胰岛素及cAMP对培养人动脉平滑肌细胞HDL受体功能的影响

对胰岛素ｃＡＭＰ对培养人动脉平滑肌细胞（ＳＭＣ）ＨＤＬ受体功能的影响进行了研究,结果发现：胰岛素使ＳＭＣＨＤＬ受体的结合容量Ｂｍａｘ即受体数目显著下降,而对ＳＭＣＨＤＬ受体的Ｋｄ值亲和力无影响;ｃＡＭｐ则ＳＭＣＨＤＬ受体亲和力增加,而对受体数目无影响。     

Troponin replacement in permeabilized cardiac muscle Reversible extraction of troponin I by incubation with vanadate

Calcium-dependent regulation of tension and ATPase activity in permeabilized porcine ventricular muscle was lost after incubation with 10 mM vanadate. After transfer from vanadate to a vanadate-free, low-Ca²⁺ solution (pCa> 8), the permeabilized muscle produced 84.8% ± 20.1% (± S.D., n=98) of the isometric force elicited by high Ca2²⁺ (pCa 4.5 prior to incubation with vanadate. Transfer back to a high Ca²⁺ solution elicited no additional force (83.2% ± 18.7% of control force). SDS-PAGE and immunoblot analysis of fibers and solutions demonstrated substantial extraction (>90%) of Troponin I (TnI). Calcium dependence was restored after incubation with solutions containing either whole cardiac troponin or a combination of TnI and troponin C subunits. This reversible extraction of troponin directly demonstrates the role of TnI in the regulation of striated muscle contractility and permits specific substitution of the native TnI with exogenously supplied protein.     

急诊阵发性心房颤动患者血清肌钙蛋白水平升高的相关影响因素分析

目的:通过比较急诊就诊时不同肌钙蛋白I(TnI)水平阵发性心房颤动(48小时内)患者临床特征的差异,探讨血清肌钙蛋白水平升高的相关影响因素,为临床识别高危房颤患者提供参考依据。方法:选取2016-2017年于宣武医院急诊诊断为阵发性房颤的患者110例,记录既往病史、临床症状体征、实验室检验结果及测定血清TnI水平,并根据TnI水平将患者分为TnI正常组(0.023μg/L)和TnI升高组(0.023μg/L),结合患者既往病史及用药史给予普罗帕酮或可达龙药物转复治疗。结果:急诊阵发房颤患者TnI升高发生率为11.8%,TnI峰值的平均值0.210μg/L,中位数0.067μg/L。TnI正常组和TnI升高组性别构成、血压、心室率、胸闷、头晕、乏力、高血压、糖尿病、慢性阻塞性肺部疾病、射频消融术、转复成功率及NT-proBNP比较差异均无统计学意义(P0.05);年龄、心悸、胸痛、腹背部不适感、冠心病、房颤病史、心电图ST段压低比较差异均有统计学意义(P0.05)。二元logistic回归分析显示既往冠心病史及入室心电图ST段压低为血清TnI升高的影响因素。结论:既往冠心病史及心电图ST段压低为阵发房颤患者肌钙蛋白升高的影响因素,临床上需关注高龄以心绞痛样症状就诊的首次房颤患者。     

不同浓度右美托咪啶对小儿麻醉后血清肌钙蛋白I、C反应蛋白以及补体水平的影响

目的:探讨不同浓度右美托咪啶对小儿麻醉后血清肌钙蛋白I、C反应蛋白(CRP)及补体水平的影响。方法:选取我院收治的拟行手术的患儿60例,根据应用右美托咪定的浓度随机分为A、B、C三组,每组各20例。A、B组患儿予右美托咪啶0.5、0.25μg/kg的负荷剂量静脉推注给药10分钟,然后以0.2-0.7μg/kg/h的速度持续静脉微量泵入维持患者镇静状态,C组直接以右美托咪啶0.2-0.7μg/kg/h的速度持续泵入维持镇静状态。比较三组患儿麻醉术后的苏醒情况,麻醉前后不同时点血清肌钙蛋白I、C反应蛋白及补体水平的变化。结果:治疗前,三组患儿的各项指标比较均无统计学差异(P0.05);治疗后,与A、C两组比较,B组患者自主呼吸恢复时间、气管导管拔管时间、解除监护时间显著缩短(P0.05),术后24、72 h血清CRP水平更低,术后1、6、24 h血清肌钙蛋白I水平较低(P0.05),术后1、24、72 h血清补体水平较高(P0.05)。结论:0.25μg/kg浓度的右美托咪啶对于对小儿麻醉后血清肌钙蛋白I、C反应蛋白及补体水平的影响明显,且有助于患儿术后的苏醒。     

肌钙蛋白I和肌红蛋白在急性心肌梗死患者血清中的变化特点及其诊断价值

目的:探讨肌钙蛋白I、CKMB的即时检测技术在急诊科心肌梗死患者中的应用及其临床意义。方法:研究对象为2012年10月至2013年8月于我急诊科急诊的急性心肌梗死患者,按就诊时间分为对照组和实验组。对照组患者采用常规化验室检测肌钙蛋白I、CKMB,实验组采用急诊科即时检测方法检测肌钙蛋白I、CKMB。对比两组患者从就诊到确诊的时间、住院天数、治愈率、心功能不全发生率和死亡率。结果:实验组患者的确诊时间为(25.5±5.6)min,住院天数为(9.89±1.5)天,治愈率为80.8%,心功能不全发生率为15.4%。对照组患者的确诊时间为(66.8±10.0)min,住院天数为(12.6±2.5)天,治愈率为56.0%,心功能不全发生率为32.0%P均.0.05,有统计学意义。两组患者死亡率分别为12%和3.8%,无明显差异。结论:对心肌梗死患者采用肌钙蛋白I、CKMB的即时检测对于提高患者治愈率,减少确诊时间和住院时间,降低心功能不全发生率有很大帮助。     

Troponin: regulatory function and disorders

Study of the molecular biology of the calcium regulation of muscle contraction was initiated by Professor Ebashi’s discovery of a protein factor that sensitized actomyosin to calcium ions. This protein factor was separated into two proteins: tropomyosin and a novel protein named troponin. Troponin is a Ca²⁺-receptive protein for the Ca²⁺-regulation of muscle contraction and, in association with tropomyosin, sensitizes actomyosin to Ca²⁺. Troponin forms an ordered regulatory complex with tropomyosin in the thin filament. Several regulatory properties of troponin, which is composed of three different components, troponins C, I, and T, are discussed in this article. Genetic studies have revealed that many mutations of genes for troponin components, especially troponins T and I, are involved in the three types of inherited cardiomyopathy. Results of functional analyses indicate that changes in the Ca²⁺-sensitivity caused by troponin mutations are the critical functional consequences leading to these disorders. Recent results of this pathophysiological aspect of troponin are also discussed.     

Structural and functional studies on Troponin I and Troponin C interactions

Troponin I (TnI) peptides (TnI inhibitory peptide residues 104-115, Ip; TnI regulatory peptide resides 1-30, TnI1-30), recombinant Troponin C (TnC) and Troponin I mutants were used to study the structural and functional relationship between TnI and TnC. Our results reveal that an intact central D/E helix in TnC is required to maintain the ability of TnC to release the TnI inhibition of the acto-S1-TM ATPase activity. Ca(2+)-titration of the TnC-TnI1-30 complex was monitored by circular dichroism. The results show that binding of TnI1-30 to TnC caused a three-folded increase in Ca(2+) affinity in the high affinity sites (III and IV) of TnC. Gel electrophoresis and high performance liquid chromatography (HPLC) studies demonstrate that the sequences of the N- and C-terminal regions of TnI interact in an anti-parallel fashion with the corresponding N- and C-domain of TnC. Our results also indicate that the N- and C-terminal domains of TnI which flank the TnI inhibitory region (residues 104 to 115) play a vital role in modulating the Ca(2+)- sensitive release of the TnI inhibitory region by TnC within the muscle filament. A modified schematic diagram of the TnC/TnI interaction is proposed.     

圆斑星鲽肌钙蛋白I基因的克隆和序列分析

通过建立圆斑星鲽肌肉 cDNA 文库,大规模的 EST 测序和PCR技术得到了圆斑星鲽肌钙蛋白I的完全表达序列,该序列全长845 bp,编码171个氨基酸.经同源性分析结果表明,与多种鱼类的肌钙蛋白 I 具有较高同源性,其中与鳜鱼的肌钙蛋白 I 同源性最高,为86%.序列提交 GenBank 收录,登录号为 GU229275.     

肌钙蛋白I水平检测对急诊房颤患者的临床意义

目的:探讨房颤患者急诊入院时检测肌钙蛋白I(Tn I)水平的临床作用及预测价值。方法:回顾性分析我院523例房颤患者的临床资料,将资料中数据进行统计学分析处理。患者分为三组:1组患者Tn I轻度升高;2组患者Tn I正常;3组患者未检测Tn I,分别就三组患者的基线特征以及就诊时的症状展开数据比较,并且单变量及多变量分析Tn I升高的预测因子。设置主要终点为1年内所有原因导致的死亡及心肌梗死(MI),患者从入院时起随访1年,记录期间的MI发生情况及死亡情况。结果:173例患者(33%)Tn I轻度升高(均值0.56 ng/m L),225例患者(43%)Tn I正常,125例患者(24%)未检测Tn I。住院期间1组患者(50%)的心功能检查异常明显高于组2和组3(28%和29%,P≤0.001),1组患者(22%)主要终点发生率明显高于2组(10%)和3组患者(15%)(P=0.002),有统计学意义。结论:肌钙蛋白I轻度升高与冠脉疾病(CAD)发生率增加及MI发生率增加有关,但对于总体死亡率无影响。     

Molecular Characterization and SNP Development for the Porcine Il6 and Il10 Genes

Different cytokines are secreted in response to specific microbial molecules referred to as pathogen associated molecular patterns (PAMPs). Interleukin 6 (IL6) and interleukin 10 (IL10), both secreted by macrophages and lymphocytes, play a central role in the immunological response. In this work we obtained the genomic structure and complete DNA sequence of the porcine IL6 and IL10 genes and identified polymorphisms in the genomic sequences of these genes on a panel of ten different pig breeds. Comparative intra- and interbreed sequence analysis revealed a total of eight polymorphisms in the porcine IL6 gene and 21 in the porcine IL10 gene, which include single nucleotide polymorphisms (SNPs) and insertion deletion polymorphisms (indels). Additionally, the chromosomal localization of the IL10 gene was determined by FISH and RH mapping.     

Molecular Characterization,Polymorphism, and Association of Porcine GADD45G Gene

Growth arrest and DNA-damage-inducible gamma (GADD45G) is a reproduction related gene. In this study, the full-length cDNA sequence of porcine GADD45G gene was cloned through rapid amplification of cDNA ends (RACE) method. The porcine GADD45G gene encodes a protein of 159 amino acids that shares high homology with the GADD45G of nine species: chimpanzee (97%), sumatran orangutan (97%), white-tufted-ear marmoset (97%), northern white-cheeked gibbon (97%), cattle (97%), human (97%), rhesus monkey (97%), rat (96%), and mouse (95%). This novel porcine gene was assigned to GeneID: 100152997. Phylogenetic analysis revealed that the porcine GADD45G gene has a closer genetic relationship with the GADD45G gene of cattle. Computer-assisted analysis indicated that porcine GADD45G gene is structured in four exons and three introns. PCR-Rsa I-RFLP was established to detect an A/G mutation on the position of 294-bp of coding sequence and eight pig breeds display obvious genotype and allele frequency differences at this mutation locus. Association of this SNP with litter size traits was assessed in Large White (n = 100) and Landrace (n = 100) pig populations, and result demonstrated that this polymorphic locus was significantly associated with the litter size of all parities in Large White and Landrace sows (P < 0.01). Therefore, porcine GADD45G gene could be a useful candidate gene in selection for increasing the litter size. These data serve as a foundation for further insight into this novel porcine gene.     

不同浓度七氟烷对小儿麻醉后cTn I, CRP及补体影响研究

目的:探究不同浓度七氟烷联合丙泊酚对小儿麻醉后肌钙蛋白I、C反应蛋白以及补体水平影响。方法:收集我院60例ASAⅠ级拟行全麻手术患儿,随机分为A、B、C三组,每组20例。A组给予2%浓度的七氟烷联合丙泊酚麻醉;B组2.5%浓度的七氟烷联合丙泊酚麻醉;C组3%浓度的七氟烷联合丙泊酚麻醉。检测三组患儿苏醒时间、术后情况,肌钙蛋白I(cTnI)、C反应蛋白(CRP)及补体C_3、C_4水平。结果:A组、B组自主呼吸时间、气管导管拔管时间、解除监护时间较C组相比时间明显较短(P0.05);但A组与B组比较无统计学差异(P0.05);与A组比,B组与C组术后肌钙蛋白I、CRP水平较低,C_3、C_4水平较高(P0.05),但B组与C组血清指标比较无统计学差异(P0.05)。结论:2.5%浓度的七氟烷联合丙泊酚是诱导小儿全身麻醉中的最佳浓度。     

Molecular Cloning of Porcine SUN5 Gene and Association between a SNP with Litter Size Trait

SUN domain-containing protein 5 (SUN5) is an important reproduction related gene. In this study, we cloned the full-length coding sequence of porcine SUN5 gene through RT-PCR. Sequence analysis of this gene revealed that the pig SUN5 gene encodes a protein of 383?amino acids that has high homology with the SUN5 protein of eight species: wild Bactrian camel (95%), alpaca (95%), Yangtze River dolphin (94%), sperm whale (94%), sheep (93%), black flying fox (93%), goat (92%), and horse (91%). This gene is structured into 13 exons and 12 introns as revealed by computer-assisted analysis. The prediction of transmembrane helices showed that pig SUN5 protein might be a transmembrane protein. PCR-Taq I-RFLP was established to detect the GU475008:c.138 G>A substitution of porcine SUN5 gene coding sequence and eight pig breeds displayed obvious genotype and allele frequency differences at this mutation locus. Association of this SNP with litter size traits was assessed in Large White (n?=?200) and Landrace (n?=?200) pig populations, and the results demonstrated that this polymorphic locus was significantly associated with the litter size of all parities in Large White and Landrace sows (P?相似文献   

右美托咪啶对苏醒期小儿血清肌钙蛋白I、C 反应蛋白及补体的影响

目的:探究不同浓度右美托咪啶对小儿麻醉后肌钙蛋白I、C反应蛋白以及补体变化的影响。方法:收集我院下腹股沟斜疝、急性阑尾炎以及肠套叠手术患儿75例,根据治疗方法不同分为三组,每组25例,其中A组实施0.5μg/kg负荷剂量右美托咪啶治疗,B组实施0.25μg/kg负荷剂量右美托咪啶治疗,C组实施右美托咪啶0.2~0.7μg/kg/h持续泵入。对比三组患者苏醒情况、血清肌钙蛋白I、C反应蛋白、补体水平、镇定及躁动评分。结果:手术后,B组自主呼吸恢复时间、气管导管拔管时间、解除监护时间明显缩短,且与其他两组比较,差异有统计学意义(P0.05);与A、C两组相比,B组手术后C反应蛋白及血清肌钙蛋白I水平较低,而C3及C4水平较高,差异具有统计学意义(P0.05);B组镇静、躁动评分明显降低,差异有统计学意义(P0.05)。结论:0.25μg/kg负荷剂量右美托咪啶能够有效减轻患儿术后炎性反应,缓解麻醉后神经损伤,有助于脏器功能的恢复。     

Removal of the Cardiac Troponin I N-terminal Extension Improves Cardiac Function in Aged Mice

The cardiac troponin I (cTnI) isoform contains a unique N-terminal extension that functions to modulate activation of cardiac myofilaments. During cardiac remodeling restricted proteolysis of cTnI removes this cardiac specific N-terminal modulatory extension to alter myofilament regulation. We have demonstrated expression of the N-terminal-deleted cTnI (cTnI-ND) in the heart decreased the development of the cardiomyopathy like phenotype in a β-adrenergic-deficient transgenic mouse model. To investigate the potential beneficial effects of cTnI-ND on the development of naturally occurring cardiac dysfunction, we measured the hemodynamic and biochemical effects of cTnI-ND transgenic expression in the aged heart. Echocardiographic measurements demonstrate cTnI-ND transgenic mice exhibit increased systolic and diastolic functions at 16 months of age compared with age-matched controls. This improvement likely results from decreased Ca²⁺ sensitivity and increased cross-bridge kinetics as observed in skinned papillary bundles from young transgenic mice prior to the effects of aging. Hearts of cTnI-ND transgenic mice further exhibited decreased β myosin heavy chain expression compared to age matched non-transgenic mice as well as altered cTnI phosphorylation. Finally, we demonstrated cTnI-ND expressed in the heart is not phosphorylated indicating the cTnI N-terminal is necessary for the higher level phosphorylation of cTnI. Taken together, our data suggest the regulated proteolysis of cTnI during cardiac stress to remove the unique cardiac N-terminal extension functions to improve cardiac contractility at the myofilament level and improve overall cardiac function.     

血清肌钙蛋白I、N端前体脑钠肽和高敏C反应蛋白水平在充血性心力衰竭中的临床意义

目的：探讨血清肌钙蛋白I（cTnI）、N端前体脑钠肽（NT-proBNP）和高敏c反应蛋白（hs—CRP）水平在充血性心力衰竭（CHF）中的临床意义。方法：选取2010年1月-2012年12月在我院诊治的CHF患者210例（CHF组）,心功能按纽约心脏病协会（NYHA）分级：Ⅱ级88例、Ⅲ级96例和Ⅳ级26例。同时选取同期在我院行健康体检的自愿者30例为健康对照组。观察和比较两组血清hs．CRP、cTnI和NT．proBNP水平的差异及其与CHF患者心功能的心功能分级的相关系。结果：CHF组患者的血清hs-CRP．cTnI和NT．proBNP水平均较对照组明显增高,差异均有统计学意义（P〈0．0t）,且CHF患者的血清hs—CRP、cTnI和NT—proBNP水平随着心功能分级的增加而升高,差异有统计学意义（P〈0．01）。血浆hs—cI冲（F0．694,P〈0．01）水平,cTnI（r=0．782,P〈0．01）水平均与NT—proBNP水平呈显著正相关,血浆hs．CRP和cTnI水平也呈显著正相关（r==0．649,P〈0．01）。结论：CHF患者的血清hs—CRP、cTnI和NT-proBNP水平与其心功能分级显著相关,有助于评估CHF患者心功能不全的严重程度,可能与CHF患者的预后相关。     

我国巴马小型猪SLA-2基因克隆及分子特征

为研究我国巴马小型猪SLA-Ⅰ分子特征,设计引物克隆了SLA-Ⅰ类分子SLA-2基因(SLA-2bm),并通过分子生物学软件分析其分子特征。经克隆及序列测定分析,SLA-2bm为1119bp,其中3~1097为ORF区,共编码364个氨基酸,分别在第125、188、227和283位置出现半胱氨酸残基,含有两对链内二硫键。氨基酸同源性分析显示SLA-2bm与其它SLA-2、SLA-3和SLA-1序列的同源率分别为88.4%~96.4%、88.3%~90.5%和87.7%~92.7%。系统进化树显示SLA-2bm与其它SLA-2等位基因在遗传关系上相对独立,进化程度较低;各功能区分析,SLA-2bm与人HLA-A2和小鼠H-2K分子结构相似,且保留了人HLA-A2基因的部分功能位点。结果表明,SLA-2bm属于一个新的等位基因,巴马小型猪是保留原始基因特征的品种。     

黑鲷转铁蛋白基因的分子特征研究

目的:克隆黑鲷转铁蛋白全基因并分析其分子特征。方法:采用同源克隆的方法,对黑鲷转铁蛋白全基因编码序列进行克隆,在cDNA获得了部分转铁蛋白基因的同源片段。经RACE PCR方法,分别对该基因的3’和5’末端进行扩增,获得的扩增片段经拼接后得到全基因片段。结果:黑鲷转铁蛋白全长2431bp,可编码691个氨基酸,分子量(MW)约为74.3kDa,等电点(PI)为5.63。它与鱼类转铁蛋白的同源性最高,约为65%-89%;与其它动物(哺乳类、两栖类等)也有一定的相识性。进化分析表明黑鲷转铁蛋白与其它鱼类和哺乳类的转铁蛋白是由早期转铁蛋白共同的祖先进化来的。结论:黑鲷转铁蛋白主要在肝脏中成组成型表达,在大脑等器官中也有少量表达。该基因的表达受病原刺激的影响,表现为经病原刺激后转铁蛋白基因的组织分布显著增多。