黄芪注射液对地塞米松诱导小鼠胸腺细胞凋亡的调节作用

目的:研究黄芪注射液对地塞米松诱导小鼠胸腺细胞凋亡的影响。方法:采用体内腹腔注射地塞米松诱导小鼠胸腺细胞凋亡,观察对小鼠胸腺指数、脾指数的影响和胸腺形态学的改变以及采用体外细胞培养的方法,以MTT法、流式细胞仪分析技术进行观测。结果:地塞米松组(5 mg/mL)出现典型的胸腺萎缩表现,黄芪注射液高剂量组(2000μg/mL)较地塞米松组有一定程度的恢复。结论:黄芪注射液对地塞米松诱导的胸腺细胞凋亡具有一定的抑制作用。

甲状腺肿瘤原位细胞凋亡检测与PTEN表达研究

目的:研究原位细胞凋亡指数(AI)及抑癌蛋白(PTEN)在原发性甲状腺肿瘤中表达及其与病变的关系。方法:对87例原发性甲状腺肿瘤分别应用原位细胞凋亡检测(TuNEL)试剂盒测定AI及应用免疫组化方法检测PTEN表达情况。结果:AI及PTEN蛋白在各甲状腺肿瘤组织均存在表达,AI在腺癌表达高于腺瘤,PTEN在乳头状癌及腺瘤表达显著高于滤泡性癌及未分化癌;AI及PTEN有正相关性。结论:AI及PTEN的表达与甲状腺肿瘤显著相关,检测AI及PTEN水平对于鉴别原发性甲状腺滤泡癌具有重要参考意义。

葛根素对缺氧缺血性脑损伤新生大鼠神经元凋亡的影响

目的:研究葛根素对新生鼠缺氧缺血性脑损伤(HIBD)后神经元凋亡的影响及其神经保护机制。方法:96只新生7d龄SD大鼠随机分为3组:葛根素治疗组、缺氧缺血性脑损伤组(HIBD组)和假手术对照组。按经典方法制作新生大鼠H IBD模型,并于脑缺氧缺血后6h,24h,48h,72h留取脑组织标本。观察各组新生大鼠海马CA 1区神经细胞形态学变化及葛根素对HIBD后海马CA1区神经细胞凋亡的影响。结果:HIBD后6h海马CA1区凋亡细胞增多,24h达高峰,48h阳性细胞开始减少,凋亡细胞多位于病变较重的皮质的边缘内带和海马CA 1区;而假手术对照组仅偶见到阳性细胞;葛根素治疗组细胞凋亡的时程与HIBD组一致,分布极其相似,但海马CA1区的凋亡细胞数在24h后各时间点较HIBD组均明显减少。结论:葛根素可通过抑制神经细胞凋亡而对HIBD后的脑组织起到保护作用。

Iressa抑制胃癌SGC-7901细胞生长的研究

目的：探讨iressa对胃癌SGC.7901细胞增殖，细胞周期和细胞凋亡的影响。方法：用四氮甲唑蓝（MTT）法检测不同浓度iressa对胃癌SGC-7901细胞增殖的影响；用流式细胞仪检测不同浓度iressa对胃癌SGC-7901细胞周期分布和细胞凋亡的影响。结果：在5～20μmol／L浓度范围内，iressa对SGC-7901的生长具有明显的抑制作用，并呈时间和剂量依赖性。10μmol／L和20uol／L的iressa可以导致SGC-7901细胞G1期阻滞，并引起细胞凋亡。结论：Iressa可改变SGC-7901细胞周期分布，诱导细胞凋亡，从而抑制细胞增殖。

Expression of TRAIL and TRAIL receptors in normal and malignant tissues

TRAIL, tumor necrosis factor-related apoptosis-inducing ligand, is a member of the TNF family of proteins.Tumour cells were initially found to have increased sensitivity to TRAIL compared with normal cells, raising hopes thatTRAIL would prove useful as an anti-tumor agent. The production of reliable monoclonal antibodies against TRAIL andits receptors that can stain fixed specimens will allow a thorough analysis of their expression on normal and malignanttissues. Here we report the generation of monoclonal antibodies against TRAIL and its four membrane-bound receptors(TR1–4), which have been used to stain a range of normal and malignant cells, as routinely fixed specimens. Low levelsof TRAIL expression were found to be limited mostly to smooth muscle in lung and spleen as well as glial cells in thecerebellum and follicular cells in the thyroid. Expression of the TRAIL decoy receptors (TR3 and 4) was not aswidespread as indicated by Northern blotting, suggesting that they may be less important for the control of TRAILcytotoxicity than previously thought. TR1 and TR2 expression increases significantly in a number of malignant tissues,but in some common malignancies their expression was low, or patchy, which may limit the therapeutic role of TRAIL.Taken together, we have a panel of monoclonal antibodies that will allow a better assessment of the normal role ofTRAIL and allow assessment of biopsy material, possibly allowing the identification of tumors that may be amenable toTRAIL therapy.

Systematic Investigation of Berberine for Treating Hepatocellular Carcinoma Based on Network Pharmacology

Objective Liver cancer is the 4th leading cause of cancer death worldwide,and hepatocellular carcinoma(HCC)accounts for the largest proportion of these deaths.Berberine is a quaternary amine compound extracted from plants such as Coptidis Rhizoma(Huang Lian,黄连)and Phellodendri Chinensis Cortex(Huang Bo,黄柏)and is considered as a potential candidate for treating HCC.This study used network pharmacology methods,reveal the core mechanism of action of berberine in the treatment of HCC,clarify its medicinal value,and locate the anti-tumor mechanism of berberine.Methods Structural information of Berberine(PubChem CID:2353)was obtained from the NCBI PubChem;ADME parameter were obtained from the Traditional Chinese Medicine Systems Pharmacology(TCMSP)database;Berberine prediction targets were collected from symmap,stitch and targetnet databases;HCC significant targets were retrieved from OncoDB.HCC and Liverome;A PPI network was established at STRING,Prediction target of berberine therapy for HCC are collected by gene mapping;The core target,pathway,biological process(BP),cellular component(CC),and molecular function(MF)of berberine in the treatment of HCC were predicted by topological analysis and enrichment analysis;the visualized"target pathway"network diagram of berberine in the treatment of HCC was established by the software of Cytoscape.Results Through PubChem and tcmsp databases,the good drugforming properties of berberine were identified;32 prediction targets of berberine were collected in symmap,stitch and targetnet databases;566 related targets of HCC were collected in oncodb.hcc and liverome databases;10 targets of berberine treatment for HCC were predicted by gene mapping,and a PPI with 10 nodes and 34 edges was established Through topological analysis and enrichment analysis,6 topologically important targets,6 related pathways and 16 BP,6 cc and 7 MF involved in Berberine treatment of HCC were obtained.Conclusions The anticancer effect of berberine is mainly involved in the regulation of cells of hepatoma cells through complex interactions between the TB52,MAPK1,CCND1,PTGS2,ESR1 that act on Hub nodes and their associated 6 pathways.The cycle is related to the immune inflammatory response,including biological processes such as proliferation and apoptosis of liver cancer cells.

Glutamine depletion induces murine neonatal melena with increased apoptosis of the intestinal epithelium

AIM:To investigate the possible biological outcome and effect of glutamine depletion in neonatal mice and rodent intestinal epithelial cells.METHODS:We developed three kinds of artificial milk with different amounts of glutamine;Complete amino acid milk (CAM),which is based on maternal mouse milk,glutamine-depleted milk (GDM),and glutaminerich milk (GRM).GRM contains three-fold more glutamine than CAM.Eighty-seven newborn mice were divided into three groups and were fed with either of CAM,GDM,or GRM via a recently improved nipple-bottle system for seven days.After the feeding period,the mice were subjected to macroscopic and microscopic observations by immunohistochemistry for 5-bromo-2'deoxyuridine (BrdU) and Ki-67 as markers of cell proliferation,and for cleaved-caspase-3 as a marker of apoptosis.Moreover,IEC6 rat intestinal epithelial cells were cultured in different concentrations of glutamine and were subject to a 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)2H-5-tetrazolio]-1,3-benzene disulfonate cell proliferation assay,flow cytometry,and western blotting to examine the biological effect of glutamine on cell growth and apoptosis.RESULTS:During the feeding period,we found colonic hemorrhage in six of 28 GDM-fed mice (21.4%),but not in the GRM-fed mice,with no differences in body weight gain between each group.Microscopic examination showed destruction of microvilli and the disappearance of glycocalyx of the intestinal wall in the colon epithelial tissues taken from GDM-fed mice.Intake of GDM reduced BrdU incorporation (the average percentage of BrdU-positive staining;GRM:13.8%,CAM:10.7%,GDM:1.14%,GRM vs GDM:P < 0.001,CAM vs GDM:P < 0.001) and Ki-67 labeling index (the average percentage of Ki67-positive staining;GRM:24.5%,CAM:22.4% GDM:19.4%,GRM vs GDM:P=0.001,CAM vs GDM:P =0.049),suggesting that glutamine depletion inhibited cell proliferation of intestinal epithelial cells.Glutamine deprivation further caused the deformation of the nuclear membrane and the plasma membrane,accompanied by chromatin degeneration and an absence of fat droplets from the colonic epithelia,indicating that the cells underwent apoptosis.Moreover,immunohistochemical analysis revealed the appearance of cleaved caspase-3 in colonic epithelial cells of GDM-fed mice.Finally,when IEC6 rat intestinal epithelial cells were cultured without glutamine,cell proliferation was significantly suppressed after 24 h (relative cell growth;4 mmol/L:100.0% ± 36.1%,0 mmol/L:25.3% ± 25.0%,P < 0.05),with severe cellular damage.The cells underwent apoptosis,accompanied by increased cell population in sub-G0 phase (4 mmol/L:1.68%,0.4 mmol/L:1.35%,0 mmol/L:5.21%),where dying cells are supposed to accumulate.CONCLUSION:Glutamine is an important alimentary component for the maintenance of intestinal mucosa.Glutamine deprivation can cause instability of the intestinal epithelial alignment by increased apoptosis.

In vitro effect of p2l^(WAF-1/CIP1) gene on growth of human glioma cells mediated by EGFR targeted non-viral vector GE7 system

Objective: To construct the EGFR targeted non-viral vector GE7 system and explore the in vitro effect of p21WAF-1/CIPI gene on growth of human glioma cells mediated by the GE7 system. Methods: The EGFR targeted non-viral vector GE7 gene delivery system was constructed. The malignant human glioma cell line U251MG was transfected in vitro with β-galactosidase gene ( reporter gene) and p21WAF-1/CIPI gee (therapeutic gene) using the GE7 system. By means of X-gal staining, MTS and FACS, the transfection efficiency of exogenous gene and apoptosis rate of tumor cells were examined. The expression of p21WAF-1/ CIPI gene in transfected U251MG cell was examined by immunohistochemis-try staining. Results: The highest transfer rate of exogenous gene was 70% . After transfection with p21WAF-1/CIPI gene, the expression of WAF-1 increased remarkably and steadily; the growth of U251MG cells were inhibited evidently. FACS examination showed G1 arrest. The average apoptosis rate was 25.2%. Conclusion: GE7 system has the ability to transfer exogenous gene to targeted cells efficiently, and expression of p21WAF-1/CIPI gene can induce apoptosis of glioma cell and inhibit its growth.

Effect of trkA kinase inhibitor on apoptosis of human pterygium fibroblasts

Objective： The aim of this study was to investigate the growth inhibition effects and the mechanisms of trkA kinase inhibitor on human pterygium fibroblasts（HPF）. Methods： Three normal conjunctivas and eleven pterygiums were surgically collected, trkA and p75 was examined in different tissues by immunohistochemistry, trkA and p75 expression was detected in HPF by immumofluorescence method. After being treated with 10-80 nmol/L trkA kinase inhibitor for 24-96 h, the growth activities of HPF were studied by MTT colorimetry. Caspase-3 was inspected in HPF by spectrophotometric method. The apoptosis of HPF was detected by flow cytometry. Results： Expression of trkA and p75 was significantly observed in epithelium and fibroblast of pterygium tissues. Expression of trkAwas observed in epithelium and fibroblast of normal conjunc- tiva. Expression of p75 was only observed in epithelium of normal conjuncUva, trkA kinase inhibitor could effectively inhibit the growth of human pterygium fibroblasts in vitro in time and dose dependent manners. Caspase-3 expression was increased. The rates of apoptosis were 8.26%-29.62% （P 〈 0.01）. Conclusion： TrkA and p75 possibly participate in genesis and development of pterygium, trkA kinase inhibitor could induce apoptosis of human pterygium fibroblasts. Inducing apoptosis through changing the ratio of trkA and p75 in fibroblasts and up-regulate Caspase-3 was probably one of its molecular mechanisms.

Expression of Smac and the relationship with cell apoptosis and proliferation in hepatocarcinoma tissues

Objective:The aim of the study was to investigate the expression of Smac protein in human hepatocarcinoma and their relationship with cell apoptosis and proliferation.Methods:The expressions of Smac and the proliferating cell nuclear antigen (PCNA) in 41 cancer tissues,41 adjacent cirrhosis tissues and 9 normal control tissues in hemangioma were assessed by two-step immunohistochemical method and apoptosis was detected by TUNEL method.Results:Smac protein was expressed in 14 (34.14%) of the 41 cases of hepatocarcinoma,in 23 (56.10%) of the 41 cases of the adjacent cirrhosis tissues,and in 7 (77.8%) of the normal tissues in hemangioma.Smac protein positive expression rate in hepatocarcinoma was significantly lower than that in the adjacent cirrhosis tissues and the normal control tissues,χ2 were 3.989 and 4.115,respectively,and P were 0.046 and 0.042,respectively.Smac protein expression in cancer was significantly correlated with the ratio of apoptotic index to proliferative index,t'=2.260,P<0.05,but was not with the clinicopathological indicators such as the age and the histological grade,P>0.05.Conclusion:The relatively lower level of the expression of Smac may in a certain extent break the dynamic balance between apoptosis and proliferation of hepatocarcinoma cells,and then plays an important role in the pathogenesis of hepatocarcinoma.