基质金属蛋白酶-2在胰腺癌组织中的表达及其临床意义

Objective To investigate the expression of matrix metalloproteinase-2 (MMP-2)in pancreatic carcinoma and the relationship between MMP-2 with tumour clinicopatholngical features. Methods The expression of MMP-2 was detected by S-P immunohistochemistry in 36 cases with pancreatic carcinoma and 14 normal pancreat-ic tissues. Results The positive expression rate of MMP-2 was 66.7% (24/36) in pancreatic carcinoma tissue and 14.3% (2/14) in normal pancreatic tissues (χ2 = 3. 587, P < 0.01 ) ;The expression rate of MMP-2 in pancreatic carcinoma tissue with positive-node was 86.7% ( 13/15 ) ,which was higher than that with negative-node,which was 52.4% ( 11/21 ) ( P < 0.05 ) ; As to TNM staging in pancreatic carcinoma, the expression rate of MMP-2 was 41.2% (7/17) with Ⅰ,Ⅱ staging and 89.5% (17/19) with Ⅲ,Ⅳ staging(χ2=9.418,P <0. 01 ) ;The expression rate of MMP-2 was 50.0% (5/10) ,66.7% (10/15) and 72.8% (8/11) in highly,moderately and poorly differentiated pancreatic carcinoma(P > 0.05 ). Conclusions The expression of MMP-2 is strengthened significantly in pancreatic carcinoma tissue and involved in turnout invasion and metastasis features; MMP-2 might be regarded as one more marker for the invasive properties of pancreatic carcinoma.     

HuR与环氧化酶-2在卵巢上皮性癌表达及相关性研究

Objective To detect the expressions of HuR and COX-2 in epithelial ovarian carcinoma,and investigate the correlation of HuR and COX-2 expression. In addition, we attempt to seek the pathway to prevent the occurrence and development of epithelial ovarian cancer by combined analysis of various clinicopathologic characteristics. Methods The expressions of HuR and COX-2 in 68 epithelial ovarian carcinoma, 10 borderline ovarian tumors and 5 normal ovarian tissues were examined by S-P immunohistochemical method. The relationship of HuR and COX-2 expressions with clinicopathologic parameterwere evaluated by correlation analysis. Results(1) The expression of HuR in epithelial ovarian cancer tissue (76. 47% ,52/68) was significantly higher than that in borderline epithelial ovarian tumor tissues (30. 00% ,3/10) and normal ovarian tissues (0, x2 = 18. 873, Ps ＜ 0. 05), but there were no significant differences betweenthe expressions of HuR in borderline epithelial ovarian tumor and normal ovarian tissue(P ＞ 0. 05).(2) The positive expression rate of cytoplasmic HuR in epithelial ovarian carcinoma and borderline epithelial ovarian tumor were 45.60% (31/68) and 10. 00% (1/10) respectively,but normal ovarian tissues showed no staining of HuR. We found no significant differences between the expression of cytoplasmic HuR in epithelial ovarian carcinoma and borderline epithelial ovarian tumor or normal ovarian tissue(x2 = 7. 999 ,P =0. 018).(3) The positive expression rate of cytoplasmic HuR in epithelial ovarian carcinoma of FIGO stage Ⅲ - Ⅳ was significantly higher than that of stage Ⅰ - Ⅱ(56. 09% vs. 29. 63%, x2 = 4. 598, P = 0. 032). The positive expression rates of cytoplasmic HuR in epithelial ovarian carcinoma of histological grade 1,2,3 were 10. 00%,46. 67% ,57. 14% respectively, which showed significant difference in the comparison among the three groups (x2 =6. 627 ,P =0. 036). (4) The positive expression rates of COX-2 in epithelial ovarian cancer (67. 64%)and borderline epithelial ovarian tumor tissues (60. 00%) were significantly higher than that in normal ovarian tissues (0, Ps ＜ 0. 05), but we found no significant difference in the comparison between the expression of malignant and borderline ovarian tumors. Statistical analysis showed that the positive expression rate of COX-2 in epithelial ovarian carcinoma was correlated with FIGO stage and lymph node metastasis. (5)There was a significantly positive correlation between cytoplasmic HuR and COX-2 expressions in epithelial ovarian carcinoma. Conclusion The expressions of HuR and COX-2 increased in the epithelial ovarian carcinoma, and the cytoplasmic expression of HuR was significantly correlated with the expression of COX-2. These results suggested that increased cytoplasmic expression of HuR and COX-2 expression might play important roles in the initiation and development of epithelial ovarian carcinoma.     

脓毒症大鼠肺基质金属蛋白酶-2/9表达与血必净干预

Objective To detect the expression of MMP-2/9 and TIMP-1/2 in the lung of Vibrio vulnificus sepsis rats and observe the intervention of Xuebijing injection. Method One hundred and ten SD rats of clean grade were randomly(random number) divided into normal control group (group A, n = 10),Vibrio vulnificus sepsis group (group B, n = 50. Sepsis was reproduced in rats with subcutaneous injection in left lower limb with Vibrio vulnificus) and Xuebijin intervention group ( group C, n = 50. Rats were intraperitoneal(ip) with the dose of Xuebijing 4mL/kg at the time of 30 min later after infection). The rats in group B and C were sacrificed at 1 h, 6 h, 12 h, 24 h, 48 h after infection, the expression of MMP-2/9 and TIMP-1/2 were examined by PCR, Immunohistochemistry or ELISA methods, the lung permeability were measured by Coomassie Brilliant Blue method. Experimental data used single factor analysis of variance, and between groups by LSD method for pairwise comparison,P ＜0.05 statistically significant difference. Results The lung permeability increased both in group B and C compared with group A,and in group B were relatively higher. The lung MMP-2/9, TIMP-1/2mRNA expression in groups B and C compared with in group A was markedly higher, and reached the peak at 6 h(0. 344 ± 0. 108 ),6 h ( 1. 230 ± 0.377 ), 12 h (0.523 ±0.098),12 h(0.280±0.070) (P＜0.05) in group B while at 12 h(0.256 ±0.074),6 h(0.968±0.225) ,12 h(0.746 ±0. 316) ,12 h(0.356 ±0.035) (P ＜0. 05) in group C; the MMP-2/9mRNA expression in group C decreased(P＜0. 05) compared with the group B while the TIMP-1/2mRNA expression increased(P＜0. 05). The lung MMP-2/9, TIMP-1/2 protein expression in groups B and C compared with the group A(0.345±0.109) also increased, and the peak was at 12 h (0. 692 ± 0. 191 ), 12 h (0. 061 ±0.017) ,24 h(1384.42 ±91) ,24 h(41.04 ±3.60)in group B while at 24 h(0. 217 ±0.065) ,12 h(0. 045± 0. 013 ) ,24 h ( 1617.22 ± 103 ) ,24 h (47.66 ± 3.58 )in group C, the MMP-2/9 protein expression in group C was lower than in group B(P＜0.05), the TIMP-1/2 protein expression in group C was similar to in group B early while marked increased(P＜0.05)later. Conclusions MMP/TIMP imbalance was one of the mechanisms of the lung injury in the rats with Vibrio vulnificus sepsis, Xuebijing could restore the balance of MMP/TIMP ratio.     

Effects of rosiglitazone on the expression of nuclear factor-κB P65 and metalloproteinase-9 mRNA in peripheral blood monocyte-derived macrophages in patients with coronary artery disease

Objective To investigate the effects and mechanisms of rosiglitazone on the expressions of nuclear factor-κB and matrix metalloprotease (MMP-9) in peripheral blood monocyte-derived macrophages (MDMs) in patients with coronary heart disease. Method This was a clinical case-control study. Forty-eight actue coronary symdrome (ACS) patients (ACS group), and 20 patients with stable angina (SA) (control group) were collected. They were performed coronary arteriography in the Department of Cardiology of the Second Xiangya Hospital from March to April in 2007. Exclusion criteria included acute infection, trauma or surgery patients within four weeks, cerebral vascular accident, liver and kidney dysfunction, cancer, and so on. The peripheral blood mononuclear cells were isolated and transformed into MDMs with macrophage colony-stimulating factor treatment. The transformed MDMs were randomly assigned into subgrougs and incubated with 0 /μmol/L, 1 μmol/L, 10 μmol/L, 20 μmol/L of rosiglitazone respectively. The expressions of PPAR-γ mRNA, MMP-9 mRNA were determined by RT-PCR and nuclear factor-κB P65 (NF-KB P65) expression by immunohistochemistry. Multiple comparisons were examined for significant differences using analysis of variance (ANOVA). Results The basal expression of PPAR-y mRNA was lower, in contrast, the levels of NF-KB P65 and MMP-9 mRNA were higher in ACS group than control group. PPAR-γ mRNA expression were significantly upregulated in both ACS and control groups with rosiglitazone treatment. PPAR-γ mRNA expression was positive correlation, while the expressions of MMP-9 mRNA were negative correlation with the rosiglitazone concentration in the ACS group. Rosiglitazone inhibited the expression of NF-KB in a concentration-independent manner in ACS and control groups. Conclusions The expression of PPAR-y mRNA is inhibited, while the activity of NF-KB and expression of MMP-9 mRNA are enhanced in MDMs of ACS cases. Rosiglitazone intervention may inhibit NF-KB activity and MMP-9 expression by upregulation of PPAR-y expression in MDMS of patiens with ACS.     

Effects of rosiglitazone on the expression of nuclear factor-κB P65 and metalloproteinase-9 mRNA in peripheral blood monocyte-derived macrophages in patients with coronary artery disease

Objective To investigate the effects and mechanisms of rosiglitazone on the expressions of nuclear factor-κB and matrix metalloprotease (MMP-9) in peripheral blood monocyte-derived macrophages (MDMs) in patients with coronary heart disease. Method This was a clinical case-control study. Forty-eight actue coronary symdrome (ACS) patients (ACS group), and 20 patients with stable angina (SA) (control group) were collected. They were performed coronary arteriography in the Department of Cardiology of the Second Xiangya Hospital from March to April in 2007. Exclusion criteria included acute infection, trauma or surgery patients within four weeks, cerebral vascular accident, liver and kidney dysfunction, cancer, and so on. The peripheral blood mononuclear cells were isolated and transformed into MDMs with macrophage colony-stimulating factor treatment. The transformed MDMs were randomly assigned into subgrougs and incubated with 0 /μmol/L, 1 μmol/L, 10 μmol/L, 20 μmol/L of rosiglitazone respectively. The expressions of PPAR-γ mRNA, MMP-9 mRNA were determined by RT-PCR and nuclear factor-κB P65 (NF-KB P65) expression by immunohistochemistry. Multiple comparisons were examined for significant differences using analysis of variance (ANOVA). Results The basal expression of PPAR-y mRNA was lower, in contrast, the levels of NF-KB P65 and MMP-9 mRNA were higher in ACS group than control group. PPAR-γ mRNA expression were significantly upregulated in both ACS and control groups with rosiglitazone treatment. PPAR-γ mRNA expression was positive correlation, while the expressions of MMP-9 mRNA were negative correlation with the rosiglitazone concentration in the ACS group. Rosiglitazone inhibited the expression of NF-KB in a concentration-independent manner in ACS and control groups. Conclusions The expression of PPAR-y mRNA is inhibited, while the activity of NF-KB and expression of MMP-9 mRNA are enhanced in MDMs of ACS cases. Rosiglitazone intervention may inhibit NF-KB activity and MMP-9 expression by upregulation of PPAR-y expression in MDMS of patiens with ACS.     

MMP-9和TIMP-1在不明原因不孕症患者子宫内膜的表达

Objective To study the expression situation of matrix metalloproteinase-9(MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the endometrium of women with unexplained infertility and normal endometrium, and to explore the relationship between MMP-9/TIMP-1 expres-sion and unexplained infertility. Methods Immunohistochemieal assay(SP method) was employed to deteet the expression of MMP-9/TIMP-1 in 20 cases of impaired endometrium(unexplained infertility group,endometrial plantation window phase)and 20 cases of normal endometrium(heahhy control group). Results There were different levels of MMP-9/TIMP-1 expression in kytoplasms of glandular epicytes and stromal cells of all endometrial samples. The expression of MMP-9/TIMP-1 was signifi-cantly weaker in unexplained infertility group than that in healthy control group(P＜0.05). Conclusion Low expression of MMP-9/TIMP-1 in the endometrial plantation window phase may be one of impor-tant factors for unexplained infertility.     

脓毒症大鼠肺基质金属蛋白酶-2/9表达与血必净干预

目的 观察创伤弧菌脓毒症大鼠肺基质金属蛋白酶(MMP)-2/9和组织型金属蛋白酶抑制剂(TIMP)-1/2的动态表达及血必净干预.方法 温州医学院生命科学院实验室,清洁级SD大鼠110只,随机(随机数字法)分为正常对照组(A组,n=10)、创伤弧菌脓毒症组(B组,n=50,采用大鼠左下肢皮下注射创伤弧菌悬液制作创伤弧菌脓毒症大鼠模型)及血必净干预组(C组,n=50,感染后0.5 h腹腔注射血必净4 mL/kg).B、C组大鼠于染菌后1,6,12,24,48 h麻醉后活杀,留取右肺标本.观察大鼠行为学变化,采用考马斯亮蓝法测肺通透性,采用逆转录-聚合酶链式反应(RT-PCR)法测肺MMP-2/9,TIMP-1/2 mRNA的表达,免疫组化法和双抗体夹心酶联免疫吸附法(ELISA)测肺MMP-2/9和TIMP-1/2的表达.数据采用单因素方差分析,并用LSD-t法进行组间两两比较,以P＜0.05为差异有统计学意义.结果 B组和C组肺通透性显著高于A组,C组显著低于B组.B组和C组MMP-2/9,TIMP-1/2mRNA显著升高,B组分别于6 h(0.344±0.108),6 h(1.230±0.377),12 h(0.523±0.098),12 h(0.280±0.070)达高峰(P＜0.05),C组分别于12 h(0.256±0.074),6 h(0.968±0.225),12 h(0.746±0.316),12 h(0.356±0.035)达高峰(P＜0.05),C组MMP-2/9mRNA升高趋势显著低于B组(P＜0.05),TIMP-1/2mRNA显著高于B组(P＜0.05).B组和C组MMP-2/9,TIMP-1/2蛋白也升高,B组分别于12 h(0.692±0.191),12 h(0.061±0.017),24 h(1384.42±91),24 h(41.04±3.60)达高峰(P＜0.05);C组分别于24 h(0.217±0.065),12 h(0.045±0.013),24 h(1617.22±103),24 h(47.66±3.58)达高峰(P＜0.05);C组MMP-2/9蛋白升高趋势低于B组(P＜0.05),TIMP-1/2蛋白早期与B组差别不大,后期显著高于B组(P＜0.05).结论 MMP/TIMP比例失衡是创伤弧菌脓毒症大鼠肺损伤机制之一,血必净可促进MMP/TIMP比例恢复平衡,对创伤弧菌脓毒症大鼠肺损伤具有保护作用.

Abstract：

Objective To detect the expression of MMP-2/9 and TIMP-1/2 in the lung of Vibrio vulnificus sepsis rats and observe the intervention of Xuebijing injection. Method One hundred and ten SD rats of clean grade were randomly(random number) divided into normal control group (group A, n = 10),Vibrio vulnificus sepsis group (group B, n = 50. Sepsis was reproduced in rats with subcutaneous injection in left lower limb with Vibrio vulnificus) and Xuebijin intervention group ( group C, n = 50. Rats were intraperitoneal(ip) with the dose of Xuebijing 4mL/kg at the time of 30 min later after infection). The rats in group B and C were sacrificed at 1 h, 6 h, 12 h, 24 h, 48 h after infection, the expression of MMP-2/9 and TIMP-1/2 were examined by PCR, Immunohistochemistry or ELISA methods, the lung permeability were measured by Coomassie Brilliant Blue method. Experimental data used single factor analysis of variance, and between groups by LSD method for pairwise comparison,P ＜0.05 statistically significant difference. Results The lung permeability increased both in group B and C compared with group A,and in group B were relatively higher. The lung MMP-2/9, TIMP-1/2mRNA expression in groups B and C compared with in group A was markedly higher, and reached the peak at 6 h(0. 344 ± 0. 108 ),6 h ( 1. 230 ± 0.377 ), 12 h (0.523 ±0.098),12 h(0.280±0.070) (P＜0.05) in group B while at 12 h(0.256 ±0.074),6 h(0.968±0.225) ,12 h(0.746 ±0. 316) ,12 h(0.356 ±0.035) (P ＜0. 05) in group C; the MMP-2/9mRNA expression in group C decreased(P＜0. 05) compared with the group B while the TIMP-1/2mRNA expression increased(P＜0. 05). The lung MMP-2/9, TIMP-1/2 protein expression in groups B and C compared with the group A(0.345±0.109) also increased, and the peak was at 12 h (0. 692 ± 0. 191 ), 12 h (0. 061 ±0.017) ,24 h(1384.42 ±91) ,24 h(41.04 ±3.60)in group B while at 24 h(0. 217 ±0.065) ,12 h(0. 045± 0. 013 ) ,24 h ( 1617.22 ± 103 ) ,24 h (47.66 ± 3.58 )in group C, the MMP-2/9 protein expression in group C was lower than in group B(P＜0.05), the TIMP-1/2 protein expression in group C was similar to in group B early while marked increased(P＜0.05)later. Conclusions MMP/TIMP imbalance was one of the mechanisms of the lung injury in the rats with Vibrio vulnificus sepsis, Xuebijing could restore the balance of MMP/TIMP ratio.     

Association between matrix metalloproteinase-9 polymorphism (-1562C ＞ T/R279Q) and acute coronary syndrome in Uygur nationality of Xinjiang Autonomous Region of China

Objective To investigate the association between matrix metalloproteinase-9 (MMP-9) gene polymorphism (-1562C ＞ T/R279Q) and acute coronary syndrome (ACS) in Uygur nationality of Xinjiang Autonomous Region of China. Methods A total of 352 patients with ACS including 213 patients with unstable angina pectoris and 139 patients with acute myocardial infarction evidenced by using coronary arteriography and 421 control subjects were recruited in this study. The MMP-9-1562C ＞ T and R279Q genotypes were detemined by using PCR-RFLP method. The relationship between the polymorphism in the MMP-9 gene and the severity of coronary arterial stenosis was analyzed. All polymorphisms were determined for confimation with Hardy-Weinberg expectations in both groups separately. Differences in distributions of genotypes and alleles between two groups were analyzed with x2 test. The association between the MMP-9 polymorphisms and the risk of ACS was estimated by odds ratio(Ors) and their 95% confidence intervals (CIs), and the comprehensive evaluation of the factors associated with ACS was determined by using multifactor logistic regression. P ＜ 0. 05 was considered to be statistically significant. Results The genotype frequencies for CT + TT genotypes and T allele were 25.9 and14.5 percent in ACS subjects and 15.7 and 8.4 percent in control subjects, respectively. The genotype frequencies were different significantly between the two groups (x2 = 12.26,P ＜ 0.01;x2 = 14.15,P ＜ 0.01, respectively). No relationship between R279Q polymorphism and ACS was found in this study ( P ＞ 0.05). The multifactor logistic regression analysis showed that the T allele carrier (CT + TT) significantly increased the risk of ACS compared with the CC genotype ( OR = 1.791,95 % CI: 1. 088 - 2.951, P = 0.022) after adjustment for tradition risk factors. The frequencies for CT + TT and CC genotypes of the -1562C ＞ T polymorphism were not statistically different among ACS patients with one, two and three or more significantly diseased vessels ( x2 = 1.15, P = 0.56). Conclusions The findings suggest that the polymorphism in MMP-9 gene promoter (-1562C ＞ T) is associated with the susceptibility to the ACS. The T allele might be an independent risk factor for the ACS. But the -1562C ＞ T polymorphism may not be useful as a predictor of the severity of coronary arterial stenosis. The R279Q polymorphism of MMP-9 gene was not significantly associated with ACS in this studied population.     

CD147、基质金属蛋白酶-9及其抑制物-1在不同级别胶质瘤中的表达及相关性研究

目的 探讨CD147、基质金属蛋白酶(MMP)-9和基质金属蛋白酶抑制物(TIMP)-1在胶质瘤中的表达及其相关性.方法 采用免疫组织化学法对78例石蜡包埋的胶质瘤组织和12例正常脑组织进行标记和分析,应用实时荧光定量RT-PCR方法检测CD147 mRNA的相对含量.结果 CD147、MMP-9、TIMP-1的阳性率分别为62%(48/78)、71%(55/78)、59%(46/78),采用Spearman进行相关性分析,其与胶质瘤的恶性程度均呈正相关(rs=0.2671～0.5631,P＜0.01),CD147与MMP-9呈正相关(rs=0.3576,P＜0.01);CD147、MMP-9、TIMP-1在胶质瘤的临床分级中的表达(低级别组分别为47%、56%和49%;高级别组为80%、89%和71%)差异有统计学意义(x2值分别为9.510、10.702、4.138,P均＜0.05);在胶质瘤的病理分级中,CD147 mRNA相对含量分别为Ⅰ级0.15,Ⅱ级0.27,Ⅲ级0.46,Ⅳ级0.78,随着胶质瘤的级别越高,CD147 mRNA相对含量越高.结论 CD147、MMP-9和TIMP-1可以作为临床判断胶质瘤预后的分子生物学标志.

Abstract：

Objective To investigate the expressions of CD147,MMP-9 and TIMP-1 in human gliomas and analyze the correlations.Methods Expressions of CD147,MMP-9 and TIMP-1 were assessed in paraffin-embedded specimens collected from 78 gliomas and 12 benign brain lesion tissues by immunohistochemistry.Real time PCR was performed to detect CD147 mRNA expression.Results The positive rates of CD147,MMP-9 and TIMP-1 expression were 62%(48/78),71%(55/78),59%(46/78) respectively.We found a significant positive correlation between CD147,MMP-9,TIMP-1 expressions and poor gliomas differentiation by Spearman analysis(rs=0.2671-0.5631,Ps＜0.01).There was also a significant positive correlation between CD147 and MMP-9 expression(rs =0.3576,P＜0.01).In addition,the expressions of CD147(47% vs.80%,x2=9.510),MMP-9(56% vs.89%,x2=10.702),and TIMP-1(49% vs.71%,x2=4.138) were significantly higher in advanced gilomas than early gliomas(Ps＜0.05).The relative expression levels of CD147 mRNA in gliomas of Ⅰ to Ⅳ pathological grades were 0.15,0.27,0.46,0.78 respectively.Conclusion The expressions of CD147,MMP-9 and TIMP-1 were important characteristic of gliomas,which may serve as biomarkers in the glioma prognostic prediction.     

ezrin在乳腺癌中的表达及其临床意义

Objective To study the expression and clinical significance of ezrin in breast carcinoma. Methods Immunohistuchemieal staining(S-P) was used to detect the expression of ezrin in 63 cases of breast car-cinoma samples. Results Among these 63 samples,the total expression rates of ezrin were 55.56% (35/63). With the increasing of axillary lymph nodes metastasis and clinical staging,the rates of expressing of ezrin elevated (P<0.05),which was remarkably lower in patients whose disease-free survival(DFS) was > 5 years,or those whose DFS≤5 years or died within five years(P<0.05) ,but the expression of ezrin was not correlated with tumor size,age and menopausal status (P>0.05). Conclusion The expression of ezrin may contribute to prognostic e-valuation for breast carcinoma.     

ezrin在乳腺癌中的表达及其临床意义

Objective To study the expression and clinical significance of ezrin in breast carcinoma. Methods Immunohistuchemieal staining(S-P) was used to detect the expression of ezrin in 63 cases of breast car-cinoma samples. Results Among these 63 samples,the total expression rates of ezrin were 55.56% (35/63). With the increasing of axillary lymph nodes metastasis and clinical staging,the rates of expressing of ezrin elevated (P<0.05),which was remarkably lower in patients whose disease-free survival(DFS) was > 5 years,or those whose DFS≤5 years or died within five years(P<0.05) ,but the expression of ezrin was not correlated with tumor size,age and menopausal status (P>0.05). Conclusion The expression of ezrin may contribute to prognostic e-valuation for breast carcinoma.     

ezrin在乳腺癌中的表达及其临床意义

Objective To study the expression and clinical significance of ezrin in breast carcinoma. Methods Immunohistuchemieal staining(S-P) was used to detect the expression of ezrin in 63 cases of breast car-cinoma samples. Results Among these 63 samples,the total expression rates of ezrin were 55.56% (35/63). With the increasing of axillary lymph nodes metastasis and clinical staging,the rates of expressing of ezrin elevated (P<0.05),which was remarkably lower in patients whose disease-free survival(DFS) was > 5 years,or those whose DFS≤5 years or died within five years(P<0.05) ,but the expression of ezrin was not correlated with tumor size,age and menopausal status (P>0.05). Conclusion The expression of ezrin may contribute to prognostic e-valuation for breast carcinoma.     

ezrin在乳腺癌中的表达及其临床意义

Objective To study the expression and clinical significance of ezrin in breast carcinoma. Methods Immunohistuchemieal staining(S-P) was used to detect the expression of ezrin in 63 cases of breast car-cinoma samples. Results Among these 63 samples,the total expression rates of ezrin were 55.56% (35/63). With the increasing of axillary lymph nodes metastasis and clinical staging,the rates of expressing of ezrin elevated (P<0.05),which was remarkably lower in patients whose disease-free survival(DFS) was > 5 years,or those whose DFS≤5 years or died within five years(P<0.05) ,but the expression of ezrin was not correlated with tumor size,age and menopausal status (P>0.05). Conclusion The expression of ezrin may contribute to prognostic e-valuation for breast carcinoma.     

ezrin在乳腺癌中的表达及其临床意义

Objective To study the expression and clinical significance of ezrin in breast carcinoma. Methods Immunohistuchemieal staining(S-P) was used to detect the expression of ezrin in 63 cases of breast car-cinoma samples. Results Among these 63 samples,the total expression rates of ezrin were 55.56% (35/63). With the increasing of axillary lymph nodes metastasis and clinical staging,the rates of expressing of ezrin elevated (P<0.05),which was remarkably lower in patients whose disease-free survival(DFS) was > 5 years,or those whose DFS≤5 years or died within five years(P<0.05) ,but the expression of ezrin was not correlated with tumor size,age and menopausal status (P>0.05). Conclusion The expression of ezrin may contribute to prognostic e-valuation for breast carcinoma.     

ezrin在乳腺癌中的表达及其临床意义

Objective To study the expression and clinical significance of ezrin in breast carcinoma. Methods Immunohistuchemieal staining(S-P) was used to detect the expression of ezrin in 63 cases of breast car-cinoma samples. Results Among these 63 samples,the total expression rates of ezrin were 55.56% (35/63). With the increasing of axillary lymph nodes metastasis and clinical staging,the rates of expressing of ezrin elevated (P<0.05),which was remarkably lower in patients whose disease-free survival(DFS) was > 5 years,or those whose DFS≤5 years or died within five years(P<0.05) ,but the expression of ezrin was not correlated with tumor size,age and menopausal status (P>0.05). Conclusion The expression of ezrin may contribute to prognostic e-valuation for breast carcinoma.     

ezrin在乳腺癌中的表达及其临床意义

Objective To study the expression and clinical significance of ezrin in breast carcinoma. Methods Immunohistuchemieal staining(S-P) was used to detect the expression of ezrin in 63 cases of breast car-cinoma samples. Results Among these 63 samples,the total expression rates of ezrin were 55.56% (35/63). With the increasing of axillary lymph nodes metastasis and clinical staging,the rates of expressing of ezrin elevated (P<0.05),which was remarkably lower in patients whose disease-free survival(DFS) was > 5 years,or those whose DFS≤5 years or died within five years(P<0.05) ,but the expression of ezrin was not correlated with tumor size,age and menopausal status (P>0.05). Conclusion The expression of ezrin may contribute to prognostic e-valuation for breast carcinoma.     

多器官功能障碍综合征大鼠肺组织血栓调节蛋白和基质金属蛋白酶-9的表达

Objective To determine the expressions of thrombomodulin (TM) and matrix metalloproteinase-9(MMP-9) in the lung of rats with multiple organ dysfunction syndrome (MODS) and to investigate the mechanism of lung injury in MODS. Method Forty adult mule Sprague-Dawley (SD) rats were randomly divided into two groups,namely the normal control group and the MODS model group. The rats of model group were further divided into four subgroups as per different intervals (6 h, 12 h, 24 h and 48 h) ,and there were 8 rats in each groups. The animal models of MODS were estabhshed by two hits,the left eyeball of each model rat was removed to bleed to 2 mL/100g,and four hours later, lipopolysaccharide (LPS 5 mg/kg) was injected into intraperitoneal cavity of model rats. The same volume of saline was injected intraperitoneally into rats of control group instead of LPS. All rats were sacrificed at various intervals. The histological changes in lung tissue were observed by naked eye and light microscope. The expressions of TM and MMP-9 proteins were deteceted by using immunohistechemistry. One-way ANOVA was used for comparison among multiple group. Results (1)There were no histopathological changes in lung of rats of control group, and the lung injury was serious in MODS rots. (2) Compared with the rots of con-trol group, the expression of TM in lung tissue of MODS rats increased 6 hours after LPS, reached peak 12 hours later(P <0.01),and then decreased during 24～48 period.There was no significant difference in expression of TM between two groups 48 hours later. Compared with control group, the expressions of MMP-9 in lung tissue of MODS rats didn't significantly increase 6 ～ 48 hours after LPS (P < 0.01). Conclusions There are endothelium damage and extracellular matrix damage found in the lung tissue of rats at the early phase of MODS. TM and MMP-9 are good biomarkers of endothelium and extracellular matrix damage, and they can be used for diagnosing and es-timating the severity of injury lungs at the early phase of MODS.     

基质金属蛋白酶家族的表达对急性单核细胞白血病细胞体外侵袭力的影响

目的 研究基质金属蛋白酶家族(TIMP-2、MT1-MMP、MMP-2)的表达对急性单核细胞白血病细胞体外侵袭力的影响.方法 以急性单核细胞白血病细胞株SHI-1细胞为研究对象,用定量PCR、Western blot法分别检测TIMP-2、MT1-MMP、MMP-2 mRNA和蛋白表达,并以NB4、K562、THP-1等人类其他白血病细胞株细胞为对照,进行比较.构建TIMP-2逆转录病毒载体,转染SHI-1细胞,G418筛选,有限稀释法挑选出单克隆S1、S2、S3细胞.RNA干扰(RNAi)法干扰SHI-1细胞TIMP-2、MT1-MMP、MMP-2表达.明胶酶谱法测定不同细胞和骨髓基质细胞(BMSC)共培养24 h后上清中MMP-2的表达,细胞侵袭实验测定SHI-1细胞侵袭人工基质膜的能力.结果 SHI-1细胞的TIMP-2、MT1-MMP、MMP-2 mRNA表达和蛋白表达均显著高于其他细胞(P＜0.05).SHI-1细胞和BMSC共培养上清中的MMP-2酶原和活化的MMP-2含量及细胞体外侵袭率均高于其他细胞(P＜0.05).单克隆S1、S2、S3细胞TIMP-2 mRNA表达水平分别是SHI-1细胞的3.0倍、2.0倍和1.5倍(P＜0.05),蛋白表达水平分别上调2.6倍、1.5倍和1.3倍(P＜0.01),体外侵袭率增加1.5～2.5倍(P＜0.05),活化的MMP-2含量增加1.5～3.0倍(P＜0.01).RNA干扰基因沉默效率为85%～98%.SHI-1细胞TIMP-2、MMP-2、MT1-MMP表达沉默后,细胞侵袭率分别下降60%～70%、50%～60%、40%～50%(P＜0.05).RNA干扰后的细胞培养上清中检测不到活化的MMP-2.结论 SHI-1细胞在mRNA水平和蛋白水平均高表达TIMP-2、MT1-MMP、MMP-2 mRNA,SHI-1细胞和BMSC共培养后这些分子的高表达促进MMP-2的活化,增强细胞的体外侵袭力.TIMP-2表达增加1.5～2.5倍对SHI-1细胞MMP-2的活化和细胞的体外侵袭力发挥的是增强作用,而不是抑制作用.

Abstract：

Objective To study the effect of matrix metalloproteinase 2 ( MMP-2), membrane type 1 MMP (MT1-MMP) and tissue inhibitor of metalloproteinase 2 (TIMP-2) expressions on the in vitro invasive capacity of acute monocytic leukemia SHI-1 cells. Methods SHI-1, NB4, K562, M937 and THP-1 human leukemia cell lines were cultured in vitro. The mRNA and protein expressions of TIMP-2, MMP-2 and MT1-MMP in different cells were detected by quantitative RT-PCR and western blot. A retroviral vector carrying human TIMP-2 cDNA was constructed and transfected into SHI-1 cells. Three subclone cells (S1, S2 and S3) were screened by G418 and selected by limiting dilution. RNA interference (RNAi) was used to knock down the expression of MMP-2, MT1-MMP and TIMP-2. Cell invasion capacity was performed through a reconstituted human basement membrane assays. Zymography was used to analyze the expression of MMP-2 in the supernatant of co-culture. Results The expressions of MMP-2, MT1-MMP and TIMP-2 in SHI-1 cells were higher than that in other leukemic cells at both mRNA and protein levels (P ＜ 0. 05 ). The amount of proMMP-2 and activated MMP-2 in the conditioned media from SHI-1 cells co-cultured with bone marrow stromal cells (BMSCs) was more than that from other cells (P ＜ 0. 05 ). The in vitro invasive capacity of SHI-1 cells were higher than that of other cells( P ＜ 0.05 ). The mRNA levels of TIMP-2 were increased by about 3 fold, 2 fold and 1.5 fold in S1, S2 and S3 cells, respectively( P ＜ 0.05 ), while the protein levels were by about 2.6 fold, 1.5 fold and 1.3 fold than that of SHI-1 cells, respectively(P ＜ 0.01 ). The invasion rates of subclone cells demonstrated a 1.5 - 2.5 fold' elevation ( P ＜ 0.05 ) and activated MMP-2 from their supernatants increased by 1.5 -2.0 fold(P＜0.01 ). The knock-down efficiency of siRNA was 85% to 98%. The down-regulation of TIMP-2, MMP-2 and MT1-MMP decreased the invasion rates of SHI-1 cells by 60% -70%, 50% - 60% and 40% - 50%, respectively ( P ＜ 0. 05 ). No activated MMP-2 in the supernatants from any knock-down cells could be found. Conclusions SHI-1 cells constitutively overexpress MMP-2,MT1-MMP and TIMP-2 at both mRNA and protein levels. After co-cultured with BMSCs the SHI-1 cells increased MMP-2 activation and cell invasion. An increase of TIMP-2 expression in SHI-1 cells reflects an activating effect on cells invasion and MMP-2 activation.     

针刺联合微创血肿抽吸术对家兔急性期脑出血灶周围脑组织损伤的影响

目的 观察针刺联合微创血肿抽吸术对家兔急性期脑出血灶周围脑组织损伤的影响.方法 共选取健康新西兰大白兔65只,将其随机分为假手术组、脑出血组、血肿抽吸组及血肿抽吸+针刺组.将脑出血组、血肿抽吸组及血肿抽吸+针刺组制成脑出血模型,制模后血肿抽吸组给予血肿抽吸治疗,血肿抽吸+针刺组则给予针刺及血肿抽吸联合治疗.分别于制模后6 h、24 h、3 d及7 d时称量不同组别脑组织干湿重并计算脑含水量,采用免疫组化法检测各组家兔血肿周围脑组织基质金属蛋白酶-9(MMP-9)表达,采用明胶酶谱技术检测各组家兔血肿周围脑组织MMP-9活性.结果 脑出血组、血肿抽吸组及针刺+血肿抽吸组脑含水量均较假手术组显著增高,随着时间进展,血肿抽吸组及针刺+血肿抽吸组脑含水量较脑出血组均有明显降低(P＜0.05或0.01),其中以制模后第3天时针刺+血肿抽吸组的下降幅度尤为显著(P＜0.01);免疫组化及明胶酶谱检测结果显示,各组家兔MMP-9阳性细胞表达及活性均随时间进展呈下降趋势,其中以制模后第3天时针刺+血肿抽吸组的下降幅度最为显著(P＜0.01).结论 针刺联合血肿抽吸术可显著减轻家兔急性期脑出血灶周围脑组织损伤,其治疗机制可能与抑制MMP-9表达有关.

Abstract：

Objective To investigate the effect of acupuncture combined with minimally invasive hematoma aspiration on a rabbit model of brain injury in the acute stage of intracerebral hemorrhage (ICH). Methods A total of 65 healthy New Zealand white rabbits were randomly divided into a sham operation (SO) group, an ICH group,a hematoma aspiration (HA) group, and an acupuncture and hematoma aspiration (AHA) group. Models of ICH were established in the latter three groups. The HA group was treated with minimally invasive hematoma aspiration and the AHA group was treated with both acupuncture and minimally invasive hematoma aspiration. At 6 hours and 1, 3 and 7 days after the ICH models were established, brain water content (BWC) was measured, and the expression of matrix metalloproteinase-9 ( MMP-9 ) in the rabbits' brains was detected by immunohistochemistry. MMP-9activity was detected by gelatin zymography. Results The BWCs of the ICH group, HA group and AHA group rabbits were significantly higher than those of the SO group. The BWCs of the HA group and AHA group animals descended significantly more than those in the ICH group as time went on, especially on the 3rd day in the AHA group. Immunohistochemistry and gelatin zymography showed that the expression and activity of MMP-9 in these test groups decreased with time, especially on the 3rd day in the AHA group. Conclusions Acupuncture combined with hematoma aspiration can reduce injury in the acute stage of ICH, and inhibition of the expression of MMP-9 may be the mechanism.     

荧光定量RT-PCR检测Rac1 mRNA基因及其与胃癌转移的关系

Objective To investigate the relationship between tumor metastasis-related Rac1 mRNA expression levels and gastric carcinomas metastasis, and to investigate the significance of Rac1 as a tumor marker for the evaluation of gastric carcinomas metastasis and distinguish between benign and malignant lesions. Methods This experiment used fluorescence quantitative RT-PCR TaqMan probe technology, chose Rac1 target gene fragment, which was cloned into the pET-20b (+) vector, constructed recombinant plasmid, and established the Rac1 mRNA fluorescence quantitative RT-PCR standard curve. And then the Rac1 mRNA levels were detected in 52 cases of gastric carcinoma tissues,52 cases of para-carcinoma tissues and 12 cases of benign gastric disease tissues. Its association with the metastasis of gastric carcinomas was analyzed. Results The positive rates and levels (median, P25-P75) of Rac1 mRNA were 100. 0% ,7.41 ×105 (3. 50 × 105-4. 36 × 106) copies/μl in gastric carcinomas, 46. 2% ,0(0-1.73 × 104) copies/μl in parscarcinoma tissues, 33. 3%, 0(0-3.55 × 103) copies/μl in benign tissues. The positive rates and leves(x2 =43.16,x2Xk-w = 64. 19, P ＜0. 01)among the three groups were significantly different. When the critical value of Rac1 mRNA level was 1.73 × 104 copies/μl determined by ROC, the sensitivity and specificity were 88. 5% and 100% respectively. When the critical value of Rac1 level was 7.49 × 105 copies/μl, the sensitivity and specificity were 57.9% and 78. 6% respectively. ConclusionThe Rac1 mRNA level detected with fluorescence quantitative RT-PCR has reference value to distinguish between benign and malignant lesions and evaluate gastric carcinoma metastasis.