Tissue inhibitor of metalloproteinase-3 expression affects clinicopathological features and prognosis of aflatoxin B1-related hepatocellular carcinoma

BACKGROUND The dysregulation of tissue inhibitor of metalloproteinase-3(TIMP3)was positively correlated with the progression of hepatocellular carcinoma(HCC).However,it is not clear whether TIMP3 expression is associated with the clinico-pathological features and prognosis of aflatoxin B1(AFB1)-related HCC(AHCC).A retrospective study,including 182 patients with AHCC,was conducted to explore the link between TIMP3 expression in cancerous tissues and the clinico-pathological characteristics and prognosis of AHCC.TIMP3 expression was detected by immunohistochemistry and its effects on the clinicopathological features and prognosis of AHCC were evaluated by Kaplan-Meier survival analysis and Cox regression survival analysis.Odds ratio,hazard ratio(HR),median overall survival time(MST),median tumor recurrence-free survival time(MRT),and corresponding 95%confidential interval(CI)was calculated to RESULTS Kaplan-Meier survival analysis showed that compared with high TIMP3 expression,low TIMP3 expression in tumor tissues significantly decreased the MST(36.00 mo vs 18.00 mo)and MRT(32.00 mo vs 16 mo)of patients with AHCC.Multivariate Cox regression survival analysis further proved that decreased expression of TIMP3 increased the risk of death(HR=2.85,95%CI:2.04-4.00)and tumor recurrence(HR=2.26,95%CI:1.57-3.26).Furthermore,decreased expression of TIMP3 protein in tissues with AHCC was significantly correlated with tumor clinicopatho-logical features,such as tumor size,tumor grade and stage,tumor microvessel density,and tumor blood invasion.Additionally,TIMP3 protein expression was also negatively associated with amount of AFB1-DNA adducts in tumor tissues.CONCLUSION These findings indicate that the dysregulation of TIMP3 expression is related to AHCC biological behaviors and affects tumor outcome,suggesting that TIMP3 may act as a prognostic biomarker for AHCC.

Long noncoding RNAs HAND2-AS1 ultrasound microbubbles suppress hepatocellular carcinoma progression by regulating the miR-873-5p/tissue inhibitor of matrix metalloproteinase-2 axis

BACKGROUND Increasing data indicated that long noncoding RNAs(lncRNAs)were directly or indirectly involved in the occurrence and development of tumors,including hepatocellular carcinoma(HCC).Recent studies had found that the expression of lncRNA HAND2-AS1 was downregulated in HCC tissues,but its role in HCC progression is unclear.Ultrasound targeted microbubble destruction mediated gene transfection is a new method to overexpress genes.AIM To study the role of ultrasound microbubbles(UTMBs)mediated HAND2-AS1 in the progression of HCC,in order to provide a new reference for the treatment of HCC.METHODS In vitro,we transfected HAND2-AS1 siRNA into HepG2 cells by UTMBs,and detected cell proliferation,apoptosis,invasion and epithelial-mesenchymal transition(EMT)by cell counting kit-8 assay,flow cytometry,Transwell invasion assay and Western blotting,respectively.In addition,we transfected miR-837-5p mimic into UTMBs treated cells and observed the changes of cell behavior.Next,the UTMBs treated HepG2 cells were transfected together with miR-837-5p mimic and tissue inhibitor of matrix metalloproteinase-2(TIMP2)overexpression vector,and we detected cell proliferation,apoptosis,invasion and EMT.In vivo,we established a mouse model of subcutaneous transplantation of HepG2 cells and observed the effect of HAND2-AS1 silencing on tumor formation ability.RESULTS We found that UTMBs carrying HAND2-AS1 restricted cell proliferation,invasion,and EMT,encouraged apoptosis,and HAND2-AS1 silencing eliminated the effect of UTMBs.Additionally,miR-873-5p targets the gene HAND2-AS1,which also targets the 3’UTR of TIMP2.And miR-873-5p mimic counteracted the impact of HAND2-AS1.Further,miR-873-5p mimic solely or in combination with pcDNA-TIMP2 had been transformed into HepG2 cells exposed to UTMBs.We discovered that TIMP2 reversed the effect of miR-873-5p mimic caused by the blocked signalling cascade for matrix metalloproteinase(MMP)2/MMP9.In vivo results showed that HAND2-AS1 silencing significantly inhibited tumor formation in mice.CONCLUSION LncRNA HAND2-AS1 promotes TIMP2 expression by targeting miR-873-5p to inhibit HepG2 cell growth and delay HCC progression.

LOW DOSE PIRFENIDONE SUPPRESSES TRANSFORMING GROWTH FACTOR BETA-1 AND TISSUE INHIBITOR OF METALLOPROTEINASE-1, AND PROTECTS RATS FROM LUNG FIBROSIS INDUCED BY BLEOMYCIN

Objective To investigate the optimal dosage of pirfenidone for the treatment of pulmonary fibrosis induced by bleomycin in Wistar rats, and the alteration of expressions of transforming growth factor beta-1 （ TGF-β1 ）, tissue inhibitor of metalloproteinase-1 （ TIMP-1 ）, and matrix metalloproteinase-13 （ MMP-13 ） in lung tissue. Methods Male Wistar rats were endotracheally instilled with bleomycin or normal saline. Pirfenidone （25-800 mg · kg^-l · d^-1 ）, dexamethasone （3 mg/kg）, or 1% carboxymethylcellulose sodium were given daily by feed 2 days before instillation of bleomycin. Groups T7 and T14 were fed pirfenidone 50 mg · kg^-1 · d^-1 at 7 days or 14 daYs after bleomycin instillation. Lungs were harvested at 28 days after bleomycin instillation. Patholological changes in luffg tissues were evaluated with HE staining. Lung collagen was stained by sirius red and measured by content of hydroxypro- line. Expression of proteins of TGF-β1 TIMP-1, and MMP-13 were detected by Western blotting. Results At doses of 25, 50, and 100 mg· kg^- 1 · d ^- 1, pirfenidone had significant anti-fibrotic effects for bleomy- cin-induced rat pulmonary fibrosis, and these effects were most significantly attenuated at the dosage of 50 mg · kg^-1 ·d^ -1（ HE： P 〈 0. 01, P 〈 0.01, and P = 0.064; sirius red： P 〈0.05, P 〈 0.01, and P 〈 0.05 ; hydroxyproline： P = 0.595, P 〈 0.01, and P = 0.976）. Pirfenidone at a dosage of 50 mg · kg^- l · d^-1 inhibited protein expression of TGF-131 and TIMP-1 in lung tissue in the early phase （0.79 and 0.75 times of control group）, but had no effect on ex- nr^eelnn nf MMP-13. Conclusion Low dose pirfenidone, especially at dosage of 50 mg · kg^-1 · d^-1, has significant anti-fibrotic effects on bleomycin-induced rat pulmonary fibrosis. Pirfenidone partially inhibits the enhancement of the expression of TGF-131 and TIMP-β1 in lung tissue.

Expression of matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 in ulcerative colitis

AIM: To examine the expression of metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the colonic mucosa of patients with ulcer- ative colitis (UC). METHODS: Reverse transcription-polymerase chain re- action (RT-PCR) and immunohistochemistry were used to study the expression of MMP-1 and TIMP-1 at both mRNA and protein levels in patients with UC and con- trols. The relationship between MMP-1 mRNA, TIMP-1 mRNA, MMP-1 mRNA/TIMP-1 mRNA ratio and the sever- ity of clinical symptoms of the patients with UC were also analyzed. RESULTS: The expression of MMP-1 mRNA and TIMP-1 mRNA in the ulcerated and inflamed colonic mucosa was signifi cantly higher than that in the non-inflamed colonic mucosa (P < 0.001), but there was no statistically signif i- cant difference in the non-inflamed colonic mucosa of UC patients and normal controls (P > 0.05). The mRNA ex- pression of MMP-1 and TIMP-1 in ulcerated colonic mu- cosa of UC patients was increased by 80-fold and 2.2-fold, respectively when compared with the normal controls. In the inflamed colonic mucosa, the increase was 30-fold and 1.6-fold, respectively. Immunohistochemical analy- sis showed that among the ulcerated, inflamed, and non-inflamed colonic mucosae of UC patients and the normal controls, the positive rate of MMP-1 expression was 87%, 87%, 40% and 35% respectively, and the positive rate of TIMP-1 expression was 89%, 89%, 80% and 75%, respectively. Furthermore, the expression of MMP-1 mRNA, TIMP-1 mRNA and the MMP-1 mRNA/ TIMP-1 mRNA ratio were correlated with the severity of clinical symptoms (P <0.05).CONCLUSION: Excessive expression of MMP-1 in the diseased colonic mucosa causes excessive hydrolysis of the extracellular matrix (ECM) and ulceration in UC pa-tients. MMP-1 mRNA, TIMP-1 mRNA and MMP-1 mRNA/ TIMP-1 mRNA ratio can be used as biomarkers to judge the severity of clinical symptoms in patients with UC. Exogenous TIMP-1 or MMP-1 inhibitor therapy is a novel treatment for patients with UC.

Plasma matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 as biomarkers of ulcerative colitis activity

AIM:Overexpression of mucosal metalloproteinases(MMP) has been demonstrated recently in inflammatory bowel disease.Their activity can be counterbalanced by the tissue inhibitor of metalloproteinases(TIMP).The aim of this study was to evaluate the effect of ulcerative colitis(UC)on MMP- 1 and TIMP-1 plasma concentrations,as two possible biomarkers of the disease activity. METHODS:MMP-1 and TIMP-1 plasma concentrations were measured with an enzyme immunoassay in 16 patients with endoscopically confirmed active UC. RESULTS:Plasma concentrations of both MMP-1(13.7±0.2 ng/ml)and TIMP-1(799±140 ng/ml)were significantly elevated in UC patients in comparison to healthy controls (11.9±0.9 ng/ml and 220±7 ng/ml respectively).There was no correlation between TIMP-1 and MMP-1 concentrations (r=0.02).TIMP-1 levels revealed significant positive correlations with scored endoscopic degree of mucosal injury, disease activity index and clinical activity index values as well as C-reactive protein concentration.There was no correlation between MMP-1 and laboratory,clinical or endoscopic indices of the disease activity.CONCLUSION: These results confirm the role of both MMP- 1 and TIMP-1 in the pathogenesis of ulcerative colitis. However only TIMP-1 can be useful as a biomarker of the disease activity, demonstrating association with clinical and endoscopic pictures.

Interleukin-1 beta up-regulates tissue inhibitor of matrix metalloproteinase-1 mRNA and phosphorylation of c-jun N-terminal kinase and p38 in hepatic stellate cells

AIM： To study the relationship between interleukin-lbeta （IL-1β） up-regulating tissue inhibitor of matrix metalloproteinase-1 （TIMMP-1） mRNA expression and phosphorylation of both c-jun N-terminal kinase （INK） and p38 in rat heffatic stellate cells （HSC）. METHODS： RT-PCR was performed to measure the expression of TIMMP-1 mRNA in rat HSC. Western blot was performed to measure IL-1β-induced JNK and p38 activities in rat HSC. RESULTS： TIMMP-1 mRNA expression （1.191± 0.079） was much higher after treatment with IL-1β （10 ng/mL） for 24 h than in control group （0.545±0.091） （P〈0.01）. IL-1β activated INK and p38 in a time-dependent manner. After stimulation with IL-1β for 0, 5, 15, 30, 60 and 120 min, the INK activity was 0.982±0.299, 1.501±0.720, 2.133±0.882, 3.360±0.452, 2.181±0.789, and 1.385 ± 0.368, respectively. There was a significant difference in JNK activity at 15 min （P〈 0.01）, 30 min （P〈 0.01） and 60 min （P〈0.01） in comparison to that at 0 min. The p38 activity was 1.061±0.310, 2.050±0.863, 2.380±0.573, 2.973±0.953, 2.421±0.793, and 1.755 ± 0.433 at the 6 time points （0, 5, 15, 30, 60 and 120 min） respectively. There was a significant difference in p38 activity at 5 min （P〈0.05）, 15 min （P〈0.01）, 30 min （P〈0.01） and 60 min （P〈0.01） compared to that at 0 min. TIMMP-1 mRNA expression trended to decrease in 3 groups pretreated with different concentrations of SP600125 （10 μmol/L, 1.022±0.113; 20 μmol/L, 0.869±0.070; 40 μmol/L, 0.666±0.123）. Their decreases were all significant （P〈0.05, P〈0.01, P〈0.01） in comparison to control group （without SP600125 treatment, 1.163±0.107）. In the other 3 groups pretreated with different concentrations of SB203580 （10 μmol/L, 1.507±0.099; 20 μmol/L, 1.698±0.107; 40 μmol/L, 1.857±0.054）, the expression of TIMMP-1 mRNA increased. Their levels were higher than those in the control group （without SB203580 treatment, 1.027 ± 0.061） with a significant statistical significance （P〈 0.01）. CONCLUSION： IL-1β has a direct action on hepatic fibrosis by up-regulating TIMMP-1 mRNA expression in ratessionin in rate HSC.JNK and p38 mitogen-activated protein kinases （MAPKs） are involved in IL-1β-induced TIMMP-1 gene expression, and play a distinct role in this process, indicating that p38 and .INK pathways cooperatively mediate TIMP-1 mRNA expression in rat HSC.

Matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 expression in early focal cerebral infarction following urokinase thrombolysis in rats

Activity of matrix metalloproteinase-9 increases following cerebral ischemia/reperfusion, and is associated with cerebral microvascular permeability, blood-brain barrier destruction, inflammatory cell infiltration and brain edema. Matrix metalloproteinase-9 also likely participates in thrombolysis. A rat model of middle cerebral artery infarction was established by injecting autologous blood clots into the internal carotid artery. At 3 hours following model induction, urokinase was injected into the caudal vein. Decreased neurological severity score, reduced infarct volume, and increased expression of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 were observed in the cerebral cortex 24 hours after urokinase thrombolysis. These results suggest that urokinase can suppress damage in the acute-early stage of cerebral infarction.

Preparation and in vitro studies of microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2

Objective： To prepare microencapsulated cells releasing human tissue inhibitor ofmetalloproteinase-2 （TIMP-2）, and investigate their biological characteristics in vitro. Methods： Chinese hamster ovary （CHO） cells were stably transfected with a human TIMP-2 expression vector, encapsulated in barium alginate microcapsules and cultured in vitro. Morphological appearance of the microcapsules was observed under a light microscope. Cell viability was assessed using MTT （3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide） assay. Enzyme linked immunosorbent assay （ELISA） and reverse zymography were used to confirm the release of biologically active TIMP-2 from the microcapsules. Cryopreservation study of the microencapsulated cells was carried out using dimethyl sulfoxide （DMSO） as preservative agent. Results： The microcapsules appeared like a sphere with diameter of 300-600 ~tm. The surface of the capsule wall was clearly smooth. The microencapsulated cells survived well and kept proliferating over the 6 weeks observed. No significant difference in TIMP-2 secretion was found between encapsulated and unencapsulated cells. Reverse zymography confirmed the bioactivity of MMP （matrix metalloproteinase） inhibition of TIMP-2. The cryopreservation process did not damage the microcapsule morphology nor the viability of the cells inside. Conclusion： Microencapsulated engineered CHO cells survive at least 6 weeks after preparation in vitro, and secrete bioactive TIMP-2 freely from the microcapsules.

Expression of tissue inhibitor of metalloproteinase-1 in progression muscular dystrophy

Objective Tissue inhibitor of metalloproteinase-1 （TIMP-1） is a muhifunctional protein that has thc capacity to modify cellular activities and to modulate matrix turnover. This paper revealed the contributive role of TIMP-1 in progressive muscular dystrophy （PMD）. Methods We examined the expression and cellular localization of TIMP-1 protein using biopsied frozen muscle from patients with Duchenne muscular dystrophy （DMD）, Becker muscular dystrophy （BMD） , congenital muscular dystrophy （CMD） by immunohistochemistry, double immunofluorescence and Western blot analysis. Results The results of immunohistochemistry and double immunofluorescence showed that TIMP-1 was positive only in vascular endothelial cells of normal muscles. Immunohistochemistry and Western blot analysis showed that the staining intensity was distinctly increased in some dystrophic muscles of PMD for TIMP-1. Double immunofluorescence revealed that TIMP-1 strongly expressed in the regenerating muscle fibers, macrophages and macrophage infiltrating necrotic fibers. Some activated fibroblasts in endomysium and perimysium of DMD and CMD muscles were also positive for TIMP- 1. Conclusion The functional consequence of overexpression of TIMP-1 in the dystrophic muscles is unknown, but the elevated local expression of TIMP-1 in diseased muscles of PMD and their distinct distribution pattern provide evidence that TIMP-1 may participate in the pathogenesis of PMD.

Expressions of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in malignant peripheral nerve sheath tumor

BACKGROUND： Matrix metalloproteinase-9 （MMP-9） can degrade collagen IV （the main structural ingredient of basilar membrane）, and it also plays an important role in tumor vascularization, tumor cell progression, formation of metastatic focus, etc. Tissue inhibitor of metalloproteinase-1 （T1MP-1） can bind with MMP-9 to form 1 ： 1 compound and inhibit its activity, and can negatively regulate the tumor progression and metastasis. OBJECTIVE： To analyze the relationship of MMP-9 and T1MP-1 expressions with the pathological grade, metastasis and prognosis of malignant peripheral nerve sheath tumor （MPNST）. DESIGN： An observational comparative experiment. SETTING： Heze Medical College. PARTICIPANTS： Fifty-eight surgical pathological samples, which were clearly diagnosed to be MPNST, were collected from the pathological laboratory archives in the Department of Pathology, Heze Municipal Hospital from January 1988 to December 2003. The MPNST pathological types were common tumor in 53 cases, malignant triton tumor in 2 cases, epithelial MPNST in 2 cases and MPNST with gland differentiation in 1 case. The pathological grade was grade 1 in 11 cases, grade 2 in 24 cases and grade 3 in 23 cases. Besides, the resected tumor samples of 20 patients with benign peripheral nerve tumor （10 cases of nerve sheath tumor and 10 cases of neurofibromatosis） and the normal peripheral nerves （by-products of some surgeries） of 5 patients were also collected. The samples were used with the approval of the patients. Rat-anti-human MMP-9, TIMP-1 monoclonal antibody and S-P kit were purchased from Fuzhou Maixin Biotechnology, Co.,Ltd. METHODS： The documented paraffin blocks were again prepared to sections of 5 lJ m. The expressions of MMP-9 and TIMP-1 in the samples were detected with mmunohistochemical S-P method. The relationships of the MPNST severity, recurrence, metastasis and survival rate with the expressions of MMP-9 and TIMP-1 were analyzed. MAIN OUTCOME MEASURES： Relationships of MMP-9 and TIMP-1 expressions with the MPNST severity and prognosis. RESULTS： ①Expressions of MMP-9 and TIMP-1 in three tissues： MMP-9 and TIMP-1 stainings were mainly observed in cytoplasm. Among the 58 MPNST patients, the MMP-9 expression was significantly higher than those in normal peripheral nerve and benign tumor （P 〈 0.05）, while the TIMP-1 expression in MPNST was lower than those in normal peripheral nerve and benign tumor （P 〈 0.05）. ②Relationship of MMP-9 and TIMP-1 expressions with the severity and prognosis of MPNST： The expressions of both proteins were observed in the four subtypes. The positive expression of MMP-9 in the MPNST patients of grades 2 - 3 was significantly higher than that in the MPNST patients of grade 1 （P 〈 0.05）, while the expression of MMP-9 was significantly lower than that in the MPNST patients of grade 1 （P 〈 0.05）. The metastatic rate was positively correlated with MMP-9 expression （r =1.696, P 〈 0.05）, but negatively correlated with TIMP-1 expression （r = - 2.125, P 〈 0.05）. CONCLUSION： MMP-9 and TIMP-1 are associated with MPNST pathological grades and metastasis, and can be used as the indicators for judging the severity and orognosis of MPNST.

血清TLR4、TIMP-1水平与小儿热性惊厥临床特征的关系及对继发癫痫的预测价值

目的分析血清Toll样受体4(TLR4)、基质金属蛋白酶组织抑制剂(TIMP)-1水平与小儿热性惊厥(FC)临床特征的关系及对FC继发癫痫的预测价值。方法选取2019年1月至2022年6月320例FC患儿作为研究组,另选取同期发热无惊厥儿童150例作为发热组,体检健康儿童150例作为对照组。根据FC患儿是否继发癫痫分为癫痫组和无癫痫组。采用酶联免疫吸附试验检测血清TLR4、TIMP-1水平,采用Pearson相关分析TLR4、TIMP-1水平及与临床指标间的相关性。采用受试者工作特征(ROC)曲线分析血清TLR4、TIMP-1预测FC继发癫痫的价值。采用Logistic回归分析FC患儿继发癫痫的影响因素。结果FC患儿、发热无惊厥儿童、体检健康儿童血清TLR4、TIMP-1水平依次降低,且两两比较,差异均有统计学意义(P<0.05)。研究组与发热组围生期异常发生情况、肿瘤坏死因子α(TNF-α)、C-反应蛋白(CRP)、白细胞介素-1β(IL-1β)水平和振幅整合脑电图(AEEG)评分比较,差异均有统计学意义(P<0.05)。癫痫组患儿血清TLR4、TIMP-1水平明显高于无癫痫组(P<0.05)。癫痫组和无癫痫组患儿首次惊厥次数、惊厥持续时间、首次惊厥前发热时间、围生期异常发生情况、TNF-α、CRP、IL-1β水平和AEEG评分比较,差异均有统计学意义(P<0.05)。血清TLR4水平与TIMP-1呈正相关(P<0.05);血清TLR4、TIMP-1水平与TNF-α、CRP、IL-1β呈正相关(P<0.05),与AEEG评分呈负相关(P<0.05)。TLR4、TIMP-1联合预测FC患儿继发癫痫的曲线下面积(AUC)明显高于单项检测的AUC(Z_(TLR4-联合)=3.016,P=0.003;Z_(TIMP-1-联合)=2.232,P=0.026)。Logistic回归分析结果表明,TLR4、TIMP-1、TNF-α、CRP、IL-1β水平升高,AEEG评分降低均为FC继发癫痫的危险因素(P<0.05)。结论血清TLR4、TIMP-1与FC患儿临床特征密切相关,TLR4、TIMP-1可能是FC继发癫痫的影响因素。

声带癌前病变组织中基质金属蛋白酶抑制剂-1、果蝇母亲DDP同源物4表达水平与术后复发和恶变的相关性研究

目的探讨声带癌前病变组织中基质金属蛋白酶抑制剂-1(tissue inhibitor of metalloproteinases 1,TIMP-1)、果蝇母亲DDP同源物4(drosophila mothers against DDP homolog 4,Smad4)表达水平与术后复发和恶变的相关性。方法回顾性分析2018年8月~2021年8月郑州大学第一附属医院收治的162例声带癌前病变患者的临床和病理资料,收集手术切除癌前病变组织(癌前病变组)及病变旁正常黏膜组织(对照组),采用免疫组织化学法检测组织中TIMP-1、Smad4表达情况。分析TIMP-1、Smad4阳性率与临床病理特征的关系,并采用Kaplan-Meier法和Cox回归分析法分析其对术后复发和恶变的影响。结果与对照组正常黏膜组织比较,癌前病变组的TIMP-1阳性率较高,Smad4阳性率较低(P<0.05)。不同病变范围、是否累及前连合、不同程度上皮异常增生患者的TIMP-1、Smad4阳性率存在差异(P<0.05)。术后随访时间24~60个月,中位随访时间36个月,随访期间失访患者6例,随访率96.30%(156/162),随访期间术后复发35例(21.60%),术后恶变16例(9.88%);Kaplan-Meier生存分析显示,TIMP-1阳性患者术后复发率和恶变率高于TIMP-1阴性患者(P<0.05);Smad4阴性患者术后复发率和恶变率高于Smad4阳性患者(P<0.05)。多因素Cox回归分析显示,喉咽反流、病变范围>1/2、中/重度异型增生、TIMP-1阳性、Smad4阴性是复发的独立危险因素(P<0.05),年龄>60岁、累及前连合、TIMP-1阳性、Smad4阴性是恶变的独立危险因素(P<0.05)。结论声带癌前病变组织中TIMP-1高表达、Smad4低表达,且TIMP-1阳性、Smad4阴性表达者术后复发和恶变风险较高。

精神分裂症患者并发糖尿病的危险因素分析及RBP4、Vaspin对糖尿病的预测价值

目的分析精神分裂症患者并发糖尿病的危险因素分析及视黄醇结合蛋白4(RBP4)、脂肪特异性丝氨酸蛋白酶抑制剂(Vaspin)对糖尿病的预测价值。方法选取2018年6月至2023年6月于我院治疗的90例精神分裂症患者为研究对象,根据患者糖尿病发生情况分为未并发糖尿病组67例,并发糖尿病组23例,分析精神分裂症患者并发糖尿病的危险因素,采用ROC曲线评价RBP4、Vaspin对糖尿病的预测价值。结果单因素分析显示,精神分裂症患者糖尿病的发生与性别、居住地、职业、文化程度、婚姻状况、具有精神病家族史无关,P>0.05;年龄>60岁、体质量为肥胖、病程>10年、住院时间>6个月、使用氯氮平、具有糖尿病遗传史、存在高血脂症、胆固醇偏高患者糖尿病发生率较高(P<0.05)。多因素Logistics回归分析显示,年龄、体质量、病程、住院时间、抗精神病药物、糖尿病遗传史、高血脂症、胆固醇为影响精神分裂症患者并发糖尿病的危险因素(P<0.05)。与未并发糖尿病组对比,并发糖尿病组患者的RBP4、Vaspin表达水平升高(P<0.05)。ROC曲线显示,与RBP4、Vaspin单项预测对比,两项联合对糖尿病具有较高的预测价值(P<0.05)。结论年龄>60岁、体质量为肥胖、病程>10年、住院时间>6个月、使用氯氮平、具有糖尿病遗传史、存在高血脂症、胆固醇偏高为精神分裂症患者并发糖尿病的危险因素,检测机体内RBP4、Vaspin表达水平,可用于糖尿病的预测。

心力衰竭患者外周血BNP、TIMP-4水平及二者交互作用对临床预后的影响

目的探究心力衰竭患者外周血脑钠肽(BNP)、金属蛋白酶组织抑制因子-4(TIMP-4)及二者交互作用对临床预后的影响。方法选取2020年10月—2022年10月200例心力衰竭作为观察组,另选取同期体检健康者200例作为对照组。比较2组外周血BNP、TIMP-4水平,分析不同水平BNP、TIMP-4对心力衰竭发病风险的影响,比较不同预后患者临床资料及外周血BNP、TIMP-4水平,分析心力衰竭患者预后影响因素及BNP、TIMP-4交互作用对心力衰竭患者预后的影响,评价BNP、TIMP-4对心力衰竭患者预后的预测价值。结果观察组BNP水平较对照组高,TIMP-4水平较对照组低(P<0.05);外周血BNP、TIMP-4高水平患者心力衰竭发病风险分别是低水平患者的3.348倍、0.409倍;预后不良患者左心室射血分数(LVEF)、TIMP-4较预后良好患者低,BNP较预后良好患者高(P<0.01);LVEF及外周血BNP、TIMP-4为心力衰竭患者预后影响因素(P<0.01);心力衰竭患者外周血BNP、TIMP-4存在相加交互作用,且预后不良风险中有32.44%归因于二者交互作用;外周血BNP、TIMP-4联合预测心力衰竭患者预后不良的ROC曲线下面积(AUC)为0.929,二者联合预测的AUC明显较单独指标高(P<0.05)。结论心力衰竭患者外周血BNP水平升高,TIMP-4水平下降,且二者存在交互作用,共同参与心力衰竭病情发展,与患者预后不良密切相关,检测其水平有助于临床医师预判患者预后。

2型糖尿病合并冠心病患者TIMP-4、FFA水平和EAT厚度与颈部血管病变相关性分析

【目的】探讨2型糖尿病(T2DM)合并冠心病(CHD)患者组织型金属蛋白酶抑制剂4(TIMP-4)、游离脂肪酸(FFA)水平及心外膜脂肪组织(EAT)厚度与颈部血管病变相关性。【方法】选取2019年5月至2022年5月在本院行冠脉造影检查诊断为T2DM合并CHD的158例患者,根据是否存在颈部血管病变分为非颈部血管病变组(n=90)、颈部血管病变组(n=68)。收集并比较两组患者临床资料;采用酶联免疫吸附法(ELSA)检测两组患者血清TIMP-4、FFA水平;使用超声心动图测量EAT厚度;采用多因素Logistic回归分析T2DM合并CHD患者发生颈部血管病变的危险因素;采用受试者工作特征(ROC)曲线评估血清TIMP-4、FFA水平及EAT厚度对T2DM合并CHD患者发生颈部血管病变的诊断价值。【结果】与非颈部血管病变组比较,颈部血管病变组患者血清TIMP-4水平降低(P<0.05),FFA水平及EAT厚度升高(P<0.05);多因素Logistic回归分析显示:TIMP-4水平降低、FFA水平及EAT厚度升高是T2DM合并CHD颈部血管病变的危险因素(OR=3.968、2.587、4.125,均P<0.05);血清TIMP-4、FFA水平及EAT厚度联合诊断T2DM合并CHD患者发生颈部血管病变的AUC分别为0.753、0.506、0.856、0.915,灵敏度分别为83.10%、70.69%、83.1%、85.77%,特异度分别为85.60%、62.87%、89.6%、82.96%。【结论】血清TIMP-4、FFA水平及EAT厚度联合检测对T2DM合并CHD患者发生颈部血管病变有一定的诊断价值,可为临床早期诊治T2DM合并CHD颈部血管病变提供一定参考。

Effect of a nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester on invasion of human colorectal cancer cell line SL-174T

AIML To investigate the effect and mechanism of action of the nitric oxide synthase （NOS） inhibitor NG-nitro-L-arginine methyl ester （L-NAME） on invasion and metastasis of human colorectal cancer cell line SL-174T. METHODS： Human colorectal cancer cel4 line SL-174T was cultured and treated separately with four different dosages of L-NAME for 72 h, Nitric oxide （NO） production was measured with Griess reagent, The effect of L-NAME on invasion and migration of SL-174T cells were evaluated by using Transwell chambers attached with polycarbonate filters and reconstituted basement membrane （Matrigel）, RT-PCR was performed to determine the mRNA levels of matrix metalloproteinase-2 （MMP-2） and tissue inhibitor metalloproteinase-2 （TIMP-2）,RESULTS： L-NAME could significantly inhibit NO production of SL-174T in a dose-dependent manner. After being treated for 72 h with 0.2, 0.4, 0.8, and 1.0 mmol/L L- NAME, respectively, the ability of the L-NAME treated SL- 174T cells to invade the reconstituted basement membrane decreased significantly （t = 8.056, P〈0.05; t= 14.467, P〈0.01; t= 27.785, P〈0.01; and t= 29.405, P〈0.01, respectively） and the inhibition rates were 10.29%, 19.62%, 34.08%, and 42.23%, respectively. Moreover, L-NAME could inhibit migration of SL-174T cells, and the inhibition rates were 20.76%, 24.95%, 39.43%, and 46. 85% for L-NAME at 0.2, 0.4, 0.8, and 1.0 mmol/L, respectively （t = 15.116, P〈0.01）. In addition, after treatment with L-NAME, expression of MMP-2 mRNA was significantly decreased （t = 71.238, P〈0.01） and that of TIMP-2 mRNA was markedly increased （t = -13.020, P〈0.01）. CONCLUSION： L-NAME exerts anti-invasive and anti- metastatic effects on SL-174T cell line via downregulating MNP-2 mRNA expression and upregulating TIMP-2 mRNA expression.

血清Syndecan-4、TIMP-1与STEMI患者冠状动脉病变血管支数、超声心动图参数的关系

目的 分析多配体蛋白聚糖4(Syndecan-4)、基质金属蛋白酶抑制剂-1(TIMP-1)与ST段抬高型心肌梗死(STEMI)患者冠状动脉病变血管支数、超声心动图参数的关系。方法 选取2021年1—12月海军军医大学第一附属医院心内科诊治的STEMI患者67例作为STEMI组,将同期确诊为慢性心绞痛的72例患者作为慢性心绞痛组,另选取同期冠状动脉造影结果正常的50例患者作为对照组。依据冠状动脉造影检查Gensini积分结果将STEMI患者再分为A亚组25例(0~20分,1支血管病变)、B亚组23例(21~40分,2支血管病变)和C亚组19例(40分以上,3支及以上血管病变)。比较各组血清Syndecan-4和TIMP-1水平,超声心动图参数[左心室射血分数(LVEF)、左心室舒张末期内径(LVEDD)、左心室收缩末期内径(LVESD)、左心室舒张末期容积(LVEDV)、左心室收缩末期容积(LVESV)、舒张早期二尖瓣血流速度/舒张晚期二尖瓣血流速度(E/A)]、心肌缺血指标[心肌肌钙蛋白I(cTnI)、肌酸激酶同工酶MB(CK-MB)]、心力衰竭指标[N末端脑钠肽前体(NT-proBNP)],Pearson法分析STEMI患者血清Syndecan-4、TIMP-1水平与Gensini积分和超声心动图参数之间的关系,受试者工作特征曲线(ROC)分析血清Syndecan-4、TIMP-1预测STEMI的价值。结果 血清Syndecan-4、TIMP-1及LVESD水平比较,STEMI组>慢性心绞痛组>对照组(F=253.293、67 612.917、20.563,P均<0.001);LVEF水平比较,STEMI组<慢性心绞痛组<对照组(F=28.194,P<0.001),3组LVEDD、LVEDV、LVESV和E/A比较差异无统计学意义(P>0.05);STEMI组cTnI、CK-MB和NT-proBNP水平高于慢性心绞痛组和对照组(F=1 809.343、1 352.872、2 289.851,P均<0.001),慢性心绞痛组cTnI和NT-proBNP水平高于对照组(P<0.05);Gensini积分比较,C亚组>B亚组>A亚组(F=951.801,P<0.001);A、B、C亚组患者血清Syndecan-4、TIMP-1及LVESD水平依次升高(F/P=4.405/0.016、2 965.986/<0.001、3.520/0.035),而LVEF水平依次降低(F/P=5.385/0.007),3亚组LVEDD、LVEDV、LVESV和E/A比较差异无统计学意义(P>0.05);STEMI患者血清Syndecan-4、TIMP-1水平与Gensini积分、LVESD均呈正相关(Gensini积分:r/P=0.418/<0.001,0.375/<0.001;LVESD:r/P=0.391/<0.001,0.278/0.004),与LVEF均呈负相关(r/P=-0.492/<0.001、-0.436/<0.001),与LVEDD、LVEDV、LVESV和E/A无明显相关性(P>0.05);血清Syndecan-4、TIMP-1及二者联合预测STEMI的曲线下面积(AUC)分别为0.849、0.917、0.927,二者联合预测价值高于单项指标(Z=18.623、11.736,P均<0.001)。结论 血清Syndecan-4、TIMP-1水平与冠状动脉病变血管支数和超声心动图参数高度相关,可作为STEMI患者冠状动脉病变程度和心肌损伤检测的重要生物标志物。

TIMP4表达与心房颤动患者心肌纤维化的相关性研究

目的探究基质金属蛋白酶抑制剂4(TIMP4)表达与心房颤动患者心肌纤维化的相关性。方法选取2021年6月至2022年12月在本院就诊治疗的82例心房颤动患者为研究对象,将患者分为阵发性房颤组(n=42)例和持续性房颤组(n=40);另选取同期体检健康者40例作为对照组。采用实时荧光定量PCR(qRT-PCR)测定研究对象血清TIMP4水平;采用酶联免疫吸附法(ELISA)检测研究对象血清Ⅰ型前胶原羧基端肽(PICP)、Ⅲ型前胶原羧基端肽(PⅢCP)水平;采用Pearson分析血清TIMP4与PICP、PⅢCP水平相关性;采用多因素Logistic回归分析发生持续性心房颤动的影响因素。结果对照组、阵发性房颤组、持续性房颤组间的肿瘤坏死因子-α(TNF-α)和白细胞介素-21(IL-21)水平呈逐渐升高趋势(P<0.05);与对照组和阵发性房颤组相比较,持续性房颤组血清TIMP4水平显著降低,血清PⅠCP、PⅢCP水平显著升高(P<0.05);与对照组比较,阵发性房颤组血清TIMP4水平显著降低,血清PⅠCP、PⅢCP水平显著升高(P<0.05);心房颤动患者血清TIMP4水平与心肌纤维化指标PⅠCP、PⅢCP均呈负相关(R=-0.513、-0.576,P<0.05);多因素Logistic回归分析显示,高水平TIMP4为患者发生持续性心房颤动的独立保护因素,高水平PICP、PⅢCP、TNF-α均为患者发生持续性心房颤动的独立危险因素(P<0.05)。结论心房颤动患者血清TIMP4表达水平显著降低,与患者心肌纤维化程度呈负相关。

血清TIMP-4水平在老年慢性心力衰竭早期诊断及预后评估中的价值

目的探讨血清组织型金属蛋白酶抑制剂4(TIMP-4)水平对老年慢性心力衰竭(CHF)早期诊断及预后评估的价值。方法选择老年CHF患者160例(CHF组),其中轻度68例、重度92例,另选体检健康的老年志愿者40例(对照组)。采用彩色多普勒超声诊断仪检测两组左心室舒张末期内径(LVEDD)、左心室射血分数(LVEF)。采集肘静脉血,采用双抗体夹心ELISA法检测血清TIMP-4。采用受试者工作特征(ROC)曲线分析血清TIMP-4对老年CHF的早期诊断价值。老年CHF患者出院3、6、9个月复查心脏彩超,统计心脏事件的发生情况。结果 CHF组LVEDD明显高于对照组,LVEF、TIMP-4明显低于对照组,以重度CHF患者上述指标变化更明显(P均<0.05);相关分析显示,老年CHF患者血清TIMP-4水平与LVEF呈正相关关系(r=0.816,P<0.05),与LVEDD呈负相关关系(r=-0.266,P<0.05)。ROC曲线分析显示,血清TIMP-4诊断老年CHF的曲线下面积为0.916(95%CI:0.869~0.951),其cut off值为1.41 ng/m L,此时血清TIMP-4诊断老年CHF的敏感性为77.0%、特异性为92.0%。以血清TIMP-4水平的cut off值为截断值,将老年CHF患者分为TIMP-4高水平者(TIMP-4>1.41ng/m L)46例和TIMP-4低水平者(TIMP-4≤1.41 ng/m L)114例;TIMP-4高水平者和TIMP-4低水平者出院3个月无心脏事件生存率比较P>0.05,而TIMP-4高水平者出院6、9个月无心脏事件生存率明显高于TIMP-4低水平者(P均<0.05)。结论血清TIMP-4可作为老年CHF患者早期诊断和预后判断的预测指标之一。

人子宫颈癌组织中MMP-26、MMP-9和TIMP-4的表达及其意义

目的:检测人子宫颈癌组织中基质金属蛋白酶-26(MMP-26)、基质金属蛋白酶-9(MMP-9)及基质金属蛋白酶组织抑制因子-4(TIMP-4)的表达,探讨其与临床病理特征的关系。方法:应用免疫组织化学SP法检测10例正常子宫颈组织、6例宫颈原位癌和79例宫颈浸润癌组织中MMP-26、MMP-9及TIMP-4的表达。结果:MMP-26在宫颈原位癌和浸润性癌中阳性表达率分别为83.33%和82.28%,较正常子宫颈组织中表达明显增强(P<0.05);MMP-26表达与子宫颈癌的肌层浸润深度和淋巴结转移有密切关联(P<0.05),随着子宫颈癌浸润深度的增加,MMP-26阳性表达增强;淋巴结转移组MMP-26蛋白阳性表达率明显高于无淋巴结转移组(P<0.05)。MMP-9在宫颈浸润癌组织中阳性率为68.35%,MMP-9表达与子宫颈癌病理分级有密切关联,其在Ⅲ级子宫颈癌组织中的表达明显高于Ⅰ和Ⅱ级(P<0.05)。TIMP-4在正常子宫颈、原位癌及浸润性癌组织中的表达呈下降趋势,但差异无显著性(P>0.05);TIMP-4的表达与子宫颈癌的临床分期有密切关联,其在早期癌中的表达高于晚期癌(P<0.05)。MMP-26与TIMP-4在子宫颈癌组织中的表达呈负相关关系(rs=-0.259,P<0.05);MMP-26与MMP-9蛋白在子宫颈癌组织中的表达无相关性(rs=0.140,P>0.05)。结论:MMP-26和MMP-9在宫颈癌组织中高表达,促进其侵袭和转移;TIMP-4在子宫颈癌组织中的表达下降,提示三者联合应用有望成为一项判断子宫颈癌预后的指标。