| ObjectiveTo observe the biological changes of human gastric epithelial cell line GES-1 which was stably transfected with HCCR2 gene; To study the molecular mechanism of HCCR gene upregulation in human gastric epithelial cell line GES-1 and human gastric cancer cell line SGC-7901 via PI3K/Akt and MAPK pathway treated with EGF.Methods1,GES-1 cells were transfected with HCCR2-pcDNA3.1, pcDNA3.1 by lipofectmine 2000, transfectants were selected and confirmed using Western blot technique. HCCR was measured on protein level by Western blot. The cell cycle was tested by flow cytometry. The reproductive activity was detected by MTT assay.2,Treated with increasing concentration (0, 25, 50, 100ng/ml) of EGF for 24h, or with 100ng/ml EGF at different time(0, 8, 16, 24h), the human gastric epithelial cells GES-1 and the human gastric cancer cells SGC-7901 were routinely cultured. Cells were pretreated with the specific inhibitor of EGFR (PD153035) and then cultured with EGF (100ng/ml) for 24h, the expression of HCCR protein in these cells weremeasured by Western blot.3,GES-1 and SGC-7901 cells were pretreated with the specific inhibitorof PI3K (LY294002) and then cultured with EGF (100ng/ml) for 24h, theexpression of HCCR protein in these cells were measured by Western blot.GES-1 cells were transfected with CA-Akt-pcDNA3.1 (constitutivelyactive Akt kinase), pcDNA3.1 by lipofectmine 2000. SGC-7901 cellswere transfected with DN-Akt-pcDNA3.1 (dominant negative Akt kinase),pcDNA3.1 by lipofectmine 2000, transfectants were selected andconfirmed using Western blot technique. HCCR was measured on proteinlevel by Western blot. The cell cycle was tested by flow cytometry. Thereproductive activity was detected by MTT assay.4,GES-1 and SGC-7901 cells were pretreated with the specific inhibitorof MEK1 (PD98059) and then cultured with EGF (100ng/ml) for 24h, theexpression of HCCR protein in these cells were measured by Westernblot.Results1,The overexpression of HCCR2 enhance the proliferation of GES-1GES-1 cells were stably transfected with HCCR2 gene. HCCR2 genetransfection can increase the proportion of S-phase cells which was testedby flow cytometry. MTT showed the growth of HCCR2 gene transfectedcells was faster than that of empty vecter transfected cells. HCCR2 gene transfection can enhance the proliferation ability of HCCR2 gene transfected cells.2,EGF treatment induced HCCR expression in GES-1 and SGC-7901 cellsGES-1 cells were cultured with EGF at 50ng/mlfor 24h, the HCCR protein were upregulated in a dose-dependent manner. HCCR protein expression gradually increased in a time-dependent manner after treated with EGF at 100ng/ml for 24h. SGC-7901 cells were cultured with EGF got similar results. Treatment of GES-1 and SGC-7901 cells with EGFR inhibitor (PD153035) significantly suppressed EGF induced HCCR expression. These results show that EGF can induce HCCR expression in GES-1 and SGC-7901 cells.3,EGF upregulated HCCR via a PI3K/Akt signal transduction pathway in GES-1 and SGC-7901 cellsTreatment of GES-1 and SGC-7901 cells with specific inhibitor of PI3K (LY294002) significantly suppressed EGF induced HCCR expression. HCCR protein were upregulated in constitutively active Akt transfectants. CA-Akt gene transfection can increase the proportion of S-phase cells which was tested by flow cytometry. MTT showed the growth of CA-Akt gene transfected cells was faster than that of empty vecter transfected cells. HCCR protein were downregulated in dominant negative Akt transfectants. DN-Akt gene transfection can decrease the proportion of S-phase cells which was tested by flow cytometry. MTT showed thegrowth of CA-Akt gene transfected cells was slower than that of emptyvecter transfected cells.4,EGF upregulated HCCR via a MAPK signal transduction pathwayin GES-1 and SGC-7901 cellsTreatment of GES-1 and SGC-7901 cells with specific inhibitor of MEK1(PD98059) significantly suppressed EGF induced HCCR expression.Conclusion1,The overexpression of HCCR2 enhances the proliferation of GES-12,EGF induces HCCR expression in GES-1 and SGC-7901 cells3,EGF upregulates HCCR via a PI3K/Akt signal transduction pathway inGES-1 and SGC-7901 cells4,EGF upregulates HCCR via a MAPK signal transduction pathway inGES-1 and SGC-7901 cells ObjectiveThe purpose of this meta-analysis is to evaluate the influence ofHelicobacter pylori eradication on the symptoms of Gastroesophagealreflux disease.MethodsA search of papers was conducted to Mar 2008 using PubMed, EMBasedigital dissertations database. A systematic review of all articles whichmet the inclusion criteria was performed.Results1,14 articles met the inclusion criteria and were included in themeta-analysis.2,The influence of Helicobacter pylori eradication on the heartburn ofGastroesophageal reflux disease.(1) The heartburn of GERD was improved after Hp eradication following 6 months (OR=1.95, 95%CI 1.11-3.44, P<0.05); The heartburn of GERD in placebo group had no change (OR=1.51, 95%CI 0.83-2.75, P>0.05).(2) The heartburn of GERD was improved after Hp eradication following 1 year (OR=1.76, 95%CI 1.36-2.28, P<0.0001); The heartburn of GERD in placebo group had no change (OR=1.32, 95%CI 0.89-1.95, P>0.05).(3) The heartburn of GERD had no change after Hp eradication following 2 years (OR=1.64, 95%CI 0.74-3.62, P>0.05); The heartburn of GERD in placebo group was improved (OR=2.10, 95%CI1.77-2.69, P<0.00001).3,The influence of Helicobacter pylori eradication on the reflux of Gastroesophageal reflux disease.(1) The reflux of GERD had no change after Hp eradication following 6 months (OR=22.66, 95%CI 0.15-3340.99, P>0.05); The reflux of GERD in placebo group had no change (OR=19.97, 95%CI 0.04-11156.34, P>0.05).(2) The reflux of GERD was improved after Hp eradication following 1 year (OR=3.51, 95%CI1.54-8.02, P<0.0001); The reflux of GERD in placebo group had no change (OR=2.23, 95%CI 0.82-6.07, P>0.05).(3) The reflux of GERD had no change after Hp eradication following 2 years (OR=1.94, 95%CI0.61-6.22, P>0.05); The reflux of GERD in placebo group was improved (OR=2.99, 95%CI 2.39-3.74, P<0.00001).4,The influence of Helicobacter pylori eradication on reflux esophagitis of Gastroesophageal reflux disease.(1) Reflux esophagitis of GERD had no change after Hp eradication following 6 months (OR=0.09, 95%CI 0.42-1.09, P>0.05); Reflux esophagitis of GERD in placebo group had no change (OR=1.10, 95%CI 0.51-2.37, P>0.05).(2) Reflux esophagitis of GERD had no change after Hp eradicationfollowing 1 year (OR=1.2, 95%CI0.33-4.39, P>0.05); Reflux esophagitisof GERD in placebo group had no change (OR=1.30, 95%CI0.59-3.20,P>0.05).Conclusion1,Helicobacter pylori eradication did not worsen the heartburn, refluxand reflux esophagitis of GERD after 6 months, 1 year and 2 years.2,Helicobacter pylori eradication improved the heartburn of GERD after6 months and 1 year; Helicobacter pylori eradication improved the refluxof GERD after 1 year.
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