Semi-perfusion cultures of suspension MDCK cells enable high cell concentrations and efficient influenza A virus production

Control and prevention of rapid influenza spread among humans depend on the availability of efficient and safe seasonal and pandemic vaccines, made primarily from inactivated influenza virus particles. Current influenza virus production processes rely heavily on embryonated chicken eggs or on cell culture as substrate for virus propagation. Today’s efforts towards process intensification in animal cell culture could innovate viral vaccine manufacturing using high-yield suspension cells in high cell density perfusion processes. In this work, we present a MDCK cell line adapted to grow as single cell suspension with a doubling time of less than 20 h, achieving cell concentrations over 1 × 10⁷ cells/mL in batch mode. Influenza A virus titer obtained in batch infections were 3.6 log₁₀(HAU/100 µL) for total- and 10⁹ virions/mL for infectious virus particles (TCID₅₀), respectively. In semi-perfusion mode concentrations up to 6 × 10⁷ cells/mL, accumulated virus titer of 4.5 log₁₀(HAU/100 µL) and infectious titer of almost 10¹⁰ virions/mL (TCID₅₀) were possible. This exceeds results reported previously for cell culture-based influenza virus propagation by far and suggests perfusion cultures as the preferred method in viral vaccine manufacturing.     

Effects of Single-Dose Preemptive Pregabalin and Intravenous Ibuprofen on Postoperative Opioid Consumption and Acute Pain after Laparoscopic Cholecystectomy

Purpose: Non-opioid medications as a part of multimodal analgesia has been increasingly suggested in the management of acute post-surgical pain. The present study was planned to compare the efficacy of the combination of pregabalin plus ?v ibuprofen. Methods: 58 patients were included in this prospective, randomized, double-blinded study. The pregabalin group (Group P, n = 29) received 150 mg pregabalin, the pregabalin plus ibuprofen group (Gropu PI, n = 29) received 150 mg pregabalin and 400mg ?v ibuprofen before surgery. Postoperative fentanyl consumption, additional analgesia requirements and PACU stay were recorded. Postoperative analgesia was performed with patient-controlled IV fentanyl. Results: VAS scores in the group PI were statistically lower at PACU, 1and 2 hours at rest, at PACU, 1, 2, 4, 12 and 24 hours on movement compared to the group P (P < 0.05). Opioid consumption was statistically significantly higher in the group P compared to the group PI (130.17 ± 60.27 vs 78.45 ± 60.40 μq, respectively, P < 0.001) and reduced in the 4th 24 hours by 55% in group PI. Rescue analgesia usage was statistically significantly higher in the group P than in the group PI (16/29 vs 7/29, respectively, P < 0.001). Four patient in the group PI did not need any opioid drug. Besides, PACU stay was shorter in the group PI than the group P (10.62 ± 2.38 vs 15.59 ± 2.11 min, respectively, P < 0.001). Conclusion: Preemptive pregabalin plus ?v ibuprofen in laparoscopic cholecystectomy reduced postoperative opioid consumption. This multimodal analgesic aproach generated lower pain scores in the postoperative period.     

Dorsal suspension for Morton’s neuroma: A comparison with neurectomy

BackgroundThe purpose of this study was to investigate and compare the clinical outcomes of dorsal suspension with those of neurectomy for the treatment of Morton’s neuroma.MethodsWe conducted a retrospective study of dorsal suspension and neurectomy group. The dorsal suspension was performed by dorsal transposition of neuroma over the dorsal transverse ligament after neurolysis. The visual analog scale (VAS), the Foot and Ankle Ability Measure (FAAM), postoperative satisfaction, and complications were evaluated.ResultsBoth groups reported significant pain relief, and there were no significant differences between the groups with respect to postoperative pain. The postoperative FAAM outcomes showed no significant between-group differences. Satisfaction analysis showed ‘excellent’ and ‘good’ results in the dorsal suspension and neurectomy groups (95% and 77.7%, respectively). Complications of numbness and paresthesia reported in the dorsal suspension group (5% and 5%, respectively) were significantly fewer than those of neurectomy group (61.1% and 33.3%, respectively) (both, p < .05).ConclusionsWith its favorable results, dorsal suspension can be another operative option for the treatment of Morton’s neuroma.Level of Evidence: Level III, retrospective comparative case series.     

LC-HR-MS/MS法鉴定头孢丙烯干混悬剂中的未知杂质

目的:检测头孢丙烯干混悬剂中的未知杂质,并对其进行结构鉴定。方法:采用高效液相色谱-串联高分辨质谱法检测并鉴定头孢丙烯干混悬剂中的未知杂质。色谱柱为Thermo HyPURITYTMC18,流动相为乙腈-0.013%甲酸水溶液(梯度洗脱),检测波长为230 nm,流速为1.0 m L/min,柱温为40℃,进样量为20μL;以电喷雾离子源行正离子全扫描,扫描范围为质荷比(m/z)100~1500,喷雾电压为3.8 kV,金属毛细管温度为320℃,鞘气压力为60 Arb,辅助气压力为10 Arb,喷雾温度为280℃。结果:在该色谱条件下,杂质K的检测限为0.202μg/mL,精密度、重复性试验的RSD均小于4%。杂质K附近发现3个未知杂质,且互为异构体,离子保留时间为17.83~19.31 min,二级母离子均为m/z 436.1500[M+H]+,可能为头孢丙烯开环、脱水后的产物。结论:本方法检测出头孢丙烯干混悬剂中杂质K附近的3个未知杂质。     

多索茶碱联合布地奈德混悬液雾化吸入对哮喘患者炎性因子及肺功能的影响

目的探讨多索茶碱联合布地奈德混悬液雾化吸入治疗哮喘的临床效果。方法选择我院2018年5月至2019年4月收治的哮喘患者114例,随机分为两组各57例。对照组采用布地奈德混悬液雾化吸入治疗,观察组在此基础上采用多索茶碱治疗。观察两组的临床疗效、炎性因子指标(IL-4、 IFN-γ)、肺功能指标(FVC、 FEV1)及不良反应情况。结果观察组的治疗总有效率为92.98%,高于对照组的78.95%(P <0.05)。治疗后,观察组的IL-4水平低于对照组,IFN-γ、 FEV1、 FVC水平均高于对照组,差异有统计学意义(P <0.05)。两组的不良反应发生率比较,差异无统计学意义(P>0.05)。结论多索茶碱联合布地奈德混悬液雾化吸入治疗哮喘可有效改善患者的肺功能和临床症状,调节炎性因子水平,且药物副作用少。     

悬吊运动训练系统对不随意运动型脑瘫的康复效果分析

目的 分析悬吊运动训练系统对不随意运动型脑瘫的康复效果,为临床治疗提供科学依据。方法 选取2015年8月-2017年8月荆州市妇幼保健院收治的42例不随意运动型脑瘫患儿为研究对象,采用数字随机法均分为观察组与对照组,各21例,对照组患儿接受常规康复治疗,观察组患儿在常规康复治疗基础上运用悬吊运动训练系统,连续康复治疗3个月。采用Berg平衡量表(BBS)、粗大运动功能评分(GMFM)评价两组患儿平衡及粗大运动功能,记录步长、步宽以及10 m最大步行速度(MWS)评价步行能力,并比较不同年龄、粗大运动功能分级系统(GMFCS)分级对步行能力的影响。结果 两组患儿治疗前GMFM 评分、BBS评分差异无统计学意义(P>0.05),治疗后均显著增加,且治疗后观察组均显著高于对照组,差异有统计学意义(t=4.767、3.543,P＜0.01);两组治疗前步行能力相关评分差异无统计学意义(P>0.05),治疗后步宽显著降低,步长、MWS显著增加,且治疗后观察组步长、MWS显著高于对照组,步宽显著低于对照组(t=4.767、3.985、4.221,P＜0.001);治疗后不同年龄及GMFCS分级患儿步宽、步长、MWS比较,差异具有统计学意义(年龄:t=3.986、2.878、2.986,P＜0.001;GMFCS分级:t=4.043、4.173、3.875,P＜0.001)。结论 在常规康复治疗的基础上加用悬吊运动训练系统能提升不随意运动型脑瘫患儿粗大运动及平衡能力,降低步行宽度,提升步速及步长,也能显著提升其康复效果,同时患儿年龄及GMFCS分级影响治疗效果。