洱海水华铜绿微囊藻生长特性的初步研究

目的：分离纯化出洱海主要的优势铜绿微囊藻并观察其生长规律及特性。方法：采用水滴-涂布平板法分离纯化铜绿微囊藻,采用90%的丙酮提取叶绿素,酶标仪测定其含量,用显微镜计数藻细胞的个数。结果：铜绿微囊藻细胞密度与OD665呈直线关系,R2=0.993 1,藻细胞的增长速率在第6天达到最大值为0.524,其叶绿素及类胡萝卜素的含量随时间的变化而逐步升高。结论：此铜绿微囊藻的生长符合＂S＂型曲线,藻细胞密度与吸光度成线性相关,其生长速率在第6天达到最大。     

稳恒磁场对人白血病细胞K562生长抑制作用研究初报

人白血病细胞K5 6 2经3,6 ,9,12 ,2 4 ,36 ,4 8,6 0 ,72h不同时间磁场处理后,利用MTT法检测其生长活性,台盼蓝拒染法进行活细胞计数.结果表明,K5 6 2细胞经9mT稳恒磁场处理0～72h后,从12h起磁场对细胞生长有显著的抑制效应(p <0 .0 5 ) .K5 6 2细胞生长曲线结果与MTT实验具有良好的一致性.     

鞣花酸对前列腺癌PC-3细胞生长和凋亡的影响

目的观察鞣花酸对前列腺癌PC-3细胞株的影响。方法：体外培养人前列腺癌PC-3细胞,加入0μg/ml、2．5μg/ml、5μg/ml、10μg/ml、20μg/ml的鞣花酸作用于PC-3细胞,分别作用12小时、24小时和48小时后,应用MTr法测定各浓度组鞣花酸对PC-3细胞的生长抑制作用。应用流式细胞仪检测鞣花酸作用48小时后,PC-3细胞的周期时相变化及凋亡情况。应用免疫细胞化学检测10μg/ml鞣花酸作用24小时后,实验组与对照组细胞Caspase-3蛋白表达情况。结果：鞣花酸明显抑制PC-3细胞的生长,抑制效应呈时间依赖型和浓度依赖型,与对照组比较均有统计学意义（P〈0．05）。应用流式细胞仪检测结果显示,鞣花酸可将PC-3细胞阻滞于G1／S期,并诱导PC-3细胞凋亡。免疫细胞化学结果显示实验组Ctmpase-3蛋白表达明显升高。结论：鞣花酸对前列腺癌PC-3细胞株有明显的生长抑制和诱导凋亡作用。     

蛋白核小球藻和四尾栅藻处理农村生活污水应用分析

目的:比较分析蛋白核小球藻和四尾栅藻在不同污水浓度下的生长及对污水中氮、磷的去除能力.方法:采用人工模拟农村生活污水高、低共两个污染浓度梯度,分别投放蛋白核小球藻和四尾栅藻,分析藻类生物量、水体环境指标参数以及N、P含量的动态变化.结果:蛋白核小球藻投放的水体中,光密度在前期增长幅度小而后快速增加,而四尾栅藻前期增加迅速,而后增长速度变缓.蛋白核小球藻在低浓度条件下,第0～4天的叶绿素A含量迅速增加后含量趋于稳定,而四尾栅藻在0～2d含量快速增加,而后稳定并缓慢下降.高浓度下,蛋白核小球藻投放2d后叶绿素A含量迅速增长,四尾栅藻投放0～2d增长迅速,而后稳定.与对照组比,投放藻类的水体,溶解氧含量(DO和pH值都有升高趋势.对照随处理时间的延长,水体中N、P有明显减少趋势.在低浓度污水条件下,两种藻类去除总氮和氨态氮效果差异不大,而在高浓度条件下,四尾栅藻对总氮去除率较高,最终去除率达88.7％.蛋白核小球藻对总磷去除优势明显,在低浓度和高浓度负荷下最终去除率分别达96.0％、73.6％.结论:蛋白核小球藻和四尾栅藻在2种污水浓度下生长状况良好,且2种绿藻均改善了水体环境,而水体环境变化对氮磷去除具有重要意义.蛋白核小球藻去除总磷效果优于四尾栅藻,而四尾栅藻在高浓度条件下对总氮和氨态氮去除效率更高.     

He—Ne激光诱导HL-60细胞凋亡

探讨He—Ne激光幅照诱导肿瘤细胞凋亡而抑制肿瘤的生长。采用He—Ne激光处理HL-60细胞,流式细胞仪检测DNA含量及细胞凋亡百分率。发现在57．24J／cm^2的He—Ne激光作用下,HL-60细胞的凋亡百分率为12．83％。同时细胞周期发生改变,G1期减少,S期降低,G2期增加。结果表明在一定剂量He—Ne激光照射下,导致细胞DNA断裂,诱导HL-60细胞凋亡并抑制肿瘤细胞生长。     

盐酸氨溴索联合阿米卡星对铜绿假单胞菌成熟生物膜的体外影响

目的通过在体外复制铜绿假单胞菌（P．a）的生物膜模型,研究盐酸氨溴索与阿米卡星对铜绿假单胞菌生物膜的影响。方法以聚氯乙烯吸痰管作为生物膜载体制作铜绿假单胞菌体外生物膜模型,分别在生物膜形成的不同时间加入3MIC的阿米卡星及不同浓度的盐酸氨溴索,采用连续稀释法进行生物膜内存活菌菌落计数。结果与单独应用阿米卡星组相比,阿米卡星与盐酸氨溴索合用后,生物膜中的活菌数明显减少。成熟生物膜中的活菌数随盐酸氨溴索浓度的增高而减少。结论盐酸氨溴索能增强阿米卡星对生物膜内铜绿假单胞菌的清除作用。     

基于CCS的粘连细胞分离计数实验设计

本文在CCS基础上设计了粘连细胞分离计数实验,实验用TMS320VC5509作为硬件平台,用CCS建立工程,在图像灰度化、二值化、滤波等预处理之后,进行腐蚀运算,从而将粘连细胞分离,并在此基础上用区域标记的方法对细胞计数,最终实现对细胞数目的统计。经过实验,计数结果和人工计数基本一致,说明本实验对粘连细胞进行分离和计数是可行的。     

用高倍镜观察线粒体实验的建议

线粒体是细胞中比较大的细胞器,常见的有短棒状和圆球状,直径大多0．5～1．0μm,长度一般1．0～3．0μm,因此光镜下可见。在光镜下观察,通常采用健那绿B（Janus green B）染液对线粒体进行活细胞染色,     

安徽省3种药用菊花总黄酮含量比较

比较安徽省3种药用菊花总黄酮含量差异,为药用菊花应用开发提供依据。以芦丁为标准品,检测波长为506nm,采用紫外一可见分光光度法测定总黄酮含量。结果表明,总黄酮在0mg／L-100mg／L与吸光值呈良好线性关系,回归方程A=0．0133C-0．0131,R^2=0．9993。3种药用菊花总黄酮含量滁菊〉贡菊〉亳菊。     

包虫囊液对体外培养小鼠NK细胞影响的研究

采用流式细胞仪检测NK细胞活性受体NKG2D的表达变化,乳酸脱氢酶细胞毒性检测法(LDH)分析脾细胞的杀伤活性。通过细粒棘球蚴囊液对体外BABL/c小鼠NK细胞的检测结果的分析讨论,得到细粒棘球蚴囊液可降低NKG2D的表达水平,进而使NK细胞杀伤活性降低,从而得到有利于包虫免疫逃逸的结论。     

L-苯丙氨酸对铜绿微囊藻生长的作用与机制

应用细胞计数法结合流式细胞术探讨了L-苯丙氨酸对铜绿微囊藻生长的作用与可能机制.结果表明:作用24 h后,较低浓度L-苯丙氨酸(0.078 ～0.312 μg/mL)对藻细胞生长表现为促进作用,而在较高浓度(0.625 ～ 20μg/mL)时表现为抑制作用.L-苯丙氨酸对铜绿微囊藻生长的抑制作用存在剂量效应和时间效应关...     

Magnetic cell sorting and flow cytometry sorting methods for the isolation and function analysis of mouse CD4＋ CD25＋ Treg cells

Objective: In this paper we compared the two methods of cell sorting (magnetic cell sorting and flow cytometry sorting) for the isolation and function analysis of mouse CD4~+ CD25~+ regulatory T (Treg) cells, in order to inform further studies in Treg cell function. Methods: We separately used magnetic cell sorting and flow cytometry sorting to identify CD4~+ CD25~+ Treg cells. After magnetic cell separation, we further used flow cytometry to analyze the purity of CD4~+ CD25~+ Treg cells, trypan blue staining to detect cell viability, and propidium iodide (PI) staining to assess the cell viability. We detected the immune inhibition of CD4~+ CD25~+ Treg cells in the in vitro proliferation experiments. Results: The results showed that compared to flow cytometry sorting, magnetic cell sorting took more time and effort, but fewer live cells were obtained than with flow cytometry sorting. The CD4~+ CD25~+ Treg cells, however, obtained with both methods have similar immunosuppressive capacities. Conclusion: The result suggests that both methods can be used in isolating CD4~+ CD25~+ Treg cells, and one can select the best method according to specific needs and availability of the methodologies.     

NDV感染体外诱导胃癌细胞BGC-823的凋亡

目的 :研究新城疫病毒在体外抗胃癌细胞活性及其与细胞凋亡的关系。方法 :应用倒置显微镜观察细胞形态、MTT法测NDV在体外对BGC - 82 3的抑制和杀伤作用 ,同时用流式细胞术检测胃癌细胞凋亡情况及细胞分裂周期各时象的变化。结果 :NDV在体外可使BGC - 82 3胃癌细胞形成明显的细胞病变效应、细胞生长抑制及细胞凋亡 ,且细胞凋亡率与感染时间呈正相关。G2、S期细胞减少 ,增殖指数 (PI)降低 ,与阴性对照组比较有显著性差异 (P<0 .0 1)。结论 :NDV具有显著的抗BGC - 82 3胃癌细胞活性。     

黄芪成分F_3新制剂对胃癌细胞株的抑制作用

[目的]观察黄芪成分F3新制剂对胃癌(BGC-823)细胞株体外细胞生长、抑制作用.[方法]利用基因组DNA电泳,流式细胞仪检测细胞凋亡.[结果]基因组DNA电泳出现“梯形”条带;流式细胞仪分析出现低于G1期DNA含量的亚二倍体凋亡率.[结论]黄芪成分F3新制剂对胃癌细胞有明显的抑制作用.     

川芎嗪对人乳腺癌MCF-7/ADM细胞多药耐药性的逆转及P-糖蛋白表达的影响

[目的]探讨川芎嗪(TMP)对人乳腺癌MCF-7/ADM细胞的耐药逆转及其对该细胞P-糖蛋白(P-gp)表达的影响.[方法]MTT法测定细胞的药敏性和抗药性逆转,流式细胞术检测耐药细胞P-糖蛋白的表达.[结果]非细胞毒性剂量(320μg/m l)和低毒剂量(1 250μg/m l)的川芎嗪均能显著降低MCF-7/ADM细胞的IC50(P<0.01),逆转倍数分别为2.14和2.80倍;并使该细胞P-gp的表达率由(90.60±0.40)%分别降低至(69.10±1.65)%和(60.30±1.25)%.[结论]川芎嗪可部分逆转人乳腺癌MCF-7/ADM细胞对阿霉素的耐药性,其逆转机制与抑制该细胞P-gp的表达有关.     

Artesunate inhibits growth and induces apoptosis in human osteosarcoma HOS cell line in vitro and in vivo

This paper aims to investigate the effects of artesunate (ART) on growth and apoptosis in human osteosarcoma HOS cell line in vitro and in vivo and to explore the possible underlying mechanisms. Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The induction of apoptosis was detected by light and transmission electron microscopy and flow cytometry. Western blot analysis was used to investigate the related mechanisms. Nude mice were further employed to investigate the antitumour activity of ART in vivo. MTT assay results demonstrated that ART selectively inhibits the growth of HOS cells in a dose- and time-dependent manner. Based on the findings of light and transmission electron microscopy, Hoechst 33258 staining, and fluorescein isothiocyanate (FITC)-annexin V staining, the cytotoxicity of ART in HOS cells occurs through apoptosis. With ART treatment, cytosolic cytochrome c was increased, Bax expression was gradually upregulated, Bcl-2 expression was downregulated, and caspase-9 and caspase-3 were activated. Thus, the intrinsic apoptotic pathway may be involved in ART-induced apoptosis. Cell cycle analysis by flow cytometry indicated that ART may induce cell cycle arrest at G₂/M phase. In nude mice bearing HOS xenograft tumours, ART inhibited tumour growth and regulated the expressions of cleaved caspase-3 and survivin, in agreement with in vitro observations. ART has a selective antitumour activity against human osteosarcoma HOS cells, which may be related to its effects on induction of apoptosis via the intrinsic pathway. The results suggest that ART is a promising candidate for the treatment of osteosarcoma.     

Berbamine induces apoptosis in human hepatoma cell line SMMC7721 by loss in mitochondrial transmembrane potential and caspase activation

Objective： To investigate the effect ofberbamine on human hepatoma cell line SMMC7721. Methods： The effects of 24 h and 48 h incubation with different concentrations （0-64 μg/ml） of the berbamine on SMMC7721 cells were evaluated using 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide （MTT） assay. Hoechst 33258 staining was conducted to distinguish the apoptotic cell, and the appearance of sub-G1 stage was determined by PI （propidium iodide） staining, the percentage of apoptotic cell was determined by flow cytometry following annexin V/PI staining. Flow cytometry was performed to analyze the cell cycle distribution and the mitochondrial membrane potential （△ψm）, the expression of activated caspase3 and caspase9 was analyzed by Western-blot. Results： The proliferation of SMMC7721 was decreased after treatment with berbamine in a dose- and time-dependent manner. Berbamine could induce apoptosis in SMMC7721 cells and could cause cell cycle arrest in G0/G1 phase, to induce loss of mitochondrial membrane potential （AVm） and activate caspase3 and caspase9. Berbamine-induced apoptosis could be blocked by the broad caspase inhibitor z-VAD-fmk. Conclusion： Berbamine exerts antiproliferative effects on human hepatocellular carcinoma SMMC7721 cells. The anticancer activity of berbamine could be attributed partly to its inhibition of cell proliferation and induction of apoptosis in cancer cells through loss in mitochondrial transmembrane potential and caspase activation.     

胃癌细胞SGC7901体外生长条件的优化研究

以胃癌细胞SGC7901为研究对象,用流式细胞仪和细胞总DNA分析技术讨论了不同培养基和不同保存条件对胃癌细胞体外生长的影响。     

人白血病细胞系KG1a中P2X7受体的表达和功能研究

用半定量RT-PCR和流式细胞术，研究了白血病细胞系KG1a中P2X7受体在基因和蛋白水平的表达。用荧光分光光度计，测定了用激动剂三磷酸腺苷（ATP）和苯甲酰苯甲酸ATP（BzATP）刺激前后细胞内钙离子浓度的变化，证明P2X7受体的功能。结果表明：KG1a细胞表达P2X7受体，且在激动剂的刺激下能引起KG1a细胞通过P2X7受体的胞外钙内流；去除胞外钙离子时，激动剂不能引起胞内钙浓度的升高；提示KG1a细胞表达P2X7受体的基因和功能蛋白，激活该受体引起胞外钙离子的内流。