Adaptation of Vero cells to suspension growth for rabies virus production in different serum free media

Vero cells are nowadays widely used in the production of human vaccines. They are considered as one of the most productive and flexible continuous cell lines available for vaccine manufacturing. However, these cells are anchorage dependent, which greatly complicates upstream processing and process scale-up. Moreover, there is a recognized need to reduce the costs of vaccine manufacturing to develop vaccines that are affordable worldwide. The use of cell lines adapted to suspension growth contributes to reach this objective.The current work describes the adaptation of Vero cells to suspension culture in different serum free media according to multiple protocols based on subsequent passages. The best one that relies on cell adaption to IPT-AFM an in-house developed animal component free medium was then chosen for further studies. Besides, as aggregates have been observed, the improvement of IPT-AFM composition and mechanical dissociation were also investigated.In addition to IPT-AFM, three chemically defined media (CD293, Hycell CHO and CD-U5) and two serum free media (293SFMII and SFM4CHO) were tested to set up a serum free culture of the suspension-adapted Vero cells (VeroS) in shake flasks. Cell density levels higher than 2 × 10⁶ cells/mL were obtained in the assessed conditions. The results were comparable to those obtained in spinner culture of adherent Vero cells grown on Cytodex 1 microcarriers.Cell infection with LP-2061 rabies virus strain at an MOI (Multiplicity of Infection) of 0.1 and a cell density of 8 ± 0.5 × 10⁵ cells/mL resulted in a virus titer higher than 10⁷ FFU/mL in all media tested. Nevertheless, the highest titer equal to 5.2 ± 0.5 × 10⁷ FFU/mL, was achieved in IPT-AFM containing a reduced amount of Ca⁺⁺ and Mg⁺⁺. Our results demonstrate the suitability of the obtained VeroS cells to produce rabies virus at a high titer, and pave the way to develop VeroS cells bioreactor process for rabies vaccine production.     

悬浮阵列技术在研究与临床中的应用

悬浮阵列技术是一种以荧光编码微球为核心,集流式细胞、激光分析、高速数字信号处理等多种技术于一体的多指标并行分析技术平台,可一次同时准确定量检测100种不同的生物分子;具有高通量、高灵敏度,并行检测等特点;常用于免疫分析、核酸研究、酶学分析、受体、配体识别分析等研究.     

支撑喉镜手术治疗声带息肉小结

目的:观察支撑喉镜下手术治疗声带息肉/小结的疗效.方法:对100例声带息肉/小结的患者在全麻下行支撑喉镜手术.结果:100例中,术后3d发音恢复正常31例,术后6d发音恢复正常34例,好转35例,随访3mo无复发.结论:支撑喉镜下手术治疗声带息肉/小结疗效好、创伤小、精确度较高,但同时要注意并发症的预防和处理.     

支撑喉镜下喉显微手术治疗声带息肉、声带小结86例分析

目的：观察支撑喉镜下喉显微手术治疗声带息肉、声带小结的临床疗效。方法：回顾性分析86例采用支撑喉镜下喉显微手术治疗的声带息肉、声带小结患者的临床资料。结果：86例中，治愈84例，治愈率97．67％；好转2例，总有效率100％。结论：支撑喉镜下喉显微手术治疗声带息肉、声带小结是安全、彻底、疗效显著的治疗方法。     

曲马多药树脂速释混悬剂的研制

为掩盖盐酸曲马多的苦味 ,并达到迅速镇痛的目的 ,利用盐酸曲马多与离子交换树脂结合制成药物树脂的制备方法 ,进行了曲马多药树脂速释混悬剂的研制实验。实验结果表明 ,该制备方法简单 ,研制出的曲马多药树脂速释混悬剂稳定、口味好、释放迅速     

丘陵地区氯硝柳胺悬浮剂灭螺费用及效果分析

[目的]了解25%氯硝柳胺悬浮剂(SCN)在丘陵山区果园内灭螺效果,并进行成本-效果评价。[方法]选择宜兴市山丘地区2处有螺果园为试验现场,采用灭螺机喷洒的方法,实验组用SCN按2.0g/m2、对照组用50%氯硝柳胺乙醇胺盐可湿性粉剂(WPN)按4.0g/m2喷洒灭螺,比较两组有螺面积下降率和活螺密度下降率;以有螺面积下降率和活螺密度下降率作为评价指标进行成本-效果分析。[结果]实验组用SCN喷洒4次,灭螺后30d有螺面积下降率和活螺密度下降率均为100.0%,有螺面积和活螺密度每下降1个百分点的费用均为56.00元,分别优于对照组WPN的99.00%、92.25%和92.93元、99.73元。[结论]用SCN山区灭螺,操作方便,成本低,灭螺效果好,具有良好应用前景。     

臭氧水消毒器对微生物杀灭效果的试验观察

目的：观察臭氧水消毒器产生臭氧水的杀菌效果。方法：采用悬液定量和载体定量杀菌试验进行检测。结果：该机产生含臭氧4．20mg／L的水溶液对大肠杆菌、金黄色葡萄球菌和白色念珠菌作用5min的杀灭率均为100％；当菌液中含有10％蛋白胨时，对大肠杆菌、金黄色葡萄球菌和白色念珠菌作用5min的杀灭率分别为55．67％、59．48％和37．44％。用消毒液流动浸泡载体定量杀菌试验，含臭氧4．20mg／L的水溶液作用30min对大肠杆菌、金黄色葡萄球菌和白色念珠菌的平均杀灭率分别为99．95％、99．99％和96．94％。结论：该臭氧水消毒器产生的臭氧水具有一定消毒效果。     

悬吊术治疗男性获得性尿失禁的研究进展

男性获得性尿失禁是指男性因外伤或手术导致括约肌受损或后天性神经源性疾病造成机体失去控制尿液的能力。相对于治疗金标准人工尿道括约肌植入术,悬吊术操作简单,费用低廉,疗效可靠,并发症少,在近年逐渐被医患所接受。该文就悬吊术治疗男性获得性尿失禁的研究进展予以综述。     

Synthesis and characterization of shape-memory poly carbonate urethane microspheres for future vascular embolization

Two types of shape memory poly carbonate urethanes (PCUs) microspheres were synthesized by pre-polymerization and suspension polymerization, based on Polycarbonate diol (PCDL) as the soft segment, Isophorone diisocyanate (IPDI) and 1,6-hexamethylene diisocyanate (HDI) as the hard segments and 1,4-butanediol (BDO) as the chain expanding agent. The structure, crystallinity, and thermal property of the two synthesized PCUs were characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), Differential scanning calorimetery (DSC), respectively. The results showed that the two types of PCUs exhibited high thermal stability with phase separation and semi-crystallinity. Also, the results of the compression test displayed that the shape fixity and the shape recovery of two PCUs were more than 90% compared to the originals, indicating their similar bio-applicability and shape-memory properties. The tensile strength, elongation at break was enhanced by introducing and increasing content of HDI. The water contact angles of PCUs decreased and their surface tension increased by surface modified with Bovine serum albumin (BSA). Furthermore, the biological study results of two types of PCUs from the platelet adhesion test and the cell proliferation inhibition test indicated they had some biocompatibilites. Hence, the PCU microspheres might represent a smart and shape-memory embolic agent for vascular embolization.     

自体表皮细胞混悬液移植治疗皮肤慢性溃疡临床观察

目的评估自体表皮细胞混悬液移植治疗皮肤慢性溃疡的疗效。方法 20例皮肤慢性溃疡患者,以1:1比例分为自体表皮细胞混悬液移植组(试验组)和常规外科换药组(对照组)。观察溃疡在两组的平均愈合时间。结果对照组与试验组创面的平均愈合时间分别为(0.177±0.114)cm2/d和(0.375±0.238)cm2/d,差异有统计学意义(P=0.029)。结论自体表皮细胞混悬液移植是治疗皮肤慢性溃疡的一种有效方法。