Diagnostic Value of the Padua Score Combined with Thrombotic Biomarker Tissue Plasminogen Activator Inhibitor-1 (tPAI-1) Detection for the Risk of Deep Vein Thrombosis in Patients with Pulmonary Heart Disease

This study explores the diagnostic value of combining the Padua score with the thrombotic biomarker tissue plasminogen activator inhibitor-1(tPAI-1)for assessing the risk of deep vein thrombosis(DVT)in patients with pulmonary heart disease.These patients often exhibit symptoms similar to venous thrombosis,such as dyspnea and bilateral lower limb swelling,complicating differential diagnosis.The Padua Prediction Score assesses the risk of venous thromboembolism(VTE)in hospitalized patients,while tPAI-1,a key fibrinolytic system inhibitor,indicates a hypercoagulable state.Clinical data from hospitalized patients with cor pulmonale were retrospectively analyzed.ROC curves compared the diagnostic value of the Padua score,tPAI-1 levels,and their combined model for predicting DVT risk.Results showed that tPAI-1 levels were significantly higher in DVT patients compared to non-DVT patients.The Padua score demonstrated a sensitivity of 82.61%and a specificity of 55.26%at a cutoff value of 3.The combined model had a significantly higher AUC than the Padua score alone,indicating better discriminatory ability in diagnosing DVT risk.The combination of the Padua score and tPAI-1 detection significantly improves the accuracy of diagnosing DVT risk in patients with pulmonary heart disease,reducing missed and incorrect diagnoses.This study provides a comprehensive assessment tool for clinicians,enhancing the diagnosis and treatment of patients with cor pulmonale complicated by DVT.Future research should validate these findings in larger samples and explore additional thrombotic biomarkers to optimize the predictive model.

Bax Inhibitor-1与Herp的相互作用

应用酵母双杂交技术筛选Herp的相互作用蛋白。构建编码Herp的基因HERPUD1真核表达载体HERPUD1plexA,应用MATCHMAKERLexA酵母双杂交系统筛选人胎脑cDNA文库,获得的阳性克隆的插入子为Herp的候选相互作用蛋白质,将Herp与筛选到的相互作用蛋白再一对一回复进行酵母双杂交实验,去除假阳性。对阳性克隆插入子的DNA序列测序,在GenBank中作匹配及生物信息学分析。结果得到其中1个阳性克隆的插入子序列与TEGT基因序列一致,编码蛋白为Baxinhibitor1。得出结论:Herp与Baxinhibitor1相互作用,Baxinhibitor1具有调节凋亡特性,提示Herp可能参与凋亡调节。

棉花Bax inhibitor-1影响内质网胁迫介导的细胞死亡的研究

Bax inhibitor-1(BI-1)是调控内质网胁迫(endoplasmic reticulum stress,ER stress)介导的细胞死亡的关键因子,在植物耐逆中具有重要作用。目前,棉花BI-1(GhBI-1)的耐逆功能及其调控细胞死亡的相关报道较少。在本研究中,采用ER stress专一性诱导毒素——衣霉素(tunicamycin,TM)对棉花幼苗进行胁迫处理,实时定量PCR结果表明,GhBI-1的TM诱导表达具有组织特异性,根GhBI-1对TM的响应更加强烈。通过TM抗性试验及细胞死亡的分析发现,GhBI-1的表达提高了拟南芥的TM抗性,减缓了ER stress介导的细胞死亡。此外,未折叠蛋白反应信号通路中的bZip60转录因子基因的表达受GhBI-1的调控。

靶向人Bax inhibitor-1的siRNA表达质粒的构建及鉴定

目的:构建靶向人Bax inhibitor-1的siRNA表达质粒。方法:借助siRNA设计工具,确定Bax inhibitor-1编码序列中4个可能的干扰位点,人工合成4对正反义脱氧寡核苷酸链(B1、B2、B3、B4),定向克隆至真核表达质粒pmU6,得质粒pmU6-B1、pmU6-B2、pmU6-B3、pmU6-B4。将重组质粒转染DH5α,PCR法筛选阳性克隆,DNA测序法检测插入序列的正确性。结果与结论:PCR和DNA测序结果证实各重组质粒的插入序列完全正确。靶向人Bax inhibitor-1的siRNA表达质粒构建成功。

Bax inhibitor-1基因研究进展

Bax inhibitor-1(简称BI-1)是1998年被克隆鉴定的新的凋亡抑制基因,其蛋白产物可以抑 制由 Bax 引起的凋亡。BI-1 最初以睾丸增强基因转录本而被克隆得到,位于12q12-q13,是一种高度保 守的单拷贝基因。BI-1 过表达与一些肿瘤发生及转移相关。

Association of plasminogen activator inhibitor-1 4G/5G promoter polymorphism with recurrent cerebral infarction in China’s North Jiangsu Province

BACKGROUND： Many international studies have shown that plasminogen activator inhibitor-1 （PAl-l） 4G/5G promoter polymorphism does not increase the risk for cerebral infarction. OBJECTIVE： Using PCR methodology and agarose electrophoresis to detect PAI-1 4G/5G promoter polymorphism in patients with recurrent cerebral infarction in the North Jiangsu Province of China, and to compare results with healthy subjects and patients with first-occurrence cerebral infarction in the same region. DESIGN, TIME AND SETTING： Non-randomized, concurrent, control trial. A total of 122 cerebral infarction patients were admitted to Xuzhou Medical College Hospital＇s Department of Neurology and Xuzhou Central Hospital＇s Department of Neurology between July 2003 and August 2006. PARTICIPANTS： The patients consisted of 63 males and 59 females, aged （62 ± 10） years. They were divided into first-occurrence （n = 58） and recurrence （n = 64） groups. In addition, 50 healthy subjects that underwent physical examination in the outpatient department, including 26 males and 24 females, aged （60 ±12） years, were selected as controls. METHODS AND MAIN OUTCOME MEASURES： PAl-1 4G/5G promoter polymorphism was detected and analyzed using PCR methodology and agarose electrophoresis. RESULTS： Significant differences were determined in terms of genotypic frequency and allele frequency of PAI-1 4G/5G promoter polymorphism, in patients with first-occurrence or recurrent cerebral infarction, when compared with healthy subjects （P 〈 0.05）. There was, however, no significant difference between the first-occurrence and recurrence groups （P 〉 0.05）. CONCLUSION： PAl- 1 4G/5G promoter polymorphism is genetic risk factor for cerebral infarction in China. However, it may be associated with recurrence of cerebral infarction in patients from the North Jiangsu Province of China.

Low levels of Bax inhibitor-1 gene expression increase tunicamycin-induced apoptosis in human neuroblastoma SY5Y cells

A human SH-SY5Y neuroblastoma cell line with a low level of Bax inhibitor-1 expression was established by lentivirus-mediated RNA interference and fluorescence-activated cell sorting. In control SH-SY5Y cells, tunicamycin treatment induced endoplasmic reticulum stress-mediated apoptosis; however, after Bax inhibitor-1 gene knockdown, cell survival rates were significantly decreased and the degree of apoptosis was significantly increased following tunicamycin treatment In addition, chromatin condensation and apparent apoptotic phenomena, such as marginalization and cytoplasmic vesicles, were observed. Our findings indicate that Bax inhibitor-1 can delay apoptosis induced by endoplasmic reticulum stress.

Construction of Tobacco Bax Inhibitor-1 ihpRNA Gene Silencing Vector and Transformation into Agrobacterium tumefaciens EHA105

The plasmid pGSA1285 was first modified by substituting its GUS sequence with the Chalcone synthase intron fragment from vector pFGC5941 to get the plant silencing expression vector that contained Kanamycin resistance site and was named as pGSA2285. Using PCR-based amplification, two different restriction sites at both ends of tobacco Bax inhibitor-1 （NtBI-I） gene were created, respectively, which made the construction of ihpRNA gene silencing vector more efficiently. Then, NtBI-1 genes were inserted into Multiple Cloning Site （MCS） of pGSA2285 respectively to form Bax inhibitor-1 ihpRNA gene silencing vector, named as pGSA4285, containing sense and anti-sense BI-1 sequence which was spliced by chalcone synthase intron. Combined PCR identification and enzyme restriction analyses, the results showed that Bax inhibitor-1 ihpRNA gene silencing vector had been constructed and transferred into Agrobacterium tumefaciens EHA105 successfully, which laid a foundation for the further study on the function of BI-1 in plant PCD regulation.

Study on Effect of Different Dosages of Ligustrazine on Level of Plasminogen Activator Inhibitor-1 Activity in Type 2 Diabetes Mellitus Patients

Objective: To observe the effect of different dosages of ligustrazine (LG) on the level of plasminogen activator inhibitor-1 (PAI-1) activity in patients with type 2 diabetes mellitus. Methods: Ninety cases of type 2 diabetes mellitus inpatients were selected, and randomly divided into LG small dosage group (SDG), LG large dosage group (LDG) and control group. The 120 mg LG, 400 mg LG and normal saline 250 ml were given through intravenous dripping respectively, once daily, 20 days as one treatment course. Before and after treatment, all the patients had their fasting blood taken for PAI-1 and tissue plasminogen activator (t-PA) assessment test to perform the comparative study. Results:Seventy-three out of the 90 patients completed the observation course, the PAI-1 activity of three groups after treatment all lowered compared with that before treatment, and the difference between groups was also significant (all P<0. 01). After treatment the PAI-1 level of SDG and LDG of LG were all markedly lowered (all P<0. 01), the LDG's lowering was more evident than that of SDG, and comparison between these two groups of patients showed significant difference (P<0. 01). Although in the control group there was some difference between before and after treatment, it was not so significant like the above-mentioned two groups (P = 0. 0140). No adverse reaction occurred in the 3 groups during the observation period. Conclusion:LG could safely and effectively lower type 2 diabetes mellitus patient's plasma PAI-1 activity level, and LDG of LG proved to be particularly effective.

Imbalance of matrix metalloproteinase-9 and matrix metalloproteinase tissue inhibitor-1 may contribute to hemorrhage in cerebellar arteriovenous malformations

In this study, we determined the expression levels of matrix metalloproteinase-2 and -9 and matrix metalloproteinase tissue inhibitor-1 and -2 in brain tissues and blood plasma of patients undergoing surgery for cerebellar arteriovenous malformations or primary epilepsy （control group）. Immunohistochemistry and enzyme-linked immunosorbent assay revealed that the expression of matrix metalloproteinase-9 and matrix metalloproteinase tissue inhibitor-1 was significantly higher in patients with cerebellar arteriovenous malformations than in patients with primary epilepsy. The ratio of matrix metalloproteinase-9 to matrix metalloproteinase tissue inhibitor-1 was significantly higher in patients with hemorrhagic cerebellar arteriovenous malformations compared with those with non-hemorrhagic malformations. Matrix metalloproteinase-2 and matrix metalloproteinase tissue inhibitor-2 levels were not significantly changed. These findings indicate that an imbalance of matrix metalloproteinase-9 and matrix metalloproteinase tissue inhibitor-I, resulting in a relative overabundance of matrix metalloproteinase-9, might be the underlying mechanism of hemorrhage of cerebellar arteriovenous malformations.

Research on the correlation of serum plasminogen activator inhibitor-1 level to vascular complications in type 2 diabetes mellitus patients with overweight or obesity

Objective:To explore the relationship between serum plasminogen activator inhibitor(PAI-1)level and Type 2 Diabetes Mellitus(T2DM)accompanied by overweight or obesity by observing not only the changes of PAI-1 level in T2DM patients with overweight or obesity,but also glucose and lipid metabolism related indicators,the changes of the inflammatory cytokines secreted by adipocytes,and then making an analysis on the correlation to PAI-1.Methods:36 cases of healthy examinees were selected as normal control group(NC group),and the experimental group can be divided into T2DM group(54 cases),Overweight/Obesity group(35 cases)and T2DM+Overweight/Obesity group(48 cases).Glucose and lipid metabolism related indicators such as fasting blood glucose(FBG),triglyceride(TG),total cholesterol(TC),low density lipoprotein cholesterol(LDL-C),glycated hemoglobin(HbA1c),fasting insulin(FINS),insulin resistance index(IR),body weight index(BMI)and inflammatory cytokines interleukin-6(IL-6),tumor necrosis factor(TNF-α)and PAI-1 were observed and compared between groups,and then made an analysis to explore the correlation of these factors to PAI-1.Results:(1)Compared with NC group,the levels of FBG,HbA1c,FINS and IR were increased in T2DM group,and the difference was of statistical significance.However,there was no statistically significant difference in TG,TC,LDL-C and BMI between NC group and T2DM group;the levels of FINS,IR,TG,LDL-C,TC and BMI were elevated in Overweight/Obesity group,and the difference was of statistical significance.However,there was no statistically significant difference in FBG and HbA1c;the levels of FBG,HbA1c,FINS,IR,TG,LDL-C,TC and BMI were up-regulated in T2DM+Overweight/Obesity group,and the difference was of statistical significance.Compared with T2DM group,the levels of TG,TC,LDL-C and BMI were increased in Overweight/Obesity group,and the difference was of statistical significance,however,the levels of FBG,HbA1c,FINS and IR were decreased,and the difference was statistically significant;The levels of FINS,IR,TG,TC,LDL-C and BMI were elevated in T2DM+Overweight/Obesity group,and the difference was of statistical significance,however,there was no statistically significant difference in FBG and HbA1c.Compared with Overweight/Obesity group,the levels of FBG,FINS,IR,HbA1c and LDL-C were increased in T2DM+Overweight/Obesity group,and the difference was of statistical significance.However,the difference in TG,TC and BMI was not statistically significant.(2)Compared with NC group,the levels of IL-6,TNF-αand PAI-1 were increased in T2DM group,Overweight/Obesity group and T2DM+Overweight/Obesity group,and the difference was statistically significant.Compared with T2DM group,the levels of IL-6 and TNF-αwere elevated in Overweight/Obesity group,and the difference was of statistical significance,but there was no statistically significant difference in PAI-1;the levels of IL-6,TNF-αand PAI-1 were up-regulated in T2DM+Overweight/Obesity group,and the difference was statistically significant.Compared with Overweight/Obesity group,there was no statistically significant difference in IL-6 and TNF-αbetween T2DM+Overweight/Obesity group and Overweight/Obesity group,but the level of PAI-1 was increased in T2DM+Overweight/Obesity group,and the difference was of statistical significance.(3)Multivariate Logistic Regression Analysis showed that HbA1c,IR,TG,BMI,IL-6 and TNF-αwere independently associated with the level of PAI-1(all p<.05).Conclusions:(1)The level of PAI-1 is higher in type 2 diabetes mellitus patients with overweight or obesity than that in patients only with type 2 diabetes mellitus,and it is one of causes that result in vascular complications.(2)The increase in the level of PAI-1 is considered to be associated with IL-6 and TNF-αsecreted by adipocytes.

BI-1基因的功能研究进展

BI-1(BAX inhibitor-1)基因是进化上高度保守的细胞凋亡抑制基因,广泛存在于动物、植物和微生物中。该基因参与线粒体凋亡途径与内质网应激途径。其表达也与肿瘤的发生发展密切相关,对BI-1功能研究成为近年来的研究热点。

BI-1在心肌缺血再灌注损伤中的作用及机制研究

目的探讨凋亡抑制基因BI-1(Bax inhibitor-1,BI-1)在心肌缺血再灌注损伤中的作用及机制研究。方法构建大鼠心肌缺血再灌注损伤和H9c2心肌细胞氧化应激损伤模型,在不同条件下通过实时荧光定量PCR(quantitative real-time PCR,RT-qPCR)检测BI-1的基因表达量,Western blot法检测BI-1的蛋白表达量,激光共聚焦检测BI-1的亚细胞定位。构建BI-1过表达和干扰重组质粒,转染至心肌细胞,观察BI-1表达量的变化、细胞存活率、乳酸脱氢酶活性、半胱天冬酶-3(caspase-3)、半胱天冬酶-9(caspase-9)活性及心肌细胞凋亡率。结果与假手术组比较,缺血再灌注组出现了心肌梗死区,乳酸脱氢酶水平明显升高(P=0.000),BI-1基因和蛋白表达水平随缺血时间或再灌注时间依赖性升高(P<0.01)。在心肌细胞氧化应激损伤模型中,BI-1基因和蛋白表达水平随过氧化氢刺激时间或刺激浓度依赖性升高(P<0.01),过表达BI-1使乳酸脱氢酶的活性降低(P<0.05),心肌细胞的凋亡数下降(P<0.01),抑制了氧化应激所引起的caspase-3、caspase-9活性上升,使细胞存活率升高(P<0.01)。干扰BI-1表达使乳酸脱氢酶的活性升高(P<0.01),细胞存活率下降(P<0.05),心肌细胞的凋亡数增加(P<0.01)。激光共聚焦观察BI-1定位于线粒体上。结论缺血再灌注损伤和氧化应激损伤可促进心肌细胞中BI-1的表达,BI-1能够抑制氧化应激所导致的心肌细胞凋亡,从而起到心肌保护作用。

通心络对短暂性脑缺血发作患者临床疗效及其对血浆t-PA、PAI-1的影响

通心络胶囊以其特殊通络作用,具有降纤抗凝、增强纤溶系统活性作用,还能修复受损血管内皮,改善内皮功能。关于通心络对纤溶系统影响的研究少有报道。本研究观察通心络胶囊治疗短暂性脑缺血发作（trancient ischemic attack,TIA）的临床疗效,并结合其对血浆组织型纤溶酶原激活物（tissue plas-minogen activator,t-PA）、血浆纤溶酶原激活物抑制物-1（plasminogen activator inhibitor-1,

TNF-α对人脐静脉内皮细胞t-PA、PAI-1表达的研究

目的:观察肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)对人脐静脉内皮细胞(HUVECs)组织型纤溶酶原激活物(tissue plasminogen activitor,t-PA)及其抑制剂-1(plasminogen activitor inhibitor-1,PAI-1)表达的影响。方法:原代分离HUVECs细胞并进行传代培养,分别以6个TNF-α浓度组(0、1、10、20、50、100 ng·m L-1)处理不同时间(0、1、3、6、12、24 h),酶联免疫吸附分析(ELISA)法测定t-PA、PAI-1抗原的表达;逆转-聚合酶链反应(RTPCR)检测t-PA、PAI-1基因的表达。结果:TNF-α促进PAI-1抗原的表达,并呈剂量和时间依赖关系,在TNF-α10 ng·m L-1作用6 h时最明显(P<0.01);TNF-α促进PAI-1 mRNA的表达,并呈剂量和时间依赖关系,在TNF-α10 ng·m L-1作用3 h时已非常明显(P<0.01),6 h时达到高峰(P<0.01);而TNF-α对HUVECs表达t-PA抗原、mRNA无明显影响。结论:炎症因子TNF-α可能通过上调PAI-1表达而诱发血栓相关疾病。

Bax抑制因子1可抑制体外培养的大鼠血管平滑肌细胞的钙化

目的探讨Bax抑制因子1(BI-1)对血管平滑肌细胞钙化的具体作用机制。方法使用β磷酸甘油和氯化钙诱导大鼠主动脉血管平滑肌细胞建立钙化模型。将模型分为对照组(使用常规培养基)、BI-1过表达组(过表达BI-1蛋白后使用常规培养基)、钙化组(使用钙化培养基)、钙化+BI-1过表达组(过表达BI-1蛋白后使用钙化培养基)4组。运用茜素红S染色测定血管平滑肌细胞钙沉积,酶联免疫吸附法测定细胞碱性磷酸酶活性和钙含量,Western blot法测定Runt相关转录因子2、骨形态发生蛋白2和半胱氨酸天冬氨酸蛋白酶3表达水平。结果(1)随着钙化诱导时间的延长,BI-1蛋白表达明显下降,结果具有明显差异性(P=0.001);(2)使用质粒转染方法过表达BI-1蛋白后,细胞钙含量、碱性磷酸酶活性明显降低(P=0.0006),Runt相关转录因子2和骨形态发生蛋白2表达明显下降(P=0.0001),血管钙化减轻;(3)钙化诱导后血管平滑肌细胞凋亡率增加,而过表达BI-1后细胞凋亡率明显减少(P=0.01),半胱氨酸天冬氨酸蛋白酶3水平下降(P=0.0003)。结论BI-1抑制细胞凋亡、钙磷沉积和细胞骨型分化起到减轻血管钙化作用。

Expression of Bmi-1 and PAI-1 in esophageal squamous cell carcinoma

AIM: To determine the correlation between invasiveness, migration and prognosis in esophageal squamous cell carcinoma (ESCC) and expression of the B-cell-specific Moloney leukemia virus insert site 1 (Bmi-1) and plasminogen activator inhibitor-1 (PAI-1).

Protein Microarrays for Quantitative Detection of PAI-1 in Serum

Objective： Plasminogen activator inhibitor-1 （PAI-1）, one crucial component of the plasminogen activator system, is a major player in the pathogenesis of many vascular diseases as well as in cancer. High levels of PAI-1 in breast cancer tissue are associated with poor prognosis. The aim of this study is to evaluate rigorously the potential of serum PAI-1 concentration functioning as a general screening test in diagnostic or prognostic assays. Methods： A protein-microarray-based sandwich fluorescence immunoassay （FIA） was developed to detect PAI-1 in serum. Several conditions of this microarray-based FIA were optimized to establish an efficacious method. Serum specimens of 84 healthy women and 285 women with breast cancer were analyzed using the optimized FIA microarray. Results： The median serum PAI-1 level of breast cancer patients was higher than that of healthy women （109.7 ng/ml vs. 63.4 ng/ml）. Analysis of covariance revealed that PAI-1 levels of the two groups were significantly different （P0.001） when controlling for an age effect on PAI-1 levels. However, PAI-1 values in TNM stage I？IV patients respectively were not significantly different from each other. Conclusion： This microarray-based sandwich FIA holds potential for quantitative analysis of tumor markers such as PAI-1.

RFLPs FOR HUMAN PAI-1 GENE IN CHINESE

Plasminogen activator inhibitor 1 (PAI-1) is considered as the main physiological inhibitor of plasminogen activators in plasma which plays an important regulatory role in the control of the fibrinolytic activity in blood. The human PAI-1 gene is located at q21-22 region of chromosome 7. By using human PAI-1 cDNA (a gift from Dr. D. Ginsburg) as a probe, restriction fragment length polymorphisms (RFLPs) were studied with 8 different endonucleases in 35 unrelated Chinese individuals and the results showed as follows: (1) Taq Ⅰdetected allelic 2.7 kb and 1.8 kb fragments, with the frequencies of 0.96 and 0.04 respectively, 2.7 kb homozygote and 2.7 kb/1.8 kb heterozygote appeared with frequencies of 91% and 9%. (2) Sac Ⅰidentified an invariant 4.8kb band and a two-allele polymorphism with fragments of either 23 kb or 16 kb whose frequencies were 0.96 and 0.04. 23kb homozygote and 23kb/16kb heterozygote appeared with the frequencies of 91% and 9%. (3) HindⅢrevealed a single two-allele polymorphism with bands at either 25 kb or 14 kb, with the frequencies of 0.34 and 0.66, 25 kb homozygote, 25 kb/14 kb heterozygote and 14 kb homozygote appeared with the frequencies of 23%, 46% and 31% respectively. The restriction fragment with a size of 1.6kb for TaqⅠwas first reported in the present paper. No polymorphism was observed for EcoRI, BamHI, BglⅡ, PvuⅡand Pst Ⅰ. The RFLPs for PAI-1 gene may be served as important genetic markers for study of thrombotic and other PAI-1 related disorders.

Analysis of Wheat GBSS1 Promoter:Tissue Specificity and DNA Methylation

The cis-regulatory elements of promoters regulate temporal and spatial expression of genes. DNA inethylation, histone methylation and histone acetylation are the main types of epigenetic modifications, which play important roles in plant growth and development. DNA methylation could seilenco transposons, affect gene imprinting and gene expression. In this study, we found that granule bound starch synthase 1 （GBSSI） gene is expressed specifically in wheat endosperm rath- er than in the embryo. We also analyzed the cis-elements within this promoter region and found some seed-specific elements. In order to confirm the tissue specifici- ty, we cloned 4k bp sequences upstream of GBSS1 gene to link to vector with GUS and this construct was transferred to tobacco by Agrobacterium mediated transfor- marion. The results showed that wheat GBSS1 promoter mediated the seed-specific expression of GUS gene, hut not mediated expression in embryo. In addition, we found that GBSSI promoter is methylated in wheat embryo and de-methylated in wheat endosperm. Our study might provide the molecular basis for specific expres- sion of GBSSI gene.