Human neuronal apoptosis secondary to traumatic brain injury and the regulative role of apoptosis.related genes

Objective: To observe human neuronal apoptosis secondary to traumatic brain injury, and to elucidate itsregulative mechanism and the change of expression of apoptosis-related genes. Methods: Specimens of brain were collected from cases of traumatic brain injury in humans. The histological and cellular morphology was examined by light and electron microscopy. The extent of DNA injury to cortical neurons was detected by using TUNEL. By in situ hybridisation and immunohistochemistry the mRNA changes and protein expression of Bcl-2, Bax, p53, and caspase 3 p20 subunit were observed. Results: Apoptotic neurons appeared following traumatic brain injury, peaked at 24 hours and lasted for 7 days. In normal brain tissue activated caspase 3 was rare, but a short time after trauma it became activated. The activity peaked at 20-28 hours and remained higher than normal for 5-7 days. There was no expression of Bcl-2 mRNA and Bcl-2 protein in normal brain tissue but 8 hours after injury their expression became evident and then increased, peaked at 2-3 days and remained higher than normal for 5-7 days. The primary expression of Bax-mRNA and Bax protein was high in normal brain tissue. At 20-28 hours they increased and remained high for 2-3 days; on the 7th days they returned to a normal level. In normal brain tissue, p53mRNA and P53 were minimally expressed. Increased expression was detected at the 8th hour, and decreased at 20-28 hours but still remained higher than normal on the 5th day. Conclusions- Following traumatic injury to the human brain, apoptotic neurons appear around the focus of trauma. The mRNA and protein expression of Bcl-2, Bax and p53 and the activity of caspase 3 enzyme are increased.     

三磷酸腺苷结合盒转运体成员2在人脑胶质瘤中的表达及其与神经巢蛋白表达的关系

Objective To study the expression of ATP-binding cassette superfamily G member 2 ( ABCG2) and its relationship with the malignant degree and expression of nestin in human gliomas. Methods Fifty-two gliomas of different WHO grades and 8 normal brain tissues were detected for the mRNA expression of ABCG2 by using real-time quantitive polymerase chain reaction (PCR). And the protein expression of ABCG2 and nestin was detected by immunohistochemistry. Results The mRNA of ABCG2 was overexpressed in gliomas as compared with that in normal brain tissues (P<0. 05), and up-regulated with the increase of pathologic degrees (P<0. 05 ). The positive protein expression rate of ABCG2 in 52 gliomas was 32.7%. The positive rate was significantly higher in grade Ⅲ-Ⅳ than in grade Ⅰ -Ⅱ (x2 =4. 62, P < 0. 05). And the expression level of ABCG2 was obviously related with the expression of nestin (x2= 7. 60,P < 0.05). Conclusion The expression level of ABCG2 was obviously related with glioma pathological grade and the expression of nestin. Brain tumor stem cells may have some homology with nerve stem cells.     

三磷酸腺苷结合盒转运体成员2在人脑胶质瘤中的表达及其与神经巢蛋白表达的关系

Objective To study the expression of ATP-binding cassette superfamily G member 2 ( ABCG2) and its relationship with the malignant degree and expression of nestin in human gliomas. Methods Fifty-two gliomas of different WHO grades and 8 normal brain tissues were detected for the mRNA expression of ABCG2 by using real-time quantitive polymerase chain reaction (PCR). And the protein expression of ABCG2 and nestin was detected by immunohistochemistry. Results The mRNA of ABCG2 was overexpressed in gliomas as compared with that in normal brain tissues (P<0. 05), and up-regulated with the increase of pathologic degrees (P<0. 05 ). The positive protein expression rate of ABCG2 in 52 gliomas was 32.7%. The positive rate was significantly higher in grade Ⅲ-Ⅳ than in grade Ⅰ -Ⅱ (x2 =4. 62, P < 0. 05). And the expression level of ABCG2 was obviously related with the expression of nestin (x2= 7. 60,P < 0.05). Conclusion The expression level of ABCG2 was obviously related with glioma pathological grade and the expression of nestin. Brain tumor stem cells may have some homology with nerve stem cells.     

瘦素和瘦素受体在食管癌中的表达及其意义

Objective To investigate the role of leptin and leptin receptor in carcinogensis and development of esophageal carcinoma. Methods The expression of leptin and leptin receptor was detected in 52 cases of esophageal carcinoma tissues and 49 cases of normal esophageal tissues by immunohistochemistry. The correlation between their expression and clinicopathological parameters was also analysized. Results The expression rate of leptin and leptin receptor in esophageal carcinoma was 78. 8% (41/52) and 82.7% (43/52) respectively, and the rate in normal esophagus was 58.3% (28/49) and 59.2% (29/49) respectively. The expression rate of leptin and leptin receptor both had statistically significantly differences between esophageal carcinoma and normal esophagus tissues (P ＜ 0. 05). The expression rate of leptin was associated with position, tumor size, differentiation, lymphatic metastasis and TNM stage (P ＜ 0. 05 ). Conclusion Leptin and leptin receptor were dually expressed in esophageal carcinoma.They played important roles in the process of carcinogensis and development of esophageal carcinoma.     

瘦素和瘦素受体在食管癌中的表达及其意义

Objective To investigate the role of leptin and leptin receptor in carcinogensis and development of esophageal carcinoma. Methods The expression of leptin and leptin receptor was detected in 52 cases of esophageal carcinoma tissues and 49 cases of normal esophageal tissues by immunohistochemistry. The correlation between their expression and clinicopathological parameters was also analysized. Results The expression rate of leptin and leptin receptor in esophageal carcinoma was 78. 8% (41/52) and 82.7% (43/52) respectively, and the rate in normal esophagus was 58.3% (28/49) and 59.2% (29/49) respectively. The expression rate of leptin and leptin receptor both had statistically significantly differences between esophageal carcinoma and normal esophagus tissues (P ＜ 0. 05). The expression rate of leptin was associated with position, tumor size, differentiation, lymphatic metastasis and TNM stage (P ＜ 0. 05 ). Conclusion Leptin and leptin receptor were dually expressed in esophageal carcinoma.They played important roles in the process of carcinogensis and development of esophageal carcinoma.     

Human papillomavirus and p53 protein immunoreactivity in condylomata acuminatum and squamous cell carcinoma of penis

Aim: To determine the immunoreactive pattern of human papillomavirus (HPV) antigen and p53 protein in condylomata acuminatum (CA) and squamous cell carcinoma (SCC) of penis. Methods: Immunohistochemistry for HPV and p53 were performed in 40 specimens of formalin fixed, paraffin embedded tissues using a polyclonal (rabbit) antibody against HPV and a monoclonal (mouse) antibody against human p53 protein. Twenty one cases of CA and nineteen cases of SCC were examined. Results: HPV antigen was detected in all 21 CA and 2 penile SCC. p53 protein overexpression was observed in 12 of 19 (63%) SCC in which 6 cases were strong positive. Five of 21 CA (24%) showed low-grade p53 protein overexpression. Conclusion: CA is related to HPV infection and some cases show p53 protein low-grade overexpression. In contrast, p53 protein overexpression is common in penile SCC, which is seldom related to HPV infection.     

乳腺浸润性导管癌钼靶X线表现与腋窝淋巴结转移及p53蛋白表达的关系

Objective To investigate the relationship among the mammography signs, the axillary lymph node ( LN) metastasis, and the p53 expression of infiltrating ductal breast carcinoma. Methods Mammography signs and axillary LN metastasis were analyzed in 72 cases of infiltrating ductal carcinoma. Immunohistochemistry was used to detect p53 expression in tumor specimens. The relationship among the mammography signs, the axillary LN metastasis and the p53 expression was assessed. Results The positive rate of p53 expression was 56. 7% in the patients with axillary LN metastasis, which was higher than 31.4% in the patients without axillary LN metastasis ( P ＜ 0. 05 ). The rate of axillary LN metastasis and p53 expression were 66. 7% and 70. 0% in the tumors larger than 2 cm, 56. 4% and 66. 7% in the tumors with more vascular signs, 28. 6% and 38. 1% in the tumors less than 2 cm, 30. 3% and 33. 3% in the tumors with normal vascular signs (P ＜0. 01) , respectively. The rate of axillary LN metastasis in the patients with spinulation was 70. 8% , which was higher than 41.1% of patients without spinulation (P ＜0.05). Conclusion Mammography signs of breast cancer can evaluate the axillary LN metastasis and p53 expression, and also can provide objective reference for clinical diagnosis, treatment and prognosis of breast carcinoma.     

乳腺浸润性导管癌钼靶X线表现与腋窝淋巴结转移及p53蛋白表达的关系

Objective To investigate the relationship among the mammography signs, the axillary lymph node ( LN) metastasis, and the p53 expression of infiltrating ductal breast carcinoma. Methods Mammography signs and axillary LN metastasis were analyzed in 72 cases of infiltrating ductal carcinoma. Immunohistochemistry was used to detect p53 expression in tumor specimens. The relationship among the mammography signs, the axillary LN metastasis and the p53 expression was assessed. Results The positive rate of p53 expression was 56. 7% in the patients with axillary LN metastasis, which was higher than 31.4% in the patients without axillary LN metastasis ( P ＜ 0. 05 ). The rate of axillary LN metastasis and p53 expression were 66. 7% and 70. 0% in the tumors larger than 2 cm, 56. 4% and 66. 7% in the tumors with more vascular signs, 28. 6% and 38. 1% in the tumors less than 2 cm, 30. 3% and 33. 3% in the tumors with normal vascular signs (P ＜0. 01) , respectively. The rate of axillary LN metastasis in the patients with spinulation was 70. 8% , which was higher than 41.1% of patients without spinulation (P ＜0.05). Conclusion Mammography signs of breast cancer can evaluate the axillary LN metastasis and p53 expression, and also can provide objective reference for clinical diagnosis, treatment and prognosis of breast carcinoma.     

p16/p38 MAPK/p53/Wip1通路在乳腺癌发生、发展中的作用及其临床意义

目的 探讨p16/p38 MAPK/p53/Wip1通路在乳腺癌发生、发展中的作用及其临床意义.方法 应用免疫组织化学方法检测70例乳腺癌组织、癌旁组织、20例正常乳腺组织中Wip1、p53、p38 MAPK、p16蛋白的表达,并对Wip1蛋白高表达与p53、p38、p16蛋白表达进行相关分析.结果 3种组织中Wip1蛋白高表达率分别为62.9%(44/70)、2.9%(2/70)、0(0/20).乳腺癌组织比癌旁组织、正常乳腺组织明显升高(P＜0.01).Wip1蛋白高表达与p53、p38、p16蛋白表达呈负相关(P＜0.01,等级相关系数rs分别为-0.529、-0.626、-0.499).结论 p16/p38 MAPK/p53/Wip1是负反馈通路,它可能在乳腺癌发生发展中起重要作用.     

食管鳞癌与细胞因子信号转导负调控因子3及其DNA甲基化的关系

目的 检测食管鳞癌(ESCC)组织中细胞因子信号转导负调控因子3(SOCS3)的DNA甲基化、mRNA及蛋白表达水平,探讨其在食管鳞癌发生、发展、浸润和转移中的作用.方法 采用甲基化特异性聚合酶链反应(MSP)、Real-Time聚合酶链反应(PCR)和Western blot法分别检测43例食管鳞癌组织中SOCS3的DNA甲基化、mRNA和蛋白表达水平,并与相应的癌旁正常食管组织进行对照研究,分析其与临床病理参数的关系.结果 (1)食管鳞癌组织SOCS3 DNA甲基化的阳性率(79.1%)明显高于癌旁组织(14.0%,P＜0.01);(2)食管鳞癌组织SOCS3 mRNA相对表达强度比值(0.53±0.30)明显低于癌旁组织(1.15±0.44,P＜0.01),食管鳞癌组织中甲基化组的SOCS3 mRNA表达(0.45±0.24)显著低于非甲基化组(0.86±0.29,P＜0.05);(3)食管鳞癌组织SOCS3蛋白表达(1.66±0.22)显著低于癌旁组织(1.83±0.15,P＜0.01),食管鳞癌组织中甲基化组SOCS3蛋白表达(1.61±0.21)显著低于非甲基化组(1.87±0.15,P＜0.01);(4)在TNM分期中Ⅲ期组表达均低于Ⅰ～Ⅱ期组(P＜0.05),伴有淋巴结转移组表达也都低于无淋巴结转移组(P＜0.05),未发现其在性别、年龄、家族史、吸烟史中有明显差异(P＞0.05);(5)食管鳞癌组织中SOCS3mRNA表达及其蛋白表达水平与肿瘤分化级别呈正相关(0.301＜r＜1,P＜0.05),与TNM分期、淋巴结转移呈负相关(-1＜r＜-0.301,P＜0.05).结论 食管鳞癌组织中SOCS3 DNA甲基化阳性率高,导致SOCS3基因表达下调,与食管鳞癌的分化、浸润和转移密切相关.     

慢病毒介导的RNA干扰沉默Wip1基因对人脑胶质母细胞瘤细胞生长的影响

目的 构建人Wip1基因的RNA干扰(RNAi)慢病毒载体,有效沉默人胶质瘤U251细胞的Wip1基因并观察其对细胞生长的影响.方法 设计并合成3条Wip1基因特异性RNAi靶序列,构建到慢病毒pFU-GW-iRNA载体中.慢病毒包装转染HEK293T细胞,获得病毒上清并测定其滴度;感染U251细胞,实时定量聚合酶链反应(PCR)及Western blot鉴定RNA干扰效率;筛选出基因沉默效率最高的慢病毒感染U251细胞,CCK-8法检测细胞的增殖,Western blot检测RNA干扰后的bcl-2蛋白表达.结果 PCR扩增和测序表明成功构建Wip1慢病毒干扰载体,病毒载体包装获得的病毒上清滴度在3×10~8～8×10~8 TU/ml.可以有效地沉默U251细胞中Wip1基因的表达,构建的RNAi慢病毒载体感染U251细胞后Wip1基因的mRNA表达量同对照组比较分别为36.3%、32.9%、23.8%.稳定Wip1 RNA干扰后的U251细胞4 d后细胞增殖能力下降35.1%.结论 慢病毒介导的RNA干扰可以高效稳定地沉默基因表达,Wip1基因促进了U251细胞的增殖.     

甲状腺乳头状癌中p16/p53/p38MAPK/Wip1通路的改变及意义

目的研究甲状腺乳头状癌中p16/p53/p38MAPK/Wip1通路的改变及临床意义。方法应用免疫组织化学方法,检测70例PTC组织和20例正常甲状腺组织中Wip1,p38MAPK,p53及p16蛋白的表达,并将Wip1蛋白高表达与p38MAPK,p53,p16蛋白表达进行相关性分析。结果 PTC组织中Wip1蛋白高表达率为64.3%(45/70),与正常甲状腺组织(0/20)有明显差异(P<0.01);Wip1高表达率在年龄、性别、肿瘤大小、淋巴结转移、病理分期各因素分组间差异无统计学意义(均P>0.05);Wip1蛋白高表达与p3 8 MAPK,p5 3,p1 6蛋白表达呈负相关(r=-0.6 2 0,r=-0.3 5 6,r=-0.550,均P<0.0 1)。结论PTC中存在p1 6/p5 3/p3 8 MAPK/Wip1通路异常,其机制可能与Wip1异常上调后抑制p3 8 MAPK,p5 3,p1 6有关。Wip1可能为甲状腺癌治疗的潜在靶点。     

脑胶质细胞瘤低密度脂蛋白受体活性研究

目的 观察人脑胶质瘤组织的低密度脂蛋白受体(LDLR)活性和其mRNA水平,探讨其与胶质瘤肿瘤级别的关系.方法 以125I-LDL为配体,采用受体-放射性配体结合分析法测定63例肿瘤及其周边脑组织的LDLR活性;逆转录-聚合酶链反应(RT-PCR)技术检测LDLR蛋白mRNA在瘤组织的表达.结果 胶质瘤组织LDLR活性为(129.43±48.04)μmol/g组织,其周边正常脑组织LDLR为(34.06±27.62)μmol/g组织,两者比较差异有统计学意义(P＜0.01);高级别胶质瘤组织的LDLR为(154.59±31.68)μmol/g组织,低级别胶质瘤组织LDLR活性为(92.32±24.37)μmol/g组织,高级别胶质瘤LDLR活性高于低级别(P＜0.05);胶质瘤组织LDLR蛋白mRNA水平均增高,但组间差异无统计学意义(P＞0.05).结论 脑胶质瘤组织LDLR活性明显增高,肿瘤级别越高,LDLR活性越高;因此,LDLR有可能成为脑胶质瘤治疗的潜在靶点.     

MDR1、MRP基因产物在星形细胞瘤中的表达及其意义

目的　探讨星形细胞瘤中MDR1及MRP基因表达及其临床意义。方法　用免疫组织化学SP法检测 5 8例星形细胞瘤中MDR1及MRP基因产物的表达。结果　MDR1、MRP在星形细胞瘤中高度表达 ,阳性表达率分别为 84.5 %和 60 .3 %;两者在正常脑组织中无阳性表达 ,差异有非常显著性 (P <0 .0 1)。两者在低度恶性星形细胞瘤 (包括Ⅰ级和Ⅱ级 )和高度恶性星形细胞瘤 (包括Ⅲ级和Ⅳ级 )中的表达相互比较 ,差异有非常显著性 (P <0 .0 1)。结论　MDR1、MRP基因编码的P gp和MRP蛋白在星形细胞瘤多药耐药机制中起重要作用。通过抑制P gp和MRP蛋白的表达可能找到逆转脑胶质瘤化疗多药耐药的方法。     

alpha-1抗胰蛋白酶在升主动脉夹层及瘤样扩张血管组织中的表达

目的 观察正常升主动脉、升主动脉瘤样扩张及Stanford A型夹层动脉瘤(TAAD)患者升主动脉血管组织中alpha-1抗胰蛋白酶表达的差异,探讨alpha-1抗胰蛋白酶对于维持升主动脉血管结构完整性中的作用.方法 收集28例升主动脉血管组织标本,TAAD 14例,升主动脉瘤样扩张7例,心脏移植供体心脏升主动脉7例.采用组织学、逆转录-聚合酶链反应(RT-PCR)法及Western blot 法对标本组织中的alpha-1抗胰蛋白酶进行检测分析.结果 在基因水平上,alpha-1抗胰蛋白酶在升主动脉瘤样扩张中表达最高,正常血管组织中表达次之,TAAD患者升主动脉中表达最低(2.192±0.133、1.213±0.156、0.672±0.101,P<0.05).在蛋白水平上,alpha-1抗胰蛋白酶在升主动脉瘤样扩张中表达明显升高,TAAD患者升主动脉中表达次之,正常血管组织中最低(0.285±0.010、0.153±0.011、0.102±0.010,P<0.05).结论 alpha-1抗胰蛋白酶在正常人、升主动脉瘤样扩张及TAAD患者升主动脉血管组织均有表达,表达差异有统计学意义.alpha-1抗胰蛋白酶对于维持升主动脉血管结构的完整性具有潜在的保护作用.