Taura 综合征病毒主要结构蛋白的表达与纯化

根据Taura综合征病毒(TSV)基因组,设计特异性引物,从感染病毒组织中提取组织总RNA后扩增,分别将3个主要结构蛋白基因VP1、VP2和VP3克隆到pGEM TEasyVector.与表达载体连接后,导入大肠杆菌中诱导表达,并纯化目的蛋白.诱导表达的融合蛋白分子量分别为54.2×103、43×103和57.1×103,在变性条件下过柱纯化VP1和VP2,一次可以纯化10mg以上纯度较高的蛋白.     

湿热证动物模型微量元素、维生素E变化及清香散的调节作用

采用多因素造模方法复制成湿热证动物模型，观察了动物模型微量元素Zn、Cu、Fe、Se和维生素E代谢水平的变化。结果显示，模型动物血清Zn下降（P＜0．05），Cu升高（P＜0．05或P＜0．01），Fe变化不大（P＞0．05），血Se水平下降（P＜0．05），血浆维生素E含量减少（P＜0．01）。经清热祛湿的经验方清香散治疗后，治疗I组动物血清Zn、血Se、血浆维生素E明显升高（P＜0．05），血清Cu下降（P＜0．05）；治疗Ⅱ组血浆血清Cu有变化（P＜0．05）外，其余变化不大。     

A molecular docking model of SARS-CoV S1 protein in complex with its receptor, human ACE2

The exact residues within severe acute respiratory syndrome coronavirus (SARS-CoV) S1 protein and its receptor, human ACE2, involved in their interaction still remain largely undetermined. Identification of exact amino acid residues that are crucial for the interaction of S1 with ACE2 could provide working hypotheses for experimental studies and might be helpful for the development of antiviral inhibitor. In this paper, a molecular docking model of SARS-CoV S1 protein in complex with human ACE2 was constructed. The interacting residue pairs within this complex model and their contact types were also identified. Our model, supported by significant biochemical evidence, suggested receptor-binding residues were concentrated in two segments of S1 protein. In contrast, the interfacial residues in ACE2, though close to each other in tertiary structure, were found to be widely scattered in the primary sequence. In particular, the S1 residue ARG453 and ACE2 residue LYS341 might be the key residues in the complex formation.     

Genomic variation and point mutations analysis of Indian COVID-19 patient samples submitted in GISAID database

Corona virus disease 2019 (COVID-19) endemic has havoc on the world; the causative virus of the pandemic is SARS CoV-2. Pharmaceutical companies and academic institutes are in continuous efforts to identify anti-viral therapy or vaccines, but the most significant challenge faced is the highly evolving genome of SARS CoV-2, which is imparting evolutionary selective benefits to the virus. To understand the viral mutations, we have retrieved nine hundred and thirty-four samples from different states of India via the GISAID database and analyzed the frequency of all types of point mutation in all structural, non-structural proteins, and accessory factors of SARS CoV-2. Spike glycol protein, nsp3, nsp6, nsp12, N and NS3 were the most evolving proteins. High frequency point mutations were Q496P (nsp2), A380V (nsp4), A994D (nsp3), L37F (nsp6), P323L & A97V (nsp12), Q57H (ns3), D614G (S), P13L (N), R203K (N), G204R (N) and S194L (N).     

治疗小儿厌食症中药强壮灵冲剂微量元素测定及其与疗效关系探讨

已证明小儿厌食症除与缺乏微量元素锌关系密切外,还与其他多种微量元素和宏量元素的缺乏或低下也有关。本院以自行研制的中药强壮灵冲剂治疗小儿厌食症疗效显著。其于浸膏粉经２０余种微量元素测定表明含有锌、铁、钴、铜、镍、锰、铬、锗、锶、铝、钡、硒、硅、钛等多种人体必需且品种齐全,含量适度的微量元素。上述元素又恰是患儿体内所缺乏和低下的品种,无疑对患儿全面适度的补充大有裨益．中药强壮灵与西药硫酸锌临床对比研究表明,无论比发锌回升数值以及从厌食等全面疗效统计,中药组均明显优于西药组,并且发现疗效与药物的补锌量不成正相关。说明中药的调理脾胃功能及综合整体治疗作用是其突出优点,值得推广．     

Intra-abdominal fat area is a predictor for new onset of individual components of metabolic syndrome: MEtabolic syndRome and abdominaL ObesiTy (MERLOT study)

Objective: To investigate the significance of intra-abdominal fat area (IAFA) on new onset of individual components of the metabolic syndrome: high blood pressure, dyslipidemia, or hyperglycemia. Methods: We conducted a longitudinal study using checkup data of a hospital from 1994 to 2010. Of 25,255 subjects, we examined 1,380 Japanese, who underwent computed tomography to measure IAFA and had no metabolic syndrome components at baseline. Results: During 3.6 years of the mean follow-up period, one of metabolic syndrome components occurred in 752 subjects. Of three components, high blood pressure was more prevalent. The multiple Cox regression analysis disclosed that IAFA is significantly associated with onset of metabolic syndrome components (HR: 1.05 per 10 cm², 95%CI: 1.03–1.07). This finding was independent of BMI, and significant even in non-obese individuals with body mass index <25 kg/m². Conclusions: MERLOT study demonstrates that IAFA is an independent predictor for new onset of individual components of the metabolic syndrome, even in non-obese healthy Japanese.     

Separation and Analysis of o.p'-DDD and its Lipophilic Metabolite,O.P'-DDE,in Tissue,Faeces and Plasma

Abstract

An analytical procedure based on electron capture gas chromatography for the determination of o,p′-DDD, a drug used for the treatment of Cushing′s syndrome, is presented. Detection of a metabolite, identified as o,p′-DDE, is also described.     

探究多囊卵巢综合征患者冻融胚胎移植子宫内膜准备方案的选择

目的探讨多囊卵巢综合征患者冻融胚胎移植子宫内膜准备的两种方案的效果。方法选择2014年3月到2016年3月间在深圳武警医院选择行冻融胚胎移植子宫内膜准备的700例多囊卵巢综合征患者作为研究对象,对所有患者的临床资料进行回顾性分析,根据患者子宫内膜准备方案分为人工组(n=350)和促排卵组(n=350),对比两组患者冻融胚胎移植效果。结果人工组患者胚胎复苏率、优质胚胎率、移植胚胎数、子宫内膜厚度、胚胎种植率、妊娠率与促排卵组相比均无差异,P0.05。结论人工子宫内膜与促排卵内膜效果相似,均可推广运用。     

Validation of a HPLC/MS method for simultaneous quantification of clonidine,morphine and its metabolites in human plasma

A high‐performance liquid chromatography–tandem mass spectrometry method was developed and validated for the simultaneous quantification of morphine, morphine's major metabolites morphine‐3‐glucuronide and morphine‐6‐glucuronide, and clonidine, to support the pharmacokinetic analysis of an ongoing double‐blinded randomized clinical trial that compares the use of morphine and clonidine in infants diagnosed with neonatal abstinence syndrome. Plasma samples were processed by solid‐phase extraction and separated on an Inertsil ODS‐3 (4 μm) column using an 0.1% formic acid in water–0.1% formic acid in methanol gradient. Detection of the analytes was conducted in the positive multiple reaction monitoring mode. The range of quantitation was 1–1000 ng/mL for morphine, morphine‐3‐glucuronide and morphine‐6‐glucuronide, and 0.25–100 ng/mL for clonidine. Intra‐day and inter‐day accuracy and precision were ≤15% for all analytes across the quantitation range. Extraction recovery rates were ≥94% for morphine, ≥90% for M3G, ≥87% for M6G and ≥ 79% for clonidine. Matrix effect ranged from 85–94% for clonidine to 101–106% for M3G. The method fulfilled all predetermined acceptance criteria and required only 100 μL of starting plasma volume. Furthermore, it was successfully applied to 30 clinical trial plasma samples.     

钙蛋白酶-10 基因SNP-19 与多囊卵巢综合征相关性研究

目的 探讨钙蛋白酶-10 基因（CAPN-10）单核苷酸多态性-19（SNP-19）与多囊卵巢综合征（PCOS）的相关性。 方法 采用聚合酶链反应- 限制性片段长度多态性（PCR-RFLP）技术,对107 例PCOS患者（PCOS 组）及111 例健康体检者（对照组）的CAPN-10 SNP-19 进行分析,并测定其性激素和生化指标。PCOS组再根据BMI与胰岛素抵抗（IR）稳态模型指数（HOMA-IR）分为肥胖组与非肥胖组、IR 组与非IR 组,分析CAPN-10 SNP-19 与其关系。结果 与对照组相比,PCOS 组BMI、空腹血糖（FPG）、TG、TC、LDL、黄体生成素（LH）、黄体生成素/ 卵泡刺激素（LH/FSH）、睾酮（T）均升高（均P＜0.01）。PCOS 组CAPN-10 SNP-19 1等位基因及1/2基因型频率均高于对照组（均P＜0.05）;且CAPN-10 SNP-19 1/2基因型PCOS 发病风险较1/1、2/2 基因型增加约2倍（OR=1.9,95％CI=1.1~3.3,P＜0.05）。CAPN-10 SNP-19 1、2等位基因频率PCOS 肥胖组与非肥胖组、IR 组与非IR组比较均无统 计学差异（均P＞0.05）;PCOS 肥胖组和IR组CAPN-10 SNP-19 1/2基因型频率分别高于非肥胖组和非IR 组,但差异均无统计学意义（均P＞0.05）。结论 CAPN-10 SNP-19 与PCOS 遗传易感性有关,可能与PCOS 肥胖及IR 无关。