Clinical significance of expression of tissue factor and tissue factor pathway inhibitor in ulcerative colitis

AIM: To investigate the clinical significance of expression of tissue factor (TF) and tissue factor pathway inhibitor (TFPI) in ulcerative colitis (UC).

Testosterone alleviates tumor necrosis factor-alpha-mediated tissue factor pathway inhibitor downregulation via suppression of nuclear factor-kappa B in endothelial cells

We have observed earlier that testosterone at physiological concentrations can stimulate tissue factor pathway inhibitor(TFPI)gene expression through the androgen receptor in endothelial cells.This study further investigated the impact of testosterone on TFPI levels in response to inflammatory cytokine tumor necrosis factor-alpha(TNF-α).Cultured human umbilical vein endothelial cells were incubated in the presence or absence of testosterone or TNF-α.TFPI protein and mRNA levels were assessed by enzyme-linked immunosorbent assay and quantitative real-time reverse transcription polymerase chain reaction.To study the cellular mechanism of testosterone’s action,nuclear factor-kappa B(NF-κB)translocation was confirmed by electrophoretic mobility shift assays.We found that after NF-κB was activated by TNF-α,TFPI protein levels declined significantly by 37.3%compared with controls(P<0.001),and the mRNA levels of TFPI also decreased greatly(P<0.001).A concentration of 30 nmol L-1 testosterone increased the secretion of TFPI compared with the TNF-α-treated group.NF-κB DNA-binding activity was significantly suppressed by testosterone(P<0.05).This suggests that physiological testosterone concentrations may exert their antithrombotic effects on TFPI expression during inflammation by downregulating NF-κB activity.

Expressions of Tissue Factor and Tissue Factor Pathway Inhibitor in Patients with Acute Graft-versus-host Disease after Allogeneic Hematopoietic Stem Cell Transplantation

This study examined the expressions of human serum tissue factor （TF） and tissue factor pathway inhibitor （TFPI） in patients with acute graft-versus-host disease （aGVHD） after allogeneic hematopoietic stem cell transplantation （allo-HSCT） and their clinical significance. The serum TF and TFPI levels were detected by ELISA in 28 allo-HSCT recipients before and after the transplanta-tion and the changes of TF and TFPI levels were dynamically monitored at different phases of the disease. No significant differences in the serum TF and TFPI levels were found in allo-HSCT recipi-ents in the absence of aGVHD or with gradeⅠaGVHD before and after the transplantation. The lev-els of serum TF and TFPI were substantially increased in the patients with gradeⅡ aGVHD at the peak of aGVHD （P〈0.05） and they were even higher in the patients with grade Ⅲ–Ⅳ aGVHD （P〈0.01）. When the conditions became stable after treatment with immunosuppressive agents, the serum TFPI level was decreased to the baseline level （P〉0.05） and the TF level was lowered but still higher than the baseline level （P〈0.05）. It was concluded that the levels of serum TF and TFPI were increased significantly in the patients with grade Ⅱ–Ⅳ aGVHD after allo-HSCT and decreased markedly after the treatment. Monitoring the levels of serum TF and TFPI in the patients with allo-HSCT is important to predict the occurrence, outcome and prognosis of aGVHD.

Oxidized low density lipoprotein inhibited tissue factor pathway inhibitor mRNA expression in human endothelial cells

To understand the role of oxidized low density lipoprotein (OX LDL) in the pathogenesis of thrombotic complications in atherogenesis Methods Low density lipoprotein was isolated from normal heparinized blood by density gradient ultracentrifugation and oxidized by CuCl 2 Total RNA was extracted from human umbilical vein endothelial cells (HUVECs) exposed to LDL or OX LDL, using the guanidinium isothiocyanate method The quantification of tissue factor pathway inhibitor (TFPI) mRNA in HUVECs was carried out by reverse transcriptase polymerase chain reaction (RT PCR) Results HUVECs were able to express TFPI mRNA constitutively The expression was not affected by LDL but was effectively inhibited by OX LDL in a time and dose dependent manner Conclusions The results suggest that oxidized LDL may play an important role in inducing coagulation in atherosclerotic lesions by the inhibition of expression of TFPI in vascular endothelial cells

Preparation and characterization of monoclonal antibody against recombinant human tissue factor pathway inhibitor

:Objective To prepare and identify monoclonal antibody (McAb) against recombinant human tissue factor pathway inhibitor (rhTFPI) and to use it for measurement of TFPI by ELISA, and to evaluate the effects of the McAb on dilute prothrombin time (PT) and activated partial thromboplastin time (APTT).Methods After intrasplenic immunization of Balb/c mouse with TFPI, hybridoma technique was used to raise monoclonal antibody against rhTFPI. The McAb was wellcharacterized and labelled with horseradish peroxidase (HRP) by using assay of TFPI in ELISA. Furthermore, the McAb was added to normal and factor Ⅸ deficient plasma for observation of dilute PT and APTT.Results Two hybridomas (4F4, 4F8) secreting McAb against TFPI were established. The Ig class and subclass of the McAb purified from 4F8 was IgG1. Immunoblotting results indicated that the McAb4F8 only recognized a single band of TFPI with molecular weight of 34.8 KD. The results of Sandwich enzymelinked immunosorbent assay (ELISA) by using the HRP labelled McAb4F8 showed that the mean of TFPI in normal human plasma is 103.2±11.5 μg/L. The McAb 4F8 was also proved to shorten markedly dilute prothrombin time of factor Ⅸ deficient plasma and normal plasma.Conclusions We established two hybridomas cell lines (4F4, 4F8) and obtained the McAb4F8 against TFPI and reported the levels of TFPI in healthy adult human plasma by Sandwich ELISA with HRP labelled McAb4F8 in Chinese.

Modulation of Matrix Metalloproteinase and TIMP-1 Expression by TGF-β_1 in Cultured Human RPE Cells

In order to investigate the effects of TGF-β1 on the expression of MMP-2, -9 and TIMP- 1 in human retinal pigment epithelial （RPE） cells, the third-sixth passage cultured RPE cells were treated with TGF-β1 at different concentrations （0.01, 0. 1, 1.0, 10 ng/mL）, the expression of MMP-2, -9 and TIMP-1 mRNA was detected by semi-qudntitative RT-PCR assays. MMP-2, -9 and TIMP-1 mRNA were expressed in the cultured RPE cells. The values of MMP-2/β-actin in the cells treated with 0.1, 1.0, 10 ng/mL TGF-β1 were 1.04±0.04, 1.07±0.02 and 1.11±0.03, respectively, significantly higher than in the control group （0.96±0.03, P〈0. 05-0.01）. The expression of MMP-2 mRNA could be up-regulated by TGF-β, , in a dose-dependent manner. The expression of MMP-9 mRNA in the cultured RPE cells was slightly up-regulated by various TGF-β1 concentrations treatment. The values of TIMP-1/β-actin in the cells treated with 0.01 and 0.1 ng/ mL TGF-β1 were 0.85 ±0.01 and 0.97 ± 0.02 respectively, significantly lower than in the control group （1.07±0.04, P〈0.01）, indicating that the expression of TIMP-1 mRNA was down-regulated by TGF-β1 at low concentrations. But along with the increase of TGF-β1 concentrations （1.0 and 10 ng/mL）, the expression of TIMP-1 mRNA was slightly up-regulated, not significantly different from that in the control group （P〉0.05）. It was concluded that TGF-β1 might play an important role in the up-regulation of the expression of MMP-2 in RPE cells and result in a directional shift in the balance between MMP and TIMP. This may be facilitated for RPE cells to migrate in the pathogenesis of vitreoretinopathy.

Comparison of Anticoagulant Effects on Vein Grafts between Human TFPI Gene Transfection and Aspirin Oral Administration

To develop a more efficient antithrombotic way after coronary artery bypass grafting （CABG）, the anticoagulant effects were compared of human tissue factor pathway inhibitor （TFPI） gene transfection and aspirin oral administration （traditional method） on vein grafts. An eukaryotic expression plasmid pCMV-（Kozak） TFPI was prepared. Animal model of carotid artery bypass grafting was constructed. In operation, endothelial cells of vein grafts in TFPI group and empty plasmid control group were transfected with pCMV-（Kozak） TFPI and empty plasmid pCMV respectively, while no transfection was conducted in aspirin control group. After operation, aspirin （2 mg·kg^-1·d^-1） was administered （i.g.） in aspirin control group. Three days later, grafts （n=10） were harvested for RT-PCR, Western blotting and immunohistochemical analyses of exogenous gene expression and for pathological, scanning electron microscopic observation of thrombus. Thirty days later, the patency rates of remnant grafts （n= 10） were recorded by vessel Doppler ultrasonography. Human TFPI gene products were detected in gene transferred vein grafts. Three days later, thrombi were found in 7 animals of aspirin control group and in 8 animals of empty plasmid control group, but in only 1 of TFPI group （P〈0.01）. Thirty days later, 5 grafts were occluded in empty plasmid control group, but none of grafts was occluded in the other groups （P〈0.05）. The endothelial surfaces of grafts in both of the control groups were covered with aggregated erythrocytes and platelets, and it were not seen in TFPI group. It was suggested that the anticoagulant effects on vein grafts of human TFPI gene transfection are better than those of aspirin.

Endogenous tissue factor pathway inhibitor in vascular smooth muscle cells inhibits arterial thrombosis

Tissue factor pathway inhibitor （TFPI） is the main inhibitor of tissue factor-mediated coagulation. TFPI is expressed by endothelial and smooth muscle cells in the vasculature. Endothefium-derived TFPI has been reported to play a regulatory role in arterial thrombosis. However, the role of endogenous TFPI in vascular smooth muscle cells （VSMCs） in thrombosis and vascular disease development has yet to be elucidated. In this TFPI^Flox mice crossbred with Sma-Cre mice were utilized to establish TFPI conditional knockout mice and to examine the effects of VSMC-directed TFPI deletion on development, hemostasis, and thrombosis. The mice with deleted TFPI in VSMCs （TFP^Sma） reproduced viable offspring. Plasma TFPI concentration was reduced 7.2% in the TFPIsma mice compared with TFPI^Flox littermate controls. Plasma TFPI concentration was also detected in the TFPI^Tle2 （mice deleted TFPI in endothefial ceils and cells of hematopoietic origin） mice. Plasma TFPI concentration of the TFPI^Tle2 mice was 80.4% lower （P 〈 0.001） than that of the TFPI^Flox mice. No difference in hemostatic measures （PT, APTT, and tail bleeding） was observed between TFPIsma and TFPI^Flox mice. However, TFP^Sma mice had increased ferric chloride-indueed arterial thrombosis compared with TFPI^Flox littermate controls. Taken together, these data indicated that endogenous TFPI from VSMCs inhibited ferric chloride-induced arterial thrombosis without causing hemostatic effects.

Insulin-like growth factor binding protein related protein 1 knockdown attenuates hepatic ?brosis via the regulation of MMPs/TIMPs in mice

Background: Previous research suggested that insulin-like growth factor binding protein related protein 1(IGFBPrP1), as a novel mediator, contributes to hepatic fibrogenesis. Matrix metalloproteinases(MMP) and tissue inhibitors of metalloproteinases(TIMP) play an essential role in hepatic fibrogenesis by regulating homeostasis and remodeling of the extracellular matrix(ECM). However, the interaction between IGFBPrP1 and MMP/TIMP is not clear. The present study was to knockdown IGFBPrP1 to investigate the correlation between IGFBPrP1 and MMP/TIMP in hepatic fibrosis. Methods: Hepatic fibrosis was induced by thioacetamide(TAA) in mice. Knockdown of IGFBPrP1 expression by ultrasound-targeted microbubble destruction-mediated CMB-shRNA-IGFBPrP1 delivery, or inhibition of the Hedgehog(Hh) pathway by cyclopamine treatment, was performed in TAA-induced liver fibrosis mice. Hepatic fibrosis was determined by hematoxylin and eosin and Sirius red staining. Hepatic expression of IGFBPrP1, α-smooth muscle actin( α-SMA), transforming growth factor β 1(TGF β1), collagen I, MMPs/TIMPs, Sonic Hedgehog(Shh), and glioblastoma family transcription factors(Gli1) were investigated by immunohistochemical staining and Western blotting analysis. Results: We found that hepatic expression of IGFBPrP1, TGF β1, α-SMA, and collagen I were increased longitudinally in mice with TAA-induced hepatic fibrosis, concomitant with MMP2/TIMP2 and MMP9/TIMP1 imbalance and Hh pathway activation. Knockdown of IGFBPrP1 expression, or inhibition of the Hh pathway, reduced the hepatic expression of IGFBPrP1, TGF β1, α-SMA, and collagen I and re-established MMP2/TIMP2 and MMP9/TIMP1 balance. Conclusions: Our findings suggest that IGFBPrP1 knockdown attenuates liver fibrosis by re-establishing MMP2/TIMP2 and MMP9/TIMP1 balance, concomitant with the inhibition of hepatic stellate cell activation, down-regulation of TGF β1 expression, and degradation of the ECM. Furthermore, the Hh pathway mediates IGFBPrP1 knockdown-induced attenuation of hepatic fibrosis through the regulation of MMPs/TIMPs balance.

Clinical Research on the Changes of Plasma TFPI and uPA System in Malignant Tumor

Objective: To investigate the expression levels and significance of TFPI, uPA, uPAR and PAI in malignant patients. Methods: The levels of TFPI, uPA and uPAR were measured by using ELISA and the level of PAI was determined by method of chromogenic substrates in 44 patients with malignant solid tumors (group A1) and 30 patients with acute leukemia (AL, group A2). Results: The levels of TFPI, uPA, and uPAR in group A1 were higher than those in normal control group (group B). TFPI, uPAR levels in group A2 were higher than those in group B, while the level of PAI in group A2 was lower than that in group B. Among the groups, TFPI was increased in the combined infection group; PAI decreased in the hemorrhage group; TFPI, uPA, uPAR and PAI increased in relapsing and metastasis group; TFPI decreased in one-week dead group, while uPA and uPAR increased. Conclusion: The patients with malignant solid tumor and AL had different anticoagulation or fibrinolysis states. TFPI, uPA, Upar and PAI can be used to evaluate the disease condition and the prognosis.

遗传性非息肉病性大肠癌中hMSH2，hMLH1，TβRⅡ，MMP-7及TIMP-2的表达和其特殊生物学行为间的关系

目的:探讨遗传性非息肉病性大肠癌(HNPCC)中错配修复基因hMSH2、hMLH1、转化生长因子βⅡ型受体(TβRⅡ)、基质金属蛋白酶-7(MMP-7)、组织抑制因子-2(TIMP-2)表达的相互关系及其与HNPCC特殊生物学行为的关系.方法:应用免疫组织化学染色法检测HNPCC和散发性大肠癌(SCRC)肿瘤组织石蜡标本各30例、正常大肠黏膜石蜡标本8例.观察其hMSH2,hMLH1,TβRⅡ,MMP-7,TIMP-2的表达,并结合临床病理资料综合分析.结果:在HNPCC和SCRC中,hMSH2,hMLH1,TβRⅡ,MMP-7,TIMP-2均与患者的性别、肿瘤大小和部位无关;而与肿瘤的侵犯深度和是否转移密切相关,阳性表达率差异显著(P<0.05,HNPCC vs sporadic CRC).在HNPCC中,hMSH2和hMLH1(r=0.835,P= 0.000),TβRⅡ与hMSH2(r=0.592,P=0.001),hMLH1(r=0.472,P=0.009)和MMP-7(r= 0.735,P=0.000)表达均呈明显正相关;而TIMP-2与TβRⅡ(r=-0.582,P=0.001),MMP-7(r=-0.421,P=0.008)表达呈明显负相关.结论:由于hMSH2,hMLH1突变引起TβRⅡ的失活而诱导的MMP表达减弱和TIMP的下调可能是HNPCC特殊生物学行为的一个原因.

彩色多普勒超声联合母体血浆游离胎盘mRNA对产前胎盘植入的诊断价值

目的:探讨彩色多普勒超声联合母体血浆游离胎盘人类胎盘催乳素(HPL)、人绒毛膜促性腺激素β亚单位(β-hCG)、组织因子途径抑制物2(TFPI2)mRNA诊断产前胎盘植入价值。方法:选择2017年8月—2019年6月本院产前检查疑似胎盘植入孕妇131例为研究对象,根据分娩后病理诊断结果分为胎盘植入组(45例)和对照组(86例)。均行经腹彩色多普勒超声检查,应用实时定量PCR检测母体血浆游离胎盘HPL mRNA、β-hCG mRNA、TFPI2mRNA表达。以术后病理结果为准,分析各方法检测胎盘植入的价值。结果:彩色多普勒超声检出胎盘植入32例,诊断胎盘植入的灵敏度71.1%,特异度87.2%。胎盘植入组母体血浆游离胎盘HPL mRNA、β-hCG mRNA、TFPI2mRNA表达均高于对照组(P<0.05),ROC分析HPL mRNA、β-hCG mRNA、TFPI2mRNA诊断胎盘植入的AUC分别为0.634、0.582、0.639,超声+HPL mRNA+β-hCG mRNA6+TFPI2mRNA诊断胎盘植入的灵敏度、特异度、阳性预测值、阴性预测值分别为89.7%、93.2%、92.2%、95.6%,高于单独诊断。结论:彩色多普勒超声联合母体血浆游离胎盘mRNA检测,可提高对产前胎盘植入的诊断价值。

人CTGF和TIMP1基因重组腺相关病毒双表达质粒的构建及其活性检测

目的构建人结缔组织生长因子(connective tissue growth factor,CTGF)和基质金属蛋白酶组织抑制剂1(tissue inhibitor of metalloproteinases1,TI MP1)基因腺相关病毒(adeno-associated virus,AAV)表达质粒,并观察其在HEK293细胞中的表达,检测目的蛋白的生物学活性。方法采用PCR技术,分别设计引物扩增所需序列并进行PCR鉴定。再采用分子克隆的方法,以内部核糖体进入位点(internal ribosome entry sites,IRES)序列连接CTGF和TI MP1并克隆到AAV表达质粒pSNAV2.0构建重组质粒pSNAV2-CTGF-IRES-TI MP1。把重组质粒转染HEK293细胞,用双重免疫荧光和Western blot的方法对重组质粒的表达进行研究,MTT法检测细胞培养上清液中CTGF蛋白的生物学活性,ELISA法检测细胞培养上清液中TI MP1蛋白的含量。结果PCR、酶切、DNA测序等方法均证实成功构建了含有完全正确的CTGF和TI MP1基因序列的AAV表达质粒。双重免疫荧光和Western blot检测到目的蛋白的表达,转染后的细胞上清具有促使成纤维细胞增殖的生物学活性,ELISA法结果证实重组载体能够高表达TI MP1蛋白。结论成功构建了双表达质粒pSNAV2-CTGF-IRES-TI MP1,转染HEK293细胞后能够表达具有生物学活性的目的蛋白,为基因治疗椎间盘退变的研究工作奠定了基础。

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。

Anticoagulant modulation of inflammation in severe sepsis

Inflammation and coagulation are so tightly linked that the cytokine storm which accompanies the development of sepsis initiates thrombin activation and the development of an intravascular coagulopathy. This review examines the interaction between the inflammatory and coagulation cascades, as well as the role of endogenous anticoagulants in regulating this interaction and dampening the activity of both pathways. Clinical trials attempting to improve outcomes in patients with severe sepsis by inhibiting thrombin generation with heparin and or endogenous anticoagulants are reviewed. In general, these trials have failed to demonstrate that anticoagulant therapy is associated with improvement in mortality or morbidity. While it is possible that selective patients who are severelyill with a high expected mortality may be shown to benefit from such therapy, at the present time none of these anticoagulants are neither approved nor can they be recommended for the treatment of sepsis.

人脂肪干细胞来源外泌体对四氯化碳诱导肝纤维化模型大鼠的治疗作用（英文）

背景:肝纤维化具有较高的发病率和死亡率,肝星状细胞的活化和增殖是肝纤维化进程中的关键环节。目前还没有针对单一环节或靶点的有效抗纤维化药物。目的:分析人脂肪干细胞来源外泌体对四氯化碳诱导的大鼠肝纤维化的影响。方法:①通过酶溶解法获取健康人群来源脂肪中干细胞,体外培养获取一定数量细胞后通过多重超滤法获取外泌体。体外培养的肝星状细胞经转化生长因子β1活化后利用不同浓度外泌体进行处理,通过定量PCR检测细胞内α-平滑肌动蛋白的表达明确其活化程度,以及分别使用CCK-8及流式细胞术检测各组外泌体处理后活化肝星状细胞的生长率及凋亡率。②通过腹腔注射四氯化碳构建肝纤维化大鼠动物模型,尾静脉注射外泌体进行治疗。检测各组动物的肝功能及血清Ⅲ型前胶原、Ⅳ型胶原,肝组织Ishak评分及肝纤维化半定量,以及通过免疫荧光法检测肝组织内基质金属蛋白酶组织抑制剂1、基质金属蛋白酶9及α-平滑肌动蛋白的表达。实验方案于2017年1月经同济大学动物实验伦理委员会以及医学伦理学委员会批准。结果与结论:人脂肪干细胞来源外泌体可抑制活化的肝星状细胞增殖,其可能的机制为抑制活化巨噬细胞的增殖,减少胶原纤维、α-平滑肌动蛋白及基质金属蛋白酶组织抑制剂1的表达,并促进基质金属蛋白酶9的表达。提示外泌体可治疗四氯化碳诱导肝纤维化。

辛伐他汀对肿瘤坏死因子α诱导的人腹膜间皮细胞组织型纤溶酶原激活物和纤溶酶原激活物抑制因子1表达的影响

目的探讨辛伐他汀对体外培养的人腹膜间皮细胞（human peritoneal mesothelial cells，HPMCs）在肿瘤坏死因子α（tumor necrosis factor-α，TNF-α）诱导后组织型纤溶酶原激活物（tis-sue-type plasminogen activator，t-PA）和纤溶酶原激活物抑制因子1（plasminogen activator inhibitor-1，PAId）表达的影响。方法应用胰蛋白酶消化法分离HPMCs进行原代培养并传代，将第三代分为5组（每组设3个样本）：①正常对照组：将HPMCs置于完全培养液中，置于37℃、5％CO2培养箱培养24h；②单纯TNF-α（1μg／L）诱导组：将HPMCs置于浓度为1μg／L的TNF-α诱导缓冲液中，环境温度及处理时间如前；③TNF-α诱导＋辛伐他汀（2．5μmol／L）组：将HPMCs置于含有TNF-α（1μg／L）诱导液＋辛伐他汀（2．5μmol／L）缓冲液中，余同前；④TNF-α诱导＋辛伐他汀（5．0μmol／L）组：将HPMCs置于含有TNF-α（1μg／L）诱导液＋辛伐他汀（5．0μmol／L）缓冲液中，余同前；⑤TND-α诱导＋辛伐他汀（10．0μmol／L）组：将HPMCs置于含有TNF-α（1μg／L）诱导液＋辛伐他汀（10．0μmol／L）缓冲液中，余同前。采用半定量逆转录聚合酶链反应（reverse transcription-polymerase chain reaction，RT-PCR）法检测细胞内t-PA和PAI-1 mRNA表达；酶联免疫吸附法（enzyme-linked immunosorbent assay，ELISA）检测细胞上清液中t-PA和PAI-1的蛋白质水平，用二喹啉甲酸（bicinchoninic acid，BCA）蛋白检测法测定细胞中蛋白质含量，用以校正ELISA结果。结果与正常对照组比较，TNF-α诱导能抑制HPMCS中t-PA表达，增加PAI-1的表达（P〈0．01）；与单纯TNF-α诱导组比较，辛伐他汀（2．5μmol／L）能显著增加TNF-α诱导HPMCs中t-PA表达，抑制PAI-1的表达（P〈0．05），t-PAmRNA表达水平显著上升和PAI-1mRNA表达水平显著降低（P〈0．01）；辛伐他汀（5．0μmol／L及10．0μmol／L）亦能明显增加TNF-α诱导的HPMCs中t-PA表达，抑制PAI-1的表达（P〈0．01），两者在蛋白质和基因水平均呈量效关系。结论辛伐他汀干预后能促进HPMCs在炎症状态下增加t-PA生成并抑制PAI-1的表达，从而为腹膜透析患者腹膜透析相关腹膜炎发生时使用他汀类药物以预防腹膜纤维化提供理论依据。

重组人表皮生长因子凝胶、负压封闭引流联合削痂植皮治疗深度烧伤患者的效果

目的 探究重组人表皮生长因子(rhEGF)凝胶、负压封闭引流联合削痂植皮治疗深度烧伤患者的效果及对炎症因子、血管内皮生长因子(VEGF)、基质金属蛋白酶-9(MMP-9)、基质金属蛋白酶组织抑制剂-1(TIMP-1)水平的影响,为临床治疗该疾病提供依据。方法 选取2020年2月至2021年12月张家港澳洋医院收治的深度烧伤患者70例,按照随机数字表法分为两组,各35例。对照组患者采用创面削痂植皮手术进行治疗,在此基础上,观察组患者联合rhEGF凝胶、负压封闭引流治疗。均连续治疗7 d。对比两组患者临床指标,术前、术后7 d血清炎症因子、VEGF、MMP-9及TIMP-1水平,以及并发症发生情况。结果 观察组患者创面恢复时间显著短于对照组,植皮成活率显著高于对照组(P<0.05),两组患者手术时间比较,差异无统计学意义(P>0.05);与术前比,两组患者术后7 d白细胞计数(WBC)、血清白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、C-反应蛋白(CRP)及MMP-9水平均显著降低,且观察组显著低于对照组;VEGF、TIMP-1水平均显著升高,且观察组显著高于对照组;观察组患者感染、积液、弹力性水疱、出血发生率均显著低于对照组(均P<0.05)。结论 采用rhEGF凝胶、负压封闭引流联合削痂植皮术治疗深度烧伤患者,可以提升治疗效果,减轻机体炎症反应,有利于新生血管生成,促进创面愈合,且安全性较高。

盐酸氨基葡萄糖联合氟比洛芬对增生性膝骨关节炎患者高迁移率族蛋白B1人类成纤维生长因子-2组织金属蛋白酶抑制剂-1水平的影响

目的 探讨盐酸氨基葡萄糖联合氟比洛芬对增生性膝骨关节炎患者高迁移率族蛋白B1(HMGB1)、人类成纤维生长因子-2(FGF-2)、组织金属蛋白酶抑制剂-1(TIMP-1)水平的影响。方法 选取2021年6月至2023年1月陆军第七十二集团军医院确诊增生性膝骨关节炎患者100例,采用随机数字表法分为对照组和观察组各50例,对照组接受盐酸氨基葡萄糖治疗,观察组联合应用氟比洛芬治疗,2组连续治疗6周。比较2组临床疗效,比较2组视觉模拟评分法(VAS)、Lysholm膝关节评分系统(LKSS)、西安大略和麦克马斯特大学骨关节炎指数(WOMAC)评分、HMGB1、FGF-2、TIMP-1水平、不良反应发生率。结果 与对照组(74%)比较,观察组临床总有效率(90%)升高(P<0.05);与治疗前比较,治疗后2组VAS、WOMAC评分降低,LKSS评分升高,并且观察组VAS、WOMAC评分低于对照组,LKSS评分高于对照组(P<0.05);治疗后2组HMGB1水平比治疗前降低,FGF-2、TIMP-1水平升高,并且观察组HMGB1低于对照组,FGF-2、TIMP-1水平高于对照组(P<0.05);2组不良反应发生率比较,差异无统计学意义(P>0.05)。结论 盐酸氨基葡萄糖联合氟比洛芬能够有效治疗增生性膝骨关节炎,同时还能降低HMGB1水平,升高FGF-2、TIMP-1水平,且不良反应少,具有较高的临床参考价值。

基质金属蛋白酶抑制因子1和nm-23在人表皮生长因子受体2阳性乳腺癌中的表达及意义

目的探讨基质金属蛋白酶抑制因子1（TIMP-1）和nm．23在人表皮生长因子受体2（HER-2）阳性乳腺癌中的表达意义。方法采用免疫组化方法检测165例HER-2阳性乳腺癌组织中TIMP-1和nm-23的表达，分析其与乳腺癌临床病理特征的关系及在预后中的作用。结果165例HER-2阳性乳腺癌组织中，TIMP-1阳性表达率为53．3％（88／165），nm-23高表达率为72．7％（120／165）。TIMP-1阴性组和阳性组患者的10年无远处转移生存率分别为58．4％和43．2％（P=0．041），10年总生存率分别为68．8％和52．0％（P=0．020）。nm-23高表达组和低表达组患者的10年无远处转移生存率分别为54．2％和40．0％（P=0．049），10年总生存率分别为65．7％和44．1％（P=0．015）。多因素分析显示，TIMP-1表达、nm-23表达、肿瘤大小和淋巴结转移是影响HER-2阳性乳腺癌患者预后的独立因素。结论在HER-2阳性乳腺癌中，TIMP-1和nm-23的表达可以进一步区分低危和高危亚组，并作为独立因素判断预后。