Glucose-stimulated insulin release by individual pancreatic beta cells: potentiation by glyburide.

To investigate the effect of glyburide on insulin secretion by individual beta cells from normal rats, we employed a reverse hemolytic plaque assay. Pancreata were harvested from female Wistar-Furth rats, the pancreatic islets isolated, and the latter dispersed into single cells. These cells were mixed with protein A-coated ox erythrocytes, the mixture was placed in a Cunningham chamber in the presence of insulin antiserum, and the cells were exposed to the various test substances. Having developed hemolytic plaques around the insulin-secreting cells with complement, the percentage of plaque-forming cells was determined and the plaque areas (reflecting the amount of insulin secreted) were quantitated. For the purpose of validation, we demonstrated that (i) plaque-forming (but not nonplaque-forming) cells could be identified as insulin secreting by an independent immunofluorescent technique, (ii), plaques did not form if insulin antiserum was deleted from the preparation, (iii) plaques failed to develop if insulin antiserum was preabsorbed with insulin, and (iv) incubation with non-protein A-coated RBC or omission of complement resulted in no plaque formation. In addition, both the percentage of plaque-forming cells and the mean plaque are increased upon exposure to glucose (0.75-20 mM) in a concentration-dependent manner at 5- and 60-min incubation times. Moreover, somatostatin suppressed the percentage of plaque-forming cells and diminished the mean plaque area of cells which continued to secrete insulin in response to glucose. Exposure of cells to 100 nM glyburide in the presence of 5 mM or 20 mM glucose had no effect on the percentage of plaque-forming cells present at 5 min or 60 min. Similarly, glyburide did not alter mean plaque area at 5 or 60 min when cells were co-incubated with 5 mM glucose. However, mean plaque area was markedly enhanced at 5 and 60 min in response to glyburide and 20 mM glucose. These results demonstrate that glyburide (i) does appear to enhance insulin secretion by an effect directly on the pancreatic beta cell; (ii) does not act by recruiting previously noninsulin-secreting cells into a secretory pool; (iii) does not potentiate the effect of glucose, at fed concentrations, on insulin secretion by individual cells; but (iv) does augment insulin secretion by beta cells stimulated with supraphysiologic concentrations of glucose.     

不明原因男性不育人群中睾丸特异性乳酸脱氢酶基因突变的发现及其意义

为了研究睾丸特异性乳酸脱氢酶,即乳酸脱氢酶C4(LDH-C4)基因突变在男性不育发病中的作用,利用LDH-C4特异性底物对100名不明原因男性不育症患者的精子LDH-C4进行活性显色,用变性高效液相色谱(DHPLC)技术对LDH-C4活性低下的患者进行LDHC基因PCR产物的突变筛查,对DHPLC峰形异常的PCR产物进行序列测定.筛选到一组精子LDH-C4活性明显下降的患者,其中1名患者的LDHC基因PCR产物在DHPLC中呈异常洗脱峰.对这一PCR产物进行序列测定,发现患者LDHC基因第5外显子的115位碱基发生了T→A的杂合改变(GenBank登录号GU479375),该突变使LDHC基因的178位密码子由原来的TTG(编码亮氨酸)变为TAG(终止密码子),形成截短的C亚基.T克隆-测序进一步证实了该无义突变的杂合状态.这是在人类LDHC基因上发现的第一个突变,提示LDHC基因突变可能是男性不育发病的原因之一.     

汉防己甲素联合顺铂对乳腺癌细胞的作用

目的:汉防己甲素和顺铂联合作用乳腺癌细胞,来提高癌细胞对顺铂的敏感性.方法:运用原子力显微镜对乳腺癌细胞表面的超微结构进行表征;MTr法和流式细胞术检测细胞的生长抑制率和细胞周期变化.结果:顺铂和汉防己甲素分别作用乳腺癌细胞48h IC50值为26.33μmoL/l和5.6μmok/l;联合用药组的IC50值为汉防己甲素2.5μmol/L和顺铂13.32μmol/l,在低浓度的联合用药作用乳腺癌细胞24h细胞膜表面结构被破坏,产生孔洞,作用48h被严重破坏使细胞周期在S比例增加为51.7%±0.30%.结论:提高了癌细胞对抗癌药物的敏感性,联合用药通过改变了细胞膜的结构对癌细胞进行有效杀伤,抑制肿瘤细胞生长.     

浦东滩涂中型土壤动物群落结构及土质酸碱度生物评价分析

1999年,对上海浦东滩涂4类不同酸碱度土壤中的中型土壤动物进行了调查。应用物种丰富度,个体数多度,多样性指数和均匀度4个群落参数,并结合种类研究,讨论了土壤动物群落结构与不同酸碱度土壤的关系。结果表明,土壤中弹尾目和蜱螨目对不同酸碱度土壤反应敏感。弹尾目的3个群落参数和蜱螨目的4个参数均很好地反映与土壤反应敏感。弹尾目的3个群落参数和蜱螨目的4个参数均很好地反映与土壤pH的关系,相关系数分别在0．9以上和0．85左右,在pH相差较大的情况下,可以区分不同酸碱度的土壤。弹尾目的符Tao(Paranura sp.)可用于评价酸碱度较接近的土壤,球角Tao(Hypogastrura sp.）可用于评价酸碱度相差较大,高pH或环境条件较恶劣的土壤。     

三种植物与紫红链霉菌组合灭螺效果

探讨了不同植物与紫红链霉菌组合的灭螺效果,为研制高效低毒植物灭螺剂提供依据。将天名精（Carpesium abrotanoides）、苍耳子（Xanthium sibiricum）、青蒿（Artemisia carvifolia）3种植物水浸液分别与紫红链霉菌液（Streptomyces violaceoruber）组合后对钉螺进行浸杀对比实验,并比较了相同植物与紫红链霉菌液的复合颗粒灭螺剂的灭螺效果。结果表明：天名精、苍耳子与紫红链霉菌复合颗粒灭螺剂7 d钉螺死亡率分别比其1%混合液提高17.6%（P=0.019）和5.3%(P=0.362);青蒿与链霉菌复合颗粒灭螺剂7 d钉螺死亡率比其1%混合液降低6.7%(P=0.022);天名精、苍耳子和链霉菌复合颗粒灭螺剂较其混合液具有一定的增效作用。     

基于稀疏表示算法的蛋白质质谱数据特征选择

高维、小样本数据的特征选择方法在蛋白质质谱数据处理分析领域有着广泛应用。本文针对蛋白质质谱特征选择问题,结合稀疏表示这一新理论框架,提出了一种基于稀疏表示的特征选择算法(sparse representation based feature selection,SRFS)。该方法将稀疏表示分类的结果作为评定某一个特征子空间特征相对重要性的度量,然后通过对大量随机采样子空间计算结果的统计,得到特征空间中每个特征的排序,并进一步分析提炼出与肿瘤疾病相关的若干谱峰。通过在卵巢癌公共数据集OC-WCX2a和浙江省肿瘤医院乳腺癌数据集BC-WCX2a上的实验结果表明,SRFS算法可以有效应用于本文所使用的SELDI-TOF蛋白质质谱数据的分析。     

青冈林土壤跳虫群落结构在落叶分解过程中的变化

1993年5月至1995年4月,用落叶代法研究青冈（Cyclobalanopsis glauca）落叶分解过程中跳虫的群落结构变化。用多样性指数、演替指数、相似系数分析跳虫在落叶分解过程中群落结构及其季节变化特点。青冈落叶分解经淋洗、养分固定 养分活化3个阶段,分解常数分别为k1=9.11,k2=2.57,k3=0.43（百分比／月）。跳中心在落叶分解过程中的集聚型分为3组：A组为落叶分解前期集聚的种类,B组为后期的种类,C组为中期或全过程的种类,分析讨论了落叶分解过程与跳虫功能群及群落结构变化的关系。     

红芽芋球茎片两步法离体快繁及其再生苗生理和光合特性研究

以江西铅山红芽芋(Colocasia esculenta L.Schott var.cormosus‘Hongyayu’)试管苗为材料,建立了芋球茎片两步法离体快繁体系,并对其再生苗的形态指标、染色体数目、生理和光合特性以及叶绿素荧光特性进行了检测。结果表明:(1)红芽芋球茎片单芽诱导的最佳培养基为MS+KT 2 mg/L+6-BA 1 mg/L+NAA0.1mg/L,诱导培养30d后将单芽从球茎片上分离,再接种到生根培养基(MS+KT 2mg/L+NAA 0.1mg/L)上培养30d即可形成完整植株,移栽成活率高达98%;(2)由球茎片单芽、丛生芽、不定芽离体快繁获得的红芽芋再生苗在形态指标、叶下表皮气孔参数、染色体数目、生理生化指标以及叶片光合特性参数和叶绿素荧光特性方面均无显著差异。说明红芽芋球茎片两步法离体培养的再生苗繁殖系数高、染色体数目稳定,该离体快繁体系可应用于江西铅山红芽芋的工厂化生产。     

蔓生百部的化学成分研究

从蔓生百部(Stemona japonica)根中分离得到13个化合物,通过波谱数据,它们鉴定为β-谷甾醇(1)、豆甾醇(2)、5,11-豆甾二烯-3β-醇(3)、苯甲酸(4)、4-甲氧基苯甲酸(5)、1,8-二羟基-3-甲基蒽醌(6)、1,8-二羟基-6-甲氧基-3-甲基蒽醌(7)、氧代狭叶百部碱(8)、百部定碱(9)、异狭叶百部碱(10)、绿原酸(11)、栀子苷(12)和藏红花素A(13).所有化合物均为首次从该植物中分离得到.