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.     

皮肤瘙痒神经传导介质的研究进展

瘙痒是皮肤科的一种常见症状,是多因素相互作用的结果,其发生受到中枢和外周机制的双重调控,但具体发生机制尚不十分清楚。近年来随着研究的不断深入,多种物质如组胺、乙酰胆碱、5-羟色胺、蛋白酶和蛋白酶相关受体、细胞因子、阿片样肽等介质在瘙痒产生过程中起重要作用,这些介质通过刺激C类神经纤维或直接与皮肤感觉神经纤维受体相结合来介导瘙痒。本文就将皮肤瘙痒的神经传导及传导介质作一综述。     

杆菌肽发酵的培养基优化及影响其检测的因素

通过对影响杆菌肽抑菌圈大小的各种因素的研究，从而得到提高地衣芽抱杆菌发酵杆菌肽的产量及测量的最佳效果。研究结果表明豆芽汁培养基发酵的杆菌肽产量最高，并且在豆芽汁中加糖量为2g／dL时，抑菌圈最大。管碟法测定时，培养基的厚度影响最为显著，下层厚度10mL，上层5mL最佳，上层培养基的酸碱度为7．0，指示菌的浓度在10^8cfu／mL时，抑菌圈最清晰，直径最大，平均直径为2．52cm。     

衣原体、支原体引起的难治性支气管—肺部感染分析

目的 探讨难治性支气管—肺部感染(难治组)衣原体、支原体两种病原体的表达。方法 应用ELISA法检测了29例难治性支气管—肺部感染两种非典型病原体急性感染指标，衣原体抗体IgM、支原体抗体IgM及冷凝集实验，并常规细菌学检查，与同期住院的普通性下呼吸道感染(普通组)40例，门诊及住院的急性上呼吸道感染(上感组)30例进行比较。结果 难治组衣原体急性感染率68．9％，显著高于普通组、上感组，上感组最低；支原体急性感染率难治组34、5％，显著高于普通组、上感组；衣原体混合支原体、衣原体混合其他致病菌感染均显著高于普通组和上感组。结论 难治性下呼吸道感染中衣原体、支原体感染率较高，且易与多种病原体混合感染，衣原体的慢性感染可能是难治感染的重要原因。     

抗莱氏支原体单克隆抗体的研制

通过细胞融合技术，筛选出ＭＡＬ－１和ＭＡＬ－２两株抗莱氏支原体（Ａ．Ｌａｉｄｌａｗｉｉ，Ａ．Ｌ）单克隆抗体（单抗）。通过ＳＤＳ－ＰＡＧＥ和Ｗｅｓｔｅｒｎ印迹法测得单抗相对应的抗原分子量均为６７．４６ｋｕ。用间接免疫荧光法（ＩＦＡ）检测，单抗与细胞培养中另外３种常见污染的支原体和呼吸道肺炎支原体无交叉反应。将单抗用于检测支原体污染的细胞培养物，结果表明，此单抗能特异性地检出莱氏支原体。故可用其推断支原体污染途径，便于制定防治措施。     

3种培养基体外培养人芽囊原虫的效果观察

目的　观察人芽囊原虫在 3种培养基中的生长繁殖情况 ,以探讨最佳的培养方法。　方法　从大学生粪便中分离人芽囊原虫 ,转种至 3种不同培养基 ,定时观察人芽囊原虫的生长繁殖情况 ,根据原虫密度评价培养效果。　结果培养 96h后 ,Locke’s鸡蛋血清 (LES)双相培养基中的人芽囊原虫密度最高 ,改良Jones单相液体培养基次之 ,Locke’s琼脂血清 (LAS)双相培养基原虫密度最低 ,差异有显著性 (P <0 .0 5 ) ;观察 3种培养基中的原虫密度高峰持续时间分别为 3 60h、96h和 72h ,最长存活时间分别为 5 5 2h、3 3 6h和 168h。　结论　Locke’s鸡蛋血清双相培养基较适用于人芽囊原虫体外增殖培养。     

支原体肺炎肺外损伤患儿T细胞亚群、IFNγ、TNFα的变化及意义

目的 探讨支原体肺炎肺外损伤患儿细胞免疫、细胞因子状况和胸腺肽的治疗效果。方法 采用流式细胞仪和酶标仪检测31例支原体肺炎肺外损伤组患儿急性期、恢复期血中CD3CD4、CD8、CD4／CD8干扰素γ（IFNγ）、肿瘤坏死因子α（TNFα水平，并与支原体肺炎组比较。结果 ①急性期支原体肺炎肺外损伤组和支原体肺炎组比较，CD3、CD4显著降低（P〈0．05），TNFα显著升高（P〈0．01），CD8、IFNγ无统计学意义（P〉0．05）；②支原体肺炎肺外损伤组急性期和4周后比较，CD3、CD4、IFNγ升高（P〈0．05），TNFα显著降低（P〈0．01），CD8无变化。用胸腺肽治疗患者以上指标变化更明显。结论 支原体肺炎肺外损伤患儿细胞免疫功能低于支原体肺炎患儿；细胞因子中TNFα早期升高，而IFNγ不明显；恢复期TNFα下降，而IFNγ升高明显。用胸腺肽治疗能缩短病程。     

Social climate within secure inpatient services for people with intellectual disabilities

Background The social climate of inpatient facilities is thought to be an important contributor to treatment outcome. However, little research has focused on this construct within secure forensic services for people with intellectual disabilities (ID). Therefore, the objective of this study was to investigate the social climate of two different types of secure units (‘low’ secure vs. ‘medium’ secure) contained within the same facility for offenders with ID. Two hypotheses were generated: (1) residents would rate the social climate of the whole facility in a more negative direction than staff, and (2) residents and staff would rate the social climate of the ‘low’ secure unit in a more positive direction than that of the ‘medium’ secure unit. Method Using a 2 (factor ‘Participant’ = Staff or Resident) × 2 (factor ‘Unit’ = ‘Low’ or ‘Medium’ Secure Unit) between‐subjects design, 18 residents and 37 staff members were recruited and completed the Correctional Institutions Environment Scale (CIES), a measure of social climate. Results Residents tended to rate the units in a more positive direction than staff on some sub‐scales. Participants rated the ‘low’ secure unit in a more positive direction than the ‘medium’ secure unit on two sub‐scales of the CIES. However, on selected sub‐scales there were differences. The findings of this study suggest that the CIES may be a valid instrument for use within forensic services for people with ID, and further suggests that residents and staff have different perceptions of the shared social climate, which may have implications for service development.     

结肠CT扫描技术初探

目的 :通过分析比较结肠扫描不同充填剂与不同体位的方法 ,探讨出最佳充填剂与最佳体位以提高结肠 CT扫描质量。方法 :分别对 4 0例结肠 CT扫描病人采用不同的充填剂 ,与不同的体位 ,评价不同的充填剂与不同体位的优缺点。结果 :水作为充填剂组病灶显示情况明显优于泛影萄胺组且伪影少 ,选择不同体位组图像质量也明显优于常规体位组。结论 :结肠 CT扫描时应首选水作为充填剂 ,根据病变不同选择不同的体位 ,有利于图像质量的提高