Guobin Wang

Endoscopic minimal invasive cholecystolithotomy vs laparoscopic cholecystectomy in treatment of cholecystolithiasis in China: A meta-analysis

INTRODUCTION Endoscopic minimal invasive cholecystolithotomy (EMIC) is recently popular in China which may offer advantages over laparoscopic cholecystectomy (LC). We try to find out the most favorable treatment for the patients underwent cholecystolithiasis. METHODS Databases PubMed, Elsevier, Wiley Online Library, The Cochrane library, CNKI, WanFang Data, and Chongqing VIP were searched for randomized controlled trials (RCTs) and on EMIC vs LC from 2009 to 2013. Odds ratio (OR), risk difference (RD) and weight mean difference (WMD) were calculated with 95% confidence intervals (CI). RESULTS 14 RCTs including 2030 patients were selected. No significant difference was present in operating time between EMIC and LC. EMIC shown significant less blood lost (WMD -23.45; 95% CI -30.34, -16.55; Z=6.66; P<0.00001) compared to LC. Shortened exhaust time (WMD -14.11; 95% CI -18.34, -9.88; Z=6.53; P<0.00001) and hospital stay (WMD -1.31; 95% CI -1.91, -0.71; Z=4.29; P<0.00001) were present in EMIC group. And EMIC shown decreased complication proportion (OR -0.14, 95% CI -0.09 to -0.21; Z=8.53; P<0.00001) in comparison with LC. There is no difference present in the recurrence of stones in two procedures but a significantly decreased recurrence rate of gallstones was present in EMIC compared to conventional cholecystolithotomy. CONCLUSION Patients treated with EMIC shown faster recovery and less complication which were superior to LC.

Expression of NALP3 in the spleen of mice with portal hypertension

SummaryThis study examined the mRNA expression of NALP3 in the spleen of the mice with hypersplenism due to portal hypertension (PH). The mouse hypersplenism models were established by oral administration of tetrachloromethane (2 mL/kg/week for 12 weeks by oral gavage). All the mice were randomly divided into a control group and an experimental group. The blood routine test was conducted, spleen index was calculated and spleen was histologically examined. Portal vein sera were taken for detection of the level of uric acid. The mRNA expressions of NALP3 and IL-1β in the spleen were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). The results showed that the platelet count was significantly lower in the experimental group [(674±102)×109/L] than in the control group [(1307±181)×109/L] (P<0.05), while the spleen index was significantly higher [(9.83±1.36) μg/g] in the experimental group than in the control group [(4.11±0.47) μg/g] (P<0.05). The histopathological changes of spleen followed the pattern of congestive splenomegaly. No significant difference was found in the uric acid level in the portal vein between the control group and the experiment group. The mRNA expressions of NALP3 and IL-1β were up-regulated significantly in the spleen in the experimental group as compared with those in the control group (P<0.05). It was concluded that NALP3 and IL-1β may play important roles in the pathogenesis of hypersplenism.This study examined the mRNA expression of NALP3 in the spleen of the mice with hypersplenism due to portal hypertension (PH). The mouse hypersplenism models were established by oral administration of tetrachloromethane (2 mL/kg/week for 12 weeks by oral gavage). All the mice were randomly divided into a control group and an experimental group. The blood routine test was conducted, spleen index was calculated and spleen was histologically examined. Portal vein sera were taken for detection of the level of uric acid. The mRNA expressions of NALP3 and IL-1β in the spleen were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). The results showed that the platelet count was significantly lower in the experimental group [(674±102)×109/L] than in the control group [(1307±181)×109/L] (P<0.05), while the spleen index was significantly higher [(9.83±1.36) μg/g] in the experimental group than in the control group [(4.11±0.47) μg/g] (P<0.05). The histopathological changes of spleen followed the pattern of congestive splenomegaly. No significant difference was found in the uric acid level in the portal vein between the control group and the experiment group. The mRNA expressions of NALP3 and IL-1β were up-regulated significantly in the spleen in the experimental group as compared with those in the control group (P<0.05). It was concluded that NALP3 and IL-1β may play important roles in the pathogenesis of hypersplenism.

Induction of apoptosis of human colon cancer cells by siRNA recombinant expression vector targeting survivin gene

This study constructed siRNA recombinant expression vector targeting survivin gene and observe the apoptosis induction effect of it in human colon cancer cells. siRNA recombinant expression vector targeting survivin gene was constructed and transfected into human colon cancer cells. The effect of siRNA recombinant expression vector was detected by RT-PCR, Western blot, MTT reduction assay and flow cytometry. It was confirmed by restriction endonuclease and sequence analysis that siRNA recombinant expression vector targeting survivin gene was constructed successfully. Inhibition rate of survivin siRNA at mRNA and protein levels was 36.33% and 44.65% respectively. Growth of cancer cells was inhibited and the apoptosis rate was (17.24±2.13)%. The siRNA recombinant expression vector targeting survivin gene has been constructed successfully. It not only can inhibit the expression of survivin gene, but also can induce apoptosis in human colon cancer cells remarkably.SummaryThis study constructed siRNA recombinant expression vector targeting survivin gene and observe the apoptosis induction effect of it in human colon cancer cells. siRNA recombinant expression vector targeting survivin gene was constructed and transfected into human colon cancer cells. The effect of siRNA recombinant expression vector was detected by RT-PCR, Western blot, MTT reduction assay and flow cytometry. It was confirmed by restriction endonuclease and sequence analysis that siRNA recombinant expression vector targeting survivin gene was constructed successfully. Inhibition rate of survivin siRNA at mRNA and protein levels was 36.33% and 44.65% respectively. Growth of cancer cells was inhibited and the apoptosis rate was (17.24±2.13)%. The siRNA recombinant expression vector targeting survivin gene has been constructed successfully. It not only can inhibit the expression of survivin gene, but also can induce apoptosis in human colon cancer cells remarkably.

Culture and Identification of Monoclonal Neural Stem Cells Derived from Cerebral Cortex

To isolate and culture the purified monoclonal neural stem cells from the cerebral cortex of new born mice, new-born mice cerebral cortex was isolated and dissociated to single-cell suspension by mechanical trituration. The dissociated single cells were cultured in serum-free medium. After the formation of neurospheres, single-cell clone culture was performed by limiting dilution and the proliferated single-cell clones were harvested for subculture. Immunocytochemistry was used to dtect the specific marker of neuroepithelial stem cells (Nestin) of the primary and monoclonal neurospheres. In the differentiated cells we detected the specific antigen of NF-200 and GFAP. Our results showed that the primary neurospheres expressed Nestin antigen positively. By limiting dilution, we cultured the cell lines from single-cell clone and the monoclonal neurospheres expressed Nestin and had capabilities of self-renewal, proliferation and the potentiality of differentiation into neurons and glial cells. It is concluded that monoclonal neural stem cells which have the ability of proliferation and multi-directional differentiation can be isolated and cultured from the cerebral cortex of new-born mice by limiting dilution.

Epigenetic regulation of the ERβ gene on the estrogen signal transfection pathway in colon cancer cells

We studied the regulatory effects of the estragen receptorβ (ERβ) gene on the downstream estrogen signal transfection pathway in colon cancer cells and the possible mechanisms involved. A human ERβ gene recombinant expression plasmid, pEGFP-C1-ERβ, was constructed and transfected into the Caco-2 colon cancer cell line, a line with low ERβ gene expression. The expression of ERβ mRNA and protein was detected 72 h after transfection. RT-PCR was used to examine the expression levels of the progesterone recepror (PR) gene containing the classic estrogen response element (ERE), the C-fos oncogene containing the AP-1 site (a non-classical ER binding site), the epigenetic modifying genes, such as Dnmt1, Dnmt3a, Dnmt3b, and histone methyltransferase (HMT), and the human mismatch repair gene hMLH1. Methylation-specific PCR was used to detect the changes in the methylated sites of the CpG islands in the promoters of the ERβ, PR, and C-fos genes. The results indicated that the human ERβ gene recombinant expression plasmid pEGFP-C1-ERβ was successfully constructed and transfected into Caco-2 cells. As compared with the control group, the mRNA and protein expression of ERβ gene was increased significantly 72 h after the transfection of pEGFP-C1-ERβ into the Caco-2 cells. As compared with the control group, the mRNA expression of the PR, C-fos, Dnmt3a and Dnmt3b genes was increased significantly 72 h after the transfection of pEGFP-C1-ERβ into the Caco-2 cells, but the mRNA expression of the Dnmt1, HMT, and hMLH1 genes decreased significantly (P<0.05). As compared with the control group, different degrees of demethylation occurred in the promoters of the ERβ, progesterone receptor (PR), and C-fos oncogene 72 h after the transfection of pEGFP-C1-ERβ into the Caco-2 cells. The methylation index of the estrogen signal transfection pathway in Caco-2 cells was decreased significantly following the expression restoration of ERβ gene (P<0.05). It is concluded that the restoration or up-regulation of the ERβ gene in Caco-2 cells may significantly activate the expression of the related target genes in the downstream estrogen signal transfection pathway and may result in the demethylation changes of the pathway. During the process, the expression level and activity of the epigenetic modifying genes and the human mismatch repair gene have changed simultaneously. The regulatory effect of the ERβ gene on the estrogen signal transfection pathway to a certain extent partly involves demethylation.SummaryWe studied the regulatory effects of the estragen receptorβ (ERβ) gene on the downstream estrogen signal transfection pathway in colon cancer cells and the possible mechanisms involved. A human ERβ gene recombinant expression plasmid, pEGFP-C1-ERβ, was constructed and transfected into the Caco-2 colon cancer cell line, a line with low ERβ gene expression. The expression of ERβ mRNA and protein was detected 72 h after transfection. RT-PCR was used to examine the expression levels of the progesterone recepror (PR) gene containing the classic estrogen response element (ERE), the C-fos oncogene containing the AP-1 site (a non-classical ER binding site), the epigenetic modifying genes, such as Dnmt1, Dnmt3a, Dnmt3b, and histone methyltransferase (HMT), and the human mismatch repair gene hMLH1. Methylation-specific PCR was used to detect the changes in the methylated sites of the CpG islands in the promoters of the ERβ, PR, and C-fos genes. The results indicated that the human ERβ gene recombinant expression plasmid pEGFP-C1-ERβ was successfully constructed and transfected into Caco-2 cells. As compared with the control group, the mRNA and protein expression of ERβ gene was increased significantly 72 h after the transfection of pEGFP-C1-ERβ into the Caco-2 cells. As compared with the control group, the mRNA expression of the PR, C-fos, Dnmt3a and Dnmt3b genes was increased significantly 72 h after the transfection of pEGFP-C1-ERβ into the Caco-2 cells, but the mRNA expression of the Dnmt1, HMT, and hMLH1 genes decreased significantly (P<0.05). As compared with the control group, different degrees of demethylation occurred in the promoters of the ERβ, progesterone receptor (PR), and C-fos oncogene 72 h after the transfection of pEGFP-C1-ERβ into the Caco-2 cells. The methylation index of the estrogen signal transfection pathway in Caco-2 cells was decreased significantly following the expression restoration of ERβ gene (P<0.05). It is concluded that the restoration or up-regulation of the ERβ gene in Caco-2 cells may significantly activate the expression of the related target genes in the downstream estrogen signal transfection pathway and may result in the demethylation changes of the pathway. During the process, the expression level and activity of the epigenetic modifying genes and the human mismatch repair gene have changed simultaneously. The regulatory effect of the ERβ gene on the estrogen signal transfection pathway to a certain extent partly involves demethylation.

Cyclin E expression and chemotherapeutic sensitivity in breast cancer cells.

The effects of the cyclin E expression levels on chemotherapeutic sensitivity of breast cancer cell line were explored. After the cyclin E expression was knockdown in MDA-MB-435 by RNA interference, FACS analysis and SA-β-gal staining were used to evaluate the response sensitivity of breast cancer cells to DNA damage drugs (adriamycin, etc.). Adriamycin could induce G1 arrest in cyclin E knockdown MDA-MB-435 breast cell line and increase the percentage of cell senescence in cyclin E knockdown MDA-MB-435 cells. It was suggested that cyclin E knockdown could increase the chemotherapeutic sensitivity of breast cancer cells to DNA damage drugs.