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.     

支撑喉镜下射频加囊肿揭盖术治疗会厌囊肿的临床研究

目的探讨囊肿揭盖术、囊肿揭盖术＋射频、射频＋囊肿揭盖术治疗会厌囊肿3种术式的疗效及其优劣。方法57例随机分为3组：单纯囊肿揭盖术组、囊肿揭盖术＋射频治疗组及射频＋囊肿揭盖术组。3组均采用表面麻醉＋强化麻醉经支撑喉镜下手术。随访6-12个月，比较3组总有效率、术中出血量及手术时间。结果3种术式总有效率均为100％，差异无显著性。囊肿揭盖术＋射频治疗组、射频＋囊肿揭盖术组与对照组比较，术中出血量、手术时间的差异均有显著性；囊肿揭盖术＋射频治疗组、射频＋囊肿揭盖术组术中出血量、手术时间相比较，差异均有显著性。结论3种术式均是治疗会厌囊肿的有效方法。射频＋囊肿揭盖术具有出血更少、手术时间更短、病人痛苦更小等优点，值得临床推广应用。     

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.     

Effect of fentanyl on the circulatory responses to orotracheal fibreoptic intubation

The effectiveness of fentanyl in attenuating the pressor and heart rate response to orotracheal fibreoptic intubation under general anaesthesia was assessed in 60 healthy patients undergoing elective surgery. Patients were randomly assigned to receive either fibreoptic intubation with or without fentanyl 6 micrograms.kg-1 or traditional Macintosh intubation with fentanyl 6 micrograms.kg-1. A standardised general anaesthetic was administered which included temazepam premedication, thiopentone, atracurium, oxygen, nitrous oxide and isoflurane. The pressor response to fibreoptic intubation was suppressed in those patients who received fentanyl and was similar to that seen in the Macintosh-fentanyl group of patients. The heart rate response to fibreoptic intubation was also significantly reduced in the patients who received fentanyl, but, in contrast, was still significantly greater than that in the Macintosh-fentanyl group. Fentanyl 6 micrograms.kg-1 appears to have a useful place in attenuating the cardiovascular effects of fibreoptic intubation under general anaesthesia.     

A Comparative Study on the Close Reduction of Arytenoid Dislocation under Indirect and Direct Laryngoscope

To assess the curative effects of different reduction techniques on the dislocation of cricoarytenoid joint caused by intubation,indirect laryngoscope (IL) and direct laryngoscope (DL) were utilized for the closed reduction of the displaced arytenoid under local anesthesia.23 patients who underwent the reduction for dislocated arytenoid under IL or DL from January 1991 to June 2001 were reviewed.The data were collected on the duration of the laryngeal injury,times of receiving reduction,side-effects after the treatment and the period for voice to returen to normal.The relationship between the duration of the laryngeal lesion and the period of the voice rehabilitation was examined.13 patients received the reduction under IL and 10 patients under DL,Except the times of the reduction,which showed significant difference,no differences were found between IL group and DL group in the course and the period of voice rehabilitation,as well as sore throat after the manipulation.The patients‘ voice recovery was positively related to their course of disease in both IL and DL group.It is coucluded that the recovery of normal voice is obviously affected by the duration of arytenoid dislocation.The reduction under Il is as effective as under DL in the treatment of arytenoid dislocation.Reduction by DL is better suit the patients with long time course of disease.     

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.     

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     

Tubeless Laryngotracheal Surgery in Infants and Children via Jet Ventilation Laryngoscope

We present the first use of tubeless superimposed combined high- and low-frequency jet ventilation (SHFJV) with a jet laryngoscope in laryngotracheal surgery in infants and children. Twenty-eight patients underwent 53 operative procedures. The average age of the patients was 7.3 years. The most common diagnoses were laryngeal papillomatosis and subglottic stenosis. The duration of jet ventilation averaged 33 min. The gas exchange was sufficient in each case. The advantages of SHFJV in the surgery of the laryngotracheal area in infants and children are optimal view at the larynx and trachea, maximum space for the handling, application of the laser without risks, no time limitation, suitability for stenosis, and neither anesthetic nor surgical complications.     

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。     

全静脉麻醉用于支撑喉镜声带手术的临床观察

目的:评价全静脉麻醉方法在支撑喉镜声带手术中的应用。方法:44例在全麻下行支撑喉镜声带手术成年患者,随机分为两组。两组均采用健忘镇痛慢诱导气管插管;麻醉维持A组采用瑞芬太尼-异丙酚,B组采用芬太尼-异氟醚。观察麻醉诱导、支撑喉镜置入、麻醉恢复各时点血流动力学的变化;记录停药到拔管到苏醒时间;比较停药10 m in后患者麻醉恢复质量。结果:①血流动力学指标:两组患者在麻醉诱导后,血流动力学无明显变化,但在支撑喉镜置入后、拔气管导管时,两组血压、心率均呈升高趋势。其中A组数据无统计学意义;B组P<0.05;两个时间点血流动力学应激反应发生率,B组明显高于A组。②两组停药到拔管时间无明显差异;B组停药到苏醒时间短于A组,P<0.05。③B组麻醉恢复评分明显高于A组。结论:选择芬太尼、瑞芬太尼联合异丙酚的全静脉麻醉用于该类手术,较芬太尼-异氟醚静吸复合麻醉,具有血流动力学更加稳定,苏醒快,恢复质量高的优点。