromal diagnosis, have been developed, but they trove not been tested and are of little demand. However, simple three-channel electrocardiographs of mass- scale application are not produced by domestic manufacturers.Foreign manufacturers offer various ultrasonic scanners and sophisticated imaging systems. Domestic manufacturers produce only simple devices with manual sector-by-sector scanning and a few simplified models with linear electronic scanning.Some positive results have been achieved in the development of endoscopic devices. These achievements are mainly due to the collaboration between LOMO and some companies from Japan. However, even these devices require further improvement of quality and reliability.Although the level of production of domestic laboratory equipment has noticeably risen in recent years, it is still too little to meet the demand. The number of organizations involved in the development of such equipment has risen. However, the available devices are simple and have limited functional capacity. Many important devices (e.g., automatic analyzers and simple routine devices) are not produced at all.Devices for blood transfusion and preparing blood substitute solutions are still in short supply (40 million items have been produced, while the demand is 200 million). The demand in dialyzers and polymer infusion systems reaches 100 and 150 million items, respectively, although such systems are not produced at all.The correspondence between production and demand, quality and technical performance, and adequate testing of medical production are put in the forefront under conditions of a market economy. The problem of competition with foreign manufacturers is also quite important because of increasing import of medical equipment and reduced sale of the production of domestic manufacturers. In this connection, the following circumstances should be taken into consideration.There is a considerable disproportion between production and demand of some groups of medical devices. For example, there is :~ huge surplus of laser therapeutic devices and their excessive development. Systems for syndromal electrocardiographic diagnosis, magnetotherapy, and electrostimulation are also in excessive supply. However, simple electrocardiographs, routine laboratory equipment, and some other ordinary but necessary devices of mass-scale application are not produced by domestic manufacturers. These disadvantages cause significant economic losses and present difficulties in the development of health service. Domestic and foreign experience show that these problems can be solved by adequate marketing, but this is in its infancy in the domestic medical industry.It should be noted that foreign companies place special emphasis on marketing and market research. They evaluate actual and pending demand as well as consumer requirements. Thefeedback between consumer and manufacturer gives valuable information on the improvement of the product quality and working performance. The marketing service in most leading companies is of paramount importance. The development of a new product often starts from marketing survey rather than from engineering or design research. Many domestic organizations of medical instrument engineering require cardinal measures for increasing the level of marketing.Testing of medical devices also requires substantial improvement. Considerable experience of foreign manufacturers of medical equipment should be taken into account. It should be noted, however, that this experience is often neglected by domestic manufacturers. Technical testing of medical equipment in foreign companies is usually carried out by independent laboratories which assess performance and quality. The specialists of the laboratories may also give recommendations for further improvement of the tested equipment. The basic goal of the testing is to check if the performance of the device matches its specifications and to conclude if the device can be used in medical organizations. However, the specialists of the laboratories usually go beyond this goal and issue comparative reviews of products of different companies. Such reviews contain the following information:- description of tested device, its specifications, and price;- results of technical testing, correspondence between specifications and actual performance, advantages and disadvan- tages, recommendations for improvement (if necessary);- comparative analysis of similar devices and apparatuses produced by different manufacturers. Such analysis is usually concluded by a most preferable model, which is recommended to medical organizations on the basis of functional capacity, reliability, and economic reasons.In the USA, activity of testing laboratories is controlled by governmental, nongovernmental, and independent nonprofit organizations.In Russia, the problem of balance between the demand in medical devices, their production by domestic manufacturers, and import is of considerable importance.The opinion of the Head of the Department of Medical Industry, Russian Ministry of Health and Medical Industry, Yu. F. Doshchitsin, which was published in the weekly "Meditsinskii Biznes" (No. 9, 1995), is that the requirements of Russian medical market must be met by domestic devices, including products of high technology. Russian medicine should not rely on imported devices alone. We certainly agree with this opinion.The total volume of medical equipment purchased from abroad is presently several times greater than purchases from domestic manufacturers. This situation is definitely unacceptable. Cardinal measures are required to boost and stimulate economically domestic manufacturers of medical equipment. This is particularly important for manufacturers of life support systems and devices for military medicine.However, positive aspects of contacts with foreign manufacturers of medical equipment should not be disregarded. International cooperation is very common in foreign practice, but it is clearly insufficient in Russia.International cooperation in medical industry is particularly vital in such areas as computer technology, microprocessors, and electronic engineering. Lack of sufficiently high-quality domestic computers and microprocessors presents considerable problems in the development of sophisticated medical devices and apparatuses.In recent years a number of domestic organizations established joint ventures withleading foreign manufacturers of medical devices. These joint ventures produce high-technology devices on the basis of imported circuitry, modules, and individual finished units. For example, VNIIMP-VITA produces ultrasonic doppler scanners, Kursk Manufacturing Association Pribor in collaboration with Frezenius (Germany) produces mobile apparatuses for hemodialysis and hemosorption, LOMO and some companies from Japan established a joint venture for manufacturing flexible endoscopes of improved design, Moscow Manufac- turing Association EMA produces ultrasonic diagnostic devices, etc.It seems reasonable to continue and extend mutually profitable contacts between domestic and foreign manufacturers of medical equipment.Active participation and patronage of the Russian Ministry of Health and Medical Industry as well as the Russian Government and local authorities are needed to solve the problems of medical industry listed above and to implement programs of development and production of high-quality domestic medical devices.References[1] V. A. Viktorov，V. P. gundarov，A. P. yurkevich. Present status and problems of domestic medical instrument engineering. Biomedical Engineehng~ V oL 30, No. 1, 1996.[2]All-Russian Scientific-Research Institute for Medical Instrument Engineering, Rusaian Academy of Medical Sciences (VNIIMP-VITA Joint-Stock Company), Moscow. Translated from Meditsinskaya Tekhnika, No. 1, pp. 4-9, January-February, 1996. Original article submitted August 23, 1995.国内医学仪器工程的现状和存在的问题近年来，国内在工程医疗器械实现取得了很大进展。

食品微生物检验英文参考文献

食品微生物检验英文参考文献1．贾英民主编，食品微生物学.高等职业教育教材，北京:中国轻工业出版社，2004.91. jiayingmin, food microbiology Textbooks for higher vocational education, Beijing: China Light Industry Press, September 20042．万萍主编，食品微生物基础与实验技术.高职高专食品类教材系列，北京:科学出版社，2. edited by Wan Ping, fundamentals and experimental techniques of food microbiology Food textbook series for higher vocational colleges, Beijing: Science Press,2004two thousand and four3.翁连海主编﹐食品微生物基础高职教材，北京:高等教育出版社，2005.43. Weng Lianhai, editor in chief, higher vocational textbook of food microbiology, Beijing: Higher Education Press, April 20054．苏世彦．食品微生物检验手册．北京:中国轻工业出版社，1998.104. sushiyan. Manual for microbiological examination of food. Beijing: China Light Industry Press, 1998.105．牛天贵．食品微生物学实验技术.北京:中国农业大学出版社，2002.85. Niu Tiangui. Experimental technology of food microbiology Beijing: China Agricultural University Press, August 20026．高鼎主编，食品微生物学，北京:中国商业出版社，1996.56. Gao Ding, chief editor, food microbiology, Beijing: China business press, 1996.5 7．谢梅英主编，食品微生物学，北京:中国轻工业出版社，2004.57. Xie Meiying, food microbiology, Beijing: China Light Industry Press, May 20048.中华人民共和国国家标准.食品卫生检验方法(微生物部分).北京:中国标准出版社，8. national standards of the people's Republic of China Methods for hygienic inspection of food (microbiological part) Beijing: China Standards Press,2003two thousand and three9.杨洁彬．食品微生物学.北京:北京农业大学出版社，19959. yangjiebin. Food microbiology Beijing: Beijing Agricultural University Press, 199510.沈萍主编﹒微生物学北京:高等教育出版社，200011.高东主编﹒微生物遗传学济南:山东大学出版社，199610. microbiology, edited by Shenping, Beijing: Higher Education Press, 200011 Gao Dong. Microbial genetics. Jinan: Shandong University Press, 1996。

生物陶瓷英文文献

Fabrication and Mechanical Properties of Dense/Porous β-TricalciumPhosphate BioceramicsFaming Zhang1, a , Jiang Chang 1, b*, Jianxi Lu 1, 2, c , Kaili Lin 1, d 1 Biomaterials and Tissue Engineering Research Center, Shanghai Institute of Ceramics, ChineseAcademy of Sciences, Shanghai 200050, China 2 Shanghai Bio-lu Biomaterials Company, Shanghai 200335, China a star.zhang@, b* jchang@,c ir2bberck@,d lklsic@Keywords: Bioceramics, calcium phosphate, bone regeneration, weight bearing sitesAbstract: Attempt t o increase the mechanical properties of porous bioceramics, a dense/porous structured β-TCP bioceramics that mimic the characteristics of nature bone were fabricated. Experimental results show that the dense/porous structured β-TCP bioceramics demonstrated excellent mechanical properties with compressive strength up to 74 MPa and elastic modulus up to 960 MPa, which could be tailored by the dense/porous cross-sectional area ratio obeying the rule of exponential growth. The interface between the dense and porous bioceramics is connected compactly and tightly with some micropores distributed in the matrix of both porous and dense counterparts. The dense/porous structure of β-TCP bioceramics may provide an effective way to increase the mechanical properties of porous bioceramics for bone regeneration at weight bearing sites.IntroductionVarious methods for bone defect treatments have been developed using biological or synthetic grafts. The synthetic alternatives are promising grafts for their unlimited availability and without risk of disease transmission [1]. Calcium phosphate bioceramics, especially hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP), have been extensively explored as bone grafts due to their compositions are similar to the inorganic components of nature bone [2]. The β-TCP bioceramics is well known as a biodegradable material demonstrated clinical efficacy. The porous β-TCP bioceramics is a structurally biomimetic of the cancellous bone, whose porous network could allow tissue to ingrowth exhibiting nicer osteoconductive properties. However, the porous β-TCP shows weak mechanical properties, which limit its application as bone grafts. The macrostructure feature of nature bone is porous cancellous bone inside with dense compact bone surrounding outside, which provides excellent biomechanical properties. Carotenuto et al [3] have prepared dense/porous layered HA bioceramic for orthopedic device coating by tape casting technique, whereas the bulk dense/porous bioceramics were rarely reported. Therefore in present study, a dense/porous structured β-TCP bioceramics that mimics the characteristics of nature bone were fabricated, and the microstructure and mechanical properties of such bioceramics were studied.ExperimentalThe β-TCP powders were synthesized by chemical precipitation reaction. The dense/porous structured β-TCP bioceramics were prepared by injected molding and subsequently pressureless sintering. The shrinkage rate of both porous and dense parts during sintering process was measured at different temperatures. X-ray diffraction (XRD) with Cu K α radiation was used to characterize the phase composition of the ceramics. The microstructures observation of the bioceramic samples was performed on a scanning electron microscopy (SEM).The compressive strength was conducted with a mechanical tester at 0.5 mm/min crosshead speed. The elastic modules were reanalyzed from the slope of the compressive strength-strain curve.All rights reserved. No part of contents of this paper may be reproduced or transmitted in any form or by any means without the written permission of the publisher: Trans Tech Publications Ltd, Switzerland, . (ID: 159.226.129.129-19/09/06,02:35:46)Results and DiscussionThe major problem in preparation of the dense/porous bioceramics is the interface adhesion between the dense and porous parts because of their different shrinkage rate during sintering process. The shrinkage rate of dense and porous bioceramics at different temperatures was measured and the results are shown in Fig.1. It can be noticed that the porous β-TCP bioceramics exhibit much higher shrinkage rate than the dense counterpart. The porous bioceramics shows about 23% shrinkage in radial direction; in contrast, the dense bioceramics presents about 17% shrinkage. It can be calculated that from 850 o C to 1100 o C, the porous β-TCP bioceramics shows about 17% shrinkage rate and almost the same with that of the dense counterpart from 600 o C to 1100 o C. So as to avoiding the shrinkage differences, the porous β-TCP bioceramics were pre-sintered at 850 o C, then the dense bioceramics were injected surrounding the porous ceramics, finally the composites were pressureless sintered at 1100 o C for 5 hours and the dense/porous structured β-TCP bioceramics were obtained.Fig.1 The radial shrinkage rate of the porous and dense β-TCP bioceramicsThe phase composition of the as prepared bioceramics was analyzed by X-ray diffraction. The XRD results show that the high temperature sintered β-TCP preserved their original β phase without transform into their α-TCP phase, as shown in Fig.2. Because the α-TCP though bioactive, have proven less useful as bone regeneration materials due to their excessively high resorption rate than the β-TCP phase. And none of the other impurity phases can be detected in the XRD patterns; resultantly, high purity β-TCP bioceramics were prepared.Fig.2 X-ray diffraction pattern of the prepared bioceramics.Fig.3 shows the optical and SEM micrographs of the prepared dense/porous β-TCP bioceramics samples. It is clear to see that the inner porous structure mimics the cancellous bone to some extent, and outer side dense structure mimics the compact bone, as shown in Fig.3(a) and indicated by theS h i n k a g e (%)Temperature (o C)1020304050607080100200300400500600 2theta (deg.)I n t e n s i t y (c p s )arrows. Fig.3 (b) shows the interface of the dense/porous β-TCP bioceramic, it can be found that the interface between the dense and porous bioceramics is connected compactly and tightly. In the porous part, the macropore size is about 500 μm in diameter; the diameter of the interconnected pores is about 100 μm. Additionally, the porosity of the porous parts is about 72%, and the interconnectivity is more than 95%. The microstructure of the macroporous wall was shown in Fig.3(c); it is obvious that there are some micropores with diameter of 1 μm distributed uniformly in the porous wall. As the results, the microstructure of porous part of the bioceramics is a combination of macroporous and microporous. Contrastively, the microstructure of the dense bioceramics shows refined particle size and few micropores, as exhibited in Fig.3(d). The dense compact part is much denser than the porous cancellous part.Fig.3 The dense/porous β-TCP bioceramic sample (a), the microstructure of dense/porous interface(b), the macroporous wall (c) and dense compact bone (d).The variation of the compressive strength and Elastic modulus of the bioceramics with different dense/porous cross-sectional area ratio (S dense /S porous ) was illustrated in Fig 4. It is exhibited that the compressive strength increases from 10 MPa to 74 MPa with the dense/porous ratio from 0.1 to 4.7 obeying rule of exponential growth. And the elastic modulus has been increased form 180 MPa to 960 MPa with the dense/porous ratio increment, also following exponential growth. Evidently, the value of the porous bioceramics is only about 2.0 MPa and the elastic modulus is about 20 MPa, indicated by the square in Fig.4. It has been achieved about 5 to 37 times increment in the mechanical properties by the dense/porous structure design. The mechanical properties of the dense/porous bioceramics could be tailored by the dense/porous cross-sectional area ratio.Porous materials always have poor mechanical properties. Applications of calcium phosphates in the body have been limited by their low strength and numerous techniques have been investigated in attempts to retain their useful bioactive properties whilst providing more suitable mechanical properties for particular applications. These include the reinforcement of β-TCP using HA fiber orbioglass additives [4, 5]; however these techniques are limited for the porous calcium phosphate Compact bone Cancellousbone (b)(c) (d)using in the load bearing sites’ bone regeneration. In this study, excellent mechanical properties of the porous β-TCP bioceramics have been achieved by the dense/porous structured design. The compressive strength of human femoral cancellous bone, weight bearing sites, is in the range of 25~90 MPa, so the dense/porous structured β-TCP is comparable to the strength of human femoral cancellous bone. The high interconnective porous structure of the dense/porous β-TCP bioceramics could allow the tissue ingrowths, and the dense structure could bear the load to some extent. The dense/porous structure of β-TCP bioceramics may provide a simple but effective way to increase the mechanical properties of porous bioceramics for the bone regeneration applications at weight bearing sites.Fig.4 The variation of the compressive strength and elastic modulus of the bioceramics withdifferent dense/porous cross-sectional area ratio. ConclusionsThe dense/porous structured β-TCP bioceramics were prepared and revealed excellent mechanical properties with compressive strength from 10 to 74 MPa and elastic modulus from 180 to 960 MPa, which is 5 to 37 times higher than that of the pure porous β-TCP and comparable to the strength of human femoral cancellous bone. The interface between the dense and porous bioceramics is connected compactly and tightly. The dense/porous structure of β-TCP bioceramics may provide a simple but effective way to increase the mechanical properties of porous bioceramics for weight bearing site’s bone regeneration.AcknowledgementFinancial supports from the Shanghai Postdoctoral Scientific Key Program and the Science & Technology Commission of Shanghai Municipality of China (No.04DZ52043) are greatly acknowledged.References:[1] Niedhart C, Maus U, Redmann E, Schmidt-Rohlfing B, Niethard FU, Siebert CH: J BiomedMater Res Vol. 65A (2003), p.17[2] Hench Larry L: Journal of the American Ceramic Society Vol. 81(1998), p.1705[3] Carotenuto G: Advanced Performance Materials Vol. 5(1998), p.171[4] Hassna R. R. Ramay, Zhang M.: Biomaterials Vol. 25(2004), p.5171[5] Ashizuka M, Nakatsu M, Ishida E: Journal of the Ceramic Society of Japan, v 98(1990), p.204. 01020304050607080012345020040060080010001200E l a st i c M o d u l u s (M P a ) C o m p r e s s i v e S t r e n g h (M P a )S dense /S porous。

分子生物学英文文献6

Chapter19Detection and Quantitative Analysis of Small RNAs by PCR Seungil Ro and Wei YanAbstractIncreasing lines of evidence indicate that small non-coding RNAs including miRNAs,piRNAs,rasiRNAs, 21U endo-siRNAs,and snoRNAs are involved in many critical biological processes.Functional studies of these small RNAs require a simple,sensitive,and reliable method for detecting and quantifying levels of small RNAs.Here,we describe such a method that has been widely used for the validation of cloned small RNAs and also for quantitative analyses of small RNAs in both tissues and cells.Key words:Small RNAs,miRNAs,piRNAs,expression,PCR.1.IntroductionThe past several years have witnessed the surprising discovery ofnumerous non-coding small RNAs species encoded by genomesof virtually all species(1–6),which include microRNAs(miR-NAs)(7–10),piwi-interacting RNAs(piRNAs)(11–14),repeat-associated siRNAs(rasiRNAs)(15–18),21U endo-siRNAs(19),and small nucleolar RNAs(snoRNAs)(20).These small RNAsare involved in all aspects of cellular functions through direct orindirect interactions with genomic DNAs,RNAs,and proteins.Functional studies on these small RNAs are just beginning,andsome preliminaryﬁndings have suggested that they are involvedin regulating genome stability,epigenetic marking,transcription,translation,and protein functions(5,21–23).An easy and sensi-tive method to detect and quantify levels of these small RNAs inorgans or cells during developmental courses,or under different M.Sioud(ed.),RNA Therapeutics,Methods in Molecular Biology629,DOI10.1007/978-1-60761-657-3_19,©Springer Science+Business Media,LLC2010295296Ro and Yanphysiological and pathophysiological conditions,is essential forfunctional studies.Quantitative analyses of small RNAs appear tobe challenging because of their small sizes[∼20nucleotides(nt)for miRNAs,∼30nt for piRNAs,and60–200nt for snoRNAs].Northern blot analysis has been the standard method for detec-tion and quantitative analyses of RNAs.But it requires a relativelylarge amount of starting material(10–20μg of total RNA or>5μg of small RNA fraction).It is also a labor-intensive pro-cedure involving the use of polyacrylamide gel electrophoresis,electrotransfer,radioisotope-labeled probes,and autoradiogra-phy.We have developed a simple and reliable PCR-based methodfor detection and quantiﬁcation of all types of small non-codingRNAs.In this method,small RNA fractions are isolated and polyAtails are added to the3 ends by polyadenylation(Fig.19.1).Small RNA cDNAs(srcDNAs)are then generated by reverseFig.19.1.Overview of small RNA complementary DNA(srcDNA)library construction forPCR or qPCR analysis.Small RNAs are polyadenylated using a polyA polymerase.ThepolyA-tailed RNAs are reverse-transcribed using a primer miRTQ containing oligo dTsﬂanked by an adaptor sequence.RNAs are removed by RNase H from the srcDNA.ThesrcDNA is ready for PCR or qPCR to be carried out using a small RNA-speciﬁc primer(srSP)and a universal reverse primer,RTQ-UNIr.Quantitative Analysis of Small RNAs297transcription using a primer consisting of adaptor sequences atthe5 end and polyT at the3 end(miRTQ).Using the srcD-NAs,non-quantitative or quantitative PCR can then be per-formed using a small RNA-speciﬁc primer and the RTQ-UNIrprimer.This method has been utilized by investigators in numer-ous studies(18,24–38).Two recent technologies,454sequenc-ing and microarray(39,40)for high-throughput analyses of miR-NAs and other small RNAs,also need an independent method forvalidation.454sequencing,the next-generation sequencing tech-nology,allows virtually exhaustive sequencing of all small RNAspecies within a small RNA library.However,each of the clonednovel small RNAs needs to be validated by examining its expres-sion in organs or in cells.Microarray assays of miRNAs have beenavailable but only known or bioinformatically predicted miR-NAs are covered.Similar to mRNA microarray analyses,the up-or down-regulation of miRNA levels under different conditionsneeds to be further validated using conventional Northern blotanalyses or PCR-based methods like the one that we are describ-ing here.2.Materials2.1.Isolation of Small RNAs, Polyadenylation,and Puriﬁcation 1.mirVana miRNA Isolation Kit(Ambion).2.Phosphate-buffered saline(PBS)buffer.3.Poly(A)polymerase.4.mirVana Probe and Marker Kit(Ambion).2.2.Reverse Transcription,PCR, and Quantitative PCR 1.Superscript III First-Strand Synthesis System for RT-PCR(Invitrogen).2.miRTQ primers(Table19.1).3.AmpliTaq Gold PCR Master Mix for PCR.4.SYBR Green PCR Master Mix for qPCR.5.A miRNA-speciﬁc primer(e.g.,let-7a)and RTQ-UNIr(Table19.1).6.Agarose and100bp DNA ladder.3.Methods3.1.Isolation of Small RNAs 1.Harvest tissue(≤250mg)or cells in a1.7-mL tube with500μL of cold PBS.T a b l e 19.1O l i g o n u c l e o t i d e s u s e dN a m eS e q u e n c e (5 –3 )N o t eU s a g em i R T QC G A A T T C T A G A G C T C G A G G C A G G C G A C A T G G C T G G C T A G T T A A G C T T G G T A C C G A G C T A G T C C T T T T T T T T T T T T T T T T T T T T T T T T T V N ∗R N a s e f r e e ,H P L CR e v e r s e t r a n s c r i p t i o nR T Q -U N I r C G A A T T C T A G A G C T C G A G G C A G GR e g u l a r d e s a l t i n gP C R /q P C Rl e t -7a T G A G G T A G T A G G T T G T A T A G R e g u l a r d e s a l t i n gP C R /q P C R∗V =A ,C ,o r G ;N =A ,C ,G ,o r TQuantitative Analysis of Small RNAs299 2.Centrifuge at∼5,000rpm for2min at room temperature(RT).3.Remove PBS as much as possible.For cells,remove PBScarefully without breaking the pellet,leave∼100μL of PBS,and resuspend cells by tapping gently.4.Add300–600μL of lysis/binding buffer(10volumes pertissue mass)on ice.When you start with frozen tissue or cells,immediately add lysis/binding buffer(10volumes per tissue mass)on ice.5.Cut tissue into small pieces using scissors and grind it usinga homogenizer.For cells,skip this step.6.Vortex for40s to mix.7.Add one-tenth volume of miRNA homogenate additive onice and mix well by vortexing.8.Leave the mixture on ice for10min.For tissue,mix it every2min.9.Add an equal volume(330–660μL)of acid-phenol:chloroform.Be sure to withdraw from the bottom phase(the upper phase is an aqueous buffer).10.Mix thoroughly by inverting the tubes several times.11.Centrifuge at10,000rpm for5min at RT.12.Recover the aqueous phase carefully without disrupting thelower phase and transfer it to a fresh tube.13.Measure the volume using a scale(1g=∼1mL)andnote it.14.Add one-third volume of100%ethanol at RT to the recov-ered aqueous phase.15.Mix thoroughly by inverting the tubes several times.16.Transfer up to700μL of the mixture into aﬁlter cartridgewithin a collection bel theﬁlter as total RNA.When you have>700μL of the mixture,apply it in suc-cessive application to the sameﬁlter.17.Centrifuge at10,000rpm for15s at RT.18.Collect theﬁltrate(theﬂow-through).Save the cartridgefor total RNA isolation(go to Step24).19.Add two-third volume of100%ethanol at RT to theﬂow-through.20.Mix thoroughly by inverting the tubes several times.21.Transfer up to700μL of the mixture into a newﬁlterbel theﬁlter as small RNA.When you have >700μL of theﬁltrate mixture,apply it in successive appli-cation to the sameﬁlter.300Ro and Yan22.Centrifuge at10,000rpm for15s at RT.23.Discard theﬂow-through and repeat until all of theﬁltratemixture is passed through theﬁlter.Reuse the collectiontube for the following washing steps.24.Apply700μL of miRNA wash solution1(working solu-tion mixed with ethanol)to theﬁlter.25.Centrifuge at10,000rpm for15s at RT.26.Discard theﬂow-through.27.Apply500μL of miRNA wash solution2/3(working solu-tion mixed with ethanol)to theﬁlter.28.Centrifuge at10,000rpm for15s at RT.29.Discard theﬂow-through and repeat Step27.30.Centrifuge at12,000rpm for1min at RT.31.Transfer theﬁlter cartridge to a new collection tube.32.Apply100μL of pre-heated(95◦C)elution solution orRNase-free water to the center of theﬁlter and close thecap.Aliquot a desired amount of elution solution intoa1.7-mL tube and heat it on a heat block at95◦C for∼15min.Open the cap carefully because it might splashdue to pressure buildup.33.Leave theﬁlter tube alone for1min at RT.34.Centrifuge at12,000rpm for1min at RT.35.Measure total RNA and small RNA concentrations usingNanoDrop or another spectrophotometer.36.Store it at–80◦C until used.3.2.Polyadenylation1.Set up a reaction mixture with a total volume of50μL in a0.5-mL tube containing0.1–2μg of small RNAs,10μL of5×E-PAP buffer,5μL of25mM MnCl2,5μL of10mMATP,1μL(2U)of Escherichia coli poly(A)polymerase I,and RNase-free water(up to50μL).When you have a lowconcentration of small RNAs,increase the total volume;5×E-PAP buffer,25mM MnCl2,and10mM ATP should beincreased accordingly.2.Mix well and spin the tube brieﬂy.3.Incubate for1h at37◦C.3.3.Puriﬁcation 1.Add an equal volume(50μL)of acid-phenol:chloroformto the polyadenylation reaction mixture.When you have>50μL of the mixture,increase acid-phenol:chloroformaccordingly.2.Mix thoroughly by tapping the tube.Quantitative Analysis of Small RNAs3013.Centrifuge at10,000rpm for5min at RT.4.Recover the aqueous phase carefully without disrupting thelower phase and transfer it to a fresh tube.5.Add12volumes(600μL)of binding/washing buffer tothe aqueous phase.When you have>50μL of the aqueous phase,increase binding/washing buffer accordingly.6.Transfer up to460μL of the mixture into a puriﬁcationcartridge within a collection tube.7.Centrifuge at10,000rpm for15s at RT.8.Discard theﬁltrate(theﬂow-through)and repeat until allof the mixture is passed through the cartridge.Reuse the collection tube.9.Apply300μL of binding/washing buffer to the cartridge.10.Centrifuge at12,000rpm for1min at RT.11.Transfer the cartridge to a new collection tube.12.Apply25μL of pre-heated(95◦C)elution solution to thecenter of theﬁlter and close the cap.Aliquot a desired amount of elution solution into a1.7-mL tube and heat it on a heat block at95◦C for∼15min.Open the cap care-fully because it might be splash due to pressure buildup.13.Let theﬁlter tube stand for1min at RT.14.Centrifuge at12,000rpm for1min at RT.15.Repeat Steps12–14with a second aliquot of25μL ofpre-heated(95◦C)elution solution.16.Measure polyadenylated(tailed)RNA concentration usingNanoDrop or another spectrophotometer.17.Store it at–80◦C until used.After polyadenylation,RNAconcentration should increase up to5–10times of the start-ing concentration.3.4.Reverse Transcription 1.Mix2μg of tailed RNAs,1μL(1μg)of miRTQ,andRNase-free water(up to21μL)in a PCR tube.2.Incubate for10min at65◦C and for5min at4◦C.3.Add1μL of10mM dNTP mix,1μL of RNaseOUT,4μLof10×RT buffer,4μL of0.1M DTT,8μL of25mM MgCl2,and1μL of SuperScript III reverse transcriptase to the mixture.When you have a low concentration of lig-ated RNAs,increase the total volume;10×RT buffer,0.1M DTT,and25mM MgCl2should be increased accordingly.4.Mix well and spin the tube brieﬂy.5.Incubate for60min at50◦C and for5min at85◦C toinactivate the reaction.302Ro and Yan6.Add1μL of RNase H to the mixture.7.Incubate for20min at37◦C.8.Add60μL of nuclease-free water.3.5.PCR and qPCR 1.Set up a reaction mixture with a total volume of25μL ina PCR tube containing1μL of small RNA cDNAs(srcD-NAs),1μL(5pmol of a miRNA-speciﬁc primer(srSP),1μL(5pmol)of RTQ-UNIr,12.5μL of AmpliTaq GoldPCR Master Mix,and9.5μL of nuclease-free water.ForqPCR,use SYBR Green PCR Master Mix instead of Ampli-Taq Gold PCR Master Mix.2.Mix well and spin the tube brieﬂy.3.Start PCR or qPCR with the conditions:95◦C for10minand then40cycles at95◦C for15s,at48◦C for30s and at60◦C for1min.4.Adjust annealing Tm according to the Tm of your primer5.Run2μL of the PCR or qPCR products along with a100bpDNA ladder on a2%agarose gel.∼PCR products should be∼120–200bp depending on the small RNA species(e.g.,∼120–130bp for miRNAs and piRNAs).4.Notes1.This PCR method can be used for quantitative PCR(qPCR)or semi-quantitative PCR(semi-qPCR)on small RNAs suchas miRNAs,piRNAs,snoRNAs,small interfering RNAs(siRNAs),transfer RNAs(tRNAs),and ribosomal RNAs(rRNAs)(18,24–38).2.Design miRNA-speciﬁc primers to contain only the“coresequence”since our cloning method uses two degeneratenucleotides(VN)at the3 end to make small RNA cDNAs(srcDNAs)(see let-7a,Table19.1).3.For qPCR analysis,two miRNAs and a piRNA were quan-titated using the SYBR Green PCR Master Mix(41).Cyclethreshold(Ct)is the cycle number at which theﬂuorescencesignal reaches the threshold level above the background.ACt value for each miRNA tested was automatically calculatedby setting the threshold level to be0.1–0.3with auto base-line.All Ct values depend on the abundance of target miR-NAs.For example,average Ct values for let-7isoforms rangefrom17to20when25ng of each srcDNA sample from themultiple tissues was used(see(41).Quantitative Analysis of Small RNAs3034.This method ampliﬁes over a broad dynamic range up to10orders of magnitude and has excellent sensitivity capable ofdetecting as little as0.001ng of the srcDNA in qPCR assays.5.For qPCR,each small RNA-speciﬁc primer should be testedalong with a known control primer(e.g.,let-7a)for PCRefﬁciency.Good efﬁciencies range from90%to110%calcu-lated from slopes between–3.1and–3.6.6.On an agarose gel,mature miRNAs and precursor miRNAs(pre-miRNAs)can be differentiated by their size.PCR prod-ucts containing miRNAs will be∼120bp long in size whileproducts containing pre-miRNAs will be∼170bp long.However,our PCR method preferentially ampliﬁes maturemiRNAs(see Results and Discussion in(41)).We testedour PCR method to quantify over100miRNAs,but neverdetected pre-miRNAs(18,29–31,38). 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关于乳酸菌的英文文献及翻译.

2.4. Chemical and microbial analyses Analysis of DM and CP concentration in the experimental diets, excreta and probiotic products was done according to AOAC (1990 methods (930.05 and 976.05, respectively. The GE was measured by using the bomb calorimeter (model 1261, Parr Instrument Co., Moline, IL, and chromium concentration was determined with an automated spectrophotometer (Jasco V-650, Jasco Corp., Tokyo, Japan according to the procedure of Fenton and Fenton (1979. The microbiological assay of faecal samples (d 14 and 28 and intestinal digesta (d 28 was conducted by culturing in different media for the determination of total anaerobic bacteria (Tryptic soy agar, Bifidobacterium spp. (MRS agar, Lactobacillus spp. (MRS agar+0.02% NaN3+0.05% L-cystine hydrochloride monohydrate, Clostridium spp. (TSC agar and coliforms (violet red bile agar. The microbiological assay of probiotic products was also carried out by culturing technique. The L. acidophilus was enumerated using MRS agar+0.02%NaN3+0.05% L-cystine hydrochloride monohydrate, B. Subtilis by using plate count agar, S. cerevisiae and A. oryzae by potato dextrose agar. The anaerobic conditions during the assay of anaerobic were created by using gas pack anaerobic system (BBL, No. 260678; Difco, Detroit, MI. The tryptic soy agar (No. 236950, MRS agar (No. 288130, violet red bile agar (No. 216695, plate count agar (No. 247940, and potato dextrose agar (No. 213400 used were purchased from Difco Laboratories (Detroit,MI, and TSC agar(CM0589 was purchased from Oxoid (Hampshire, UK. The pH of probiotic products was determined by pH meter (Basic pH Meter PB-11, Sartorius, Germany.2.5. Small intestine morphology Three cross-sections for each intestinal sample were prepared after staining with azure A and eosin using standard paraffin embedding procedures. A total of 10 intact, welloriented crypt-villus units were selected in triplicate for each intestinal cross-section as described previously (Jin et al., 2008. Villus height was measured from the tip of the villi to the villus crypt junction, and crypt depth was defined as the depth of the invagination between adjacent villi. All morphological measurements (villus height and crypt depth were made in 10-μm increments by using animage proce ssing and analysis system (Optimus software version 6.5, Media Cybergenetics, North Reading, MA.2.6. Statistical analysesAll the data obtained in the current study were analyzed in accordance with a rand omized complete block design using the GLM procedure of SAS (SAS Inst. Inc., C ary, NC. In Exp. 1, one-way analysis of variance test was used and when signific ant differences (Pb0.05 were determined among treatment means, they were separ ated by using Duncan's multiple range tests. In Exp. 2, the data were analyzed as a 2×2 factorial arrangement of treatments in randomized complete block design. T he main effects of probiotic products (LF or SF, antibiotic (colistin or lincomycin, a nd their interaction were determined by the Mixed procedures of SAS. However, as the interaction (probiotic x antibiotic was not statistically significant (Pb0.05, it wa s removed from the final model. The pen was the experimental unit for all analysis in both experiments. The bacterial concentrations were transformed (log before st atistical analysis.3.1. Experiment 13.1.1. Growth performance and apparent total tract digestibilityDietary treatments had no effect on the performance of pigs during phase I (Table 3. However, during phase II and the overall experimental period, improved (Pb0.05 ADG, ADFI and G:F were observed in pigs fed PC, LF and SF dietswhen compared with pigs fed NC diet. Moreover, pigs fed PC and SF diets had hi gher (Pb0.05 ADG and better G:F than pigs fed LF diet during phase II and the o verall experimentalperiod. The dietary treatments had no influence on the ATTDof DM and GE; however, pigs fed PC and SF diets had greater ATTD of CP whe n compared with pigs fed NC and LF diets (Table 4.3.1.2. Bacterial population in faecesDietary treatments had no effect on the faecal total anaerobes and Bifidobacterium spp. population at d 14 and 28, and Lactobacillus spp. at d 14 (Table 5. However, pigs fed PC (d 14 and 28 and SF (d 28 diets had less (Pb0.05 faecal Clostridium spp. and coliforms than pigs fed NC diet.Moreover, pigs fed SF diet had greater (Pb0.05 faecal Lactobacillus spp. populatio n (d 28 than pigs fed NC, PC and LF diets.3.2. Experiment 23.2.1. Growth performance and apparent total tract digestibilityDuring phase I, pigs fed SF diet consumed more feed than pigs fed LF diet, wher eas the ADG and ADFI were similar between pigs fed LF and SF diets (Table 6. During phase II and the overall experimental period, pigs fed SF diet showed better ADG(Pb0.01, ADFI (Pb0.01 and G:F (Pb0.05 thanpigs fed LF diet. Howev er, different antibiotics had no effect on the performance of pigs. Pigs fed SF diet had greater ATTD of DM and CP during phases I and II (Pb0.01 and 0.001, respe ctively when compared with pigs fed LF diet (Table 7.However, different antibiotics had no effect on the ATTD of DM, CP and GE.3.2.2. Bacterial population in intestinePigs fed SF diet had greater (Pb0.05 Lactobacillus spp. And less Clostridium spp. (Pb0.01 and coliform (Pb0.05 population in the ileum than pigs fed LF diet (Table 8. Additionally,higher (Pb0.05 caecal Bifidobacterium spp. Population was observed in pigs fed SF diet. Antibiotics had no effect on the ileal microbial population; however, pigs fed colistin diet had less number of Bifidobacterium spp. (Pb0.05 and coliforms (Pb0.01 inthe cecum, whereas, feeding of lincomycin diet resulted in reduced (Pb0.05 caecal Clostridium spp.population.3.2.3. Small intestinal morphologyThe different probiotic products and antibiotics had no influence on the morphology of different segments of the small intestine, except for the greater (Pb0.05 villus height:crypt depth at the jejunum and ileum noticed in pigs fed lincomycin diet (Table 9.4.DiscussionPrevious studies on probiotics lack information on the method of production used, however, the preparation of probiotics by LF method is fairly common (Patel et al., 2004. The probiotic products used in the present study differedfrom the previous reports in that harvested probiotic microbes were added directly to the diets. In this study, the microbial biomass grown on the CB was directly sprayed onthe carrier (corn and soybean meal to obtain LF probiotic product. In case of the SF probiotic product, corn and soybean meal was used as a substrate during fermentation and as a carrier of probiotic microbes. We have reported previously that multi-microbe probiotic product prepared by SF method was better than the probiotic product prepared by submerged liquid fermentation in improving performance, nutrient retention and reducing harmful intestinal bacteria in broilers (Shim et al., 2010. In the current study, LF and SF method was used and corn–soybean meal was used as a substrate forthe growth of potential probiotic microbes under optimum conditions.2.4 化学和微生物分析在试验日粮干物质和粗蛋白含量的分析中,排泄物和益生菌产品是根据AOAC(1990方法(分别为930.05和976.05 分析。

党参属植物化学物、生物活性及质量控制综述(2015年英文文献)

REVIEWThe genus Codonopsis (Campanulaceae):a review of phytochemistry,bioactivity and quality controlJing-Yu He •Na Ma •Shu Zhu •Katsuko Komatsu •Zhi-Yuan Li •Wei-Ming FuReceived:7May 2014/Accepted:18July 2014/Published online:7August 2014ÓThe Japanese Society of Pharmacognosy and Springer Japan 2014Abstract Codonopsis ,in the family Campanulaceae,is a genus containing 42species of dicotyledonous herbaceous perennial plants,predominantly found in Central,East and South Asia.Several Codonopsis species are widely used in traditional medicine and are considered to have multiple medicinal properties.Among the Codonopsis species,Codonopsis pilosula (Franch.)Nannf.and nceolata (Sieb.et Zucc.)Benth.&Hook.f.ex Trautv.are more popular than others according to the ﬁndings,especially phytochemical and bioactive studies.Phytochemical research shows that Codonopsis species contain mainly polyacetylenes,phenylpropanoids,alkaloids,triterpenoids and polysaccharides,which contribute to multiple bioac-tivities.However,the mechanisms of their bioactivities need to be further elucidated.The less popular Codonopsis species remain to be studied and exploited.In addition,although a series of methods for the quality evaluation of Codonopsis species have been developed,a feasible and reliable approach to the efﬁcacious and safe use of various Codonopsis species is still needed,with considering botanical origin,chemical constituents and bioactive effects.This review aims to provide up-to-date and com-prehensive information on the phytochemistry,bioactivity and quality control of medicinal plants in the genus Codonopsis and to highlight current gaps in knowledge,which is useful for the wider development of the Codon-opsis genus.Keywords Codonopsis ÁPhytochemistry ÁBioactivity ÁQuality controlIntroductionCodonopsis ,which is a dicotyledonous genus of herba-ceous perennial plants in the family Campanulaceae,has 42species predominantly distributed in Central,East and South Asia;40Codonopsis species can be found in China [1].However,Codonopsis pilosula (Franch.)Nannf.,C.pilosula Nannf.var.modesta (Nannf.)L. D.Shen, C.tangshen Oliv.and nceolata (Sieb.et Zucc.)Benth.&Hook.f.ex Trautv.are the only species widely used,of which the fresh or dried roots are collectively regarded as famous herbal medicines and have been used in folk medicine for hundreds of years.Codonopsis Radix is pre-scribed as the dried roots of C.pilosula ,C.pilosula var.modesta and C.tangshen in the Chinese Pharmacopoeia (2010edition)[2].It is called ‘‘Dangshen’’in Chinese and ‘‘Tojin’’in Japanese,and has been used in traditional Chinese medicine for replenishing qi (vital energy)deﬁ-ciency,strengthening the immune system,improving poor gastrointestinal function,gastric ulcer and appetite,decreasing blood pressure,etc.,and is sometimes used as a substitute for Ginseng (Panax ginseng C.A.Mey.)[3,4].The roots of other Codonopsis species,including C.tu-bulosa ,C.subglobosa ,C.clematidea and nceolata ,are reported to be used as substitutes for Codonopsis Radix in some regions [3]. nceolata ,commonly called bonnet bellﬂower,is a herb with high value in traditional Chinese medicine and its root is also becoming popular as aJ.-Y.He ÁN.Ma ÁZ.-Y.Li ÁW.-M.Fu (&)Guangzhou Institute of Advanced Technology,Chinese Academy of Sciences,1121Haibin Rd.,Nansha Dist.,511-458Guangzhou,People’s Republic of China e-mail:wm.fu@S.Zhu ÁK.KomatsuDivision of Pharmacognosy,Department of MedicinalResources,Institute of Natural Medicine,University of Toyama,2630Sugitani,Toyama 930-0194,JapanJ Nat Med (2015)69:1–21DOI10.1007/s11418-014-0861-9special vegetable in some Asian countries[5]nceo-lata has been used for the treatment of bronchitis,asthma, cough,tuberculosis,dyspepsia and psychoneurosis[6–8]. Phytochemical researches have revealed that the roots of Codonopsis species contained alkaloids,phenylpropanoids, triterpenoids,polyacetylenes,ﬂavones,organic acid,poly-saccharides,etc.[9–54].Among them,polyacetylenes, phenylpropanoids,alkaloids,triterpenoids and polysac-charides are considered to be the major constituents and responsible for most of the activities found in the plants of this genus.The chemical proﬁle varied greatly between species and sample collections may cause these Codon-opsis species to possess diverse bioactivities.Some com-pounds belonging to theseﬁve chemotypes have been evaluated for potential biological activity and pharmaco-logical mechanisms.However,the pharmacological mechanisms of these Codonopsis species related to bio-logical activity and clinical application remain largely unexplained.Additionally,the toxicity of Codonopsis has not been reported in the scientiﬁc literature.As many investigations indicated that a variety of chemical constituents contributed to the effects of Codon-opsis species,the quantitation of bioactive components becomes urgent for ensuring the efﬁcacy of Codonopsis species.In the Chinese Pharmacopoeia(2010edition),only lobetyolin is used as the chemical marker for identiﬁcation of Codonopsis Radix,which seems useless for many Codonopsis species involved[2].Hence,a number of studies have attempted to develop accurate,sensitive and selective analytical methods for qualitative and quantitative evaluation of Codonopsis materials.To provide information beneﬁting traditional uses and scientiﬁc studies,this review summarizes and evaluates the available phytochemical and bioactive properties of Codonopsis genus reported by the literature.In addition, for the efﬁcacious and safe uses of Codonopsis,the pro-gress of research on quality evaluation of various Codon-opsis species is also presented.Chemical constituents of genus Codonopsis Phytochemical studies have been performed on Codonopsis species plants over the last30years all around the world. Only some of the different Codonopsis species plants have been explored for obtaining information on chemo-taxo-nomical identiﬁcation,isolation and identiﬁcation of vari-ous important chemicals from this genus and comparison of the chemicals in different plants or species.C.pilosula,C.tangshen,nceolata and C.clemati-dea have been widely investigated in their phytochemistry; more than100compounds have thereby been isolated and identiﬁed.On the other hand,few compounds in C.cordifolioidea,C.nervosa,C.thalictrifolia,C.xundian-ensis and C.tubulosa are reported because they are found only in selected regions.The components in other Codonopsis species have not yet been reported since these Codonopsis species may be difﬁcult to collect and/or be scarce.To date,polyacetylenes,phenylpropanoids,alka-loids,triterpenoids,etc.have been isolated and character-ized from the different parts of these Codonopsis species plants.The names of these constituents,the plant and the parts from which they are derived are summarized in Table1.The structures of the compounds are shown in Figs.1,2,3,4,5,6and7.AlkaloidsThe pyrrolidine alkaloids codonopsine(1),codonopsinine (2),codonopsinol(3)and radicamine A(4)were isolated from the aerial parts of C.clematidea[9,10].Two pyr-rolidine alkaloids,codonopyrrolidiums A(5)and B(6), were isolated from the roots of C.tangshen[13],and were also found in the roots of C.pilosula and C.pilosula var. modesta[12,14].In addition,the pyrrolidine alkaloids codonopsinols A(7),B(8),C(9)and the glycoside,cod-onopiloside A(10)were obtained from the roots of C. pilosula[11].Codotubulosine B(11)was found in the roots of C.tubulosa[15].Other alkaloids,n-9-formyl harman(12),norharman (13),1-carbomethyl carboline(14),1,2,3,4-tetrahydro-b-carboline-3-carboxylic acid(15)and tryptophan(16),were isolated from the roots of nceolata[16,17,19]. Tryptophan(16),perlolyrine(17)and nicotinic acid(18) were obtained from the roots of C.pilosula[18,20,21]. The common compounds uracil(19)and adenosine(20) were found in the roots of C.pilosula and the roots of C. pilosula and C.tangshen,respectively[13,18,22]. PhenylpropanoidsThe phenylpropanoids tangshenosides I(21),II(22),III (23)and IV(24)wereﬁrst isolated from C.tangshen[23, 25].Tangshenoside V(25),tangshenoside VI(26)and codonosides A(27)and B(28),considered to be the characteristic components,were isolated from C.tangshen [13,26].Tangshenoside VI(26)was also isolated from the aerial parts of C.nervosa[27].Recently,tangshenoside VIII(29)has been obtained from the roots of nceolata [24].In addition,12phenylpropanoids,cordifoliketones A (30)and B(31),sinapinaldehyde(32),coniferaldehyde (33),coniferoside(34),isoconiferin(35),nervolans B(36) and C(37),dillapiole(38),1-allyl-2,6-dimethoxy-3,4-methylenedioxybenzene(39),4-allyl-2-(3-methylbut-2-en-1-yl)phenol(40)and sachaliside(41),were isolated from the roots of C.cordifolioidea[28,29].Syringin(42)has been commonly found in5Codonopsis species[21,23,30–32].TriterpenesThree new triterpenyl esters,codonopilates A(43),B(44) and C(45),together with seven known triterpenoids, 24-methylenecycloartanyl linolate(46),24-methylene-cycloartan-3-ol(47),friedelin(48),1-friedelen-3-one(49), stigmast-7-en-3-one(50),taraxerol(51)and stigmast-7-en-3-ol(52),were isolated from the CHCl3-soluble fraction of the methanol extract of C.pilosula[14].Additionally,a-spinasterol(53)was obtained from C.pilosula,C.tang-shen,nceolata and C.thalictrifolia[32–35],and tar-axeryl acetate(54)was obtained from C.pilosula, C. tangshen and C.clematidea[10,34,35].The oleanan-type bisdesmoside with sugars at C-3and C-28,codonolaside (55),codonolasides I(56),II(57)and III(58),and their prosapogenins,eclalbasaponin XIII(59)and echinocystic acid3-O-b-D-glucuronopyranoside(60),were isolated from the roots of nceolata[36].The triterpene saponins, lancemasides A(61),B(62),C(63),D(64),E(65),F(66) and G(67),have also been isolated from the roots of C. lanceolata cultivated in Korea[19].Codonolaside IV(68), codonolaside V(69),foetidissimoside A(70),aster saponin Hb(71),oleanolic acid(72),echinocystic acid(73)and stigmasterol(74)were found in the roots of nceolata [19,30,34,37–39].Foetidissimoside A(70)and rubiprasin B(75)were isolated from the aerial parts of C.clematidea [10].For the aerial parts of C.thalictrifolia,isolation of a-spinasterol(53)and b-amyrin acetate(76)was reported [32].Zeorin(77)and lupeol(78)were isolated from the whole plants of C.nervosa[40].PolyacetylenesIsolation and identiﬁcation of lobetyolin(79),lobetyolinin (80)and lobetyol(81)from the roots and aerial parts of plants belonging to the genus Codonopsis have also been reported[10,12,18,25,27,29,32,34,41,42].Three new polyacetylene glucosides,cordifolioidynes A(82),B(83) and C(84),were isolated from a95%ethanol extract of the roots of C.cordifolioidea[29].Recently,cordifoli-oidynes B(83)has also been found in C.pilosula,C. pilosula var.modesta and C.tangshen,which are the botanical sources of Codonopsis Radix[12].FlavonesChrysoeriol(85),tricin(86),wogonin(87)and luteolin (88)were isolated from the roots of C.xundianensis Wang ZT and Xu GJ,which grows in Yunnan Province,China [43].Luteolin(88),kaempferol(89),luteolin-5-O-b-D-glucopyranoside(90),luteolin-7-O-b-D-gentiobioside(91), apigenin-7-O-b-D-glucopyranoside(92)and luteolin-7-O-Table1Compounds in Codonopsis speciespound names Species Part of the plant ReferencesAlkaloids1Codonopsine C.clematidea Aerial parts[9]2Codonopsinine C.clematidea Aerial parts[9]3Codonopsinol C.clematidea Aerial parts[10]4Radicamine A C.pilosula Roots[11]C.clematidea Aerial parts[10]5Codonopyrrolidium A C.pilosula Roots[12]C.pilosula var.Roots[12]modestaC.tangshen Roots[13]6Codonopyrrolidium B C.pilosula Roots[14]C.pilosula var.Roots[12]modestaC.tangshen Roots[13]7Codonopsinol A C.pilosula Roots[11]8Codonopsinol B C.pilosula Roots[11]9Codonopsinol C C.pilosula Roots[11]10Codonopiloside A C.pilosula Roots[11]11Codotubulosine B C.tubulosa Roots[15]12n-9-Formyl harman nceolata Roots[16]13Norharman nceolata Roots[16]141-Carbomethyl carboline nceolata Roots[16] 151,2,3,4-Tetrahydro-b-carboline-3-carboxylic acid nceolata Roots[17]16Tryptophan C.pilosula Roots[18]nceolata Roots[19]17Perlolyrine C.pilosula Roots[20]18Nicotinic acid C.pilosula Roots[21]19Uracil C.pilosula Roots[18]20Adenosine C.pilosula Roots[22]C.tangshen Roots[13]Phenylpropanoids21Tangshenoside I C.pilosula Roots[12]Roots[12]C.pilosula var.modestaC.tangshen Roots[23]nceolata Roots[24]22Tangshenoside II C.tangshen Roots[23]nceolata Roots[24]23Tangshenoside III C.tangshen Roots[25]nceolata Roots[24]24Tangshenoside IV C.tangshen Roots[25]nceolata Roots[24]25Tangshenoside V C.tangshen Roots[26]26Tangshenoside VI C.tangshen Roots[26]C.nervosa Aerial parts[27]27Codonoside A C.tangshen Roots[13]28Codonoside B C.tangshen Roots[13]29Tangshenoside VIII nceolata Roots[24]30Cordifoliketone A C.cordifolioidea Roots[28]31Cordifoliketone B C.cordifolioidea Roots[28] 32Coniferaldehyde C.cordifolioidea Roots[29] 33Sinapinaldehyde C.cordifolioidea Roots[29] 34Coniferoside C.cordifolioidea Roots[29] 35Isoconiferin C.cordifolioidea Roots[29] 36Nervolan B C.cordifolioidea Roots[28] 37Nervolan C C.cordifolioidea Roots[28] 38Dillapiole C.cordifolioidea Roots[28] 391-Allyl-2,6-dimethoxy-3,4-methylenedioxybenzene C.cordifolioidea Roots[28] 404-Allyl-2-(3-methylbut-2-en-1-yl)phenol C.cordifolioidea Roots[28] 41Sachaliside C.cordifolioidea Roots[29] 42Syringin C.pilosula Roots[21]C.tangshen Roots[23]nceolata Roots[30]C.nervosa Aerial parts[31]C.thalictrifolia Aerial parts[32]Triterpenes43Codonopilate A C.pilosula Roots[14] 44Codonopilate B C.pilosula Roots[14] 45Codonopilate C C.pilosula Roots[14] 4624-Methylenecycloartanyl linolate C.pilosula Roots[14] 4724-Methylenecycloartan-3-ol C.pilosula Roots[14] 48Friedelin C.pilosula Roots[14]C.tangshen Roots[12]nceolata Roots[33] 491-Friedelen-3-one C.pilosula Roots[14] 50Stigmast-7-en-3-one C.pilosula Roots[14] 51Taraxerol C.pilosula Roots[14]C.tangshen Roots[34] 52Stigmast-7-en-3-ol C.pilosula Roots[14] 53a-Spinasterol C.pilosula Roots[35]C.tangshen Roots[34]nceolata Roots[33]C.thalictrifolia Aerial parts[32] 54Taraxeryl acetate C.pilosula Roots[35]C.tangshen Roots[34]C.clematidea Aerial parts[10] 55Codonolaside nceolata Roots[36] 56Codonolaside I nceolata Roots[36] 57Codonolaside II nceolata Roots[36] 58Codonolaside III nceolata Roots[36] 59Eclalbasaponin XIII nceolata Roots[36] 60Echinocystic acid-3-O-(60-O-methyl)-b-D-glucuronopyranoside nceolata Roots[36] 61Lancemaside A nceolata Roots[19] 62Lancemaside B nceolata Roots[19] 63Lancemaside C nceolata Roots[19] 64Lancemaside D nceolata Roots[19] 65Lancemaside E nceolata Roots[19]66Lancemaside F nceolata Roots[19] 67Lancemaside G nceolata Roots[19] 68Codonolaside IV nceolata Roots[37] 69Codonolaside V nceolata Roots[38] 70Foetidissimoside A nceolata Roots[19]C.clematidea Aerial parts[10] 71Aster saponin Hb nceolata Roots[39] 72Oleanolic acid nceolata Roots[30] 73Echinocystic acid nceolata Roots[30] 74Stigmasterol C.tangshen Roots[34]nceolata Roots[33] 75Rubiprasin B C.clematidea Aerial parts[10] 76b-Amyrin acetate C.thalictrifolia Aerial parts[32] 77Zeorin C.nervosa Whole plants[40] 78Lupeol C.nervosa Whole plants[40] Polyacetylenes79Lobetyolin Codonopsis pilosula Roots[18]Roots[12]C.pilosula var.modestaC.tangshen Roots[25]nceolata Roots[41]C.clematidea Aerial parts[10]C.cordifolioidea Roots[29]C.nervosa Whole plants[27]C.thalictrifolia Aerial parts[32] 80Lobetyolinin C.pilosula Roots[18]Roots[12]C.pilosula var.modestaC.tangshen Roots[12]C.clematidea Aerial parts[10] 81Lobetyol C.pilosula Roots[42]Roots[12]C.pilosula var.modestaC.tangshen Roots[34]C.cordifolioidea Roots[29] 82Cordifolioidyne A C.cordifolioidea Roots[29] 83Cordifolioidyne B C.pilosula Roots[12]Roots[12]C.pilosula var.modestaC.tangshen Roots[12]C.cordifolioidea Roots[29] 84Cordifolioidyne C C.cordifolioidea Roots[29] Flavones85Chrysoeriol C.xundianensis Roots[43] 86Tricin C.xundianensis Roots[43] 87Wogonin C.xundianensis Roots[43]88Luteolin C.nervosa Whole plants[40]C.thalictrifolia Aerial parts[32]C.clematidea Aerial parts[10]C.xundianensis Roots[43] 89Kaempferol C.nervosa Whole plants[40] 90Luteolin-5-O-b-D-glucopyranoside C.nervosa Aerial parts[27]C.thalictrifolia Aerial parts[32] 91Luteolin-7-O-b-D-gentiobioside C.nervosa Aerial parts[27]C.thalictrifolia Aerial parts[32] 92Apigenin-7-O-b-D-glucopyranoside C.nervosa Aerial parts[31]93Luteolin-7-O-b-D-glucopyranosyl(1?6)-[(6000-O-caffeoyl)]-b-D-glucopyranoside C.nervosa Whole plants[40] C.clematidea Aerial parts[10]94Hesperidin C.pilosula Roots[35] Organic acids95Succinic acid C.pilosula Roots[44]C.nervosa Aerial parts[31] 963-O-caffeoylquinic acid C.nervosa Aerial parts[31]C.thalictrifolia Aerial parts[32] 975-O-caffeoylquinic acid C.nervosa Aerial parts[31] 984-(b-D-Glucopyranosyl)-benzoic acid C.nervosa Aerial parts[31] 99Caffeic acid C.thalictrifolia Aerial parts[32] 100Linoleic acid C.thalictrifolia Aerial parts[32] 1019,10,13-Trihydroxy-(E)-octadec-11-enoic acid C.pilosula Roots[35] 102Shikimic acid nceolata Roots[33] 103Vanillic acid C.tangshen Roots[34] Other compounds104Atractylenolide III C.pilosula Roots[21] 1055-Hydroxymethyl-2-furaldehyde C.pilosula Roots[14]C.tangshen Roots[34] 106Angelicin C.pilosula Roots[44] 107Psoralen C.pilosula Roots[44] 108Emodin C.pilosula Roots[18] 109Geniposide C.pilosula Roots[17] 110Hexyl-b-D-glucopyranoside C.pilosula Roots[18] 111Butyl-b-D-fructournanoside C.pilosula Roots[18] 112b-Sitosterol C.pilosula Roots[35]C.nervosa Whole plants[40] 113b-Daucosterol C.pilosula Roots[35] 114Hexyl-b-gentiobioside C.tangshen Roots[25] 115Hexyl-b-sophoroside C.tangshen Roots[25] 116(E)-2-hexenyl-b-sophoroside C.tangshen Roots[25] 117(E)-2-hexenyl-a-L-arabinopyranosyl(1?6)-b-D-glucopyranoside C.tangshen Roots[25]C.clematidea Aerial parts[10] 118Cordifoliﬂavane A C.cordifolioidea Roots[45] 119Cordifoliﬂavane B C.cordifolioidea Roots[45] 120Lanceolune A nceolata Roots[46] 121Lanceolune B nceolata Roots[46] 122Lanceolune C nceolata Roots[46]b-D-glucopyranosyl(1?6)-[(6000-O-caffeoyl)]-b-D-gluco-pyranoside(93)were obtained from C.nervosa[27,31,40],and luteolin(88),luteolin-5-O-b-D-glucopyranoside(90)and luteolin-7-O-b-D-gentiobioside(91)were alsofound in the aerial parts of C.thalictrifolia[32].In addi-tion,luteolin(88)and luteolin-7-O-b-D-glucopyrano-syl(1?6)-[(6000-O-caffeoyl)]-b-D-glucopyranoside(93)were isolated from the aerial parts of C.clematidea[10].Hesperidin(94)was only isolated from the roots of C.pilosula[35].Organic acidsTo date,succinic acid(95),3-O-caffeoylquinic acid(96),5-O-caffeoylquinic acid(97)and4-(b-D-glucopyranosyl)-benzoic acid(98)have been found in C.nervosa[31].Caffeic acid(99),linoleic acid(100)and3-O-caffeoyl-quinic acid(96)were isolated from C.thalictrifolia[32].Succinic acid(95)and9,10,13-trihydroxy-(E)-octadec-11-enoic acid(101)were isolated from C.pilosula[35,44].Shikimic acid(102)and vanillic acid(103)were onlyobtained from the roots of nceolata and C.tangshen,respectively[33,34].Other compoundsAtractylenolide III(104),5-hydroxymethyl-2-furaldehyde(105),angelicin(106),psoralen(107),emodin(108),ge-niposide(109),hexyl-b-D-glucopyranoside(110),butyl-b-D-fructournanoside(111),b-sitosterol(112)and b-dau-costerol(113)were isolated from the roots of C.pilosula[14,17,18,21,35,44].Hexyl-b-gentiobioside(114),hexyl-b-sophoroside(115),(E)-2-hexenyl-b-sophoroside(116),(E)-2-hexenyl-a-L-arabinopyranosyl(1?6)-b-D-glucopyranoside(117)and5-hydroxymethyl-2-fur-aldehyde(105)were isolated from the roots of C.tangshen[25,34].Cordifoliﬂavanes A(118)and B(119)were iso-lated from the roots of C.cordifolioidea[45].Three newbenzofuranylpropanoids,lanceolunes A(120),B(121)andC(122),as well as a new cerebroside,codonocerebrosideA(123),have been isolated from the roots of nceolata[41,46].(E)-2-Hexenyl-a-L-arabinopyranosyl(1?6)-b-D-glucopyranoside(117),3-oxo-a-ionol-b-D-glucopyranoside (124)and1,6-hexanediol,3,4-bis(4-hydroxy-3-methox-yphenyl)(125)were isolated from the aerial parts of C. clematidea[10].In addition,sweroside(126)and b-sitos-terol(112)were obtained from the whole plants of C. nervosa[40].Nutritive constituents including amino acids and trace elements in C.pilosula have been reported[47]. Essential oilsAs one of the important compositions,essential oils of several Codonopsis species have been reported.In the essential components from C.pilosula,50of66separated components were identiﬁed by GC–MS,mainly containing 1,2-benzonedicarboxylic acid dibatyl-ester(12.45%), heptedecanoic acid(8.10%)and2,4,5-triisopropyl styrene (7.62%)[48].Using the GC–MS method,54peaks were separated and37of them were identiﬁed in the essential components extracted from C.clematidea,in which the most abundant component was methyl hexadecanoate(30.40%)[49].The essential oils from the whole plants ofC.thalictrifolia,as a traditional Tibetan medicine,were analyzed by GC–MS,and45of60separated components were identiﬁed by comparing their mass spectra,in which the main principles were palmitic acid(43.5%)and linolic acid(18.3%)[50].In the essential oils extracted from the fresh and dried roots of C.cordifolioidea,63compounds were identiﬁed by GC–MS analysis,indicating that linolic acid(21.9%),retene(11.4%),pentadecane(7.4%), methyl9,12,15-octadecatrienoate(6.8%)and heneicosyl-cyclopentane(3.8%)were the main components[51]. PolysaccharidesLarge-molecule components in Codonopsis species were also studied.A water-soluble polysaccharide with a molecular mass of1.19104Da was obtained from the roots of C.pilosula and its structure investigation revealed that this polysaccharide had a backbone consisting of (1?3)-linked-b-D-galactopyranosyl,(1?2,3)-linked-a-D-galactopyranosyl and(1?3)-linked-b-D-rhamnopyr-anosyl residues and were branched with two glycosyl123Codonocerebroside A nceolata Roots[41] 1243-Oxo-a-ionol-b-D-glucopyranoside C.clematidea Aerial parts[10] 1251,6-Hexanediol,3,4-bis(4-hydroxy-3-methoxyphenyl) C.clematidea Aerial parts[10] 126Sweroside C.nervosa Whole plants[40]residues composed of a-L-arabinose-(1?5)-a-L-arabi-nose,whose C-1linked residues at the O-2position of galactosyl along the main chain in the ratio of1:1:2:1:1[52].Another polysaccharide with a molecular mass of 7.49104Da was isolated from C.pilosula and its com-ponents were galactose,arabinose and rhamnose in themolar ratio of1.13:1.12:1.Its main chain was shown to be (1?3)-linked-b-GalpNAc,(1?3)-linked-a-Rhap and (1?2,3)-b-Galp[53].Furthermore,a pectic polysac-charide with a molecular mass of1.459105Da was at ﬁrst isolated from C.pilosula,and its structural analysis revealed that this polysaccharide is composed of rhamnose, arabinose,galactose and galacturonic acid in the molar ratio of0.25:0.12:0.13:bined with chemical and spectroscopic analyses,its structure was proposed to be 1,4-linked-a-D-GalpA and1,4-linked-a-D-GalpA6Me interspersed with rare1,2-linked-b-L-Rhap,1,2,6-linked-a-D-Galp and terminal a-L-Arap[54].BioactivitiesAlthough there is information on the uses of many Codonopsis species in traditional medicine,only bioactiv-ity studies on C.pilosula and nceolata have been reported frequently,which proved their importance as medicinal plants.Bioactivity studies on other Codonopsis species such as C.clematidea and C.cordifolioidea were scarce.The studies generally referred to the bioactive effects of aqueous,methanol and ethanol extracts,as well as their further puriﬁed fractions,ﬂavones,saponins and polysaccharides.Codonopsis pilosulaAnti-tumor activityThe polysaccharide from C.pilosula(10l g/mL)was able to inhibit the activities of human gastric adenocarcinoma cells and hepatoma carcinoma cells[55].A pectic poly-saccharide(50,100,200and400l g/mL)exhibited marked cytotoxicity to human lung adenocarcinoma A549cells,in a dose-dependent manner[54].Anti-diabetic activityAfter mice were orally administered the polysaccharide from C.pilosula for a week,Fu et al.[56]found that three different doses of polysaccharide(100,200and300mg/ kg/day)could effectively decrease fasting blood glucose and insulin in serum,enhance superoxide dismutase(SOD)activity and reduce the content of malondialdehyde(MDA) in serum.It was therefore considered to possess a signiﬁ-cant hypoglycemic effect in diabetic mice by improving insulin resistance.He et al.[57]showed that the aqueous extract of the roots of C.pilosula(equal to4.5g raw material/kg/day)might retard the progression of diabetesby reducing the blood glucose level and preventing the increase of aldose reductase activity in streptozotocin-induced diabetic mice after3days of oral administration.Anti-aging activityXu et al.[58]found that after mice were orally administrated the polysaccharide from C.pilosula for8weeks,the poly-saccharide(50and150mg/kg/day)was able to increase the thymus index and spleen index as well as the activities of SOD in serum and liver,glutathione peroxidase and nitric oxide synthase particularly in kidney,while decreasing MDA in serum and liver and lipofuscin in brain.Its post-ponement of senility might be related to raising immunity, eliminating free radicals and anti-lipoperoxidation. Effects on gastric mucosaLiu et al.[59,60]found that the water-soluble fraction from the roots of C.pilosula(equal to10g raw material/ kg)had a signiﬁcant protective effect on gastric mucosal damage caused by alcohol,0.6N HCl and0.2N NaOH, and suggested that the pharmacological mechanism was related to the synthesis and/or release of prostaglandins in gastric mucosa.To date,Song et al.[61]found that lob-etyolin at the oral dose of1.5mg/kg had an effect on decreasing the ulcer index and the level of gastrin and increasing the level of6-keto-prostaglandin F1a in rats with gastric ulcer induced by ethanol,and suggested that lobetyolin played a protective role in gastric mucosa injury. Effects on blood systemAqueous extracts of C.pilosula(500l g/mL)potently inhibited erythrocyte hemolysis[62].In addition,after ischemia–reperfusion injury rats received8mg/100g body weight of a solution of saponins via intraperitoneal injec-tion,the results showed that the increase in SOD levels was accompanied by decreases in MDA,serum creatinine and blood urea nitrogen levels;bcl-2mRNA and protein levels were raised in transplanted kidneys from treated animals, while bax mRNA and protein levels were reduced.The apoptosis index was signiﬁcantly decreased in transplanted kidneys from treated animals relative to untreated controls. These results clearly demonstrated protective effects on ischemia–reperfusion injury after kidney transplantation, which might be explained by decreasing lipid peroxidation and inhibition of apoptosis[63].Effects on immunityZhang and Wang[64,65]found that6days of oral administration of the polysaccharide from C.pilosula (800mg/kg/day)had effects on immunosuppressed mice induced by cyclophosphamide,including increasing the thymus and spleen index and the phagocytic activity of peritoneal macrophages and recovering the activity of a-naphthyl-acetate esterase in peripheral lymphocytes.In an immunological study in vitro,a water-soluble polysaccha-ride(50,100and200l g/mL)could stimulate concanavalin A-or lipopolysaccharide(LPS)-induced lymphocyte pro-liferation in a dose-dependent manner[52].In addition,the methanol extract of C.pilosula(1mg/mL)inhibited inducible nitric oxide synthase and protein oxidation in LPS-stimulated murine RAW264.7macrophage cells[66].Effects on nervous systemTotal alkaloids(30l g/mL)caused a signiﬁcant enhancement of nerve growth factor-induced neurite outgrowth in PC12 cells as well as an increase in the phosphorylation of mitogen-activated protein kinase[67].Moreover,Pan et al.[68]orally administered alkaloids from C.pilosula(1mg/kg/day)to mice for4days after they suffered from amnesia by scopol-amine,and found that the alkaloids were effective against the decrease in acetylcholine.Other chemicals,saponins from Codonopsis Radix,were reported to have protective effects on the damage to astrocytes induced by hypoxia/hypoglycaemia and reoxygenation,and were able to inhibit the necrosis of astrocytes at three different concentrations(5.2,52and 520l g/mL)[69].Additionally,the polysaccharide from C. pilosula(1.1mmol/mL)also had marked protective effect on neural stem cell injury induced by sodium thiosulphate[70]. Other bioactivitiesThe extract of C.pilosula(20,40and60l g/mL)signiﬁ-cantly attenuated angiotensin II(AngII)-induced insulin-like growth factor II receptor(IGFIIR)promoter activity.C.pilosula also reversed Ca2?inﬂux,mitochondrial outer-membrane permeability and apoptosis increased by AngII plus Leu27-IGFII which was applied to enhance the AngII effect.Molecular markers in the IGFIIR apoptotic pathway and IGFIIR-Gaq association were down-regulated by C. pilosula.However,p-BadSer136and Bcl-2were increased. The results suggested that C.pilosula could suppress the AngII plus Leu27-IGFII-induced IGFII/IGFIIR pathway in myocardial cells[71].Codonopsis lanceolataAntioxidant activityThe water-soluble fraction and the n-butanol-soluble frac-tion of ethanol extract of nceolata showed。

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Application of α-amylase and Researchα-amylase to be widely distributed throughout microorganisms to higher plants. The International Enzyme classification number is EC. 3.2.1.1, acting on the starch from the starch molecules within the random cut α a 1,4 glycosidic bond to produce dextrin and reducing sugar, because the end product of carbon residues as Α configuration configuration, it is called α-amylase. Now refers to α-amylase were cut from the starch molecules within the α-1,4 glycosidic bond from the liquefaction of a class of enzymes.α-amylase is an important enzyme, a large number of used food processing, food industry, brewing, fermentation, textile industry and pharmaceutical industries, which account for the enzyme about 25% market share. Currently, both industrial production to large-scale production by fermentation α-amylase. α-amylase in industrial applications1.1 The bread baking industry, as a preservative enzymes used in baking industry, production of high quality products have been hundreds of years old. In recent decades, malt and microbial α-amylase, α-amylase is widely used in baking industry. The enzymes used for making bread, so that these products are much larger, better colors, more soft particles.Even today, baking industry have been α-amylase from barley malt and bacterial, fungal leaf extract. Since 1955 and after 1963 in the UK GRAS level validation, fungal amylase, has served as a bread additive. Now, they are used in different areas. Modern continuous baking process, add in f lour α-amylase can not only increase the fermentation rate and reduce dough viscosity (improving product volume and texture) to increase the sugar content in the dough, improved bread texture, skin color and baking quality, but also to extend the preservation time for baked goods. In the storage process, the bread particles become dry, hard, not crisp skin, resulting in deterioration of the taste of bread. These changes collectively referred to as degenerate. Each year simply because the losses caused by deterioration of bread more than 100 million U.S. dollars. A variety of traditional food additives are used to prevent deterioration and improve the texture and taste of baked goods. Recently, people started to pay attention enzyme as a preservative, preservative agent in improving the role of the dough, as amylopectin, amylase enzyme and a match can be effectively used as a preservative. However, excessive amylase causes a sticky bread too. Therefore, the recent trend is the use of temperature stability (ITS) α a amylase activity are high in starch liquefaction, but the baking process is completed before the inactivation. Despite the large number of microbes have been found to produce α-amylase, but with the temperature stability of the nature of the α-amylase only been found in several microorganisms.1.2 starch liquefaction and saccharification of the main α-amylase starch hydrolysis product market, such as glucose and fructose. Starch is converted into high fructose corn syrup (HFCS). Because of their high sweetness, are used in the soft drink beverage industry sweeteners. The liquefaction process is used in thermal stability at high temperature α-amylase. α-amylase in starch liquefaction ofthe application process is already quite mature, and many relevant reports.1.3 fiber desizing modern fiber manufacturing process in knitting yarn in the process will produce large amounts of bacteria, to prevent these yarn faults, often increase in the surface layer of the yarn can remove the protective layer. The surface layer of the material there are many, starch is a very good choice because it is cheap and easy to obtain, and can be easily removed. Starch desizing α-amylase can be used, it can selectively remove the starch without harming the yarn fibers, but also random degradation of starch dextrin soluble in water, and are easily washed off. 1.4 Paper Industry amylase used in the paper industry mainly to improve the paper coating starch. Paste on the paper is primarily to protect the paper in the process from mechanical damage, it also improved the quality of finished paper. Paste to improve the hardness and strength of paper, enhanced erasable paper, and is a good paper coating. When the paper through two rolls, the starch slurry is added the paper. The process temperature was controlled at 45 ~ 6O ℃, need a stable viscosity of starch. Grinding can also be controlled according to different grades of paper starch viscosity. Nature of the starch concentration is too high for the sizing of paper, you can use part of α-amylase degradation of starch to adjust.1.5 Application of detergents in the enzyme is a component of modern high-efficiency detergents. Enzymes in detergents in the most important function is to make detergents more modest sound. Automatic dishwasher detergents early is very rough, easy to eat when the body hurts, and on ceramics, wood tableware can also cause damage. α-amylase was used from 1975 to washing powder. Now, 90% of the liquid detergents contain an amylase, and automatic dishwasher detergents α-amylase on demand is also growing. α 1 amylase ca2 + is too sensitive to low ca2 + in the stability of poor environment, which limits an amylase in the remover in. And, most of the wild-type strains produced an amylase on raw materials as one of the oxidants detergents are too sensitive. Keep household detergents, this limitation by increasing the number of process steps can be improved. Recently, two major manufacturers of detergents NovozymesandGcncncoreInternational enzyme protein technology has been used to improve the stability of amylase bleaching. They leucine substitution of Bacillus licheniformis α-amylase protein in the first 197 on the methionine, resulting in enzymes of the oxidant component of resistance increased greatly enhanced the oxidation stability of the enzyme stability during storage better. The two companies have been pushing in the market these new products.1.6 Pharmaceutical and clinical chemical analysis with the continuous development of biological engineering, the application of amylase involved in many other areas, such as clinical, pharmaceutical and analytical chemistry. Have been reported, based on the liquid α-amylase stability of reagents have been applied to automatic biochemical analyzer (CibaComingExpress) clinical chemistry system. Amylase has been established by means of a method of detecting a higher content of oligosaccharides, is said this method is more than the effective detection method of silver nitrate.2.1 Research amylase α-amylase enzymes in domestic production and application in 1965, China began to apply for a 7658 BF Bacillus amyloliquefaciens amylase production of one, when only exclusive manufacturing plant in Wuxi Enzyme. 1967 Hangzhou Yi sugar to achieve the application of α-amylase production of caramel new technology can save 7% ~ 10% malt, sugar, increase the rate of 10%.1964, China began a process of enzymatic hydrolysis of starch production of glucose. In September l979 injection of glucose by the enzyme and identification of new technology and worked in North China Pharmaceutical Factory, Hebei Dongfeng Pharmaceutical Factory, Zhengzhou Songshan applied pharmaceutical units and achieved good economic benefits. Compared with the traditional acid to improve the yield of 10% Oh, cost more than 15%. In addition to enzyme for citric acid production in China, glutamic acid fermentation system for beer saccharification, fermentation, rice wine, soy sauce manufacture, vinegar production also has been studied and put into production successfully.2.2 Overseas Researc h α-amylase, present, and in addition a large number of T for conventional mutation breeding, the overseas production has been initially figure out the regulation of α-amylase gene, the transduction of the transformation and gene cloning techniques such as breeding. The Bacillus subtilis recombinant gene into the production strain to increase α-amylase yield of 7 to 10 times and has been used in food and the wine industry, for breeding high-yield strains of α-amylase to create a new way.2.3 domestic and foreign research institutions and major research direction as α-amylase is an important value of industrial enzymes, weekly discussion group and outside it was a lot of research. Representative of the domestic units: Sichuan University, major research produc tion of α-amylase strains and culture conditions; Jiangnan University, the main research structure of α-amylase and application performance, such as heat resistance, acid resistance; Northwest universities, major research denatured α-amylase and the environment on the mechanism of α-amylase; South China University of Technology, the main α-amylase of immobilization and dynamic nature; there Huazhong Agricultural University, Chinese Academy of Sciences Institute of Applied Ecology in Shenyang, Tianjin University, Nankai University, College of Life Sciences, Chinese Academy of Agricultural Sciences, Chinese Academy of Sciences Institute of Microbiology and a number of research institutions on a variety of bacterial α-amylase production of a amylase gene cloning and expression. Representative of foreign research units are: Canada UniversityofBritishColumbia, they were a pancreatic amylase structure and mechanism of in-depth research; Denmark's Carlsberg Research Laboratory of the main structure of barley α-amylase domain and binding sites; U.S. WesternRegionalResearchCenter major study α-amylase in barley and the role of antibiotics and the barley α-amylase active site.3, α-amylase conclusion has become the industrial application of one of the most important enzymes, and a large number of micro-organisms can be used for efficient production of amylase, but large-scale commercial production of the enzyme is still limited to some specific fungi and bacteria. For the effective demandfor α-amylase more and more, this enzyme by chemical modification of existing or improved technology through the white matter are. Benefit from the development of modern biotechnology, α an amylase in the pharmaceutical aspects of growing importance. Of course, the food and starch indust ries is still the main market, α amylase in these areas, a demand is still the largest.Journal of Southeast University(English Edition)2008 24（4）。