对3种虫生真菌菌丝进行游离氨基酸的分析,了解其游离氨基酸的种类及含量,为进一步食用、药用的研究和开发应用提供信息和技术资料.采用安捷伦1200 HPLC C18反相柱梯度进行分离,邻苯二甲醛-芴甲酯(OPA-FMOC)柱前自动衍生化,利用已知浓度的氨基酸标准品的峰面积与样品峰面积之比计算出游离氨基酸的含量.结果表明,蝙蝠蛾拟青霉(Paecilomyces hepiah)、中国弯颈霉(Tolypocladium sinense)、中国被毛孢(Hirsutella sinensis)分加含20、23、22种游离氨基酸.3种真菌菌丝的游离氨基酸含量和种类不同,各有特异性,但有互补作用,游离氨基酸总量相差较大,在食品药品开发应用中建议配合使用.

了解不同地区的冬虫夏草种内的遗传分化及变异情况,进行18S rDNA ITS区的序列分析,以期为它们的差别建立分子标记,也为不同地区来源的商品冬虫夏草的鉴定提供分子依据.冬虫夏草样品应用DNA提取试剂盒所提供的方法(CTAB法)提取基因组DNA,1％琼脂糖凝胶电泳检测DNA纯度,DNA经PCR扩增,PCR扩增产物在DNA全自动测序仪上进行序列测定.测序结果获得6个不同地域的冬虫夏草18S rDNAiTS区基因序列图谱和GC含量(％).实验结果表明,6个不同地区的冬虫夏草基因序列,反映了冬虫夏草种内的遗传分化和变异,其遗传分化与变异的程度与海拔高度和纬度呈正相关性,这些分化可应用于不同地区冬虫夏草的鉴定.

Objective To analyze the contents of amino acids in Paecilomyces hepiali (P.hepiali) and Tolypocladium sinenis (T.sinensis) and their nutritive values,so as to provide a basis for related researches and applications of the two kinds of fungi.Methods The mycelia of P.hepiali and T.sinensis were hydrolyzed with 6 mol · L-1 HC1,and the amino acids from the mycelia were separated by C18 reversed-phase column with gradient elution and o-phthalaldehyde-fluorenylmethyl chloroformate (OPA-FMOC) pre-column derivation using Agilent 1200 high-performance liquid chromatograph.The content of sample amino acids was determined by the peak areas ratio between the samples and standard amino acids.Results A total of 17 proteolytic amino acids from P.hepiali and T.sinensis were determined in this study.The contents of 4 kinds of amino acids (aspartic acid,valine,isoleucine and leucine) in P.hepiali were higher than those in T.inensis,while the contents of 11 kinds of amino acids (glutamic acid,serine,glycine,threonine,arginine,alanine,tyrosine,cystine,methionine,lysine and proline) in T.inensis were higher than those in P.hepiali.The total contents of amino acids,essential amino acids and non-essential amino acids in T.Sinensis were higher than those in P.hepiali,and the content of branched chain amino acids in P.hepiali was higher that in T.sinensis.World Health Organization/Food and Agriculture Organization (FAO/WHO) reference model of essential amino acids was used to evaluate the nutritional values of amino acids in P.hepiali and T.sinensis,and the results showed that the scores of ratio coefficient of amino acids in P.hepiali and T.sinensis were 92.06 and 88.17,respectively.Conclusion The contents of protein amino acids in P.hepiali and T.sinensis are different,but they both have relatively high nutritional values.

Objective To establish a method of microbial limit tests for compound Hongjingtian oral liquid and to per-form the validation of the method.Methods The validation on the microbial limit test method was conducted according to the counting method of bacteria,mycetes and yeasts and the control bacteria test method stated in the appendix of China pharmaco-poeia (2010 edition).Results The recoveries were over 70% when culture diluting method was used for staphylococcus aureus and routine test method was used for Escherichiacoli,Bacillus subtilis,Candida albicans and Aspergillus niger.In the test of positive control bacteria,the growth was detected,while the negative control was no growth detected.Conclusion The micro-bial limit test for compound Hongjingtian oral liquid is validated in this study.The amount of staphylococcus aureus can be de-termined using diluted culture method ;while the amounts of Escherichiacoli,Bacillus subtilis,Candida albicans,Aspergillus niger can be determined by conventional method.

目的 建立高效液相色谱法(HPLC)测定冬虫夏草中虫草素含量的方法.方法 色谱柱为Poroshell 120SB-C8柱(4.6 mm×250 mm,5 μm);流动相为乙腈-水系统,梯度洗脱,流速1.0 mL/min;检测波长260nm;柱温35℃;进样量10μL.结果 虫草素检测的浓度范围为0.0476～9.520 μg/mL,r=1.0000,线性关系良好.冬虫夏草中虫草素的平均加样回收率为93.26％,RSD为2.07％.结论 方法简便可靠,精密度和回收率良好,可为提高冬虫夏草的质量标准提供参考和依据.

冬虫夏草是冬虫夏草菌（麦角菌科真菌）寄生在蝙蝠蛾科昆虫幼虫上的子座和幼虫尸体的复合体。在我国传统医学中，冬虫夏草以其调节阴阳、肺肾双补之特效，广泛用于免疫、心血管、呼吸、泌尿生殖、消化、神经系统等疾病的治疗。

Objective To investigate the inhibitive effect of total alkaloids from Pinellia ternate( TAPT) on the proliferation of human hepatic carcinoma strain QJY-7703. Methods The proliferation inhibition of human hepatic carcinoma cells treated with TAPT was measured with methyl thiazolil tetracolium colorimetic( MTT) method and plate clone formation assay to detect the inhibitive effect of different concentrations of TAPT on the growth of QJY-7703 strains. Single cell gel electrophoresis( SCGE) was used to detect QJY-7703 cell DNA damage induced by TAPT. Results When compared with the control group,in vitro proliferation of QJY-7703 cells treated with TAPT was decreased significantly,and it was positively correlated with drug dosage and medication time,and statistical significance could be seen,when comparisons were made( P 0. 01,P 0. 05). In SCGE detection,significant differences could be seen in cell tail DNA content,tail length and tail movement in the TAPT experimental group,when compared with those of the control group( P 0. 01),and a dose-dependent relationship was quite marked. Conclusion In vitro,a certain dosage of TAPT could obviously inhibit the proliferation of QJY-7703 cells,the mechanism of which might be correlated to DNA damage induced by TAPT.

目的 对天然冬虫夏草、冬虫夏草菌丝及发酵液样品进行24种游离氨基酸的分析;进一步了解三者游离氨基酸的种类和含量,为它们的应用研究提供可用的信息及参考.方法 采用安捷伦1200 HPLC,C18反相柱梯度分离,邻苯二甲醛-芴甲酯(OPA-FMOC)柱前自动衍生化,利用已知浓度氨基酸标准品进行外标法定量,根据已知浓度氨基酸标准品的峰面积与样品峰面积之比计算出游离氨基酸的含量.结果 分析结果表明在24种游离氨基酸的测定中,天然冬虫夏草含有19种,冬虫夏草菌丝含22种,发酵液含23种,三者所缺少的游离氨基酸种类互不相同.三者游离氨基酸的含量有着明显差别和变化,其原因是由于天然冬虫夏草是在野外天然生长,冬虫夏草菌丝和发酵液是人工发酵培养,处于不同的温度和环境条件的原故.结论 天然冬虫夏草,冬虫夏草菌丝及发酵液的游离氨基酸的种类和含量并不等同,建议临床配合使用.

Objective: In order to research on the active ingredient and medicinal value of Cordyceps sinensis,free amino acids and trace elements of C.sinensis oral solution were determined.Methods: A C18 column was used with gradient elution by RP-HPLC and OPA-FMOC pre-column derivatization.The external standard method was used to quantify the amino acids by the peak areas ratio between the sample and the standard.Results: The anal ysis showed that C.sinensis oral solution contained 23 kinds of free amino acids and 18 kinds of trace elements.The total amount of amino acid in the oral solution was 0.892%,and the essential amino acids to total amino ac ids was 53.1%.Content of lysine was the most high,followed by leucine and arginine,among all kinds of amino acids in the oral solution.Conclusion: Free amino acids are bioactive ingredients of C.sinensis oral solution,which supply basic data for the efficacy of C.sinensis oral solution.

目的研究冬虫夏草子实体提取物(CSE)对小鼠抗疲劳和耐缺氧能力的影响。方法连续给予小鼠灌胃0.25 g.kg-1、0.50 g.kg-1、1.00 g.kg-1冬虫夏草子实体7 d,进行游泳实验和耐缺氧实验。观察小鼠的常压耐缺氧、亚硝酸钠中毒、急性脑缺血性缺氧存活时间和负重游泳时间,以及90 min游泳运动后全血乳酸、血尿素氮和肝糖元含量的变化。结果冬虫夏草子实体提取物可显著延长小鼠耐缺氧存活时间(P<0.01),延长负重游泳时间达2.95倍,使游泳后小鼠的全血乳酸含量下降51.18%,血尿素氮含量降低19.50%,而肝糖元含量增加28.95%。结论冬虫夏草子实体提取物具有提高小鼠耐缺氧能力和抗疲劳的作用。

Objective To study the protective effects of planting Dendrobii caulis and tissue culture planting Dendrobium officinale Kimura et Migo on glutamate damage induced in cultured SH-SYSY cells.Methods The sera were obtained from rats by using the serum pharmacology method,and cell damage model was established by treating SH-SY5Y cells with glutamate.the proliferation of damaged cells were observed by 72 h.MTT assay was used to test lactate dehydrogenase(LDH),malondialde-1yde(MDA)and superoxide dismutase(SOD) in the culture medium.Results Both planting and tissue culture Dendrobii caulis could promote the cell proliferation of glutamate damaged SH-SY5Y cells.They also could decrease LDH and MDA content and increase SOD level significantly. Tissue cultured Dendrobii caulis preceded planting ones in antioxidant potential.Conclusion Dendrobii caulis had protective effect on glutamate damaged SH-SY5Y cells.The protective effect of tissue culture one,while,was more distinctive than planting one,which could prove that officinal effect of tissue culture Dendrobium huoshanense was preceded to Dendrobium plants.

目的研制一种适合舰船环境条件下使用的伤病员生命体征支持与监护系统,满足伤病员现场急救和转运的需要。方法采用单片机技术、射流技术、微电子技术、电磁兼容技术、综合集成技术,将生命支持功能和生命体征监护功能有机结合。结果该系统在对伤病员进行心电、血氧、体温、脉率、血压监护功能的同时根据需要可进行间歇正压供氧和加压输液,并能够对生命体征数据进行存储、回放,对异常数据进行判断和报警。结论该系统体积小,操作简单,智能化程度高,适合船用的同时,兼顾车载和机载,在伤病员现场急救和转运过程中起到至关重要的作用。

据统计,尽管我国每年有上万项成果获得省部级以上科技奖励,但成果转化率仅为25%左右,真正能够实现商品化的不足15%(7.2%～14.8%)[1],与发达国家50%以上的转化率存在着巨大的差距,军队医学科技成果转化率更低,在一定程度上造成军队医学科研成果的"浪费"和"贬值".