Investigation of the sensitivities of distinct gastric cancer cells to parvovirus H‐1 induced cytotoxicity

OBJECTIVE: To investigate the sensitivities of distinct gastric cancer cells to parvovirus H‐1 induced cytotoxicity and the possible mechanism(s). METHODS: There were six distinct differentiated gastric cancer cell lines: HGC27 (undifferentiated), BGC823 (undifferentiated), MKN45 (poorly differentiated), AGS (poorly differentiated), SGC7901 (moderately differentiated) and MKN28 (well differentiated). The cell cycle distributions were measured by flow cytometry and the differential sensitivities of the six distinct gastric cancer cells after H‐1 virus infection were detected by MTT assay. RT‐PCR was used to detect viral NS1 gene expression in all six gastric cancer cell lines. RESULTS: The S phase ratios of HGC27, BGC823, MKN45, AGS, SGC7901 and MKN28 were 24.72%, 30.15%, 27.10%, 29.03%, 31.82% and 33.73%, respectively. HGC27 cells were sensitive to H‐1 virus induced cytotoxicity, followed by SGC7901 cells. MKN45 and AGS cells were moderately sensitive and MKN28 cells were insensitive. However, BGC823 cells were resistant to H‐1 virus induced cytotoxicity. The expressions of viral NS1 were higher in HGC27, BGC823, MKN45 and SGC7901 cells, and lower in AGS and MKN28 cells. CONCLUSIONS: The sensitivities of the distinct gastric cancer cells to H‐1 virus induced cytotoxicity were markedly different. In general, the poorly differentiated cells showed an enhanced sensitivity to H‐1 virus attack compared with well‐differentiated ones. The enhanced sensitivity of poorly versus well‐differentiated gastric cancer cells to H‐1 virus is related in part to the enhanced capacity of the former for NS1 protein production and accumulation. The undifferentiated BGC823 cells were resistant to H‐1 virus triggered cytotoxicity. It may further verify that not all tumor cells are sensitive to H‐1 virus lytic effects.     

A method for enzymatic dissociation of cells from isolated pancreatic islets

Summary An enzymatic method for isolation of single cells from the islets of Langerhans is described. The isolated cells appeared well preserved and survived for at least 7 days when maintained in culture. The dry mass of the isolated islet cells was found to be decreased 30 min after administration of alloxan to obese-hyperglycemic mice. Isolated individual islet cells from obese-hyperglycemic mice had a higher dry mass than those from their lean litter mates. Traduzione a cura di G. U.     

白色念珠菌对人口腔黏膜上皮角质细胞的作用研究

目的研究白色念珠菌对人口腔黏膜上皮角质细胞的作用机制。方法将培养的口腔黏膜上皮角质细胞分别加入孢子型白色念珠菌(孢子组)和菌丝型白色念珠菌(菌丝组),另将单独培养的口腔黏膜上皮角质细胞作为阴性对照组。比较各组对口腔黏膜上皮角质细胞的黏附作用,并观察白色念珠菌对口腔黏膜上皮角质细胞的凋亡和增殖作用。结果菌丝组黏附指数(7.63±1.39)显著高于孢子组(3.47±1.04),差异有统计学意义(P<0.05);菌丝组凋亡率[(3.49±0.4)%]显著高于孢子组[(2.07±0.15)%]和阴性对照组[(1.98±0.07)%],差异有统计学意义(P<0.05),而孢子组和阴性对照组凋亡率相比差异无统计学意义(P>0.05)。菌丝组G0/G1期细胞比例[(38.17±7.83)%]显著低于孢子组[(49.47±11.41)%]和阴性对照组[(57.71±9.39)%],差异有统计学意义(P<0.05);S期[(8.54±4.03)%]、G2/M期[(20.18±9.59)%]细胞比例上升,且显著高于孢子组[(6.47±2.73)%、(10.71±3.19)%]和阴性对照组[(5.35±2.11)%、(12.38±4.35)%],差异有统计学意义(P<0.05);PI[(42.79±18.73)%]明显高于孢子组[(28.67±8.79)%]和阴性对照组[(26.87±7.64)%],差异有统计学意义(P<0.05),而孢子组和阴性对照组PI相比差异无统计学意义(P>0.05)。结论白色念珠菌能够导致口腔黏膜上皮角质细胞发生凋亡和增殖周期改变。     

抗MDA5抗体对成年及幼年型皮肌炎合并间质性肺病诊断效能的meta分析

目的通过meta分析探究抗黑色素瘤分化相关基因5(MDA5)抗体对不同类型皮肌炎(DM)并发快速进展型间质性肺病(RPILD)及慢性间质性肺病(ILD)的诊断效能。方法检索Pub Med、Embase、Cochrane library、中国知网、万方、维普和中国生物医学文献数据库,时间起止为建库至2017年5月。运用Meta-disc1.4行异质性检验,计算汇总敏感度、特异度、诊断比值比、阳性阴性似然比和SROC曲线。采用QUADAS-2和stata 12.0进行质量评价和发表偏倚。结果共纳入32篇研究,文献质量较高,各部分不存在高偏倚风险,为中等异质性。抗MDA5抗体对成年DM合并RPILD的诊断效能(AUC=0.927,Q*=0.862)优于对成年DM合并慢性ILD(AUC=0.717,Q*=0.667)、幼年型皮肌炎(JDM)合并RPILD(AUC=0.836,Q*=0.768)的诊断效能。抗MDA5抗体对DM合并RPILD诊断准确性:临床无肌病型皮肌炎(CADM)(AUC=0.942,Q*=0.880)高于DM(AUC=0.926,Q*=0.860),亚洲(AUC=0.960,Q*=0.891)优于中国(AUC=0.925,Q*=0.859)及欧美人群(AUC=0.928,Q*=0.863),ELISA法(AUC=0.929,Q*=0.864)不劣于免疫沉淀法(AUC=0.927,Q*=0.859)。Deek's漏斗图提示不存在发表偏倚。结论抗MDA5抗体诊断成年及幼年型DM合并ILD有较高敏感性和特异性。     

人嗅黏膜间充质干细胞的生物学特性

目的：观察及研究人嗅黏膜间充质干细胞的生物学特性。方法：分离、培养、鉴定人嗅黏膜间充质干细 胞,在透射电子显微镜、扫描电子显微镜下观察其超微结构,并在体外诱导其向脂细胞、骨细胞、神经干细胞球和神 经元分化。结果：人嗅黏膜间充质干细胞生长以梭形细胞为主,呈放射状排列,高表达表面标志CD73和CD90,不表 达CD34和CD45;在扫描电子显微镜下可见细胞表面有短而粗的微绒毛突起,在透射电子显微镜下可见到两种不同的 细胞形态;具有成脂、成骨、成神经元干细胞球和神经元分化的能力。结论：人嗅黏膜间充质干细胞具有间充质干细 胞的一般生物学特性,经诱导培养后具有多向分化潜能,可作为组织工程修复的理想的种子细胞。     

CCK-8法与Alamar blue法对RKO细胞株增殖活力测试条件的对比分析

目的 以RKO细胞株为增殖活力测试的研究对象,比较CCK-8法与Alamar blue法在检测中波长、孵育时间、细胞密度等方面的差异性.方法 取体外培养对数生长期的RKO细胞株,分别比较两者在不同检测波长(380、430、480、530、580、630 nm)、不同孵育时间(1、2、3、4、5、6 h)、不同细胞密度(2×104、5×103、1.25×103 CUF·mL-1)下所测的光密度值(OD值),并比较两者在相同条件下同一标本重复检测时的平衡性.结果 ALamar blue法在不同细胞密度中的最佳检测波长皆为580 nm,而CCK-8法的最佳检测波长会随着细胞密度的变化而有所改变;1～6 h时,CCK-8法适合的孵育时间在1h以后Alamar blue为2h以后,且CCK-8法的灵敏度、平衡性都强于Alamar blue法.结论 CCK-8法在细胞增殖活力检测中的灵敏性、平衡性、快捷性等都略强于ALamar blue法.     

Murrayafoline A Induces a G0/G1-Phase Arrest in Platelet-Derived Growth Factor-Stimulated Vascular Smooth Muscle Cells

The increased potential for vascular smooth muscle cell (VSMC) growth is a key abnormality in the development of atherosclerosis and post-angioplasty restenosis. Abnormally high activity of platelet-derived growth factor (PDGF) is believed to play a central role in the etiology of these pathophysiological situations. Here, we investigated the anti-proliferative effects and possible mechanism(s) of murrayafoline A, a carbazole alkaloid isolated from Glycosmis stenocarpa Guillamin (Rutaceae), on PDGF-BB-stimulated VSMCs. Murrayafoline A inhibited the PDGF-BB-stimulated proliferation of VSMCs in a concentration-dependent manner, as measured using a non-radioactive colorimetric WST-1 assay and direct cell counting. Furthermore, murrayafoline A suppressed the PDGF-BB-stimulated progression through G₀/G₁ to S phase of the cell cycle, as measured by [³H]-thymidine incorporation assay and cell cycle progression analysis. This anti-proliferative action of murrayafoline A, arresting cell cycle progression at G₀/G₁ phase in PDGF-BB-stimulated VSMCs, was mediated via down-regulation of the expression of cyclin D1, cyclin E, cyclin-dependent kinase (CDK)2, CDK4, and proliferating cell nuclear antigen (PCNA), and the phosphorylation of retinoblastoma protein (pRb). These results indicate that murrayafoline A may be useful in preventing the progression of vascular complications such as restenosis after percutaneous transluminal coronary angioplasty and atherosclerosis.