不同组织来源透明质酸的分离纯化及其鉴定

以猪牛眼玻璃体、鸡冠、人脐带为原料,利用水浸提,乙醇沉淀的方法,提取透明质酸粗品,经ＤＥＡＥ－纤维素纯化,获得高纯度的透明质酸．经测定,其蛋白含量０．３８％～０．６３％,分子量７．９×１０~４～４．７×１０~５,收率０．６４％～２．４％,红外扫描图谱与文献报道一致     

巴拉斯自鹤芋的组织培养和快速繁殖

１植物名称巴拉斯白鹤芋（Ｓｐａｔｈｉｐｈｙｌｌｕｍｐａｌａｓ）。２材料类别小苗茎尖或茎段。３培养条件以ＭＳ为基本培养基：（１）诱导分化与生长培养基为ＭＳ＋ＢＡ２ｍｐ·Ｌ－１（单位下同）＋ＮＡＡ０．２＋３％蔗糖;（２）生根培养基为１／２ＭＳ＋ＩＢＡ０．２＋１．５％蔗糖。以上培养基均加０．７５％琼脂,ｐＨ５．８,培养温度２６～２８℃,每日光照１０～１２ｈ,光照度１５００～２５００ｌｘ。４生长与分化情况剪取顶芽或茎段约０．５～１．０ｃｍ长,用７０％～７５％乙醇消毒３０～６０ｓ,以０．１％ＨｇＣｌ２溶液…     

不同温湿度组合对甜菜夜蛾生长发育及繁殖力的影响

不同温湿组合对甜菜夜蛾各虫态生长发育和繁殖力有明显的影响,适宜该虫生长发育的温湿度为２６℃与ＲＨ８０％或ＲＨ９４％两个组合．在这种条件下,其卵孵化率、１～５龄幼虫存活率、蛹羽化率、雌虫比率、产卵的雌虫比率和平均产卵量分别为８２．６３％～８３．７０％、８１．９０％～８２．４７％、８８．００％～８８．０３％、９０．４０％～９１．００％、９３．３６％～９４．１７％、９５．９７％～９６．２６％、９２．７７％～９４．１６％、０．４２～０．４３、０．９８和６０５～６０６粒／雌,且明显高于其它温湿组合．在温湿度为３２℃与ＲＨ９４％条件下,该虫的内禀增长能力最大（０．２２５）．     

非OI群霍乱弧菌在不同理化环境中产生色素的研究

本文报道了理化因素对非ＯＩ群霍乱弧菌的生长及产生色素的影响。实验结果表明,２５℃～３６℃、０％－３％ＮａＣｌ、ｐＨ７．４～９．０;４２℃、１％～３％ＮａＣｌ、ｐＨ５．０—１０．０为最佳生长环境,适宜生长,并有利于色素的产生。     

镧对马尾松苗的根系生长和IAA含量的影响

在０．１～１．０ｍｇ／Ｌ硝酸镧的Ｈｏａｇｌａｎｄ培养液中,马尾松（ＰｉｎｕｓｍａｓｏｎｉａｎａＬａｍｂ．）截根苗的侧根数目明显增加,为对照的１．７５～３．７５倍;根的鲜重、干重显著提高,增幅达３３．７％～４６．４％;根系还原力也提高１３．５％～２８．５％。根长仅在０．０５～０．５ｍｇ／Ｌ的低浓度下比对照有所增加,在浓度高于１．０ｍｇ／Ｌ时,根长只有对照的５３．３％。侧根内的游离态ＩＡＡ含量随溶液中镧的浓度增加而相应增加,ＩＡＡ氧化酶的活性则相应降低。硝酸镧处理对该苗地上部的促进效应不如根部显著。     

角质蛋白酶固态发酵工艺及酶解条件的研究

曲霉Ａ２８－８是一株优良的角蛋白酶分泌突变菌株。其最佳固体发酵培养基为：２０％羽毛粉,８０％麸皮和微量无机氮（每克培养基中加０．５ｍｇ）;最适产酶条件为：起始ｐＨ７．５～８．０,温度２８℃～３０℃,时间为６０～７０小时,酶活高达２５００ＫＵ／ｇ曲;最适酶解条件为ｐＨ７．０～９．０,温度４５℃～５０℃。     

藏红花愈伤组织诱导及其细胞培养的研究

藏红花幼叶愈伤组织的诱导频率在ＭＳ,Ｂ５和Ｗｈｉｔｅ三种培养基上均高达９８％;幼叶和芽的诱导率差异不大,高达９９％;不同时期的叶片差异较大,以幼叶诱导为佳;球茎诱导率为近８０％;激素配比以２．４－Ｄ２．０ｍｇ／Ｌ,ＢＡＰ０．１～０．５ｍｌ／Ｌ为宜。在继代培养阶段,ＭＳ比Ｂ５和Ｗｈｉｔｅ更适合细胞的快速生长繁殖;并且以叶片的愈伤组织生长较快,芽次之,球茎最慢,适合于细胞生长的激素配比为ＮＡＡ２．０～３．０ｍｇ／Ｌ,ＢＡＰ０．５～１．０ｍｇ／Ｌ为宜。     

黄樟油素新资源植物狭叶桂的研究

狭叶桂〔Ｃｉｎｎａｍｏｍｕｍｈｅｙｅａｎｕｍ（Ｎｅｅｓ）Ｈ．Ｗ．ＬｉｅｔＢ．Ｑ．Ｃｈｅｎｇ〕是一种新的资源植物,全株可提取芳香油,含黄樟油素６３％～９９％;鲜枝叶出油率为０．５１％～０．９６％,黄樟油素含量为９７％～９９％。用种子繁殖可保持母本优良性状,是一种具有发展价值的新香料植物     

广西亚热带主要林型的树干茎流

１９８０～１９９１年对广西亚热带主要林型的树干茎流进行定位观测,结果表明：广西亚热带主要林型的年树干茎流量为６．２～６６．７ｍｍ,分别占林外降雨量０．５～３．３％;不同林型的年树干茎流率显示阔叶林或针阔叶混交林（占１．７～３．３％）大于针叶林（占０．５～１．４％）。树干茎流的大小主要受树皮的吸水性能、树木胸径、树干枝角与冠型结构、降雨量及降雨强度的影响。杉木林树干茎流营养元素的年平均浓度为８．７０７～１４．８４８ｐｐｍ,是林外雨年平均浓度（２．４５３～３．７５３ｐｐｍ）的３．０～６．０倍;是林内穿透雨年平均浓度（５．８６１～１０．４５４ｐｐｍ）的１．４～２．０倍。说明树干茎流具有重要的水文生态功能。     

利用固定化酵母细胞转化反式肉桂酸生产L—苯丙氨酸

研究了深红酵母（Ｒｈｏｄｏｔｏｒｕｌａｒｕｂｒａ）的培养基成分,培养固定化及转化条件,实验表明最佳基成分（％）葡萄糖０．５,胰蛋白胨０．５,酵母膏０．５,磷酸二氢钾０．０５,Ｌ－Ｐｈｅ０．０５,ｐＨ７．０,３０℃２０Ｌ发酵罐中培养１５～１７ｈ,最佳固定化条件为：用２．５％卡拉胶包埋１８％的湿菌体,最佳转化条件为：１．０％反式肉桂酸,４ｍｏｌ／Ｌ铵离子,ｐＨ０．５,３０℃,用卡拉胶固定化深红酵母（Ｒ     

Detection of endometrial cancer by flow cytometry using two monoclonal antibodies.

BACKGROUND: To develop a supplementary diagnostic method for endometrial cancer by measuring the reactivity of various endometrial lesions with two monoclonal antibodies. METHODS: We investigated the reactivity of various endometrial lesions with two monoclonal antibodies (MSN-1 and MSN-3) by flow cytometry (one-color and two-color methods). RESULTS: The two-color method appeared to be suitable for use in place of simultaneous performance of the one-color methods with MSN-1 and MSN-3. The positivity rate for normal endometrium was 16.0% with the two-color method, which was lower than the rate of 30.0% obtained with concomitant used of the one-color methods. The positivity rate for endometrial cancer was high, 84.0%, with the two-color method. The positivity rate was 85.7% for well-differentiated endometrial cancer, 71.4% for moderately differentiated cancer, and 100.0% for poorly differentiated cancer; thus, the rate was high irrespective of the cellular differentiation. CONCLUSIONS: The two-color method is more useful than the one-color method as a supplementary diagnostic procedure for endometrial cancer.     

Synchronization of 9L rat brain tumor cells by centrifugal elutriation

Asynchronous 9L cells were separated into relatively homogeneously-sized populations using centrifugal elutriation with both a conventional collection method and a long collection method. A substantial increase in the homogeneity of the volume distributions and in the degree of synchrony of the separated fractions was obtained using the long collection method. Autoradiographic data indicated that fractions containing greater than or equal to 97% G1 cells, greater than or equal to 80% S cells, and 70-75% G2 cells could be routinely recovered with this procedure. Recovery in these fractions varied from 5 to 8% of the total number of cells elutriated. The colony forming efficiency (CFE) of cells from fractions representing each phase of the cell cycle was a constant 60-70%, which was comparable to the 60-80% usually found for asynchronous 9L cells. The percentage of cells in the G1, S, and G2 phases in the elutriated fractions was more accurately determined from the volume distribution than from computer fits of the DNA histogram obtained from flow cytometry. In genereal, the degree of synchrony was related to the coefficient of variation (CV) of the volume distributions of the elutriated fractions. The CV was about 14% for all elutriated fractions. When the greater than or equal to 97% G1 population was allowed to progress to S and G2, the CVs were about 17 and 20.2%, respectively. Thus, the best nonperturbing method for obtaining synchronous 9L cells in the S or G2 phases was direct elutriation with the long collection method.     

Conventional freezing, straw, and open-pulled straw vitrification of mouse two pronuclear (2-PN) stage embryos

Little is known on the cryopreservation of mouse pronuclear (PN) stage embryos. In the present experiment the mouse 2-PN stage embryos were cryopreserved by conventional freezing, straw, or open-pulled straw (OPS) vitrificaiton methods. The conventional freezing solution was 1.5 mol/L ethylene glycol (EG), and vitrification solutions were EFS30 (30% EG, Ficoll, and sucrose), EFS40 (40% EG, Ficoll, and sucrose), EDFS30 (15% EG, 15%dimethyl sulfoxide [DMSO], Ficoll, and sucrose), or EDFS40 (20% EG, 20%DMSO, Ficoll, and sucrose). The blastocyst rate of 2-PN stage embryos cryopreserved by conventional method (30.4%) was lower than those vitrified by straw method with EDFS (56.9% to 69.1%), by OPS method (66.0% to 85.7%), and that of control (80.8%) (P < 0.05). With a given vitrificaiton solution EFS30, EFS40, EDFS30, or EDFS40, the blastocyst rate of embryos vitrified by the OPS method (66.7%, 66.0%, 85.7%, or 76.9%) was higher than that of those vitrified by the straw method (46.8%, 43.8%, 69.1%, or 56.9%) (P < 0.05). When mouse 2-PN-stage embryos were vitrified with EDFS30 by straw or OPS method, the highest blastocyst rate was achieved (69.1% or 85.7%) and was similar to that of the control, respectively. The embryos transfer results revealed that the full-term development of blastocysts derived from 2-PN stage embryos vitrified by OPS method with EDFS30 (19.9%) was similar to that of the control (23.5%), and higher than that of those cryopreserved by conventional freezing (9.3%) (P < 0.05). The present research demonstrates that the OPS method, especially with EDFS30, is more effective in cryopreserving mouse 2-PN embryos.     

不同炮制方法对斑蝥体内有效物质的影响

在采用不同炮制方法处理3种斑蝥后,测定它们体内的游离斑蝥素和结合斑蝥素的损失率.结果显示:在浅程度炮制方法的处理下,3种斑蝥体内的游离斑蝥素损失率均小于60%,而结合斑蝥素损失率均大于60%;深程度炮制方法的处理下,3种斑蝥体内的游离斑蝥素损失率均小于80%,而结合斑蝥素损失率均大于80%.结合斑蝥素的损失率及损失的量...     

龙须菜多糖脱硫酸化及免疫活性研究

考察了Miller等和Nagasawa等报道的脱硫酸方法对龙须菜多糖硫酸基脱除效果,并比较了脱硫酸前后多糖的免疫活性变化。实验结果显示,采用Miller等报道的方法时,以草酸作为催化剂对硫酸基脱除效果最好,脱除率达到71.4%,远好于其他种类酸催化,但多糖回收率却只有36.4%;采用Nagasawa等报道的方法时,向二甲基亚砜溶液中加入10%甲醇比加入2%吡啶或2%吡啶+2%三甲基氯硅烷具有更好的脱硫酸基效果,硫酸基脱除率达到72.9%,回收率48.9%,是本次实验中效果最好的。免疫活性实验表明,当降低龙须菜多糖硫酸基含量时,免疫活性相应降低。     

Role of modified bleach method in staining of acid-fast bacilli in lymph node aspirates

OBJECTIVE: To correlate acid-fast bacilli (AFB) positivity with cytomorphologic patterns of tuberculous lymphadenitis and evaluate bleach concentration method in diagnosing lymph node tuberculosis compared to Ziehl-Neelsen (ZN) method. STUDY DESIGN: One hundred cases of tuberculous lymphadenitis diagnosed by fine needle aspiration cytology (FNAC) were analyzed and classified into 6 cytomorphologic patterns and correlated with bacillary load using routine and modified bleach methods of ZN staining. Smears were graded for AFB positivity. Sensitivity of routine ZN and modified bleach concentration was compared. RESULTS: The classic cytomorphologic pattern of tuberculosis showing epithelioid granulomas, Langerhans giant cells and caseous necrosis was seen in 23% of cases. Routine ZN staining detected AFB in 27% of cases and the modified bleach method in 72%. In 58 cases the modified bleach method had a higher grade of AFB positivity than the routine method. The modified bleach method did not miss any AFB positivity detected on routine ZN staining. CONCLUSION: The modified bleach method demonstrated AFB positivity in 72% of cases. AFB positivity grade was much higher than with routine ZN staining, making bacilli easily visible, with shorter screening time. The modified bleach method is inexpensive, easily performed and more sensitive and safe than routine ZN staining.     

A HPLC-UV method for the determination of puerarin in rat plasma after intravenous administration of PEGylated puerarin conjugate

A sensitive and reproducible HPLC method for quantitative determination of puerarin (PUE) in rat plasma was developed and validated using 4-hydroxybenzaldehyde as an internal standard. The separation of PUE was performed on a CAPCELL PAK C18 column by gradient elution with 0.2% aqueous phosphoric acid and acetonitrile as the mobile phase. The method was validated and found to be linear in the range of 80-12,000ng/mL. The limit of quantification was 80ng/mL based on 100μL of plasma. The variations for intra- and inter-day precision were less than 8.3%, and the accuracy values were between 98% and 105.2%. The extraction recoveries were more than 85%. The method was successfully applied in the comparative study of pharmacokinetics of PEGylated puerarin (PEG-PUE) versus PUE in rats. Compared with PUE, PEG-PUE showed a 5.2-fold increase in half-life of PUE and a 4.7-fold increase in mean residence time. In addition, this method was also successfully applied to determine the low plasma concentration of PUE regenerated from PEG-PUE in vitro.     

用气相色谱-质谱联用比较牛耳草代谢物的提取方法

通过比较不同的提取方法对牛耳草新鲜和脱水叶片中代谢物的提取效率,旨在建立一种可以有效鉴定并分析牛耳草脱水过程中关键小分子代谢物的种类和含量变化的方法,为研究植物耐脱水分子机制提供技术方法。本研究以气相色谱-质谱联用(GC-MS)为分析方法,对复苏植物牛耳草代谢物提取方法进行比较。从提取总色谱峰数目、提取效率、代谢物保留时间和提取效率稳定性等方面比较甲醇溶液(A法)和甲醇-氯仿-水溶液(B法)两种提取方法的提取效果。对牛耳草新鲜样品提取结果表明,B法提取的总色谱峰数目多于A法;对9种共有代谢物的提取效率比较结果表明,B法的提取效率高于A法;对10种色谱峰的保留时间和提取效率的方法学考察结果表明,两者保留时间RSD(相对标准偏差)值均小于1%,A法提取效率的RSD值≤10%的比例为50%,B法的为100%。A法对干样的提取色谱峰数目远少于鲜样,而B法对干样的提取色谱峰数目和鲜样没有显著差异,保留时间RSD值均小于1%,提取效率的RSD值与鲜样没有差异,稳定性良好。     

大豆卵磷脂的提纯研究

本研究采用简便有效的方法来提纯大豆卵磷脂,产品纯度达到了90％以上。(1)用无机盐沉淀法提取卵磷脂,产品纯度达到82％,并除去主要杂质;(2)用柱层析法精制卵磷脂,纯度91％;(3)纯度鉴定用HPLC法。     

Immuno-PCR法诊断早期梅毒的方法学建立及其应用

目的建立Immuno-PCR法诊断早期梅毒的方法学,评价其灵敏度、特异性、重复性及其临床应用。方法利用基因重组TpN47抗原免疫新西兰兔,制备抗体并用Weston blotting检测;利用抗TpN47抗体作为捕获抗体与血清中TpN47抗原结合,通过链霉亲和素、生物素化抗体、生物素化DNA和PCR扩增等建立Immuno-PCR法检测梅毒螺旋体抗原TpN47体系;评价该方法的灵敏度、特异性和重复性;收集200例临床标本通过Immuno-PCR法、ELISA、TPPA和TURST法进行临床应用比较。结果 Weston blotting结果显示TpN47抗体阳性;Immuno-PCR比ELISA法敏感性强103倍,比TPPA、TURST强105倍;特异性高,重复性好。临床标本中Immuno-PCR法敏感性和特异性分别为86.00%（P〈0.05）和100.00%,ELISA法为71.00%和98.00%,TPPA法为65.00%和100.00%,TRUST法为68.00%和95.00%。结论 Immuno-PCR法检测梅毒螺旋体TpN47抗原敏感性高,特异性强,重复性好,可作为梅毒螺旋体感染的早期诊断方法 。