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.     

雾化吸入布地奈德治疗AECOPD合并2型糖尿病患者疗效及对肺通气功能和糖代谢的影响

目的分析雾化吸入布地奈德治疗慢性阻塞性肺疾病急性加重(AECOPD)合并2型糖尿病(T2DM)患者疗效及对肺通气功能和糖代谢指标的影响。 方法选择四川大学华西医院2019年1月至2020年5月诊治的120例AECOPD合并T2DM患者作为对象，根据非随机临床同期对照研究及患者自愿原则分为对照组58例和观察组62例，其中对照组给予低剂量布地奈德治疗，观察组给予高剂量布地奈德治疗。比较两组患者治疗前后肺通气功能、糖代谢指标和炎性因子水平变化，并评估疗效和安全性。 结果两组患者治疗有效率比较无显著差异(P>0.05)；与治疗前比，两组患者治疗后肺通气功能指标均显著升高，但观察组较对照组升高更为显著(P<0.05)；两组患者之间及其治疗前后糖代谢指标均无显著差异(P>0.05)；与治疗前比，两组患者治疗后炎症因子水平均显著降低，但观察组较对照组降低更为显著(P<0.05)；两组不良反应发生率比较无统计学意义(P>0.05)。 结论布地奈德雾化吸入治疗AECOPD合并T2DM安全有效，对糖代谢指标无明显影响，但高剂量的布地奈德对于患者肺通气功能和炎症水平改善效果更好。     

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.     

鼻塞式同步间歇正压通气(NIPPV)联合布地奈德雾化治疗新生儿急性呼吸窘迫综合征疾病的临床应用研究

目的:分析鼻塞式同步间歇正压通气(NIPPV)联合布地奈德雾化治疗ARDS(新生儿急性呼吸窘迫综合征)疾病的临床应用效果。方法:纳入病例是2017年5月—2019年11月收治的104例ARDS新生儿,随机平均分为两组。参照组52例采纳CPAP(持续气道正压通气通气)治疗,实验组52例采纳NIPPV+布地奈德雾化治疗,对比两组呼吸机通气时间、用氧时间、住院时间、血气指标以及并发症发生情况。结果:实验组呼吸机通气时间、用氧时间、住院时间均明显短于参照组,差异有统计学意义P<0.05;实验组治疗3 d后PaCO2明显低于参照组,实验组治疗3 d后PH以及PaO2明显高于参照组,差异有统计学意义P<0.05;实验组并发症发生率(3.85%,2/52)明显低于参照组(21.15%,11/52),差异有统计学意义P<0.05。结论:NIPPV+布地奈德雾化可有效缩短ARDS患者机械通气时间,改善血气指标,降低并发症发生率,值得借鉴。     

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。     

布地奈德联合博利康尼治疗慢性阻塞性肺疾病急性加重期疗效分析

目的观察布地奈德联合博利康尼治疗慢性阻塞性肺疾病急性加重期的临床疗效。方法选择2017年12月~2018年12月我院住院治疗的慢性阻塞性肺疾病患者80例,随机分为观察组和对照组,各40例,对照组予布地奈德雾化液1 mg+生理盐水3 m L,每天2次雾化吸入;观察组在对照组基础上加用博利康尼雾化液l mg联合布地奈德雾化液1 mg+生理盐水3 m L雾化吸入,每天2次,6 d为一个疗程。治疗后对比分析两组的临床疗效及肺功能各项指标的变化情况。结果观察组患者治疗后的总有效率达97.2%,显著高于对照组的75.0%,两组比较差异具有显著性(P0.05)。观察组患者治疗后的FEV1、PEF及FEV1/FVC分别为(1.93±0.27)L,(3.81±1.16)L/s、(90.43±12.65),均较治疗前明显提高,且观察组患者治疗后的FEV1、PEF及FEV1/FVC水平分别显著高于对照组,两组比较差异具有显著性(P0.05)。结论布地奈德联合博利康尼治疗慢性阻塞性肺疾病急性加重期疗效确切,能明显改善患者的肺功能,从而缓解患者的临床症状,提高生活质量,改善预后。     

糖皮质激素与β2-受体激动剂小剂量吸入治疗哮喘临床疗效观察

目的：评估糖皮质激素与β2－受体激动剂小剂量吸入治疗哮喘的临床疗效。方法：将我院自2012年5月至2014年3月收治的86例哮喘患者随机分为观察组与对照组，每组43例。观察组采用糖皮质激素与β2－受体激动剂（布地奈德与沙美特罗）小剂量吸入治疗；对照组则采用糖皮质激素（布地奈德）治疗，比较两种方法的疗效。结果：两组患者分别经治疗2周与4周后，FEV1均有增长趋势，但观察组增长程度明显高于对照组，两组相比，差异有统计学意义（ P＜0．05）。且治疗后观察组患者PEF变异率较对照组相比，差异有统计学意义（ P＜0．05）。对比两组患者哮喘控制测试表得分情况，观察组患者0～1分人数明显高于对照组，表明观察者患者哮喘恢复情况明显优于对照组，且观察者治疗过程中发生咽部感染，咳嗽，皮疹等不良反应发生率较对照组明显降低（ P＜0．05）。结论：糖皮质激素与β2－受体激动剂治疗哮喘较单一糖皮质激素相比，可明显缓解哮喘症状，促进患者恢复，降低不良反应发生率，具有重要的临床价值。     

清肺平喘颗粒联合布地奈德治疗支气管哮喘疗效观察

目的探讨清肺平喘颗粒联合布地奈德治疗支气管哮喘疗效及对患者肺功能改善的作用。方法 160例符合入选标准的支气管哮喘患者按治疗方案分为对照组(n=80)和观察组(n=80)。对照组给予布地奈德治疗,观察组联合应用清肺平喘颗粒治疗。比较两组患者临床疗效。结果观察组的治疗总有效率为92.5%,明显高于对照组81.2%的总有效率(P0.05)。结论清肺平喘颗粒联合布地奈德对治疗支气管哮喘疗效可靠,值得临床上进一步研究。