Objective:To observe the expression patterns of miR-195 and DLL4 in colorectal cancer, and to explore the relationship between miR-195 and DLL4 and clinicopathological parameters and prognosis of patients with colorectal cancer.Methods:The relative expression of miR-195 in 56 colorectal cancer tissues was detected by real-time fluorescent quantitative PCR the expression of DLL4 protein and tumor microvessel density (MVD) were detected by bloting and immunohistochemistry. Colon cancer cell line SW480 was treated with miR-195. The expression of DLL4, Jagged1, (the intracellular domain of notch, NICD), CyclinD1, Hes1, Bcl-2 and NF-kB were detected by bloting.Results:The expression level of DLL4 protein in colorectal cancer tissues was significantly higher than that in normal intestinal mucosa (30/56 vs.16/56, t=5.323, P=0.018). The expression level of miR-195 was significantly lower than that in normal intestinal mucosa (36/56 vs.20/56, t=2.371, P=0.008). The expression of DLL4 was negatively correlated with the expression of miR-195 ( r=- 0.881, P=0.015) , which was closely related to the differentiation degree, lymph node metastasis and TNM stage . The prognosis of patients with high expression of DLL4 was significantly worse than that with low expression of DLL4 ( P=0.013). The prognosis of patients with low expression of miR-195 was worse than that with high expression of miR-195 ( P=0.009) . Conclusion:The antagonistic expression of miR-195 and Notch might be closely related to the occurrence and development of colorectal cancer, which can be used as a new reference index for prognosis of colorectal cancer.

This study aimed to validate the high yield and soluble expression of proteins carrying the transactivator of transcription (Tat) peptide tag, and further explored the potential mechanism by which the Tat tag increases expression. Escherichia coli superoxide dismutase (SOD) proteins, including SodA, SodB and SodC, were selected for analysis. As expected, the yields and the solubility of Tat-tagged proteins were higher than those of Tat-free proteins, and similar results were observed for the total SOD enzyme activity. Bacterial cells that overexpressed Tat-tagged proteins exhibited increased anti-paraquat activity compared with those expressing Tat-free proteins that manifested as SodA>SodC>SodB. When compared with an MG1655 wild-type strain, the growth of a ΔSodA mutant strain was found to be inhibited after paraquat treatment; the growth of ΔSodB and ΔSodC mutant strains was also slightly inhibited. The mRNA transcript level of genes encoding Tat-tagged proteins was higher than that of genes encoding Tat-free proteins. Furthermore, the α-helix and turn of Tat-tagged proteins were higher than those of Tat-free proteins, but the β-sheet and random coil content was lower. These results indicated that the incorporation of the Tat core peptide as a significant basic membrane transduction peptide in fusion proteins could increase mRNA transcripts and promote the high yield and soluble expression of heterologous proteins in E. coli.
Escherichia coli* ; Escherichia* ; HIV-1* ; Membranes ; Paraquat ; RNA, Messenger* ; Solubility ; Superoxide Dismutase ; Superoxides* ; Trans-Activators

Objective To study postoperative delirium in elderly patients.Methods We investigate the morbidity of postoperative delirium in 142 elderly patients (≥ 60 years)after gastrointestinal surgery by using Confusion Assessment Method (CAM) and Delirium Rating Scale Revised-98 (DRS-R98) scores.Data were analyzed using Student's t test and Chi-squaretest respectively with SPSS 19.0.Results Of 142 patients,delirium was diagnosed in 36 patients(25.4％),delirium developed in 4,7,17,7,1 patients in posto perative 1,2,3,4-7,7 + days respectively.There were significant difference in hospital stay:17.7 ± 2.6 days (postoperative delirium) and 13.4 ± 2.3 days (no postoperative delirium),t =4.608,P =0.000 1.The postoperative complications (52.8％ / 23.6％,x2 =10.710,P =0.001) and ICU admission (22.2％/6.6％,x2 =6.939,P =0.008) significantly increased.Conclusions Postoperative delirium is recognized as one of the most common surgical complications in elderly patients with gastrointestinal surgery leading to other major postoperative complications,and prolonged hospitalization.

Objective To investigate the expression and significance of IDO and c-myc in bladder urothelial carcinoma.Methods IDO mRNA expression from fresh tumor and mucosa specimens from 20 cases were detected by fluorescence quantitative PCR,and SP was used to detect the IDO,c-myc protein in paraffin-embeded specimens from 84 cases and mucosa specimens from 22 cases.Results In bladder urothelial carcinoma,IDO,c-myc protein expression level had no relationship with age and sex.Expression of IDO protein in invasive bladder urothelial carcinoma was significantly higher than that of no-invasive (x2 =5.600,P =0.018).With the increase of the histological grade (x2 =20.268,P =0.000) and UICC stage (x2 =12.075,P =0.007),the positive expression rate of IDO protein was increased.There was not positive expression of c-myc protein in normal bladder tissue,but in bladder urothelial carcinoma,44 cases (52.4 ％) were positive expression (x2 =10.733,P =0.001).The expression of c-myc protein had no relationship with the histological classification,histological grade and UICC stage.In bladder urothelial carcinoma tissue,IDO protein expression level was positively correlated with c-myc protein expression (r =0.205,P =0.047),and was positively correlated with the histological classification,histological grade and UICC stage (r =0.258,P =0.018; r =0.491,P =0.000; r =0.365,P =0.001).Expression of IDO mRNA in bladder urothelial carcinoma (7.696 1±1.745 2) was significantly higher than that in normal bladder tissue (6.397 0±1.205 1)(t =2.367,P =0.023).The average expression level of IDO mRNA in bladder urothelial carcinoma of Ta-T1 stage was 6.803 4±1.567 5,which was significantly lower than that in T2-T4 stage (9.183 8±0.690 3) (t =4.955,P =0.000).The average expression levels of IDO mRNA in grade Ⅰ,Ⅱ,Ⅲ bladder urothelial carcinoma were 7.058 7±1.771 5,7.934 2±1.530 5,9.290 7±0.574 5,respectively,which were increased with the grade increasing (t =2.729,P =0.011).Conclusions The high expression of IDO correlates with bladder urothelial carcinoma progression,and expression of c-myc is irrelevant with bladder urothelial carcinoma progression,but maybe associate with bladder urothelial carcinoma occurrence.IDO may be a predictive factor of prognosis.IDO and c-myc may be therapeutic target in the treatment of bladder urothelial cancer.

Objective To investigate the expression and significance of bin 1,C-myc in bladder urothelial carcinoma.Methods SP method was used to detect the expression of bin 1,C-myc protein in paraffin specimens from bladder urothelial carcinoma of 84 cases and mucosa of 11 cases,and its relationship with clinical pathological charac-teristics was analyzed.Results The positive expression rate of bin1 in normal bladder tissues was 81.81%,that in bladder urothelial carcinoma was 46.43%,the difference was not significant (χ2 =4.873,P=0.051).No expression of C-myc was observed in normal bladder tissue ,the positive expression rate of C-myc in bladder urothelial carcinoma was 52.38%,the difference was statistically significant (χ2 =10.733,P=0.001).The expression of bin1 in invasive bladder urothelial carcinoma was significantly lower than the non-infiltrating type(χ2 =7.685,P =0.007),with increased histological grade and UICC stage ,the positive rate of bin1 expression decreased (χ2 =15.817,P=0.000;χ2 =11.104,P=0.010).The expression of C-myc had no relationship with the histological classification ,histological grade and UICC stage .Correlation analysis showed that the expression of bin 1 was negatively correlated with histologi-cal classification,histological grade and UICC stage (r=-0.302,P=0.005;r=-0.411,P=0.000;r=-0.302, P=0.005).With the increased expression of bin1,C-myc expression decreased,but the difference was not statistical-ly significant.Conclusion Low expression of bin1 or loss of bin1expression were associated with the progression of bladder urothelial carcinoma , the expression of C-myc was related with bladder urothelial carcinoma lesions .The results suggest that bin1 may be predictive factor for prognosis of urinary bladder urothelial carcinomas ;bin1,C-myc may be viewed as molecular targets for tumor chemotherapy .

OBJECTIVETo investigate the effect of propofol on the expression of thrombospondin-1 (THBS-1) mRNA and protein in purified newborn rat cortical astrocytes in vitro.

METHODSAstrocytes were isolated from newborn rat cortex and grown in culture before exposure to propofol at 3, 10, 30, 100 or 300 µmol/L for 6 h, 12, or 24 h. The mRNA level of THBS-1 was detected by RT-PCR, and the protein level of THBS-1 was detected by immunofluorescence cytochemistry and Western blotting.

RESULTSPropofol exposure caused significantly upregulated THBS-1 level in cultured astrocytes (P<0.05) to a level about 1.3 times higher than that in control cells. The mRNA and protein levels of THBS-1 in cultured rat cortical astrocytes were upregulated by exposures to 10, 30 and 100 µmol/L propofol (P<0.01). High expression of THBS-1 mRNA and protein was detected in the cells with exposures for different durations (P<0.05), especially in the 12 h group (P<0.01).

CONCLUSIONPropofol at clinically relevant concentrations can modulate the level of THBS-1 secreted by astrocytes of rat cerebral cortex in vitro.

Animals ; Astrocytes ; drug effects ; metabolism ; Cells, Cultured ; Cerebral Cortex ; cytology ; Propofol ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Thrombospondin 1 ; metabolism

Objective To detect the expressions of enhancer of zeste homology 2 (EZH2) and signal transducer and activator of transcription 3 (STAT3) in gastric cancer tissues,and analyze the relationship between their expressions and clinicopathological factors and prognosis of patients.Methods The clinical data of 67 patients with gastric cancer who were admitted to the People's Hospital of Peking University from May 1999 to April 2000 were retrospectively analyzed.Gastric cancer tissues and adjacent normal tissues were harvested and fixed in 4％ formaldehyde,and then to make the tissue chips.The protein expressions of EZH2 and STAT3 were detected by immunohistochemical staining,and the relationship between the expressions of EZH2 and STAT3 and the prognosis of patients was analyzed.The correlation between the protein expressions of EZH2 and STAT3 in the gastric and colorectal tissues and the clinicopathological parameters was analyzed using the paired chi-square test.The survival curve was drawn by Kaplan-Meier method,and the survival rate was analyzed by using the Logrank test.The relationship between the clinicopathological factors and the prognosis of patients was analyzed using the COX proportional hazard model.Results The positive protein expression rates of EZH2 and STAT3 in the gastric cancer tissues were 73.1％ (49/67) and 67.2％ (45/67),which were significantly higher than 32.8％ (22/67) and 37.3％ (25/67) in the adjacent normal tissues (x2=21.839,11.964,P ＜0.05).The protein expression of STAT3 was correlated with the age,TNM staging,lymph node metastatic status (x2=5.475,9.998,5.475,P ＜ 0.05).The protein expression of EZH2 was correlated with TNM staging and lymph node metastatic status (x2=11.573,5.902,P ＜ 0.05).The co-expression rate of EZH2 and STAT3 proteins was 73.1％ (49/67),EZH2 and STAT3 had the common expression trend (r =0.397,P ＜ 0.05).The co-expression rate of EZH2 and STAT3 was increased as the increase of TNM stages,and the co-expression of EZH2 and STAT3 in the gastric cancer tissue was correlated with the TNM stages (x2 =6.997,P ＜ 0.05).The 5-year survival rate of patients with low protein expression of EZH2 was significantly higher than those with high protein expression of EZH2 (x2=7.386,P ＜ 0.05).The 5-year survival rate of patients with low proteins expression of STAT3 was significantly higher than those with high proteins expression of STAT3 (x2=12.253,P ＜ 0.05).The 5-year survival rate of patients with low co-expression of EZH2 and STAT3 protein was significantly higher than those with high co-expression of EZH2 and STAT3 proteins (x2 =8.765,P ＜ 0.05).The results of univariate analysis showed that age,TNM staging,EZH2 and STAT3 expression,lymph node metastasis and distal metastasis were the factors influencing the survival of patients with gastric cancer (RR =2.136,3.452,3.179,8.341,11.773,6.873,P ＜0.05).The results of multivariate analysis showed that TNM staging and STAT3 expression were independent factors influencing the prognosis of patients with gastric cancer (RR =2.453,7.535,P ＜ 0.05).Conclusions There is significant correlation between EZH2 and STAT3 protein expressions in gastric cancer tissues,and co-expression of EZH2 and STAT3 is associated with the TNM staging and the prognosis of the patients with gastric cancer.

Objective To investigate the effect of propofol on the expression of thrombospondin-1 (THBS-1) mRNA and protein in purified newborn rat cortical astrocytes in vitro. Methods Astrocytes were isolated from newborn rat cortex and grown in culture before exposure to propofol at 3, 10, 30, 100 or 300μmol/L for 6 h, 12, or 24 h. The mRNA level of THBS-1 was detected by RT-PCR, and the protein level of THBS-1 was detected by immunofluorescence cytochemistry and Western blotting. Results Propofol exposure caused significantly upregulated THBS-1 level in cultured astrocytess (P<0.05) to a level about 1.3 times higher than that in control cells. The mRNA and protein levels of THBS-1 in cultured rat cortical astrocytes were upregulated by exposures to 10, 30 and 100 μmol/L propofol (P<0.01). High expression of THBS-1 mRNA and protein was detected in the cells with exposures for different durations (P<0.05), especially in the 12 h group (P<0.01). Conclusion Propofol at clinically relevant concentrations can modulate the level of THBS-1 secreted by astrocytes of rat cerebral cortex in vitro.

Objective To investigate the effect of propofol on the expression of thrombospondin-1 (THBS-1) mRNA and protein in purified newborn rat cortical astrocytes in vitro. Methods Astrocytes were isolated from newborn rat cortex and grown in culture before exposure to propofol at 3, 10, 30, 100 or 300μmol/L for 6 h, 12, or 24 h. The mRNA level of THBS-1 was detected by RT-PCR, and the protein level of THBS-1 was detected by immunofluorescence cytochemistry and Western blotting. Results Propofol exposure caused significantly upregulated THBS-1 level in cultured astrocytess (P<0.05) to a level about 1.3 times higher than that in control cells. The mRNA and protein levels of THBS-1 in cultured rat cortical astrocytes were upregulated by exposures to 10, 30 and 100 μmol/L propofol (P<0.01). High expression of THBS-1 mRNA and protein was detected in the cells with exposures for different durations (P<0.05), especially in the 12 h group (P<0.01). Conclusion Propofol at clinically relevant concentrations can modulate the level of THBS-1 secreted by astrocytes of rat cerebral cortex in vitro.

Objective To explore protein expression and significance of MAGE genes in colorectal carcinoma(CRC)tissues.Methods The expression of MAGE genes were studied by using tissue chip and immunochemistry methods in primary CRC tissue and paired adjacent tissue samples in 97 cases.Data were analyzed with x2-test by SPSS 16.0 software.Results The protein expression of MAGE-A3,MAGE-A4 and MAGE-A10 genes were 57％(56/97),63％(61/97)and 28％(27/97)respectively in 97 cases of primary adenocarcinoma.The protein expression frequency of MAGE-A3 in poor colorectal adenocarcinomas was significantly higher than in well-and moderately disfferentiated adenocarcinomas(x2 =9.133,P =0.010).MAGE-A10 in poor colorectal primary adenocarcinomas was significantly higher than in well and moderately adenocarcinomas(x2 =15.280,P =0.000); MAGE-A10 protein expression was significantly higher in stage TNM Ⅲ + Ⅳ than in stage TNM Ⅰ + Ⅱ(x2 =4.227,P=0.040); MAGE-A10 gene expression was higher in metastasis lymphoid node than in no metastasis lymphoid node(x2 =5.557,P =0.018),and the expression level was higher in primary lesion with the increasing of the numbers of lymphoid node metastasis(x2 =7.296,P =0.026).Conclusions The protein expression of MAGE genes is associated with the tumor differentiation,TNM stage and lymphoid node metastasis.MAGE-A3 and MAGE-A10 genes are the possible prognosis marker and potential target of immunotherapy of CRC.