肿瘤治疗的新途径：嵌合抗原受体T细胞疗法

免疫疗法已经成为癌症治疗中的重要手段。使用嵌合抗原受体（chimeric antigen receptor,CAR）修饰的T细胞治疗癌症因其显著的疗效成为了癌症治疗研究的新方向。CAR包含抗原结合部分、铰链区、跨膜结构域以及结合活化T细胞的细胞共刺激结构。对目标肿瘤的特异性清除是通过CAR-T细胞对抗原的特异性选择和共刺激信号转导,CAR-T细胞在过继后的转运、维持,以及抗肿瘤功能的保留实现的。此文对目前基于CAR-T细胞疗法的基本原理及临床应用作一阐述。     

生物制品中的残留宿主细胞蛋白及其检测方法

宿主细胞蛋白（host cell protein,HCP）是指生物制品中来自生产细胞系的宿主细胞的蛋白成分,主要包括宿主细胞分泌的蛋白以及细胞凋亡、代谢产生的部分结构蛋白。作为生物制品生产过程中的残留杂质,HCP具有潜在的安全风险,包括诱导免疫应答。建立合适的HCP残留量检测方法将有助于监测生产工艺,确保生物制品的质量和安全。此文主要对HCP的分类、质量控制标准、常见检测方法及其原理进行阐述。     

水痘-带状疱疹病毒膜抗原荧光抗体试验的建立与评价

目的  建立用于检测血清中水痘-带状疱疹病毒（varicella-zoster virus,VZV)特异性抗体的膜抗原荧光抗体（fluorescent antibody to membrane antigen,FAMA）试验并对其进行验证和评价。方法  用VZV感染细胞作为抗原制备抗原载玻片,以异硫氰酸荧光素标记的羊抗人IgG为二抗建立FAMA试验。对试验的灵敏度、特异性和重复性进行验证。用FAMA试验和市售ELISA试剂盒对200份血浆样品中的抗VZV抗体进行检测。采用Spearman秩相关检验对两种方法的检测结果进行比较。结果 FAMA试验的灵敏度为0.04 IU/ml,与常见的人疱疹病毒（单纯疱疹病毒1和2型、人巨细胞病毒）没有交叉反应且重复性良好。经FAMA试验检测,200份血浆样品的抗体阳性率为93.5%,抗体几何平均效价为1:18.1,检测结果与市售ELISA试剂盒的符合率为90.0%。两种方法的检测结果具有相关性,Spearman秩相关系数为0.49,P<0.000 1。结论  建立的方法具有良好的灵敏度、特异性和重复性,可用于血清中抗VZV抗体的检测。     

麻疹疫苗及疫苗株基因特性分析

麻疹病毒（measles virus,MV）引起的麻疹是一种急性呼吸道传染病。对麻疹疫苗株与原型野毒株Edmonston的全基因组序列分析比较发现,所有疫苗株的磷蛋白、V蛋白、C蛋白、基质蛋白和血凝素中都含有相同的氨基酸突变位点。这些位点可作为未来研究MV减毒分子机制的重要靶点,但目前尚不明确是否与MV的减毒表型相关,需要结合反向遗传学、动物模型等进一步确定。     

流感疫苗病毒株在MDCK细胞中的培养条件

 目的   研究4价流感疫苗所用病毒株在MDCK细胞中的扩增条件。方法 在6孔细胞培养板中以不同感染复数（multiplicity of infection,MOI）（0.100 0、0.010 0、0.001 0、0.000 1）和对甲苯磺酰-L-苯丙氨酸氯甲基酮（TPCK）-胰蛋白酶（胰酶）浓度（0、2、4、8 μg/ml）进行病毒接种和扩增,感染后72 h收获细胞上清液,检测病毒血凝素效价,确定病毒最佳扩增条件。随后,在搅拌瓶中进行MDCK细胞微载体悬浮培养,研究不同起始细胞接种密度（1.5×10⁵、2.0×10⁵、3.0×10⁵ 个/ml）和微载体浓度（3、5、10 g/L）对MDCK细胞生长的影响,确定细胞最佳扩增条件。根据确定的最佳扩增条件,在搅拌瓶培养体系中扩增4种病毒。结果 6孔板中4种流感病毒的最佳接种条件分别是,A/Michigan/45/2015(H1N1) pdm09：MOI 0.010 0、TPCK-胰酶浓度2 μg/ml;A/Hongkong/4801/2014(H3N2)：MOI 0.010 0、TPCK-胰酶浓度4 μg/ml;B/Brisbane/60/2008：MOI 0.001 0、TPCK-胰酶浓度4 μg/ml;B/Phuket/3073/2013：MOI 0.010 0、TPCK-胰酶浓度4 μg/ml。在搅拌瓶中以3.0×10⁵ 个/ml初始细胞密度接种,3 g/L微载体浓度培养,MDCK细胞能够实现较好的扩增,最高密度可达2.1×10⁶ 个/ml;搅拌瓶悬浮培养,以最佳接种条件接种后,4种病毒的血凝素效价为：6.75~8.42log₂ 血凝素单位/50 μl。结论   通过摸索病毒接种最佳MOI、TPCK-胰酶浓度及优化MDCK细胞微载体悬浮培养条件,能够在MDCK细胞微载体悬浮培养体系中有效扩增4价流感疫苗用病毒株,为后期工艺放大研究奠定基础。     

4株脑膜炎球菌菌株的16S rRNA、PorA和PorB基因序列分析

目的  对A、C、Y和W135 4个不同血清群脑膜炎球菌菌株的16S rRNA、PorA和PorB基因进行序列分析,从分子水平对其做出鉴定。方法  提取脑膜炎球菌基因组DNA为 模板,设计特异性引物进行16S rRNA、PorA和PorB基因的PCR扩增并测序。结果  A、C、Y和W135群4个菌株的16S rRNA基因序列与GenBank中奈瑟菌属的脑膜炎球菌的同源性最高。4个菌株的PorA基因分型分别为P1.5-2,10; P1.7-1,1; P1.5-1,2-2; P1.5,2。PorB基因分别与porB3-26、porB2-40、porB3-100和porB3-25同源。结论  从基因水平分析表明4个菌株属于脑膜炎球菌,并在PorA和PorB基因分型上有差异。     

Comparative study on the antibacterial activity of phytochemical flavanones against methicillin-resistant Staphylococcus aureus

Differently substituted flavanones were isolated from Leguminosae and their antibacterial activity was comparatively studied against methicillin-resistant Staphylococcus aureus (MRSA). The minimum inhibitory concentrations (MICs) of phytochemical flavanones to clinical isolates of MRSA were determined by a serial agar dilution method. The structure-activity relationship has indicated that 2′,4′- or 2′,6′-dihydroxylation of the B ring and 5,7-dihydroxylation of the A ring in the flavanone structure are important for significant anti-MRSA activity and that substitution with a certain aliphatic group at the 6- or 8-position also enhances the activity. Among the thirteen flavanones tested, tetrahydroxyflavanones with these structural characteristics isolated from Sophora exigua and Echinosophora koreensis showed intensive activity to inhibit the growth of all MRSA strains at 3.13-6.25 μg/ml. The present hydroxyflavanones would be useful in the phytotherapeutic strategy against MRSA infections.     

卡介菌多糖核酸治疗口腔扁平苔藓有效性和安全性的荟萃分析

目的:荟萃分析卡介菌多糖核酸治疗口腔扁平苔藓的有效性和安全性。方法:检索美国医学文献数据库、Embase、Cochrane图书馆、万方数据库、维普数据库、中国期刊全文数据库、中国生物医学文献数据库、中国临床试验注册中心、美国临床试验注册中心,搜集卡介菌多糖核酸与非卡介菌多糖核酸比较治疗口腔扁平苔藓的临床随机对照试验。筛...     

Unusual problem with pain relief in an opium addict

The intra- and postoperative anaesthetic management of an opium addict is described. What initially appeared to be a withdrawal syndrome was later concluded to be a physiological manifestation of the anaesthetic and surgical technique, as well as an effect of opium.     

Thioredoxin-1 blocks methamphetamine-induced injury in brain through inhibiting endoplasmic reticulum and mitochondria-mediated apoptosis in mice

Methamphetamine (METH) has been reported to induce endoplasmic reticulum (ER) stress and neuronal apoptosis in the central nervous system (CNS) during the development of addiction. Thioredoxin-1 (Trx-1) is a redox regulating protein and plays an important role in inhibiting apoptosis and protects neurons from cytotoxicity through ER and mitochondria-mediated pathways. Our previous study has been reported that Trx-1 protects mice from METH-induced rewarding effect. However, whether Trx-1 plays the role in resisting METH injury is still unclear. Here, we aim to investigate whether Trx-1 participates in the regulation of METH-induced CNS injury via ER stress and mitochondria-mediated pathways. Our study first repeated the conditioned place preference expression induced by METH. Then we detected and found that METH increased the expression of N-methyl-d-asparate (NMDA) receptor subunit 2B (NR2B) and the level of glutamate (Glu) in the ventral tegmental area (VTA) and nucleus accumbens (NAc), while Trx-1 overexpression suppressed the increases. We further examined ER stress-related proteins and mitochondrial apoptosis pathway in the VTA and NAc, and found that METH increased the expressions of glucose regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), and Bax, as same time decreased the expressions of procaspase12, Bcl-2, and procaspase3, while Trx-1 overexpression blocked these changes. These results indicate that Trx-1 blocks METH-induced injury by suppressing ER stress and mitochondria-mediated apoptosis in the VTA and NAc via targeting glutamatergic system.