Establishment and characterization of atypical fibroblasts from human adult liver contributing to hepatocyte cord-like arrangement

We report the phenotypic and functional characterization of fibroblasts established in culture from the non-parenchymal epithelial cell populations of adult human livers. Human liver fibroblasts (hLF) expressed mesenchymal antigens vimentin, alpha-smooth muscle actin, collagen, fibronectin, CD73, CD90, CD105, and CD166 together with non-mesenchymal antigens cytokeratins 8 and 18, glial fibrillary acidic protein, and nestin. Mixed cell lineage-specific protein expression was not associated with stem-like cell properties. Coculturing hepatocytes onto confluent hLF showed that they survived and maintained metabolic activity such as albumin, glycogen, and urea production. Moreover, hepatocytes formed cord-like arrangements resembling those established in vivo. Hepatocyte arrangement depended on cell-to-cell contact and the tissue origin of fibroblasts. Time-lapse video imaging of cocultured cells showed that hepatocyte arrangement was coordinated by the stretching and shortening of underneath hLF. Our data suggest that hLF may represent resident fibroblasts of the adult human liver, which could assume guiding functions for hepatic epithelial cells.     

麻疹、腮腺炎二联活疫苗的研制

用于生产麻诊活疫苗的“沪191”毒株和由武汉生物制品研究所引进的腮腺炎疫苗“Wms4”毒株生产各自的病毒原液,全面检定合格后,对麻疹与腮腺炎两种病毒原液进行配比试验、热稳定性试验等,最终选定联合疫苗中麻疹与腮腺炎病毒CCID50之比1：30,试制出二批麻－腮二联活疫苗,经全面检定,完全符现行规程要求,目前正在I期临床研究。     

Cyclic stretch-induced stress fiber dynamics - Dependence on strain rate, Rho-kinase and MLCK

Stress fiber realignment is an important adaptive response to cyclic stretch for nonmuscle cells, but the mechanism by which such reorganization occurs is not known. By analyzing stress fiber dynamics using live cell microscopy, we revealed that stress fiber reorientation perpendicular to the direction of cyclic uniaxial stretching at 1 Hz did not involve disassembly of the stress fiber distal ends located at focal adhesion sites. Instead, these distal ends were often used to assemble new stress fibers oriented progressively further away from the direction of stretch. Stress fiber disassembly and reorientation were not induced when the frequency of stretch was decreased to 0.01 Hz, however. Treatment with the Rho-kinase inhibitor Y27632 reduced stress fibers to thin fibers located in the cell periphery which bundled together to form thick fibers oriented parallel to the direction of stretching at 1 Hz. In contrast, these thin fibers remained diffuse in cells subjected to stretch at 0.01 Hz. Cyclic stretch at 1 Hz also induced actin fiber formation parallel to the direction of stretch in cells treated with the myosin light chain kinase (MLCK) inhibitor ML-7, but these fibers were located centrally rather than peripherally. These results shed new light on the mechanism by which stress fibers reorient in response to cyclic stretch in different regions of the actin cytoskeleton.     

应用不同代次兔肾细胞培养风疹病毒松叶株的研究

目的为提高兔肾细胞产量用于增加风疹病毒生产。方法 SPF级家兔肾细胞经传代培养10代,对不同代次兔肾细胞进行遗传稳定性、外源因子及兔脑原虫检测后接种风疹病毒松叶株(Matsuba strain)的试验。结果兔肾细胞产量得到提高,由1对兔肾平均生产1瓶细胞增加为8瓶,细胞核型检查传至第10代的细胞染色体数目与初代细胞一致,细胞培养物均一性提高。第0～第3代细胞病毒培养物滴度间无显著性差异,χ2检验返回相关性的P值均大于0.95。结论第1～第3代细胞培养风疹病毒与第0代相比,可获得相同滴度的病毒培养物,p3代兔肾细胞应用于疫苗生产可显著提高细胞及病毒产量。     

呼吸道合胞病毒疫苗及其抗体制剂的研究进展

呼吸道合胞病毒(respiratory syncytial virus,RSV)感染是一个全球性的健康问题,目前临床上仍缺乏特异的治疗手段。接种疫苗主动免疫预防或使用抗体制剂被动免疫预防是避免重症感染和减少死亡的重要措施。针对不同的人群,须研制不同类型的RSV疫苗：减毒活疫苗对婴儿来说,可能是最佳选择;亚单位疫苗有引起增强型呼吸道疾病的风险,不适合RSV血清学阴性的婴幼儿接种,主要适用于老年人和孕妇。研发安全且有效的RSV疫苗难度大,虽然已有30余种RSV疫苗进入临床研究阶段,并显示出应用潜力,其中F纳米颗粒疫苗已率先进入III期临床试验,但在老年人和孕妇中未达预期效果。在RSV流行季节前,使用特异性抗体制剂也是预防高危人群严重RSV感染性疾病的有效手段。长效单克隆抗体MEDI8897比帕利珠单抗更具成本效益,已进入III期临床试验,且获得优先研发资格。多克隆免疫球蛋白RI-002已在免疫缺陷人群中显示出较好的预防效果,具有进一步研发的现实意义。本综述针对近年来RSV疫苗及其抗体的研究进展进行阐述,期望为RSV的预防提供参考。     

核心通识课“微生物的世界”网课直播工具筛选

由于新型冠状病毒肺炎疫情的影响,本学期目前只能上网课。开学前作者测试了"微助教"、QQ群"分享屏幕"、QQ"群课堂"和"腾讯课堂"等四种直播授课工具,分析了每种直播工具的特点和利弊。本学期作者讲授核心通识课"微生物的世界",选课人数超过100人,根据本课程特点,最终选择"腾讯课堂"作为直播工具,并结合"微助教"平台授课,获得了较好的教学效果,为网课教学实践提供了参考。     

声触诊组织量化技术（VTQ）测量肝硬度与血清AST/ALT比值评价肝硬化患者严重程度及预后分析

摘要 目的：探讨VTQ测量肝硬度与血清AST/ALT比值对肝硬化患者严重程度及预后的评价价值。方法：回顾性选择2018年1月至2022年10月来我院诊治的肝硬化患者80例,根据Child-Pugh分级将80例患者分为Child-Push A级30例、B级25例、C级25级,根据是否存在并发症将80例患者分为并发症组（45例）与非并发症组（35例）,80例患者均用VTQ法检测VTQ值,检测所有患者的血清ALT、AST水平,计算ALT/AST比值。对比不同Child-Push分级患者不同部位的VTQ值,对比不同Child-Push分级患者的AST、ALT水平及AST/ALT比值,对比有无并发症组的不同部位VTQ值,对比有无并发症组患者的AST、ALT水平及AST/ALT比值,分析80例患者不同部位VTQ值与AST、ALT、AST/ALT比值的相关性。结果：C组患者不同肝脏部位的VTQ值明显较A组及B组高,B组患者不同肝脏部位的VTQ值较A组高（P<0.05）。C组的AST水平、AST/ALT值明显较A组、B组高,B组的AST水平、AST/ALT值明显较A组高（P<0.05）,C组的ALT水平较A组、B组高,B组的ALT水平较A组高,但组间对比无统计学意义（P>0.05）。并发症组不同肝脏部位的VTQ值明显较无并发症组高（P<0.05）。并发症组的AST、AST/ALT比值明显较无并发症组高（P<0.05）,并发症组的ALT较无并发症组高,但组间对比无统计学意义（P>0.05）。80例肝硬化患者的AST、AST/ALT比值与不同部位的VTQ值正相关（P<0.05）,ALT水平与不同部位的VTQ值无相关性（P>0.05）。结论：VTQ测量肝硬度与血清AST/ALT比值可用于评价肝硬化严重程度及预后。     

An apparatus for sampling gut contents of large,living fishes

Synopsis Apparatus and methods are described and illustrated for flushing and retaining gut contents from large, living fishes with water supplied by a 12 volt portable pump.     

HF10, an attenuated herpes simplex virus (HSV) type 1 clone, lacks neuroinvasiveness and protects mice against lethal challenge with HSV types 1 and 2

Herpes simplex virus (HSV), a neurotropic virus, establishes life-long and, although rare, life-threatening infection in humans, and it may precipitate substantial medical and psychosocial morbidity. Here we show that HSV-1 strain HF clone 10 (HF10) exhibits impaired neuroinvasiveness in peripheral olfactory, vomeronasal and trigeminal conduits following intranasal as well as corneal inoculation. HF10 attenuation likely arises from multiple defects of HSV genes, so that HF10 will not revert to a virulent phenotype. Intranasal vaccination of mice with HF10 conferred significant protection against lethal challenge with HSV-1 and HSV-2 via the intranasal and intravaginal routes. Thus, we propose that HF10 explicitly meets the prerequisites for a candidate live attenuated HSV vaccine.