Basic & Clinical Medicine

New thoughts on efficient endoscopic screening in high-risk areas of esophageal cancer in china

2006, 26(10): 1047-1047.

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The incidence rates of esophageal cancer have been decreasing since 1988 in Linxian and Cixian as the results of year-round preventive effort for several decades. But remarkable decreasing in incidence is confined to age groups above the age of 60 year old only. For the age group from 40 through 59, the decreasing is slight; and for those under the age of 40 years old, no decrease can be perceived. This may suggest that the primary risk of esophageal cancer still remains as strong in the high-risk areas, and the decreased incidence may be credited mainly to secondary preventive measures. Although early diagnosis is still the key to control esophageal cancer presently, the rate of early detection is very low due to the lacking of efficient screening programs in the high-risk areas. Recently, Cancer Foundation of China initiated a series of endoscopic screening programs across the country aimed to improve early detection and treatment in the field. The objective of this article is to provide clues to help improve screening efficiency in the field.

The regulatory effect of hydrogen sulfide on the content of collagen in aorta of spontaneous hypertensive rats

2006, 26(10): 1052-1052.

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Objective To study the regulatory effect of hydrogen sulfide (H2S) on the accumulation of collagen in thoracic aorta of spontaneous hypertensive rats (SHR). Methods Seventeen male SHR rats were randomly divided into SHR group (n=6), SHR+H2S group (n=6), SHR+PPG group (n=5). Five WKY rats were included as WKY group (n=5). NaHS, which is H2S donator, was injected peritoneally innto rats of SHR+H2S group with a dosage of 56mmol/kg everyday for 5 weeks. For rats in SHR+PPG group, D, L-propargylglycine (PPG), which is CSE inhibitor, was injected peritoneally with a dosage of 37.5mg/kg for 5 weeks. The other two groups of rats were injected with the same volume of sterile water. At the age of 9 weeks, the arterial blood pressure was measured by tail capacity method. Using the thoracic aorta as study object, the enzymatic activity of cystathionine g-lyase (CSE) was determined by enzyme facilitating reaction; the hydroxyproline quantification was measured by hyperthermia splitting method; the amount of collagen type Ⅰ was determined by ELISA method; the expression of transforming growth factor b3 (TGF-b3) was detected by immunohistochemistry method. Results For rats in SHR group, the blood pressure increased by 52%; the CSE activity decreased by 55% (p<0.01); the content of hydroxyproline increased by 87% (p<0.01); the amount of collagen type Ⅰ increased by 28%; the expression of TGF-b3 in the aortic tissue of SHR group increased by 10% (p<0.05) compared with rats in WKY group. Supplying with exogenous H2S significantly alleviate the changes of these parameters above. Conclusion Supplying exogenous H2S to SHR rats could decrease the blood pressure and reduce the content of hydroxyproline and collagen type I in the vessels, therefore, lessen the accumulation of collagen in the thoracic aorta of SHR.

Epidermal growth factor promotes mouse oocyte maturation in vitro:potential role of the c-erbB2 protooncogene

2006, 26(10): 1056-1056.

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objective Potential roles of the C-ERBB2 protooncogene in contributing to mouse oocyte maturation and in conducting Epidermal Growth Factor (EGF) promoteing mouse oocyte maturation in vitro were investigated. Methods This research used mouse oocyte culture model and RT-PCR method to study the effects of both C-ERBB2 and EGF on mouse oocyte maturation in vitro, and the potential role of the C-ERBB2protooncogene on EGF effect. Results C-ERBB2 ASODN inhibited the rate of germinal vesical breakdown(GVBD) and the first polar(PBⅠ) extrusion of denuded oocytes(DOs) in a dose-dependent and time-dependent way, and delayed their maturation significantly. C-ERBB2ASODN also inhibited the effect of EGF on oocyte maturation in a time-dependent way. In GVBD oocytes, RT-PCR showed that C-ERBB2mRNA was expressed in oocytes and the level of C-ERBB2mRNA was the highest in EGF treated group, midst in EGF+c-erbB2ASODN treated group, and lowest in C-ERBB2 ASODN treated group. Conclusions Our results indicate that C-ERBB2 exists in the oocyte and has promoting effect on oocyte maturation in vitro. EGF plays a role in inducing denuded oocyte maturation through up-regulating C-ERBB2expression of oocyte. Interfering C-ERBB2 expression can reduce the effect of EGF on oocyte maturation.

The changes and significanfce of urotensin II in rat nodels of chronic obstructive pulmonary disease

2006, 26(10): 1063-1063.

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Objective　To investigate the roles of urotensin II (U-II) in rat with Chronic Obstructive Pulmonary Disease(COPD). Methods Male SD rats were randomly divided into chronic bronchitis group (group B), COPD group (group C), exsmoker group (group D) and control group (group A). The concentrations of U-II in plasma, BALF and lung tissues were measured by RIA. Lung function and pathological analysis of lung specimens were performed routinely. Results　There were no significant differences of plasma U-II levels in each groups. The BALF levels of U-II were 1.7, 2.2, 4.7, 4.9 folds higher than plasma levels in group A, B, C and D respectively (all P<0.01). The BALF levels of U-II from group B,C and D were 50%, 225%, 225% higher than those in group A respectively (all P<0.01).The lung tissue levels of U-II were also higher than those from the controls respectively (all P<0.05). The BALF levels of U-II correlated positively with the neutrophils counts in the BALF, with the pathological scores of airway inflammatory cell infiltrations, airway muscles and fibrous connective tissues proliferation (all P<0.01). There was a negative correlation between the BALF levels of U-II with the peak expiratory flow (P<0.01). Conclusion In the rat model of COPD, U-II levels in BALF and lung tissues were increased, indicating that it may act locally, or, via paracrin/autocrine way, play a role in the mechanism of the airway remodeling in COPD.

Construction of a recombinant human acidic fibroblast growth factor and its expression

2006, 26(10): 1067-1067.

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Objective To investigate the feasibility of recombinant adeno-associated virus encoding secrected forms of human acidic fibroblast growth factor transfecting endothelial progenitor cells. Methods sp-haFGF was obtained through combining signal peptide sequence of FGF-4 with native aFGF gene by PCR. Sp-haFGF was cloned into AAV vector plasmid pAAV-IRES-hrGFP . Recombinant AAV encoding sp-haFGF was packaged through co-transfecting HEK293 cells with plasmid sp-haFGF-pAAV-IRES-hrGFP, pAAV-RC and pHelper. Ex vivo cultured EPCs were infected with concentrated rAAV . Expression of green fluorescent protein(GFP) in EPC infected was observed by fluorescence microscope and expression of sp-haFGF in EPCs was verified by RT-PCR and western blot analysis. Results Recombinant

Inhibition of gene expression of Nrf2 by RNAi in Caco-2 cells

2006, 26(10): 1072-1072.

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Objective To identify the RNA interference action of recombined pSUPER-NRF2 vectors for the expression of NRF2 gene in colon cancer cells. Methods Two sequences targeting the ORF of NRF2 were cloned into the RNA polymerase III based expression vector pSUPER. These recombinants were transfected into Caco-2 cells. Fluorescence microscopic examination and Flow cytometry were performed after being transfected with pEGFP-N1 plasmids to observe the lipfectin transfection efficiency. The stable cells were selected in medium 48h after co-transfected pEGFP-N1 with G418. The expression of NRF2 was assayed using RT-PCR and Western blot. Results The construction of the recombinant expression vector pSUPER-NRF2-A1、B1 and its control vector pSUPER-NRF2-A2、B2 were successfully confirmed by the results of enzyme digestion, electrophoresis and sequencing. The transfection efficiency were 45.6％、74.3％、53.0％ and 46.5％ respectively in 24、48、72 and 96h。We compared the ability of these vectors to inhibit NRF2 in a transient and stable expression experiment. Importantly, pSUPER- NRF2-B1 was able to significantly knockdown NRF2 expression. pSUPER- NRF2-A1 only had a moderate activity, whereas pSUPER- NRF2-A2、B2 were inactive in this assay. Conclusion The constructed pSUPER-NRF2-A1、B1 showed an interfering effect on the expression of NRF2 and could product the stable cells with low NRF2 expression. Therefore，the pSUPER vector constitutes a new and powerful system to analyze NRF2 gene function in colon cancer.

Proliferation of vascular smooth muscle cells regulated by NO/PKG mediated via Ca2+/calcineurin signaling pathway

2006, 26(10): 1078-1078.

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Objective Here we studied regulation of VSMC proliferation by NO/PKG mediated via modulating intracellular Ca2+/calcineurin (CaN) signaling pathway. Methods Primary VSMCs from rat aorta were used as the experimental model. CaN protein and its activity were assayed using immunoblotting and free inorganic phosphate content analysis, respectively. growth and survival rate of cells were determined by MTT assay and acridine orange and ethidium bromide staining. Results The addition of SNAP and Sp-8-pCPT-cGMPS decrease absorbance of cells stimulated by phenylephrine (PE), whereas the addition of Rp-8-pCPT-cGMPS increases it. In SMCs pretreated with Ver, absorbance of cells stimulated by PE decreased and was further inhibited by the additional treatment of SNAP and Sp-8-pCPT-cGMPS. In SMCs pretreated with CsA, absorbance of cells stimulated by PE decreased, but it could not be further altered by the additional treatment of SNAP, Sp-8-pCPT-cGMPS and Rp-8-pCPT-cGMPS. Moreover, expression and activities of CaN induced by PE was inhibited by SNAP and Sp-8-pCPT-cGMPS. Conclusions NO/PKG inhibits the proliferation of vascular SMCs without decreasing cell survival rate, which is mediated via intracellular Ca2+/CaN signaling pathway.

INhibition of human lung carcinoma-associated antigen gene expression and is effect on the growth characteristic and associated-gene expression of L78 cells

2006, 26(10): 1083-1083.

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Objective To study the effect of inhibition of human lung carcinoma-associated antigen gene (ALT04-AG) expression on the growth characteristic and associated gene expression of human lung carcinoma cells (L78). Methods L78 cells were transfected with recombinant plasmid vector expressing ALT04-AG antisense RNA mediated by liposome reagent, and treated with DFMO, respectively. The growth characteristics were studied by MTT, FCM analysis. The associated gene expression was detected by immunocytochemical staining，Northern blot and gene chip analysis. Results The recombinant plasmid expressing ALT04-AG antisense RNA（ pALT04-AG(as)） constructed. It was shown that down-regulation of ALT04-AG gene expression was observed not only in pALT04-AG(as) transfected L78 cells, but also did in DFMO-treated L78 cells, sequentially resulted in growth inhibition of these cells. Gene chip analysis indicated that inhibition of ALT04-AG gene expression by ALT04-AG antisense RNA could regulate some proliferation associated gene expression, while inhibition of polyamine biosynthesis by DFMO could regulate proliferation associated genes and also some apoptosis associated genes. Conclusions It is suggested that human lung carcinoma associated antigen gene ALT04-AG plays an essential role in maintaining malignant phenotype of cancer cells. The inhibition of ALT04-AG gene expression of L78 cells either by pALT04-AG(as) transfection or by polyamine biosynthesis inhibition with DFMO could promote the reversion of malignant phenotype of cancer cells, but the effect of treatment with DFMO on tumor-associated gene expression regulation is widespread than that of pALT04-AG(as) transfection. These results provide a significant clue to search new target for diagnosis and therapy of cancer.

Expression and clinical significance of cyclooxygenase-2 and VEGF in human astrocytomas

2006, 26(10): 1089-1089.

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Objective To investigate the expression of cyclooxygenase-2(COX-2) and vascular endothelial growth factor(VEGF) and their relationship with the clinicopathological characteristics of human astrocytomas.Methods Immunohistochemistry method was used to investigate the expression of COX-2 and VEGF among human astrocytoma samples (n=50) and normal brain tissue (n=10) respectively. Results The positive rate of COX-2 and VEGF was significantly different between the human astrocytomas and normal brain tissue（P＜0.01）.The expression of COX-2 in human astrocytomas was significantly related to the tumor's pathological grades（P＜0.05）.So was the expression of VEGF（P＜0.05）.The expression of VEGF was closely associated with COX-2（P＜0.01）.Conclusions COX-2 and VEGF may be involve in gliomas'occurance and development.Expression of COX-2 and VEGF protein was positively correlate with the malignant phenotype of gliomas. Key words Human astrocytomas; Cyclooxygenase-2; vascular endothelial growth factor ;immunohistochemistry

Induction of heme oxygenase-1 prolongs cardiac grft survivai in rats

2006, 26(10): 1093-1093.

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Objective To study the effect of heme oxygenase-1(HO-1) on cardiac graft survival in rats and its mechanism. Methods Male LEW (RT1l) and LEW.1W(RT1u) rats were used as donor and recipient of heterotopic heart transplantation,respectively.Recipient rats were treated with CoPPIX (5mg/kg/day，n=6),SnPPIX (5mg/kg/day，n=6)and control(n=6),starting from one day to rejection.Histology,Immunohistochemistry staining ,HO activity assay and Western blot of HO-1 were performed using cardiac grafts.Mixed lymphocyte cultures were completed using irradiated LEW.1W splenocytes with splenocytes derived from cardiac recipients. Grafts were excised from recipient at 5 days after transplantation. Results Induction of HO-1 with CoPPIX resulted in a significant prolongation of graft survival,the median survival time of the grafts was 13.5 days(p<0.05).The median survival time of SnPPIX treatment and control group were 6.5 and 7.0 respectively.In the grafts of control and SnPPIX treatment group,there is significant cellular infiltration.Treatment of CoPPIX induced the cellular infiltrate of CD4+,CD25+,Ki67+ cells. CoPPIX treatment significantly suppressed donor reactive spenocyte proliferation when compared to control and SnPPIX treatment(p<0.05). Conclusions Induction of HO-1 inhibits alloreactive lymphocyte activity and prolongs cardiac graft survival in rats.

Effects of NS-398 on angiogenesis of hepatocullar carcinoma cell cline of SMMC-7721 in vivo and vitro

2006, 26(10): 1099-1099.

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Objective To explore the role of NS-398 on MVD and VEGF、MMP-9 expression of hepatocullar carcinoma. Methods VEGF and MMP-9 expressions were detected by RT-PCR and flow cytometry. Microvascular density of xenograft tissue were detected by immunohistochemical staining and the serum PGE2 was detected by ELISA. Results The expressions of VEGF and MMP-9 were significantly downregulated in SMMC-7721 cells exposed to NS-398 in a dose-dependence and time-dependent manner (P<0.001). Expression of VEGF, MMP-9 mRNA and protein in xenografts treated with NS-398 were lower than those of control group (P<0.001). The density of microvessel was notably lower in xenografts treated with NS-398 than in those without treatment and serum PGs level was also significantly lower than in those without treatment (P<0.001). Conclusions Expression of COX-2 contributes to angiogenesis in hepatocullar carcinoma. NS-398 may suppress angiogenesis of hepatocellular carcinoma cell via reducing expressions of VEGF and MMP-9.

Effects of carbamazpine on morphology and GFAP expression of astrocytesin hippocampus of rats

2006, 26(10): 1104-1104.

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Obiective To observe the effects of carbamazepine on morphology and glial fibrillary acidic protein (GFAP)expression of astrocytes in hippocampus of Sprague-Dawley rats.Methods Once daily intragastrically,in therapeutical dose for 1 week,1 and 3 months, the changes of astrocytic expression were observed with GFAP immunohistochemical staining. Results Compared to control group,GFAP immunoreactivity and the number of GFAP- positive astrocytes were significantly increased after 3 months of carbamazepine administration.Conclusion It is concluded that the changes of astrocytic expression induced by carbamazepine are time-dependent .

Expression of Sterol Carrier Protein 2 and Its Correlation with the Gallstone Formation

2006, 26(10): 1108-1108.

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Objective To analyze the level of hepatic sterol carrier protein 2 (Scp2) mRNA and bile cholesterol saturation index(CSI) during gallstone information in C57BL/6 mice. Methods Twenty C57BL/6 mice were divided into two groups equally: control group and gallstone group. The gallstone group mice were fed with lithogenic diet (containing 1% cholesterol) for 4 weeks. The mRNA level of Scp2 in liver tissue was measured by RT-PCR. The content of biliary lipids and CSI were analyzed by Automatic Biochemistry Analyzer. Results The levels of Scp2 mRNA and bile CSI were significantly increased in gallstone group. Conclusion A positive correlation between the expression of liver Scp2 and bile CSI was found during gallstone formation.

Effect of small interference RNA of HPV18 E6 gene on apoptosis in Hela cells

2006, 26(10): 1112-1112.

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Objective To study the effect of small interfering RNA (siRNA) of human papillomavirus (HPV) 18 E6 gene on apoptosis in HPV-related cervical HeLa cell line. Methods siRNA targeting HPV18 E6 mRNA were designed and generated by PCR amplification. The PCR products containing U6 promoter and the siRNA sequence were then transfected into HeLa cells via LipofectamineTM2000. At different times after transfection, cell viability was determined by MTT assay. Apoptosis was detected by morphological observation and flow cytometry analysis. The expression levels of HPV18 E6 mRNA was assayed by RT-PCR. Results The cell growth and viability of siRNA transfected group were significantly inhibited（P＜0.05）. There were obvious morphological changes in HeLa cells after transfection of siRNA under optical microscopy and the apoptosis rate at 72h was 55.8%. HPV18 E6 mRNA expression in the cells of the interference group at 24、48 and 72h after transfection were significantly reduced by about 57%、78% and 40% respectively, as compared with that of the negative and blank control groups, whereas the latter two groups had similar expression levels. Conclusions Interference with siRNA against HPV18 E6 gene can effectively inhibit the specific gene expression which may play significant role of the induction of apoptosis of the corresponding Hela cells.

Effects of cardiomyopeptidin on transient outward potassium current in ventricular myocytes of rats

2006, 26(10): 1118-1118.

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Objective: To determine the effect of cardiomyopeptidin on transient outward potassium current (Ito) of rat ventricular myocytes and its action mechanism on the ion channels of myocardium. Methods: Single ventricular myocytes of rats were obtained by enzymatic dissociation method. The whole-cell patch-clamp recording technique was used to record the change of transient outward potassium current (Ito) by different dosage of cardiomyopeptidin. Results: Cardiomyopeptidin decreased Ito in a dose-dependent manner. Cardiomyopeptidin in dose of 10, 50, 100, 250 and 500 mg/L decreased Ito (%) by 4, 13, 22, 32 and 38 respectively. Cardiomyopeptidin 50 mg/L moved down the current density-voltage curve of Ito, but the shape of the curve had no changes. Cardiomyopeptidin 50 mg/L did not change the steady state activation curve of Ito. Conclusions: Cardiomyopeptidin decreases the Ito of rat ventricular myocytes, which might be one of the mechanisms of its antiarrhythmic effect.

Expression of survivin and bcl-X gene in cervical carcinoma and their relationship

2006, 26(10): 1122-1122.

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The establitshment of human mesangial cell line with high expression of type A scaenger receptor and receptor function study

2006, 26(10): 1125-1125.

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Tumor necrosis factor alpha and plasma adiponectin concentration in women with gestational diabetes

2006, 26(10): 1131-1131.

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