TLR2、TLR4、TLR5和TLR9在小鼠Hp感染模型中的表达

目的:研究BALB/c小鼠在幽门螺杆菌(Hp)感染前后和用抗生素治疗后胃黏膜组织中Toll样受体(TLR)2、TLR4、TLR5、TLR9、IL-1β的表达,探讨Toll样受体家族在Hp感染机制中的作用。方法:60只BALB/c小鼠随机分为3组,第1组不予处理(正常组),第2组予感染Hp(Hp感染组),第3组感染Hp后予抗生素治疗(抗生素治疗组)。RT-PCR和Western blotting法半定量检测小鼠胃内TLR2、TLR4、TLR5、TLR9、IL-1β的表达,Giemsa染色切片计数Hp定植数量,HE染色切片判断黏膜炎症水平。结果:(1)TLR5、TLR9在各组小鼠胃黏膜组织中均无表达。(2)TLR2在Hp感染组胃黏膜组织中表达(PCR:0.13±0.025;Western:1.32±0.27)高于抗生素治疗组(PCR:0.04±0.011;Western:0.43±0.08),正常组无表达。(3)TLR4在Hp感染组胃黏膜组织中表达(PCR:0.22±0.051;Western:0.72±0.17)高于抗生素治疗组(PCR:0.06±0.009;Western:0.21±0.04),正常组无表达。(4)IL-1β在Hp感染组胃黏膜组织中有表达(PCR:0.27±0.038;Western:0.58±0.14),抗生素治疗组和正常组无表达。结论:TLR2、TLR4可能参与了Hp的致病机制,TLR5、TLR9可能未参与Hp的致病机制。     

TLR2和TLR4激动剂对人脐血CD34+细胞迁移能力的影响

本研究旨在探讨Toll样受体2(Toll-like receptors 2,TLR2)和TLR4激动剂对人脐血CD34+细胞迁移功能的影响及其机制.采用流式细胞术检测脐血CD34+细胞表面TLR2和TLR4的表达情况;用趋化实验和黏附实验观察PAM3CSK4(TLR2激动剂)和LPS(TLR4激动剂)对人脐血CD34+细胞的迁移活性和黏附活性的影响.结果表明:人脐血CD34+细胞表面表达TLR2(14.2±3.8)%和TLR4(19.6±4.1)%.与对照组相比,LPS可明显抑制SDF-1诱导的CD34+细胞的趋化作用(p<0.01),但LPS对CD34+细胞的黏附能力,以及PAM3CSK4对CD34+细胞的趋化和黏附能力均无明显影响.进一步研究显示,LPS对CD34+细胞表面CXCR4的表达无明显影响,但可明显抑制CD34+细胞的自发迁移作用(p<0.05).结论:TLR4的活化可显著降低SDF-1诱导CD34+细胞的趋化功能,这可能与其抑制CD34+细胞的自发迁移作用具有一定的关系.     

TLR2 和 TLR4 激动剂对人骨髓来源的间充质干细胞迁移能力的影响简

本研究探讨Toll样受体2（Toll-like receptors2,TLR2）和TLR4激动剂对人骨髓来源的间充质干细胞（mesenchymal stem cells,MSC）迁移能力的影响及其机制.采用流式细胞术检测MSC表面TLR2和TLR4的表达情况,应用趋化实验和黏附实验观察PAM3CSK4（TLR2激动剂）和LPS（TLR4激动剂）对人骨髓来源的MSC的趋化活性和黏附活性的影响.结果表明,人骨髓来源的MSC表面表达TLR2 （24.5±3.2）％和TLR4（91.3±5.2）％.与对照组相比,PAM3CSK4可明显抑制SDF-1诱导的MSC的趋化作用,并增强MSC的黏附作用;而LPS对MSC的趋化和黏附能力均无明显影响.进一步研究显示,PAM3CSK4可呈剂量依赖性抑制MSC的自发迁移作用,但对MSC表面CXCR4的表达无明显影响.结论：TLR2的活化可显著抑制MSC的迁移能力,这可能与其抑制MSC的自发迁移和增强MSC的黏附作用具有一定的关系.     

The correlation between SNPs within the gene of adrenergic receptor and neuropeptide Y and risk of cervical vertigo

Background

The current investigation was aimed to explore the potential associations of SNPs within ADRB2, ADRB1, NPY, and ADRA1A with risk and prognosis of cervical vertigo.

Methods

Altogether 216 patients with cervical vertigo and 204 healthy controls were gathered, and their DNAs were extracted utilizing the whole‐blood DNA extraction kit. Besides, the PCR reactions were conducted using the TaqMan^R single nucleotide polymorphism (SNP) genotyping assays, and the SNPs were detected on the 7900HT real‐time fluorogenic quantitative polymerase chain reaction (PCR) instrument. Finally, the severity of cervical vertigo was classified according to the JOA scoring, and the recovery rate (RR) of cervical vertigo was calculated in light of the formula as:

Results

The SNPs within ADRA1A [rs1048101 (T>C) and rs3802241 (C>T)], NPY [rs16476 (A>C), rs16148 (T>C), and rs5574 (C>T)], ADRB1 [rs28365031 (A>G)] and ADRB2 [rs2053044 (A>G)] were all significantly associated with regulated risk of cervical vertigo (all P < .05). Haplotypes of ADRA1A [CT and TC] and NPY [CCT and ATT] were also suggested as the susceptible factors of cervical vertigo in comparison with other haplotypes. Furthermore, the SNPs within ADRA1A [rs1048101 (T>C)], NPY [rs16476 (A>C), rs16148 (T>C)], as well as ADRB1 [rs28365031 (A>G)] all appeared to predict the prognosis of cervical vertigo in a relatively accurate way (all P < .05). Ultimately, the haplotypes of ADRA1A (CC) and NPY (CCT) tended to decrease the RR.

Conclusions

The SNPs within ADRB2, ADRB1, NPY, and ADRA1A might act as the diagnostic biomarkers and treatment targets for cervical vertigo.

     

A population analysis of the DGAT1 inhibitor GSK3008356 and its effect on endogenous and meal‐induced triglyceride turnover in healthy subjects

Non‐alcoholic steatohepatitis (NASH) is a liver disease in which fatty infiltration is accompanied by liver inflammation. GSK3008356 is under development as a selective inhibitor of diacylglycerol acyltransferase 1 (DGAT1), a key enzyme involved in the formation of triglyceride (TG). Decreased DGAT1 activity can reduce circulating TG and liver TG, and therefore could potentially prevent or treat NASH. The aim of the current study was to develop a population pharmacokinetic–pharmacodynamic (PKPD) model that characterizes the PK disposition of GSK3008356 and its relation to the changes in blood TG. Drug concentrations were measured in 104 healthy adults receiving various single (SD) and repeat doses (RD) in a first time in human (FiH) study. A 30% fat meal was given at hour 2 postdose, and blood postprandial TG concentrations were measured at various time points. The population PKPD model consists of several parts including a PK model, drug effect model, meal effect model, and a turnover model. The pharmacokinetic data were described using a 3‐compartment model. Drug effect was described by an inhibitory sigmoidal Emax model. Since TG levels change with the introduction of a meal, a bi‐exponential meal effect model was utilized. The total change in TG was fitted using a turnover model with drug and meal effects on the TG production rate. The current analysis presents a PKPD modeling strategy of time‐varying TG data coming from both endogenous and exogenous sources. In general, the presented model could be utilized in the model‐based drug development of drugs that influence TG levels in blood.     

重症脓毒症患者外周血单核细胞人类白细胞抗原和TLR4的表达与预后关系

目的 观察重症脓毒症患者外周血单核细胞(PBMC)人类白细胞抗原(HLA-DR)、天然免疫的模式识别受体TIR4(toll like receptor 4)在不同预后的表达变化.方法 选取35例符合重症脓毒症患者为重症脓毒症组和15名健康志愿者作为对照组.35例重症脓毒症患者按预后又分为好转组(25例)与死亡组(10例),用流式细胞仪检测PBMC表面HLA-DR、TLR4表达.结果 治疗前重症脓毒症组外周血单核细胞数量[(0.39±0.30)×109/L]及其表面HLA-DR(10.25±5.35)FMI、TLR4(10.93±5.66)FMI表达均低于正常对照[(0.62±0.41)×106/L、(59.28±14.76)FMI、(39.86±8.55)FMI](t=2.423、12.52、14.12,P<0.01或P<0.05).重症脓毒症组经ICU综合治疗后,好转组,随着各项指标好转,单核细胞的数量及其表面HLA-DR、TLR4表达各观察点逐渐呈升高趋势,单核细胞数量[(1.03±0.78)×106/L]、HLA-DR[(25.12±9.29)FMI]及TLR4(36.68±16.61)FMI于第5天恢复正常人水平,与治疗前比较差异均有统计学意义(P<0.01,<0.05).死亡组单核细胞的数量及其表面HLA-DR、TLR4表达各观察点持续在较低水平,各时点比较差异均无统计学意义(P均>0.05).结论 重症脓毒症患者外周血单核细胞数量及其表面HLA-DR、TLR4表达可能与患者的免疫状态、预后情况密切相关.     

TLR4基因多态性与重症社区获得性肺炎易感性和预后的关联

目的 探讨Toll样受体4(toll-like-receptor4,TLR4)基因单核苷酸多态性(single nucleotide polymorphism,SNP)与中国汉族人群重症社区获得性肺炎(severe community-acquired pneumonia,SCAP)的易感性和预后的相关性.方法 收集2005年5月至2008年4月复旦大学附属中山医院急诊科收治的360例社区获得性肺炎(conununity-acquired pneumonia,CAP)患者进行前瞻性研究.排除既往患有自身免疫性疾病、正在应用免疫抑制剂、AIDS和肿瘤患者.本研究获得复旦大学附属中山医院伦理委员会同意.根据患者的病情程度分为SCAP组和NSCAP组,每组各180例.根据患者30 d内是否存活,分为存活组和死亡组,存活组300例,死亡组60例.利用Hapmap中国人群数据库选择SNPs和标签SNPs(TagSNPs),利用Primer 3软件设计引物;采用DNA抽提试剂盒提取外周血DNA,PER-直接测序法进行SNPs分型.基因型频率和等位基因频率在各组之间的比较采用χ2检验.结果 TLR4基因3个TagSNPs(rs2149356,rs11536879,rs1927907)等位基因频率分布在研究样本中符合Hardy-Weinberg平衡.TagSNPs等位基因频率和基因型频率在SCAP组和非重症肺炎(non-SCAP,NSCAP)组相比,差异无统计学意义;存活组和死亡组相比,差异无统计学意义(P＞0.05);研究发现由rs2149356和rs11536879组成的3种单倍型(GA,TA,TG)频率在SCAP组和NSCAP组差异无统计学意义,在死亡组和存活组中差异无统计学意义(P＞0.05).结论 TLR4基因多态性与中国汉族人群SCAP的易感性和预后无相关性.     

Early‐onset hypoparathyroidism and chronic keratitis revealing APECED

Early diagnosis of potentially life‐threatening autoimmune polyendocrinopathy‐candidiasis‐ectodermal dystrophy (APECED) is crucial, but is often delayed due to the clinical heterogeneity of the disorder. Even in the absence of the classic disease triad of chronic mucocutaneous candidiasis, hypoparathyroidism, and adrenocortical insufficiency, a diagnosis of APECED should be considered in children who have hypoparathyroidism and chronic keratitis, with a past medical history showing a mild and transient Candida infection.     

The genetic influences on oxycodone response characteristics in human experimental pain

Human experimental pain studies are of value to study basic pain mechanisms under controlled conditions. The aim of this study was to investigate whether genetic variation across selected mu‐, kappa‐ and delta‐opioid receptor genes (OPRM1, OPRK1and OPRD1, respectively) influenced analgesic response to oxycodone in healthy volunteers. Experimental multimodal, multitissue pain data from previously published studies carried out in Caucasian volunteers were used. Data on thermal skin pain tolerance threshold (PTT) (n = 37), muscle pressure PTT (n = 31), mechanical visceral PTT (n = 43) and thermal visceral PTT (n = 41) were included. Genetic associations with pain outcomes were explored. Nineteen opioid receptor genetic polymorphisms were included in this study. Variability in oxycodone response to skin heat was associated with OPRM1 single‐nucleotide polymorphisms (SNPs) rs589046 (P < 0.0001) and rs563649 (P < 0.0001). Variability in oxycodone response to visceral pressure was associated with four OPRM1 SNPs: rs589046 (P = 0.015), rs1799971 (P = 0.045), rs9479757 (P = 0.009) and rs533586 (P = 0.046). OPRM1 SNPs were not associated with oxycodone visceral heat threshold, however, one OPRD1 rs419335 reached significance (P = 0.015). Another OPRD1 SNP rs2234918 (P = 0.041) was associated with muscle pressure. There were no associations with OPRK1 SNPs and oxycodone response for any of the pain modalities. Associations were found between analgesic effects of oxycodone and OPRM1 and OPRD1 SNPs; therefore, variation in opioid receptor genes may partly explain responder characteristics to oxycodone.     

Electrocardiographic Diagnosis of Biventricular Pacing in Patients with Nonapical Right Ventricular Leads

Background: Assessment of left ventricular (LV) capture is of paramount importance in patients with biventricular (BiV) pacing. Our goal was to identify electrocardiographic features that differentiate between BiV and right ventricular (RV)‐only pacing in patients with nonapical RV leads. Methods: The study enrolled 300 consecutive patients with BiV devices and nonapical RV leads, and obtained from them 558 electrocardiograms with either BiV pacing (n = 300) or RV‐only pacing (n = 258). RV pacing served as a surrogate for loss of LV capture. Electrocardiograms from the first 150 patients were used to identify BiV‐specific features, and to construct an algorithm to differentiate between BiV and RV‐only pacing. Electrocardiograms from the second 150 patients were used to validate the algorithm. Results: The following electrocardiographic features typical of BiV pacing were identified: QS in lead V6 (specificity = 98.7%, sensitivity = 54.7%), dominant R in lead V1 (specificity = 100%, sensitivity = 23.3%), q in lead V6 (specificity = 96%, sensitivity = 22.7%), and a QRS < 160 ms (specificity = 100%, sensitivity = 66.0%). The algorithm based on those features was found to have an overall diagnostic accuracy of 95.0%, a specificity of 96.0%, and a sensitivity of 93.5%. Conclusions: The study identified QRS features that were very specific for BiV pacing in patients with nonapical RV leads. Sequential arrangement of those features resulted in an algorithm that was very accurate for differentiating between BiV pacing and loss of LV capture. (PACE 2012; 35:1199–1208)     

TLR2基因单核苷酸多态性与重症社区获得性肺炎易感性和预后的关联研究

目的 探讨Toll样受体2(toll-like receptor 2, TLR2)基因单核苷酸多态性(single nucleotide polymorphism, SNP)与中国汉族人群重症社区获得性肺炎的易感性和预后的相关性.方法 收集2005-05～2008-04中山医院急诊科收治的360例社区获得性肺炎患者的病史资料和血液样本,其中包括重症肺炎和非重症肺炎患者各180例.根据Hapmap中国人群数据库选择SNPs和标签SNPs(Tag SNPs),利用Primer 3 软件设计引物;采用DNA抽提试剂盒提取外周血的DNA,PCR-直接测序法进行SNPs分型;利用Haploview 4.1和SPSS15.0软件进行数据分析.结果 TLR2基因2个Tag SNPs 即rs1898830和rs3804099在研究样本中符合Hardy-Weinberg平衡,2个SNPs等位基因频率和基因型频率在重症肺炎和非重症肺炎组间比较差异无统计学意义,存活组与死亡组比较差异也无统计学意义(P>0.05).结论 TLR2基因单核苷酸多态性与中国汉族人群重症社区获得性肺炎的易感性和预后无明显的相关性.     

Single Transseptal Big Cryoballoon Pulmonary Vein Isolation using an Inner Lumen Mapping Catheter

Background: The single big cryoballon technique for pulmonary vein isolation (PVI) has been limited by the need for two transseptal punctures (TP). We therefore investigated feasibility and safety of a simplified approach using a single TP and a novel circumferential mapping catheter (CMC). Methods: Patients underwent 28‐mm cryoballoon PVI using a single TP. The CMC (Achieve^© Medtronic Inc., Minneapolis, MN, USA) served as (1) guidewire and (2) as a PV mapping tool. Primary endpoint was PVI without switching to a regular guidewire. Secondary endpoints included: (1) PV signal quality during freezing, (2) time to PVI, (3) classification of successful ablation technique, (4) complications, and (5) procedural data. Results: A total of 32 patients (126 PVs) were studied (mean age: 62 ± 11 years, 24 males, left atrium: 40 ± 4 mm). The primary endpoint was achieved in 29/32 patients (91%) and 123/126 PVs (98%) with a procedure and fluoroscopy time of 126 ± 26 minutes and 18.9 ± 7.5 minutes, respectively. Real‐time visualization of PVI could be observed in 61/126 (48%) PVs. Time to sustained PVI versus nonsustained PVI was 66 ± 56 seconds versus 129 ± 76 seconds (P < 0.001). One phrenic nerve palsy was observed. After a follow‐up of 250 ± 84 days 23/32 patients (72%) remained in sinus rhythm. Conclusion: The “simplified single big cryoballoon” PVI strategy appears to be safe and feasible. However, real‐time PV recording was achieved in <50% of PVs. Therefore, further catheter refinements are warranted. (PACE 2012; 35:1304–1311)