Replication of dengue,yellow fever,St. Louis encephalitis and vesicular stomatitis viruses in a cell line (TRA-171) derived fromToxorhynchites amboinensis

Summary The replication of seven arboviruses in a cell line (TRA-171) derived from a nonhematophagous mosquito was studied. Four serotypes of laboratory adapted and three serotypes of unadapted dengue viruses replicated in the TRA-171 cell line, inducing syncytia. The sensitivity of TRA-171 cells to dengue virus infection was comparable to that ofAedes albopictus orA. pseudoscutellaris cells. Yellow fever, St. Louis encephalitis, and vesicular stomatitis viruses also replicated. All four serotypes of dengue viruses could be plaque assayed with TRA-171 cell cultures. Use of trade names is for identification only and does not constitute endorsement by the Public Health Service or by the U.S. Department of Health and Human Services.     

In Vitro Selection of Variants of Herpes Simplex Virus Type 1 Which Differ in Cytopathic Changes

To analyze the mechanisms for in vitro emergence of the syncytial variants of herpes simplex virus type 1 (HSV-1), several cell lines were infected with a mixture of equal amounts of two HSV-1 variants, one syncytial and the other non-syncytial, and changes in their relative abundance were monitored during passage. With a combination of two variants of the Miyama strain of HSV-1, the syncytial variant became dominant during passage in Vero, RK-13 and FL cells. On the other hand, the ratios of the two variants remained around 1:1 during the passage in HEp-2, MGC and HEL cells. In another set of variants of the SKO strain of HSV-1, the outcomes were different from those of the Miyama strain in the FL, MGC and HEp-2 cells. The ratios of the two variants remained around 1:1 during passage in FL cells, while the non-syncytial variant became dominant during passage in MGC and HEp-2 cells. In addition, we examined the effects of a complement and interferon-β (IFN-β) on the outcome of the selection. As a result, the complement slowed the selection of a syncytial variant, whereas IFN-β facilitated it.     

98.55W/cm2脉冲微波辐照对大鼠血清生化指标影响的研究

二级雄性Wistar大鼠35只,随机分为7组,每组5只,1组为空白对照,其余6组为辐照组以98.55W/cm^2的脉冲微波辐照动物,辐照后1、6、24h和7、14、28d分别采血,用Coulter-JTIR全自动生化仪进行测定,所有数据经Spss8.0统计软件处理,辐射后1h,CRE明显增高（P＜0.01）,而TP、ALB、AST、LD1、CK、ALP及CHOL均明显降低（P＜0.05）;辐射后6h,春CRE明显增高（P＜0.05）,而TP、ALB、LD1、ALP及CHOL均明显降低（P＜0.05）;辐照后24h,其ALT、GLU、BUN、CRE、AST、LD和HBDH明显升高（P＜0.05）,而LB和CHOL则明显降低（P＜0.05）;辐照后7d,其ALT、ALB、GLU、CRE、CK－MB和GGT明显升高（P＜0.05）,而P和CHOL则明显降低（P＜0.05);辐照后14d,其CRE、LD、CK－MB和GGT明显升高（P＜0.05）,而P和CHOL则明显降低（P＜0.01）;辐照后28d,其ALT、TP、BUN和CRE明显升高（P＜0.05）,而ALB、AST、LD1、CK、ALP和CHOL则明显降低（P＜0.01）。提示HPPMW可引起大鼠血清多种酶、蛋白及代谢产物的紊乱,即有早期影响,也表现为一定的持续效应。HPPMW可能引起大鼠多系统、多脏器的损伤。     

A cytopathic effect of Trichomonas vaginalis probably mediated by a mannose/N-acetyl-glucosamine binding lectin.

The pathogenic effect of a highly pathogenic strain of Trichomonas vaginalis on McCoy cell monolayers was investigated. Specific inhibition of the cytopathic effect by monosaccharides, such as N-acetyl-glucosamine (GlcNAc) and mannose (Man), was observed. Our preliminary results suggest that the pathogenicity of T. vaginalis depends on a lectin specifically sensitive to GlcNAc and to a lesser extent to Man. Although N-acetyl-mannosamine was found to be the most efficient inhibitor, this effect seems to be unrelated to the natural biological behaviour of the infested host.     

疱疹病毒Ⅱ型感染SH-SY5Y细胞的生物学效应

目的：探讨疱疹病毒Ⅱ型（HSV-2）感染人神经母细胞瘤细胞株SH-SY5Y的生物学效应。方法：病毒液接种SH-SY5Y细胞后,用相差和电子显微镜观察感染细胞的形态变化,RT-PCR检测病毒在细胞中的增殖,MTT法检测病毒感染对细胞增殖的影响,流式细胞仪测定感染后的细胞凋亡状况。结果：相差显微镜显示细胞病变,从24～72h,细胞变性、坏死的程度和数量随感染时间延长而增加;电镜结果显示感染24h后,细胞核染色质固缩,出现多核巨细胞,线粒体内嵴紊乱、断裂,出现不同程度的自噬化、溶酶体化、空泡化,并可见大量鹰眼样已包装成熟的病毒颗粒及正在包装的病毒粒子;HSV-2LAT基因RT-PCR扩增表明,病毒能在SH-SY5Y细胞中增殖;凋亡检测显示HSV-2在体外细胞感染中并未使细胞出现凋亡现象;感染后24、48及72h,SH-SY5Y细胞的抑制率分别为11.3%、31.2%和63.1%,与对照组相比均存在显著性差异（P〈0.05）;分别用0.1、1、10MOI的病毒感染SH-SY5Y细胞,上述不同组在24、48、72h时细胞形态变化基本一致,感染结果相似,各组之间病毒毒力无明显差异（P〉0.05）。结论：初步在人神经母细胞瘤细胞株SH—SY5Y中建立了HSV-2感染的细胞模型,并研究了感染对细胞生物性状的影响,为探讨HSV-2的潜伏与激发机制、了解HSV-2的致病机制打下基础。     

Attachment characteristics of normal human cells and virus-infected cells on microcarriers

The attachment kinetics of normal and virus-infected LuMA cells were studied to improve the production of live attenuated varicella viruses in human embryonic lung (LuMA) cells. Normal LuMA cells and LuMA cells infected by varicella virus at various cytopathic effects (CPE) were grown on microcarriers. Ninety-three percent of suspended LuMA cells attached to the solid surface microcarriers within fifteen minutes and cell viability was greater than 95% when the cell suspension was stirred. Low serum levels did not affect the attachment rate of virus-infected cells in the microcarrier culture system. Kinetic studies showed that varicella infected cells had a lower attachment rate than normal LuMA cells. Virus inoculum (= infected cells) at low CPE showed a relatively better attachment rate on cell-laden microcarriers than virus inoculum at a higher CPE. Maximum titers were obtained at 2 days post-infection. Based on cell densities, the use of viral inoculum showing a 40% CPE led to an approximately 2- and 1.2-fold increase in the cell associated and in cell free viruses, respectively, than a virus inoculum with a CPE of 10%.However, the ratio of cell-free to cell-associated virus in a microcarrier culture was very low, approximately0.04–0.06. These studies demonstrate that the virus inoculum resulting in a high CPE yielded a high production of cell-associated and cell-free virus in microcarrier cultures because of the high cellular affinity of the varicella virus. This revised version was published online in August 2006 with corrections to the Cover Date.     

Zika virus induces massive cytoplasmic vacuolization and paraptosis‐like death in infected cells

The cytopathic effects of Zika virus (ZIKV) are poorly characterized. Innate immunity controls ZIKV infection and disease in most infected patients through mechanisms that remain to be understood. Here, we studied the morphological cellular changes induced by ZIKV and addressed the role of interferon‐induced transmembrane proteins (IFITM), a family of broad‐spectrum antiviral factors, during viral replication. We report that ZIKV induces massive vacuolization followed by “implosive” cell death in human epithelial cells, primary skin fibroblasts and astrocytes, a phenomenon which is exacerbated when IFITM3 levels are low. It is reminiscent of paraptosis, a caspase‐independent, non‐apoptotic form of cell death associated with the formation of large cytoplasmic vacuoles. We further show that ZIKV‐induced vacuoles are derived from the endoplasmic reticulum (ER) and dependent on the PI3K/Akt signaling axis. Inhibiting the Sec61 ER translocon in ZIKV‐infected cells blocked vacuole formation and viral production. Our results provide mechanistic insight behind the ZIKV‐induced cytopathic effect and indicate that IFITM3, by acting as a gatekeeper for incoming virus, restricts virus takeover of the ER and subsequent cell death.     

微波生物效应研究的现状及应用

着重介绍了微波对微生物、黄曲霉素、人及动物的生物效应,以及微波在浸取和医学上的应用。     

Limits and Sites of Lysinoalanine Formation in Lysozyme, α-Lactalbumin and αs1- and β-Caseins by Alkali Treatment

Lysinoalanine was determined by gas chromatography. Lysinoalanine formation in lysozyme depended on alkali concentration, pH, temperature and exposure time. The upper limits of lysinoalanine formation in lysozyme and α-lactalbumin were related to the number of lysine residues with a cystine disulfide bond in the adjacent position rather than the individual contents of these residues. In cases of α_sl- and β-caseins, the upper limits were related to the number of phosphoserine residues, regardless of their sequential relationship to lysine residues. Gel electrophoresis suggested that intermolecular cross-linking took place in the α-lactalbumin and caseins.     

过柱纯化-高效液相色谱法测定玉米黄曲霉毒素

通过硅镁净化柱纯化,利用高效液相色谱法测定了玉米（Zea mays）中的黄曲霉毒素,在短时间（10min）内实现了黄曲霉毒素主要组分B1、G1、B2和G2的分离,确定了它们的回归方程和检测限。并在此基础上,结合目前研究现状,分析了该方法在应用中存在的问题,讨论了B1、G1、B2和G24种主要组分的出峰顺序。该试验结果将有助于高效液相色谱法在检测黄曲霉毒素应用中的日臻完善。     

100～700 mL液体培养基的微波灭菌效果观察

为研究家用微波炉对少量液体培养基的灭菌效果,对100 -700 mL液体培养基采取微波分别进行灭菌,测定其灭菌时间,并对达到灭菌效果的培养基通过接种不同细菌进行质检.结果显示100、200、300、400、500、600、700 mL液体培养基分别在4.5、5.0、7.0、9.0、12.0、19.0、25.0 min达到最有效的灭菌效果.室温保存15 d仍无细菌生长.枯草杆菌黑色变种芽胞菌测试无菌生长.金黄色葡萄球菌、大肠埃希菌及普通变形杆菌在液体培养基里的生长现象、特种后细菌的菌落和形态染色特征及其生化反应均无明显差异.实验表明2 450 MHz,700 W的微波炉对100 - 700 mL的液体培养基不但能快速达到有效的灭菌效果,且营养成分不会被破坏.