Expression and distribution of C1C-3 in human glioma specimen

AIM: To investigate the expression of voltage-gated chloride channels (ClC)-3 protein and mRNA in human glioma specimen and its biological function. METHODS: The expression of C1C-3 was observed by immunohistochemical staining in 24 cases of human glioma, 4 cases of brain metastic cancer specimens and 5 cases of normal brain tissue as control; The C1C-3 mRNA expression were detected in the specimens with positive expression of ClC-3 protein by RT-PCR. RESULTS: ClC-3 protein was found negative in 4 cases of normal brain tissues and positive in 19 cases of human glioma and 4 cases of brain metastic cancer specimens. ClC-3 protein was mainly expressed in the membrane or cytoplasm of neoplastic cells and microvascular endothelial cells. The expression of ClC-3 mRNA was detected in 16 cases of human glioma and 4 cases of brain metastasis cancer specimens among the tissues with the positive expression of ClC-3 protein. The level of protein and RNA of ClC-3 in high malignant oligodendrogliomas was higher than that in low malignant ones. CONCLUSION: ClC-3 is generally expressed in human glioma and brain metastic cancer and is probably correlated with the classification of its pathological malignance.     

Expression of B7-H4 in human glioma tissues and its clinical significance

AIM: To investigate the expression of B7-H4 in human glioma tissues and its clinical significance. METHODS: The histological staging of 150 cases of human glioma tissues was determined by HE staining. Immunohistochemistry was performed to study the protein level of B7-H4 in 150 human glioma specimens. Furthermore, the relationships between the expression level of B7-H4 and clinicopathological parameters, as well as patients survival rate, were analyzed. RESULTS: HE staining result indicated that there were 12 cases staged as stage I, 50 cases stage II, 39 cases stage III and 49 cases stage IV in 150 glioma specimens. Ninety-seven cases highly expressed B7-H4 in total 150 glioma samples with a 64.7% high expression rate. The histological grade of the tissues with high B7-H4 expression was mostly III~IV. There were 53 cases with low B7-H4 expression in the total 150 glioma patients with a 35.3% low expression rate. The histological grade of the tissues with low B7-H4 expression was mostly I~II. The B7-H4 expression was related to the age of the patients (P<0.01) and the pathological grade of glioma (P<0.01), but not related to the location of glioma (P>0.05). Kaplan-Meier survival curves demonstrated that increased B7-H4 expression was associated with shorter overall survival time (P<0.01). Multivariate Cox regression analysis revealed that B7-H4, age, sex and pathological grade were independent prognostic factors in glioma patients. CONCLUSION: B7-H4 is expressed in most of glioma tissues. B7-H4 may be a novel prognostic biomarker and a new target of molecular therapy for gliomas.     

Clinical significance of immunohistochemically detecting forkhead box P3 expression in human astrocytic tumors

AIM: To investigate the expression of forkhead box p3 (FOXP3) protein in astrocytic tumors, and analyze its clinical significance.METHODS: We measured the incidence of FOXP3 in 121 astrocytomas, 5 meningiomas, 5 normal brain tissues using immunohistochemistry, analyzed the correlation of the presence of FOXP3 with histopathologic features and survival. RESULTS: FOXP3 exists in human brain astrocytic tumors, which was not found in meningioma and normal brain tissues. FOXP3 was expressed in interstitial lymphocytes mainly, and expressed in tumor cells rarely. The differences on FOXP3 expression between all grade levels were statistically significant (P<0.05). The Kaplan-Meier survival analysis showed that there was significant difference between the two survival curves, and the survival rate of the group with negative FOXP3 expression was higher than that of the positive group. Multivariate Cox proportional hazards regression analysis indicated that FOXP3 could not serve as an independent prognostic factor of astrocytic tumors survival time (P＞0.05).CONCLUSION: FOXP3 immunoreactivity is significantly associated with the age and the malignancy of human astrocytic tumors. Astrocytic tumor patients with FOXP3 expression have poorer prognosis, while FOXP3 can not serve as an independent prognostic factor of astrocytic tumors survival time. FOXP3 may provide a new target for immunotherapy of astrocytic tumors.     

Effect of down-regulation of X-box binding protein 1 gene expression on viability and apoptosis of glioma cells

AIM: To investigate the effect of down-regulation of X-box binding protein 1 (XBP1) expression on the viability and apoptosis of glioma cells. METHODS: The mRNA expression of XBP1 in the glioma tissues was detected by qPCR. Small interfering RNA (siRNA) interfering with XBP1 expression (XBP1-siRNA) was transfected into human brain glioma U251 cells. At the same time, control group (the cells without special treatment) and negative control (NC-siRNA) group (transfected with siRNA without any interference) were set up. The mRNA expression of XBP1 in the 3 groups 48 h after transfection was detected by qPCR. The protein levels of XBP1, proliferating cell nuclear antigen (PCNA), B-cell lymphoma/leukemia-2 (Bcl-2), Bcl-2-associated X protein (Bax), cyclin D1 (cyclin D1), phosphatidylinositol 3-kinase (PI3K) and phosphorylated Akt (p-Akt) were determined by Western blot. The cell viability was measured by CCK-8 assay. The cell cycle distribution and apoptosis were analyzed by flow cytometry. RESULTS: The expression level of XBP1 in the glioma tissues was significantly higher than that in the tumor adjacent tissues (P<0.05). The XBP1 expression at mRNA and protein levels was significantly decreased in the cells transfected with XBP1-siRNA (P<0.05). No statistically significant difference of the cell viability, cell cycle, apoptotic rate and the protein levels of PCNA, Bcl-2, Bax, cyclin D1, PI3K and p-Akt between NC-siRNA group and control group was observed. Compared with control group, the cell viability, S-phase cells and the protein levels of PCNA, Bcl-2, cyclin D1, PI3K, and p-Akt in XBP1-siRNA group were decreased significantly, and the apoptotic rate, G₀/G₁-phase cells and Bax protein expression were significantly increased (P<0.05). CONCLUSION: Down-regulation of XBP1 gene expression in brain glioma cells reduces the viability of cancer cells, blocks the cells in G₁ phase and promote apoptosis. The mechanism is related to the inhibition of PI3K/Akt signaling pathway.     

Expression of chemokine receptor CCR7 and VEGF-C is associated with prognosis of breast carcinoma

AIM: To explore the protein levels of chemokine receptor 7 (CCR7) and vascular endothelial growth factor (VEGF)-C in breast carcinoma, and to investigate the effects of CCR7 and VEGF-C on prognosis of breast carcinoma. METHODS: The protein expression levels of CCR7 and VEGF-C in the breast carcinoma tissues and normal breast tissues were detected by the method of immunohistochemistry. At the same time, the relationship between clinicopathologic characteristics and the protein expression of CCR7 and VEGF-C in the breast carcinoma tissues was analyzed. The relationship between the protein expression of CCR7 and VEGF-C and survival time of the breast cancer patients was estimated by Kaplan-Meier method.RESULTS: The positive expression rates of CCR7 and VEGF-C in the breast carcinoma tissues were significantly higher than those in the normal breast tissues (P<0.01). A positive correlation was observed between the protein expression of CCR7 and the protein expression of VEGF-C in the breast carcinoma tissues (r=0.613, P<0.01). The protein expression of CCR7 and VEGF-C was correlated with lymph node metastasis and TNM stage (P<0.05), but both were not related to patients' age, primary tumor size, estrogen receptor and progesterone receptor. The survival time of the patients with CCR7 and VEGF-C positive expression was significantly shorter than that of the patients without the expression (P<0.05).CONCLUSION: The positive expression of CCR7 and VEGF-C proteins is associated with the prognosis of breast cancer, and combined detection of CCR7 and VEGF-C protein expression levels may be helpful to judge the prognosis of breast cancer.     

Vasculogenic mimicry and E-cadherin expression in epithelial ovarian cancer

AIM: To explore whether vasculogenic mimicry (VM) exists in human epithelial ovarian cancer (EOC), and to elucidate the relationship between E-cadherin (E-cad) expression and VM. METHODS: The E-cad expression and VM in 80 specimens of EOC and 20 specimens of benign ovarian epithelial tumor tissues were detected by the methods of immunohistochemical and histochemical staining. RESULTS: The positive rates of VM and E-cad protein in EOC were 57.4% and 48.7%, respectively.The positive rates of VM and E-cad protein in benign epithelial tumor tissues were 0% and 75.0%, respectively.There was a significant difference between the two groups (P<0.05 or P<0.01). The E-cad expression and VM in EOC was significantly related to differentiation, metastasis to abdominal organ and lymphnode, and PTNM stage (P<0.05). A negative relationship between the expression of E-cad and VM (r=-0.578,P<0.01) was also observed. PTNM stage, metastasis to abdominal organ and lymphnode, the expression of E-cad and VM were independent prognosis factors of EOC patients after total correction (P<0.05). The five-year survival rate between VM-positive group and VM-negative group was significantly different (4.3% vs 88.2%), while the five-year survival rate was significantly lower in E-cad-negative group than that in E-cad-positive group (9.8% vs 71.8%). CONCLUSION: EOC with VM has a poor differentiation and a bad clinical prognosis. The levels of VM and E-cad correlate with the progression and prognosis of EOC.     

Clinical significance of KLF15 expression in human lung adenocarcinoma

AIM: To explore the clinical significance of Krüpple-like factor 15 (KLF15) protein expression in the patients with lung adenocarcinoma for exploring the therapeutic and prognositic biomarkers of lung cancer. METHODS: Four cases of lung adenocarcinoma tissues and matched adjacent tissues were collected from our hospital, and the expression of KLF15 protein in these tissues was analyzed by Western blot. At the same time, 72 cases of archived paraffin-embedded samples and clinical data of the patients with lung adenocarcinoma were also collected. The KLF15 protein expression in the archived paraffin-embedded lung adenocarcinoma samples was detected by immunohistochemical staining. The correlations between KLF15 protein expression and clinical characteristics of the patients including prognosis were also analyzed. In addition, the KLF15 protein was up-regulated in A549 cells, and then the effects of KLF15 protein on the viability of the cells were measured by CCK-8 assay. RESULTS: The protein expression of KLF15 in the 4 cases of lung adenocarcinoma tissues was significantly lower than that in matched paracancerous tissues. Fifty-three cases of lung adenocarcinoma specimens showed low expression or no expression of KLF15 protein in total 72 cases (73.6%). The 5-year survival rate of the patients with high expression of KLF15 protein in their specimens was higher than that of the patients with the low expression of KLF15 protein (P<0.01), and the expression of KLF15 protein was significantly correlated with the pathological staging (P<0.01) and T stage (P<0.01) of the patients with lung adenocarcinoma. Furthermore, the low expression of KLF15 protein was an important poor prognostic indicator of the patients. Up-regulation of KLF15 protein in the A549 cells significantly inhibited the growth of the cells. CONCLUSION: KLF15 inhibits the growth of lung adenocarcinoma cells. It could be used as a therapeutic target and a prognostic biomarker for the patients with lung adenocarcinoma.     

芍药切花贮藏后水分与膜脂过氧化的研究

试材为芍药‘莲台’品种 (Paeonialactiflora‘Liantai’) ,采自山东荷泽 ,在花蕾萼片疏松、外层花瓣显现真正花色时采样 ,用水洗去叶片和花蕾上的分泌物 ,采后 40h火车运到实验室。在水中剪截成花枝长 35cm ,留 2～ 3片复叶 ,基部在 15cm深水中浸 2h复水 ,10 0 0倍的百菌清浸泡 1min ,晾干 ,普通白纸包裹后封入聚乙烯塑料袋中 ,于 0～2℃干藏。定期取样测定花瓣各项指标 ,均测 3次重复。定期取 5枝花在蒸馏水中瓶插 (室内散射光 ,温度 2 5℃±3℃ ,相对湿度 40 %～ 6 0 %) ,每天称花枝和 (水 +瓶 )质量 ,计算吸水量和失水量 ,记录瓶插寿命…     

Prognostic significance of NAD(P)H-quinone oxidoreductase 1 overexpression in ovarian mucinous cystadenocarcinoma

AIM: To investigate the significance of NAD(P) H-quinone oxidoreductase 1 (NQO1) protein overexpression for prognostic evaluation of ovarian mucinous cystadenocarcinoma.METHODS: NQO1 protein was detected in 162 cases of ovarian mucinous cystadenocarcinoma, 35 cases of ovarian mucinous cystadenoma and 29 samples of normal ovarian epithelial tissues by the method of EnVision immunohistochemical staining. The correlation between high expression of NQO1 protein and clinicopathological features of ovarian mucinous cystadenocarcinoma was also evaluated. Overall survival and disease-free survival rates of ovarian mucinous cystadenocarcinoma patients were calculated by Kaplan-Meier method. RESULTS: The positive rate and strongly positive rate of NQO1 protein were 85.8% and 64.2% in ovarian mucinous cystadenocarcinoma, respectively, which are significantly higher than those in ovarian mucinous cystadenoma, and normal ovarian epithelial tissues (P<0.01). NQO1 expression was significantly correlated with the histological grade (P<0.05) and clinical stage (P<0.01) of ovarian mucinous cystadenocarcinoma. Kaplan-Meier survival analysis showed that the overall survival rate and disease-free survival rate were significantly higher in ovarian mucinous cystadenocarcinoma patients with high NQO1 expression than those with low NQO1 expression (P<0.01).CONCLUSION: NQO1 expression is closely correlated with the progression and prognosis of the patients with ovarian mucinous cystadenocarcinoma. High expression of NQO1 protein may be used as an important indicator for the patients with poor prognosis of ovarian mucinous cystadenocarcinoma.     

Expression and clinical significance of bone morphogenetic protein 3 in hilar cholangiocarcinoma tissues

AIM: To investigate the expression and clinical significance of bone morphogenetic protein 3 (BMP3) in hilar cholangiocarcinoma tissues.METHODS: Thirty cases of hilar cholangiocarcinoma specimens were collected. The expression of BMP3 at mRNA and protein levels in the tumor tissues and paracancerous tissues was detected by real-time PCR and Western blot. The hilar cholangiocarcinoma paraffin-embedded specimens (n=103) were collected. The protein expression of BMP3 was determined by immunohistochemical method, and the relationship of BMP3 protein expression with clinical pathological characteristics was evaluated.RESULTS: In the 30 patients with hilar cholangiocarcinoma, the expressions of BMP3 protein and mRNA in 22 cases of tumor tissues were significantly decreased compared with the adjacent normal tissues. The results of immunohistochemistry showed that 87 cases were negative and 16 cases were weakly positive in all 103 cases of hilar cholangiocarcinoma. The expression of BMP3 protein was associated with the tumor TNM staging, lymph node metastasis and tumor differentiation (P<0.05).CONCLUSION: BMP3 gene might be inhibited in human hilar cholangiocarcinoma. The down-regulation of BMP3 gene might be associated with the carcinogenesis and development of hilar cholangiocarcinoma.     

Effects of ginsenoside Rg1 of Panax notoginseng on brain tissue injury and Nrf2/HO-1 signal pathway after cerebral ischemia/reperfusion in mice

AIM:To observe the effects of ginsenoside Rg1 of Panax notoginseng on brain tissue injury after mouse cerebral ischemia/reperfusion(I/R), and to explore the mechanisms involving nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) signal pathway. METHODS:C57BL/6 mice were randomly divided into sham group, cerebral I/R group, ginsenoside Rg1+cerebral I/R group and edaravone+cerebral I/R group. Ginsenoside Rg1 was successively administered for 3 d. One hour after final administration, bilateral common carotid arteries were ligated to induce brain tissue injury for 20 min, and then reperfusion for 24 h. Edaravone, a drug for anti-oxidative stress injury in the treatment of ischemic cerebro-vascular disease, was used as a positive control. The brain tissues were obtained to determine the neural cellular pathology in hippocampal CA1 region. The mRNA expression of Nrf2 and HO-1 was detected by RT-PCR. The protein levels of Nrf2 in the nucleus and cytoplasm and HO-1 in the whole cells in the brain tissues were measured by Western blotting. RESULTS:After ischemia/reperfusion for 24 h, the pathological injury in the neural cells was obvious, and the cell survival rate decreased. Ginsenoside Rg1 and edaravone attenuated the neural cell injury, and significantly increased the cell survival rate. After ischemia/reperfusion for 24 h, the mRNA expression of Nrf2 and HO-1 significantly increased in the brain tissues. The protein levels of Nrf2 in the nucleus and cytoplasm in the brain tissues were increased, the nuclear translocaition rate and the protein expression of HO-1 also increased. Ginsenoside Rg1 and edaravone both decreased the protein levels of Nrf2 in the cytoplasm of the brain tissues, increased that in the nucleus, and also increased Nrf2 nuclear translocation rate and the protein expression of HO-1. The effect of edaravone was higher than that of ginsenoside Rg1, but they had no significant effect on the mRNA expression of Nrf2 in the brain tissues. CONCLUSION: Ginsenoside Rg1 has the effect of anti-brain tissue injury on cerebral ischemia/reperfusion. The mechanisms may be related to activating the Nrf2/HO-1 signal pathway, promoting Nrf2 synthesis and nuclear translocation, thus promoting the expression of downstream antioxidant protein HO-1.     

Expression and prognostic significance of cyclin D1, RBL2/p130 and MCM7 in hepatocellular carcinoma

AIM: To investigate the expression and prognostic significance of cyclin D1, retinoblastoma-like protein 2(RBL2/p130) and minichromosome maintenance protein 7(MCM7) in hepatocellular carcinoma(HCC). METHODS: The expression of cyclin D1, RBL2/p130 and MCM7 in 44 HCC specimens, 26 adjacent noncancerous cirrhotic liver specimens and 18 normal liver specimens were detected by the method of immunohistochemistry. The correlations of cyclin D1, RBL2/p130 and MCM7 with the clinical parameters of HCC patients were analyzed. RESULTS: The positive expression rates of cyclin D1 and MCM7 in HCC were 68.2% and 72.7%, higher than those in normal livers and adjacent noncancerous cirrhotic livers(P<0.01). The positive expression rate of RBL2/p130 in HCC was 34.1%, lower than that in normal livers and adjacent noncancerous cirrhotic livers(P<0.01). The expression of MCM7 in HCC was positively correlated with the expression of cyclin D1(r=0.349, P<0.05), and negatively correlated with the expression of RBL2/p130(r=-0.421, P<0.01). The expression of cyclin D1 in HCC was negatively correlated with the expression of RBL2/p130(r=-0.435, P<0.01). The expression of MCM7 and cyclin D1 was associated with the integrity of tumor capsule, differentiation degree and TNM stage(P<0.05). The expression of MCM7 was also associated with tumor size and the value of alpha fetoprotein(AFP). The tumor size, the expression levels of MCM7 and cyclin D1 were correlated with the prognosis determined by multivariable analysis. The patients with positive expression of MCM7 and cyclin D1 had lower survival rate than those with negative expression(P<0.05). The patients with positive expression of RBL2/p130 had higher survival rate than those with negative expression(P<0.05). CONCLUSION: The abnormal expression of cyclin D1, RBL2/ p130 and MCM7 plays an important role in the development of HCC, indicating that monitoring their expression in HCC patients may be helpful to the judgment of prognosis.     

Expression and significance of 4-1BB/4-1BBL and IL-15 in the experimental autoimmune myocarditis

AIM: To establish the animal model of experimental autoimmune myocarditis (EAM) in BALB/c mice and to investigate the expression and significance of 4-1BB/4-1BBL and IL-15 in mouse EAM. METHODS: Cardiac myosin was extracted from porcine ventricular myocardium. Genetically predisposed BALB/c mice were immunized with cardiac myosin covered by complete freunds adjuvant (CFA) on 1 d, 8 d and 30 d. The control mice were immunized with CFA only. On 21 d and 80 d after the first immunization, the areas of inflammation were identified by HE staining and cTnI was examined. Then 4-1BBL and IL-15 were determined by immunohistochemistry and ELISA, respectively. The mRNA levels of 4-1BB/4-1BBL and IL-15 were measured by semi-quantitative RT-PCR. RESULTS: In experimental group, histopathological examination of cardiac tissue showed an obvious inflammatory cell infiltration with myocyte necrosis at 21 d, the cTnI was higher than that in control group. On 80 d, fibrosis and a few inflammatory cells were observed. The cTnI in EAM group was still higher than that in control group (P<0.05). No inflammation and fibrosis were emerged in the myocardium of control mice. In addition, as compared with control groups, immunohistochemistry and mRNA abundance for both 4-1BB/4-1BBL and IL-15 were up-regulated at acute phase of the EAM model (P<0.01). On 80 d, the mRNA abundance was still up-regulated (P<0.05).CONCLUSION: The expressions of 4-1BB/4-1BBL and IL-15 in myocardium are up-regulated in experimental autoimmune myocarditis. The 4-1BB/4-1BBL costimulating pathway and IL-15 may play a role in the occurrence and development of autoimmune myocardtis.     

Association of miR-497 and prognosis in patients with renal cell carcinoma and its effects on proliferation,apoptosis and invasion of human renal cell carcinoma cell line 786-0

ATM: To investigate the association of microRNA-497 (miR-497) and prognosis in the patients with renal cell carcinoma (RCC) and its effects on the proliferation, apoptosis and invasion of human RCC cell line 786-0. METHODS: Paired RCC and adjacent non-tumor tissue specimens were surgically collected from 80 patients who were diagnosed with primary RCC between 2011 and 2015. The expression of miR-497 in the paired RCC and adjacent non-tumor tissue specimens was detected by real-time PCR. Recurrence-free survival was estimated using the Kaplan-Meier method and compared using the log-rank test. The 786-0 cells were transfected with miR-497 mimics or scramble control miRNA. The proliferation, apoptosis and invasion abilities of the transfected cells were assessed by MTT assay, Trypan blue exclusion, flow cytometry and Transwell chamber experiment. The protein expression of miR-497-targeted gene cyclin D1 in the transfected cells was quantified by Western blotting. RESULTS: miR-497 was down-regulated in the RCC specimens compared with the adjacent tissues. miR-497 was down-regulated in the RCC 786-0 cells compared with the HK-2 cells. By the end of the study, 74 cases were followed up. The follow-up rate was 92.5%. Median follow-up was 29 months (ranging from 2 months to 48 months). The 3-year recurrence-free survival rates of the patients with high and low miR-497 expression were 71.2% and 40.1%, respectively. Over-expression of miR-497 resulted in significant suppression effect on RCC cell proliferation, invasion and the expression of cyclin D1. CONCLUSION: Low expression of miR-497 was correlated with poor prognosis in the RCC patients. miR-497 inhibits proliferation and invasion of RCC 786-0 cells and its mechanism is associated with the down-regulation of cyclin D1.     

Effects of microRNA-483-3p on human glioma cells

AIM:To investigate the effects of microRNA (miRNA)-483-3p on the growth and migration of human glioma cell line A172 and its potential mechanisms.METHODS:The abundance of miRNA-483-3p in human embryonic kidney 293 cells and different human glioma cell lines (A172,U251 and SHG44) was measured by RT-qPCR.After down-regulation of miRNA-483-3p by transfection of inhibitor in the A172 cells,the cell viability,cell cycle distribution and cell migration were detected by CCK-8 assay,flow cytometry and Transwell assay,respectively.Furthermore,the protein levels of cell cycle-related molecules and epithelial-mesenchymal transition markers were measured by Western blot.Luciferase reporter assay was used to predict and verify the target gene of miRNA-483-3p.RESULTS:miRNA-483-3p was highly expressed in human glioma cells.Knockdown of miRNA-483-3p inhibited A172 cell viability,arrested cell cycle and decreased cell migration rate.Furthermore,the protein levels of cyclin D1,cyclin-dependent kinase 4,phoshorylated retinoblastoma protein,N-cadherin and vimentin were significantly decreased after knockdown of miRNA-483-3p,accompanied with the up-regulation of E-cadherin and β-catenin protein expression.Luciferase reporter assay demonstrated that Smad4 was a potential target gene of miRNA-483-3p.Down-regulation of Smad4 in the A172 cells transfected with miRNA-483-3p inhibitor partially reversed the effect of miRNA-483-3p on cell viability and migration.CONCLUSION:Knockdown of miRNA-483-3p restrains the growth and migration of A172 cells by targeting Smad4.     

Effects of taurine on expression of glucose transporters in rat brain with diffused brain injury

AIM: To investigate the effect of taurine on the expression of glucose transporter 1 (GLUT1) and transporter 3 (GLUT3) in rat brain with diffused brain injury (DBI).METHODS: Sixty-four male Sprague-Dawley rats were randomly divided into 4 groups: sham-operated group, DBI group, low-dose taurine group (200 mg/kg, ig) and high-dose taurine group (300 mg/kg, ig).After fed with the corresponding drugs for 7 days, the animal model of DBI was made, and the rats were executed 24 h after DBI.The expression of GLUT1 and GLUT3 in the brain was detected by the methods of immunohistochemistry and Western blotting.The pathomorphological changes of the cerebral cortex were observed under electron microscope.RESULTS: The expression of GLUT1 was detected in capillary vascular endothelial cells in each group, and cytoplasm-positive cells or the cells with buffy membrane were observed.No significant difference of the GLUT1 expression in brain tissues between DBI group and sham-operated group was detected.Compared with DBI group, the expression of GLUT1 in the brain tissues were significantly increased in low-and high-dose taurine groups (P<0.01).The expression of GLUT1 in the brain tissues in low-dose taurine group were significantly higher than that in high-dose taurine group (P<0.05).The positive staining of GLUT3 only appeared in the periphery of the third ventricle in each group in the cells with buffy membrane or positive cytoplasm.The expression of GLUT3 in the brain tissues in DBI group was significantly higher than that in sham-operated group (P<0.01).The expression of GLUT3 in the brain tissues in low-and high-dose taurine groups was significantly higher than that in DBI group (P<0.01).Compared with low dose taurine group, the expression of GLUT3 in the brain tissues were significantly increased in high-dose taurine group (P<0.01).The pathological damage of cerebral cortex in low-dose taurine group was obviously alleviated.CONCLUSION: Taurine may take part in the neuroprotective mechanisms in DBI by increasing the expression of GLUT1 and GLUT3 at protein level to maintain the energy supply in brain tissues.     

Experimental study of antitumor immunity of DC/C6 fusion vaccine

AIM:To study the anti-tumor effect and mechanism of fusion vaccine on DCs and C6 glioma cells. METHODS:PEG was used to fuse DCs with C6 glioma cells. Immunofluorescence with GFAP-FITC was used to identify the DC/C6 fusion cells. Rat brain glioma models were made by stereotactic technique. After 5 days of inoculation of C6, 107 fusion cells were injected through tail vein in group A. The same number of DCs and the same volume of PBS were used in group B and group C. The survival time of rats in these three groups was analyzed by Log-rank survival analysis. Tumor samples were checked by HE staining and immunohistochemical staining with CD8Mcab. RESULTS:Positive result of GFAP-FITC immunofluorescence was observed in DC/C6 fusion cells. The Log-rank survival analysis showed that statistically significant difference in group A was observed compared to that in group B and group C (P<0.01). Tumor sample stained with HE showed that many inflammatory cells infiltrated in tumor tissues. The result of immunohistochemical staining with CD8Mcab was positive. CONCLUSION:DC/C6 fusion cells had the ability of antigen presentation and activating T lymphocytes effectively. CD8⁺T lymphocytes play an important role in antitumor immunity.     

Expression and clinical significance of Bmi-1 in colorectal cancer

AIM:This study was to investigate the expression and significance of Bmi-1 in colorectal carcinoma (CRC), and to explore the effect of Bmi-1 on Ki67 expression in human CRC.METHODS:The samples from sixty CRC, thirty adenomas and twenty normal colorectal mucosal tissues were used in this study.The expression of Bmi-1 protein was detected by immunohistochemistry.The clinicopathological features and survival rate of patients were also analyzed.RESULTS:The overexpression of Bmi-1 was respectively 25.0％, 6.7％and 0% in CRC and adenomas as well as normal colorectal mucosal tissues.The results showed that the expression of Bim-1 was significantly higher in CRC, compared with that in adenomas and normal colorectal mucosal tissues (P<0.05).The overexpression of Bmi-1 protein in CRC was obviously associated with distant metastasis and TNM stage (P<0.05), but not with gender, age, tumor size, tumor site, histological type, differentiation degree and lymph node metastasis (P>0.05).Kaplan-Meier survival analysis showed that the overexpression of Bmi-1 reduced significantly survival of CRC patients (P<0.05).No statistical relation between expression of Bmi-1 and Ki67 in CRC was observed.CONCLUSION:The overexpression of Bmi-1 protein is significantly correlated with tumorigenesis, metastasis and prognosis of CRC.Bmi-1 might be regarded as a parameter in evaluating prognosis of CRC.     

Changes of aquaporin expression in blood-brain barrier induced by glioma cells

AIM:To investigate the effects of glioma cells on aquaporin expression in blood-brain barrier and their importance in pathophysiology.METHODS:A blood-brain barrier model was established by coculture of ECV304 and astrocytesin vitro. HPLC was used to determine the change of water transport ofin vitroblood-brain barrier model after the influence of glioma cells. The expression levels of AQP1 and AQP4 were analyzed by semiquatitative RT-PCR.RESULTS:Glioma cells decreased expression level of AQP4 of astrocytes and induced abnormal expression of aquaporin-1 in endothelial cell line. The water transport ofin vitroblood-brain barrier model from luminal side to abluminal side was increased after coculture with glioma cells.CONCLUSION:The vasogenic brain edema induced by glioma cells may not be the result of hyperpermeability of blood-brain barrier to macromolecules in plasma. The changes of aquaporin expression may be the molecular basis of brain edema induced by glioma cells.