Mendelian randomization as a tool to gain insights into the mosaic causes of autoimmune diseases

Autoimmune diseases (ADs) are a broad range of disorders which are characterized by long-term inflammation and tissue damage arising from an immune response against one's own tissues. It is now widely accepted that the causes of ADs include environmental factors, genetic susceptibility and immune dysregulation. However, the exact etiology of ADs has not been fully elucidated to date. Because observational studies are plagued by confounding factors and reverse causality, no firm conclusions can be drawn about the etiology of ADs. Over the years, Mendelian randomization (MR) analysis has come into focus, offering unique perspectives and insights into the etiology of ADs and promising the discovery of potential therapeutic interventions. In MR analysis, genetic variation (alleles are randomly dispensed during meiosis, usually irrespective of environmental or lifestyle factors) is used instead of modifiable exposure to explore the link between exposure factors and disease or other outcomes. Therefore, MR analysis can provide a valuable method for exploring the causal relationship between different risk factors and ADs when its inherent assumptions and limitations are fully considered. This review summarized the recent findings of MR in major ADs, including systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), multiple sclerosis (MS), and type 1 diabetes mellitus (T1DM), focused on the effects of different risk factors on ADs risks. In addition, we also discussed the opportunities and challenges of MR methods in ADs research.     

我国弓形虫Chinese 1优势基因型感染对小鼠脑组织铁代谢影响

目的　探讨我国弓形虫Chinese 1优势基因型感染对宿主脑组织铁代谢及脑损伤的影响。方法　将20只C57BL/6（体质量15～17 g）小鼠随机分为对照组和感染组，每组10只。感染组每只小鼠腹腔注射4 000个弓形虫Chinese 1优势基因型TgCtwh3虫株速殖子，对照组小鼠注射等量无菌PBS，饲养6 d后处死小鼠并取其脑组织。采用电感耦合等离子体质谱法（inductively coupled plasma mass spectrometry, ICP⁃MS）检测小鼠脑组织铁元素水平；采用RNA芯片检测两组小鼠脑组织差异基因数目并对功能基因表达情况进行基因本体论（Gene Ontology, GO）功能富集；采用实时荧光定量PCR（fluorescent quantitative real⁃time PCR, qPCR）技术检测小鼠脑组织中弓形虫表面抗原1（Toxoplasma gondii surface antigen 1，TgSAG1）基因及部分锌铁调控蛋白（Zrt⁃ and Irt⁃like protein, ZIP）家族mRNA表达水平；采用光镜和电镜观察小鼠脑组织海马齿轮回（dentate gyrus, DG）超微结构；采用Western blotting检测谷胱甘肽过氧化物酶4（glutathione peroxidase 4, GPx4）蛋白表达水平；采用硫代巴比妥酸（TBA）法检测丙二醛（malondialdehyde, MDA）水平；采用免疫组化检测血管内皮生长因子（vascular endothelial growth factor, VEGF）蛋白表达光密度（optical density, OD）值。结果　光镜下可见感染组小鼠脑组织海马DG区细胞坏死，电镜下见感染组小鼠脑组织海马区出现胞质空泡化、核皱缩坏死、线粒体嵴断裂消融、自噬小体增加等超微结构变化。与对照组相比，感染组小鼠脑组织中铁元素水平上调[（32.92 ± 0.90） µg/g vs.（37.72 ± 1.10） µg/g；t = 3.397, P < 0.01]；RNA芯片检测感染组小鼠脑组织发现721个基因上调、276个基因下调，差异表达基因在金属离子结合能力上有明显富集。与对照组相比，感染组小鼠脑组织金属元素转运体ZIP2 mRNA表达水平上调（t = 8.659，P < 0.05）、GPx4表达下降[（1.046 ± 0.025） vs. （0.720 ± 0.101）；t = 3.129，P < 0.01]）、MDA水平升高[（4.37 ± 0.33） nmol/mgprot vs.（5.93 ± 0.54） nmol/mgprot；t = 2.451，P < 0.05）]、VEGF蛋白平均OD值上调[（0.348 3 ± 0.017 8） vs. （0.490 6 ± 0.010 5）；t = 6.641，P < 0.01]。结论　Chinese 1优势基因型弓形虫感染后，小鼠脑组织中铁元素蓄积、抗氧化能力下调、氧化应激水平升高，提示弓形虫感染可影响宿主脑组织铁代谢而导致脑损伤。     

Dose optimization with population pharmacokinetics of ritonavir-boosted lopinavir for Thai people living with HIV with and without active tuberculosis

BackgroundPrior to dolutegravir availability, ritonavir-boosted lopinavir (LPV/r) was an alternative recommendation when first-line drugs could not be used. A high concentration of protease inhibitors was observed in the Thai people living with HIV (PLWH). Thus, dose reduction of LPV/r may be possible. However, the pharmacokinetics and dose optimization of LPV/r have never been investigated. This study aimed to develop a population pharmacokinetic model of LPV/r and provide dosage optimization in Thai PLWH.MethodsLPV and RTV trough concentrations from Thai PLWH were combined with intensive data. The data were analyzed by the nonlinear mixed-effects modeling approach. The influence of RTV concentration on LPV oral clearance (CL/F) was investigated.ResultsRifampicin (RIF) use increased LPV and RTV CL/F by 2.16-fold and 1.99-fold, respectively. The reduced dose of 300/75 and 200/150 mg twice daily provided a comparable percentage of patients achieving LPV target trough concentration to the standard dose for PI-naïve patients. For HIV/TB co-infected patients receiving RIF who could not tolerate the recommended dose, the reduced dose of 600/150 mg twice daily was recommended.ConclusionThe population pharmacokinetic model was developed by integrating the interaction between LPV and RTV. The reduced LPV/r dosage offers sufficient LPV exposure for Thai PLWH.     

Metabolism of non-steroidal anti-inflammatory drugs (NSAIDs) by Streptomyces griseolus CYP105A1 and its variants

In the field of drug development, technology for producing human metabolites at a low cost is required. In this study, we explored the possibility of using prokaryotic water-soluble cytochrome P450 (CYP) to produce human metabolites. Streptomyces griseolus CYP105A1 metabolizes various non-steroidal anti-inflammatory drugs (NSAIDs), including diclofenac, mefenamic acid, flufenamic acid, tolfenamic acid, meclofenamic acid, and ibuprofen. CYP105A1 showed 4′-hydroxylation activity towards diclofenac, mefenamic acid, flufenamic acid, tolfenamic acid, and meclofenamic acid. It should be noted that this reaction specificity was similar to that of human CYP2C9. In the case of mefenamic acid, another metabolite, 3′-hydroxymethyl mefenamic acid, was detected as a major metabolite. Substitution of Arg at position 73 with Ala in CYP105A1 dramatically reduced the hydroxylation activity toward diclofenac, flufenamic acid, and ibuprofen, indicating that Arg73 is essential for the hydroxylation of these substrates. In contrast, substitution of Arg84 with Ala remarkably increased the hydroxylation activity towards diclofenac, mefenamic acid, and flufenamic acid. Recombinant Rhodococcus erythrocyte cells expressing the CYP105A1 variant R84A/M239A showed complete conversion of diclofenac into 4′-hydroxydiclofenac. These results suggest the usefulness of recombinant R. erythropolis cells expressing actinomycete CYP, such as CYP105A1, for the production of human drug metabolites.     

Carboxylesterase-2 plays a critical role in dabigatran etexilate active metabolite formation

Dabigatran etexilate (DABE), an oral anticoagulant prodrug, is nearly completely metabolized to the dabigatran (DAB) active metabolite by carboxylesterase-1 (CES1) and carboxylesterase-2 (CES2). The high interpatient variation in DAB plasma concentrations, coupled with its low therapeutic index, emphasizes the need to understand how CES1 and CES2 impact active metabolite formation. Previous work focused on CES1 enzyme activity but the contributions of CES2 remain unclear. The purpose of this study was to determine how CES2 activity influences DAB active metabolite formation. We compared the efficiency of DAB formation from DABE when exposed sequentially to human intestinal and then human hepatic microsomes (mimicking the normal metabolic sequence) with the reverse metabolic sequence in which DABE is exposed to hepatic and then intestinal microsomes. The poor efficiency of DAB formation with reverse sequential hydrolysis indicates that CES2 activity is crucial for active metabolite formation. Thus, the decrease in DAB formation with normal sequential hydrolysis was more sensitive to CES2 inhibition by verapamil (CES2 IC50 = 3.4 μM) than CES1 inhibition by diltiazem (CES2 IC50 = 9.1 μM). These results show CES2 activity plays a crucial role in DAB formation and that variability in its activity is an important determinant of therapeutic response.     

TOPSIS法和秩和比法模糊联合在基本公共卫生服务质量综合评价中的应用研究

背景国家基本公共卫生服务项目的开展是我国新医改的重要举措，自2009年国家基本公共卫生服务项目开展后，其服务经费与服务项目不断扩增，由于涉及指标较多，覆盖面较广，探寻科学、客观、全面的基本公共卫生服务综合评价方法十分必要。目的探索适宜的基本公共卫生服务质量综合评价方法，通过质量评价为调整相关政策和提高服务质量提供依据。方法2019年2—4月，采用多阶段立意抽样方式从Z省南部、中部和北部地区共选取24家社区卫生服务中心（乡镇卫生院）作为评价对象，记为机构A~X。采用逼近理想解排序法（TOPSIS法）、秩和比法及二者模糊联合的方法对24家社区卫生服务中心（乡镇卫生院）2018年基层医疗卫生机构基本公共卫生服务质量进行综合评价（参考2018年国家基本公共卫生服务项目选取12项评价指标）。结果在TOPSIS法评价中，Ci值排名前三名的为A（0.917 4）、C（0.875 9）和G（0.787 9），Ci值排名后三名的为I（0.414 2）、W（0.413 7）和N（0.407 7）。在秩和比法评价中，RSR值排名前三名的为A（0.890 6）、G（0.765 6）和C（0.711 8），RSR值排名后三名的为V（0.381 9）、W（0.362 8）和K（0.357 6）。根据模糊集理论，将W1Ci+W2RSR值进行排序，依据"择多原则"，排名前三名的分别为A、C和G，排名后三名的分别为I、K和W，这与TOPSIS法和秩和比法的评价结果基本一致。结论TOPSIS法和秩和比法模糊联合得到的评价结果及影响因素与其他研究结果相一致，并且两者联用能克服单一使用TOPSIS法或秩和比法的局限性，适宜在基本公共卫生服务质量评价中推广应用。     

孕期亲密伴侣暴力和孕晚期就寝-晨起规律与小于胎龄儿的关系

目的:探讨孕期亲密伴侣暴力(IPV)与小于胎龄儿(SGA)的关联,并分析孕晚期就寝-晨起规律在其关联中的作用。方法:从2015年3月-2019年6月在合肥市三家医院的产科门诊招募孕妇4908名,使用WHO多国妇女健康和家庭暴力研究小组总结而来的清单式问卷进行调查,并采用多元logistic回归模型进行分析。结果:孕期精神暴力、躯体暴力、性暴力和总IPV发生率分别为9.1%、1.4%、0.9%和9.7%,SGA发生率为7.1%。回归分析结果显示,孕期IPV与SGA正向关联(OR=1.59);分别按就寝时间和晨起时间分层,7∶00后起床、22∶00-23∶00和23∶00后就寝组的IPV与SGA正向关联(OR=2.85、2.18、1.98);按就寝-晨起规律分层,非早睡早起组的IPV和IPV组的非早睡早起与SGA正向关联(OR=2.15、3.32)。结论:孕期遭受IPV(尤其是精神暴力)会增加SGA的风险,而就寝-晨起规律会使IPV与SGA的关联在不同的规律下增强或减弱。     

M-MDSC在非霍奇金淋巴瘤中表达及临床意义

目的 探讨单核细胞型髓系来源抑制性细胞(M-MDSC)在非霍奇金淋巴瘤(NHL)患者中的表达水平以及临床意义.方法 选取2017年9月至2020年1月解放军第九○一医院和安徽医科大学第二附属医院35例初诊NHL患者作为试验组,同期选取20例健康志愿者作为对照组.比较两组研究对象M-MDSC、精氨酸酶-1(Arg-1)mRNA及诱导型一氧化氮合酶(iN-OS)mRNA的表达水平.分析M-MDSC与NHL患者年龄、性别、临床分期、病理分型、乳酸脱氢酶(LDH)以及IPI评分等临床指标的相关性.结果 试验组患者M-MDSC表达水平为(32.64±11.23)％,高于对照组,差异有统计学意义(P<0.05).Ⅲ～Ⅳ期、LDH升高及IPI评分为3～5分的NHL患者,M-MDSC水平升高,差异有统计学意义(P<0.05).试验组患者Arg-1水平为(18.12±5.25)ng/mL,iNOS mRNA相对水平为(12.36±3.68),均高于对照组,差异有统计学意义(P<0.05).结论 M-MDSC参与NHL患者发病过程,与疾病进展密切相关,可能成为治疗NHL的新靶点.     

黄芪甲苷调节PI3K/Akt/FoxO1通路抑制糖尿病大鼠肝糖异生

目的:探讨黄芪甲苷对2型糖尿病(T2DM)大鼠肝损伤保护潜在机制及肝组织磷酯酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/叉头框转录因子1(FoxO1),磷酸烯醇式丙酮酸羧基酶(PEPCK)和葡萄糖6-磷酸酶(G6Pase)蛋白表达的影响。方法:6周高糖高脂饮食后,链脲佐菌素(STZ)一次性腹腔注射(0.035 g·kg^-1)建立T2DM大鼠模型,随机分为正常组、模型组、黄芪甲苷低、中、高剂量组及二甲双胍组。黄芪甲苷低、中、高剂量组灌胃黄芪甲苷0.02,0.04,0.08 g·kg^-1·d^-1,二甲双胍组灌胃二甲双胍0.2 g·kg^-1·d^-1;给药8周后,于末次灌胃24 h禁食不禁水后处死,测血清肝生化指标,肝脏指数等;采用苏木素-伊红(HE)染色观察肝脏组织病理形态学变化;马松(Masson)染色观察纤维化程度;过碘酸希夫(PAS)染色反应观察细胞内糖原等变化;免疫组化及蛋白免疫印迹法(Western blot)检测各组肝中PI3K/Akt/FoxO1信号蛋白及PEPCK,G6Pase蛋白表达水平。结果:与正常组比较,模型组肝脏指数显著升高(P<0.01),肝功能指标丙氨酸氨基转移酶(ALT),天门冬氨酸氨基转移酶(AST),总胆固醇(TC),甘油三酯(TG)的含量显著升高(P<0.01),高密度脂蛋白胆固醇(HDL-C)显著降低(P<0.01),大鼠体质量显著减轻(P<0.01),PI3K/Akt/Fox O1信号减弱(P<0.01),PEPCK,G6Pase蛋白表达水平显著增强(P<0.01);与模型组比较,黄芪甲苷中、高剂量组及二甲双胍组肝功能指标ALT,AST,TC,TG的含量明显降低(P<0.05,P<0.01),大鼠体质量明显增加(P<0.05,P<0.01),肝组织Fox O1,PEPCK及G6Pase蛋白水平明显降低(P<0.05,P<0.01),Fox O1的磷酸化水平明显增强(P<0.05,P<0.01)。结论:黄芪甲苷可能通过调节PI3K/Akt/Fox O1信号通路抑制高脂高糖加小剂量STZ诱导T2DM肝糖异生,从而起到延缓T2DM大鼠肝脏损伤作用。