Expression of Glypican-3 gene and its regulating mechanism in hepatocellular carcinoma

AIM:To investigate the expression of Glypican-3 gene and its regulating mechanism in hepatocellular carcinoma. METHODS:The expression of GPC3 mRNA and its gene mutation in 48 hepatocellular carcinoma tissues, 39 paracarcinomatous tissues and 31 normal liver tissues were detected by using RT-PCR and PCR-SSCP. The expressions of GPC3 protein, P53 and PCNA protein were detected by using immunohistochemistry S-P. RESULTS:The positive expressive rate of GPC3 mRNA was 77.1% in hepatocellular carcinoma tissue. No expression of GPC3 mRNA in paracarcinomatous tissues and normal liver tissues was observed. No gene mutation of GPC3 in hepatocellular carcinoma tissue was found. No correlation was found between GPC3 and P53 (r=-0.12574, P>0.05). The mean index of proliferating cell nuclear antigen(PCNA) in positive and negative GPC3 expression were (46.32±27.54)% and (39.83±21.47)%, respectively (P>0.05). CONCLUSION:The high expression of GPC3 mRNA in hepatocellular carcinoma is independent to the gene mutation, and to the expression of P53 and PCNA protein.     

《世界名花赏析》

AIM: To study the expression of MDM2 and mutant-type P53 proteins in primary hepatocellular carcinoma(HCC). METHODS: Using immunohistochemical staining method(SP),the expression of mutant-type P53 and MDM2 proteins was examined in 55 cases of HCC,23 cases of corresponding paracancerous tissues and 10 cases of normal hepatic tissues. RESULTS: The frequencies of MDM2 and P53 positive expression in HCC were 17/55(30.9%) and 23/55 (41.8%), respectively. There was both positive expression of MDM2 and mutant-type P53 in 11 cases (20%) with HCC. Expression of MDM2 showed a significantly positive correlation with expression of mutant-type P53 (r=0.310,P＜0.05). The frequencies of MDM2 and P53 positive expression in corresponding paracancerous tissues cells were 1/23 (4.3%) and 2/23(8.6%), respectively. Otherwise, there was a significant statistical difference in MDM2 and mutant-type P53 protein expression between HCC and paracancerous tissues(P＜0.05). There was not positive expression of MDM2 and mutant-type P53 in normal hepatic tissue. CONCLUSION: There is MDM2 and mutant-type P53 protein overexpression in HCC. The p53 gene inactivation resulting from MDM2 protein overexpression and/or p53 gene mutation may play an important role in hepatocarcinogenesis.     

Prognostic significance of FOXA1 and BRCA1 expression in triple negative breast cancer

AIM: To investigate the expression of forkhead box protein A1(FOXA1) BRCA1 protein,P53 and vascular endothelial growth factor (VEGF) in triple negative breast cancer (TNBC) and non-TNBC, and the relevance with the clinicopathological parameters for evaluating the prognosis. METHODS: The tumor samples were collected from 113 cases of breast cancer patients in the First Affiliated Hospital of Jinan University,and divided into TNBC group, luminal subtype group and HER-2 overexpression subtype group by the immunohistochemical results of estrogen receptor, progesterone receptor and HER-2. EnVision two-step method was used to detect the expression of FOXA1, BRCA1, P53 and VEGF in the tumor samples. RESULTS: Total FOXA1 positive expression rate was 63.7% (72/113), with 45.2% (19/42) in TNBC, 88.0% (44/50) in luminal subtype and 42.9% (9/21) in HER-2 overexpression subtype.The statistically sigfnificant difference among the 3 groups was observed (P<0.01). Total BRCA-1 positive expression rate was 47.8% (54/113), with 66.7% (28/42) in TNBC, 44.0% (22/50) in luminal subtype and 19.0% (4/21) in HER-2 overexpression subtype.The statisticallysignificant difference among the 3 groups was also observed (P<0.01). In the cases of clinical stages Ⅰ~Ⅱand histological grades 1~2, FOXA1 positive rate was higher than the FOXA1 negative rate (P<0.01). Negative correlations between FOXA1 positive rate and expression of P53/VEGF, and between FOXA1 positive rate and the recurrence rate were found (P<0.05). In the cases of clinical stages Ⅱ~Ⅲ and histological grades 2~3, the BRCA1 positive rate was higher than the BRCA1 negative rate (P<0.05). Positive correlations between BRCA-1 positive rate and the expression of P53/VEGF, and between BRCA1 positive rate and the recurrence rate were also observed (P<0.05). CONCLUSION: Expression of FOXA1 and BRCA1 in breast cancer is different. BRCA1 may be an adverse prognostic indicator for triple negative breast cancer.     

Molecular mechanism of miR-1246 enhancing radiosensitivity of cervical cancer cells

AIM: To investigate the molecular mechanism of microRNA-1246(miR-1246) enhancing radiosensitivity of cervical cancer cells. METHODS: Cervical cancer lines HeLa, CaSki, C33A and SiHa were transfected with miR-1246 mimic and negative control mimic (NC-mimic) using Lipofectamine 2000 kit, and the expression level of miR-1246 in cervical cancer tissue, normal tissue, cervical cancer cell lines and endometrial epithelium cell line ESC was detected by real-time PCR. The transfected cells were exposed to X-ray radiation. The cell viability and migration rate were measured respectively by MTT assay and Transwell method. The protein levels of γH2AX, ATM, p-ATM and p-p53 were monitored by immunofluorescence and Western blot. RESUITS: Higher miR-1246 level was found in normal tissue and ESC cells, while lower miR-1246 level was found in HeLa, SiHa, C33A and Caski cells and cervical cancer tissues. The expression level of miR-1246 in the cells transfected with miR-1246 mimic was significantly higher than that in the cells transfected with NC-mimic (P<0.05). The cell viability and migration rate of the cervical cancer cells with miR-1246 over-expression were notably lower than those of the cells transfected with NC-mimic (P<0.05) under the same conditions. The results of immunofluorescence indicated that the protein expression level of γH2AX significantly increased in the cervical cancer cells with miR-1246 over-expression exposed to radiation compared with the negative control (P<0.05). The protein expression level of γH2AX was significantly increased in the cervical cancer cells with miR-1246 over-expression, while the protein levels of p-ATM and p-p53 were significantly decreased as compared with the negative control group (P<0.05). CONCLUSION: miR-1246 is highly expressed in normal tissue and normal endometrial epithelial cells, while is low expressed in the cervical cancer tissues or cells. miR-1246 over-expression inhibits growth and migration, and significantly enhances radiosensitivity of cervical cancer cells. The molecular mechanism is possibly related to inhibiting ATM pathway and DNA damage repair.     

Correlation between E-cadherin and FOXO3a expression in gastric can-cer tissues and cells

AIM: To investigate the expression of E-cadherin and forkhead box protein O3a (FOXO3a) in gastric cancer tissues and cells, and its correlation with cell viability. METHODS: The expression of E-cadherin and FOXO3a was detected by immunohistochemical staining in 53 specimens of gastric cancer tissues and their adjacent tissues, and the relationship between their expression and clinicopathological characteristics were analyzed. E-cadherin-over-expressing gastric cancer AGS cells were constructed by lentivirus-mediated cell transfection, and the protein expression of E-cadherin and FOXO3a was detected by immunocytochemistry method. The expression of E-cadherin, FOXO3a, Akt, Bcl-2 and Bax was determined by Western blot. The cell viability was detected by CCK-8 assay. RESULTS: The positive expression rates of E-cadherin and FOXO3a proteins in gastric cancer tissues were both significantly lower than those in their adjacent tissues (P<0.05). E-cadherin positive expression in gastric cancer tissues was significantly related to tumor grade and TNM stage (P<0.05), but not related to age, sex, location, T stage or lymph node metastasis. FOXO3a positive expression was significantly related to tumor grade (P<0.05), but not related to age, sex, location, TNM stage, T stage or lymph node metastasis. The expression of E-cadherin was positively correlated with FOXO3a expression in gastric cancer tissues (r=0.376, P=0.003). After over-expression of E-cadherin, the viability of gastric cancer AGS cells was significantly inhibited, the expression of FOXO3a, Bcl-2 and Bax was significantly increased, and the expression of Akt was significantly decreased. CONCLUSION: E-cadherin and FOXO3a are involved in the development of gastric cancer, and E-cadherin may affect the viability of gastric cancer cells by regulating Akt/FOXO3a signaling pathway.     

Protein expression of EMMPRIN,MMP-2 and P53 detected by technique of quantum dots immunofluorescence in human lung cancer

AIM: To investigate the expression of extracellular matrix metalloproteinase inducer (EMMPRIN/CD147), matrix metalloproteinase2 (MMP-2) and P53 proteins in human lung cancer tissues and to explore the relationship between EMMPRIN protein and malignant biological behaviour of lung cancer.METHODS: Fluorescent semiconductor nanocrystals were applied, which were nanometer-sized light-emitting particles and were emerging as a new class of fluorescent probes for cancer detection due to the unique optical and electronic properties. The technique of QDs immunofluorescence histochemistry (QDs-IHC) was used to detect the protein expression of EMMPRIN, P53 and MMP-2 in the human lung cancer microarray, and co-expression of EMMPRIN/P53 proteins was also simultaneously detected by double-labeling immunofluorescence.RESULTS: Compared with non-cancerous lung tissues, the positive rates of EMMPRIN, P53 and MMP-2 proteins in the lung cancer tissues were 70.00%, 77.14% and 72.86%, respectively, and the differences were all significant (P<0.05). The positive rates of EMMPRIN, P53 and MMP-2 proteins were all significantly related with tumor staging(TNM stage) and lymph node metastasis (P<0.05). The positive correlation between EMMPRIN expression and protein levels of MMP-2 and P53 was observed (P<0.01).CONCLUSION: The protein expression of EMMPRIN, P53 and MMP-2 is correlated with the development of lung cancer. Malignant progression of lung cancer promoted by EMMPRIN may be closely related with the expression of MMP-2 and P53.     

Inhibitory effect of hypoxia preconditioning on hypoxia/reoxygenation-induced apoptosis in cardiomyocytes

AIM: To study the role of hypoxia preconditioning (HP) in hypoxia-reoxygenation (HR)-induced apoptosis in neonatal rat cardiomyocytes and the possible mechanisms. METHODS: Cultured neonatal rat cardiomyocytes were divided into three groups: normal group, HP+H/R group and H/R group. Acridine orange (AO) staining was performed to detect morphological changes of apoptotic cells. Apoptosis rates of cardiomyocytes were detected by flow cytometry. Colorimetric assay was used to detect caspase-3 activity. Expression of Bcl-2 protein was detected by immunohistochemistry combined with computer image analysis. RESULTS: Apoptotic cells were detected by AO staining after hypoxia of 6 h followed by 3 h-reoxygenation. The hypodiploid apoptotic peak was detected by flow cytometry with the apoptotic rates of (29.7±5.4)%. A significantly reduced apoptotic rates of (7.8±1.3)% was detected in HP group（P＜0.01）. The caspase-3 relative activity of cardiomyocytes induced by H/R was 5.9±0.8, significantly higher than that of control group. HP markedly reduced caspase-3 relative activity to 2.6±0.5 in contrast with H/R group （P＜0.01）. Bcl-2 protein was positive in normal cardiomyocytes with an A value of 119.4±7.1. The A value of H/R group was 99.6±5.0, significantly lower than that in normal group （P＜0.01）. The A value of HP+H/R group was 126.5±6.2, significantly higher than that in H/R group（P＜0.01）. CONCLUSION: HP inhibits H/R-induced apoptosis of cardiomyocytes by improving the expression of Bcl-2 and reducing caspase-3 activity.     

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…     

Expression of GATA3 in human breast cancer tissues and its clinical significance

AIM: To investigate the expression of GATA3 in human breast carcinoma and its clinical significance. METHODS: The expression level of GATA3 in breast cancer tissues from 124 patients was detected by the method of immunohistochemistry and the relationships between GATA3 expression and other clinicopathological factors were analyzed. RESULTS: Low expression of GATA3 in breast cancer tissues was associated with estrogen receptor (ER)/progesterone receptor (PR) negative, high histological tumor grade, p53 mutation and vascular invasion (P<005), but not with age, tumor size,human epidermal growth factor receptor 2 (HER-2) expression and lymph node metastasis (P>005). In all breast cancer tissues, the positive expression rate of GATA3 was 56.4%. The positive expression rate of GATA3 in luminal breast cancer is 684%, higher than that in non-luminal breast cancer (326%, P<005). In all breast cancer tissues, the expression of GATA3 in middle recurrence risk group was higher than that in high recurrence risk group (P<005). CONCLUSION: GATA3 expression in breast cancer is related to differentiation and biological characteristics of the tumor, which can be a factor for evaluation of the treatment and prognosis.     

Effect of siRNA-mediated Smad3 silence on proliferation and apoptosis in activated hepatic stellate cells

AIM: To investigate the effects of siRNA-mediated Smad3 silence on proliferation and apoptosis in activated hepatic stellate cells (HSCs).METHODS: HSCs-T6 cells were divided into 3 groups: blank group, negative control group and siRNA-Smad3 transfection group. The siRNA-Smad3 was transfected into HSCs-T6 cells. At different time points after transfection, cell proliferation was measured by CCK-8, cell apoptosis was detected by flow cytometry, and protein levels of P53 and Bcl-2 were determined by immunocytochemistry.RESULTS: HSCs proliferation was significantly inhibited at the time points of 24 h, 48 h and 72 h after transfection. Meanwhile, the apoptosis of HSCs was significantly increased in siRNA-Smad3 transfection group (P<0.01). Compared to the control cells, the protein expression of P53 was significantly increased while Bcl-2 protein was significantly decreased 48 h after transfection in siRNA-Smad3 transfection group (P<0.01).CONCLUSION: The siRNA-mediated Smad3 silence significantly inhibits HSCs proliferation and induces apoptosis by up-regulating the P53 expression and down-regulating the Bcl-2 expression in HSCs.     

Expression of oxygenase-1 protein and mRNA on mouse acute viral myocarditis caused by coxsackie viruses B3

AIM: To observe the dynamic variation of oxygenase-1 protein and mRNA on mouse acute viral myocarditis caused by coxsackie viruses B3.METHODS: A total 72 inbred male BALB/c mice of 4-6 weeks were divided randomly into 2 groups as follows: 32 mice were inoculated intraperitoneally (ip) with virus free 1640 culture solution 0.1 mL on day 0 as blank group (C); 40 mice were ip 0.1 mL tissue culture infectious dose 50 (TCID50 is 10-4.36/mL) coxsackie viruses B3 (CVB3) on day 0 as VMC group (V), then each mouse in both groups was ip 0.1 mL NS every day. 8 mice in each of C group and V group were sacrificed on 4, 8, 15, and 21 d respectively after infections. The blood specimens gathered by taking out the eyeballs of mice were tested for the content of carboxyhemoglobin (COHb) using spectrophotometer method.The heart tissue slides were also stained by immunohistochemistry (IHC) for HO-1 and in situ hybridization (ISH) for HO-1 mRNA. The histological and ultrastructure changes were observed under light microscope and electron microscope.RESULTS: (1)The histopathological changes of myocardial cells: many inflammatory cells were found in the heart and large area myocardial cells necrosis was observed under light microscope. The inflammatory area was reduced at late stage in the heart of mouse in group V, while the myocardium in group C was normal. (2) The myocardial observation by electron microscopy: the myofibril in group V was dissolved and mitochondrial membrane disappeared, mononuclear cell infiltration was also observed under electron microscopy, which contained many lysosomes. The myocardial cells in group C were normal. (3) The changes of blood COHb level: compared with group C, the group V COHb level showed significantly higher on the day 8 and day 15 after CVB3 innoculation (0.047±0.005 vs 0.031±0.004; 0.076±0.006 vs 0.030±0.005, P<0.01). No obvious change in group C was observed. (4) The result of HO-1 IHC staining: myocardial cells had positive expression in group V, group C was negative. The absorbance (A) values in group V was significantly higher than that in group C (P<0.01) at different time points. (5) The result of HO-1 ISH was similar to HO-1 IHC. The A values in group V was all higher than that in group C (P<0.01).CONCLUSION: Viral myocarditis caused by coxsackie viruses B3 induces the expression of HO-1 mRNA and protein, and these expressions may play self-protection in inflammation injury in myocardial cells.     

NET-1 promotes metastasis of lung squamous-cell carcinoma by activating EMT

AIM: To explore the effect of neuroepithelial cell transforming gene-1 (NET-1) expression on the metastasis of lung squamous-cell carcinoma (LSC) and the underlying molecular mechanism. METHODS: Immunohistochemistry was used to detect the expression of NET-1 protein in 53 cases of lung squamous-cell carcinoma (LSC group), 24 cases of normal lung epithelium (NLE group) and 27 cases of lung squamous intraepithelial lesions (SIL group). The correlation of clinical and pathological factors was analyzed. The protein expression of NET-1 in human lung squamous-cell carcinoma cell lines H226, H1703, H2170, SK-MES-1, H520 and YTMLC-90 was determined by Western blot. The RNA interference recombinant adenovirus against NET-1 gene (Ad-NET-siRNA) and Ad-control with control sequence were constructed and infected with human lung squamous cell carcinoma cell YTMLC-90 to silence the expression of NET-1 gene. The protein expression of NET-1, E-cadherin, vimentin and Snail1 in the BEAS-2B cells and the YTMLC-90 cells was determined by Western blot. The mRNA expression of E-cadherin and vimentin in each group of the cells was detected by qPCR. The invasive ability of the cells in each group was detected by Transwell chamber assay. RESULTS: The positive expression rate of NET-1 in LSC group was significantly higher than that in NLE group and SIL group(P<0.05). The distribution of NET-1 protein positive expression population was correlated with histological grade, lymph node metastasis, and TNM stage. The NET-1 expression rate of LSC with lymph node metastasis was significantly higher than that without lymph node metastasis. Over-expression of NET-1 protein in YTMLC-90 cells was observed. The expression of E-cadherin was decreased, and the protein expression of vimentin and Snail1 was increased in YTMLC-90 cells. Knock-down of NET-1 expression increased the expression of E-cadherin, and decreased the expression of vimentin and Snail1 in the YTMLC-90 cells. CONCLUSION: The expression of NET-1 promotes the lymphatic metastasis of lung squamous-cell carcinoma. This promotion may be achieved through the activation of epithelial-mesenchymal transition (EMT) by NET-1 expression.     

Expression and significance of thrombospondin-1 in myocardium of type 2 diabetic cardiomyopathy rats

AIM: To investigate the expression and significance of thrombospondin-1 (TSP-1) in left ventricular myocardium of type 2 diabetic cardiomyopathy (DCM).METHODS: The rat model of DCM was established by eating a high-fat diet together with injection of low dose streptozocin (30 mg/kg) intrapertoneally.After 12 weeks,the content of collagen was quantified by Masson staining.The mRNA level of TSP-1 was determined by quantification real-time RT-PCR,while the protein level of TSP-1 was analyzed by Western blotting and immunohistochemistry.RESULTS: Compared with the control group,the content of collagen in the DCM group was increased greatly (11.01±3.05 vs 16.92±3.18,P<0.01).The mRNA and protein expressions of TSP-1 were significantly higher than those in control group (0.0089±0.0034 vs 0.0141±0.0037,P<0.05；96.38±16.80 vs 129.98±16.96,P<0.05).In DCM group,the mRNA and protein expressions of TSP-1 showed significantly positive correlations with the levels of fasting blood glucose and collagen (r=0.762,P<0.01; r=0.717,P<0.05; r=0.735,P<0.01; r=0.750,P<0.01).There was a significantly positive correlation of TSP-1 mRNA level with LVEDP (r=0.658,P<0.05).In contrast,there was a significantly negative correlation of TSP-1 protein with LVSP and -dp/dtmax (r=-0.605,P<0.05; r=-0.694,P<0.05).There was a significantly positive correlation of TSP-1 protein with LVEDP (r=0.716,P<0.05).There was a significantly negative correlation of TSP-1 protein with LVSP and -dp/dtmax (r=-0.633,P<0.05; r=-0.669,P<0.05).CONCLUSION: The increased expression of TSP-1 may play an important role in the development of myocardial interstitial fibrosis in DCM.     

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 of WNT5B in breast cancer and its clinical significance

AIM: To study the expression of WNT5B in the breast cancer and further to discuss the correlation between WNT5B and clinicopathologic characteristics of breast cancer. METHODS: The expression of WNT5B at mRNA and protein levels was measured by real-time PCR and Western blot in 67 cases of breast cancer and the tissue adjacent to carcinoma. In addition, the immunohistochemical method was used to detect the expression of WNT5B in the breast cancer and the tissue adjacent to carcinoma. The relationships between WNT5B expression and clinicopathologic indexes were also analyzed. RESULTS: The expression of WNT5B in the breast cancer was obviously lower than that in the tissue adjacent to carcinoma (P<0.05). The expression of WNT5B at mRNA and protein levels in 67 samples of breast cancer was in various degrees. The expression of WNT5B in T≤20 mm group of human breast cancer was obviously higher than that in T>20 mm group (P<0.05). The expression of WNT5B had no obvious correlation with axillary lymph node metastasis, histological grade and immunohistochemical indexes of ER, PR, c-ErBb-2, p53 and Ki67 (P>0.05) in the breast cancer. CONCLUSION: The expression of WNT5B decreases obviously in breast cancer. The expression of WNT5B is related to primary tumor size, which provides new ideas for the diagnosis and treatment of breast cancer, suggesting that WNT5B may be a new molecular marker for prognosis of breast cancer.     

Mesenteric lymph drainage alleviates spleen injury in hemorrhagic shock rats

AIM：To observe the effects of post-shock mesenteric lymph (PSML) drainage on histopathology, apoptosis, cell cycle and proliferation of the spleen in rats with hemorrhagic shock. METHODS：Eighteen Wistar rats were randomly divided into sham, shock and shock+drainage groups (n=6 in each group). The hemorrhagic shock model was established in the shock and shock+drainage groups. Fluid resuscitation for 30 min was performed 1.5 h after hypotension, and PSML was drained in the rats in shock+drainage group from 1 h after hypotension to 3 h after resuscitation finished. The fixed spleen tissue was harvested from each rat for histological observation with HE staining. The apoptosis of splenocytes was observed by Hoechst 33258 staining. The expression of Bcl-2 and Bax proteins was detected by immunohistochemical staining. The cell cycle and the expression of p53 protein were measured by flow cytometry, and the proliferation index (PI) was calculated. RESULTS：Compared with sham group, splenic tissue injury appeared in the shocked rats. The apoptotic cells and the expression of Bax and p53 in shock group were increased, while Bcl-2 expression was decreased. The percentage of G2/M cells in shock group was decreased. Compared with shock group, the splenic tissue damage in shock+drainage group was significantly attenuated. Moreover, the number of apoptotic cells, the percentage of G0/G1 cells, and the expression of Bax and p53 were obviously decreased, and the G2/M cells, Bcl-2 protein expression and PI were significantly increased in shock+drainage group. CONCLUSION: PSML drainage alleviates splenic injury in hemorrhagic shock rats, which may be related to reducing the apoptosis of splenocytes.     

Mortalin expression in cervical squamous-cell carcinoma and its effect on tumor metastasis

AIM: To investigate the significance of mortalin expression in clinical pathology of cervical squamous-cell carcinoma. METHODS: Immunofluorescence staining was used to detect the location of mortalin in human cervical squamous-cell carcinoma SiHa cells. The protein expression of mortalin was detected in 59 cases of normal cervical epithelial tissues and 93 cases of cervical squamous-cell carcinoma tissues by immunohistochemical staining, and its correlation with clinicopathological features of cervical squamous-cell carcinoma was also analyzed. MTT assay was used to evaluate the optimal concentration and dosing time of mortalin inhibitor MKT-077. After the protein expression of mortalin in SiHa cells was inhibited, wound-healing and migration assays were performed. The protein expression of epithelial-mesenchymal transition (EMT)-related molecules was determined by Western blot. RESULTS: Immunofluorescence staining showed that mortalin was located in the cytoplasm of SiHa cells. The positive rate and strongly positive rate of mortalin in the cervical squamous-cell carcinoma patients were 88.7% (55/62) and 61.3% (38/62), respectively, and they were significantly higher than those in normal cervical epithelial tissues (23.7% and 5.1%, P < 0.01). Additionally, mortalin expression was statistically correlated with the histological grade, clinical stage and lymph node metastasis. After inhibiting the expression of mortalin in the SiHa cells by MKT-077, the results of wound-healing and migration assays showed that the migration ability of SiHa cells was down-regulated. The protein expression of E-cadherin was up-regulated, and vimentin and Snail were significantly down-regulated. CONCLUSION: Mortalin over-expression is an effective biomarker for prediction of malignant potential and poor prognosis of cervical squamous-cell carcinoma.     

Protein and mRNA expression of KAI1 metastasis suppressor gene and its clinical significance in cervical carcinoma

AIM: To study the mRNA and protein expression of Kang ai1 (KAI1) tumor suppressor gene and to determine the relationship between KAI1 and invasiveness and metastasis of cervical cancer. METHODS: The expression of KAI1 metastasis suppressor was detected by immunohistochemistry in paraffin slides and by real-time quantitative polymerase chain reaction (RT-PCR) in fresh tissue. The samples included 20 cases of normal cervical tissues, 20 cases of cervical intraepithelial neoplasia (CIN) and 40 cases of cervical carcinoma. The results of the gene expression combined with the pathological and clinical data were also analyzed. RESULTS: The expression of KAI1 protein and mRNA was related to the tissue differentiation of cervix. The positive rates of KAI1 expression were the highest in the normal cervical tissue, the middle in CIN and the lowest in cervical carcinoma with significant difference among three groups (P<0.01). The expression of KAI1 protein was not related with the grade of CIN (P>0.05). However, both mRNA and protein expression of KAI1 were related to the differentiation and the clinical stages of cervical cancer (P<0.01) and also related to the metastasis of the cancer. The positive rates between the non-lymphatic metastasis and lymphatic metastasis (P<0.05) were significant different. Cox regression and logistic regression showed that the tissue differentiation, clinical stages, lymphatic metastasis and expression of KAI1 were all related factors with recurrence and prognosis of cervical cancer. CONCLUSION: The down-regulation of KAI1 tumor suppressor gene at both mRNA and protein levels is related to the differentiation, clinical stages and metastasis of cervical cancer, indicating that the expression of KAI1 is a prognostic factor for cervical cancer.