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ObjectiveTo observe the effects of bifidobacterial sectory adhesin on NF-κB DNA binding activity,cytoplasm IκB,nuclei NF-κBp65,TNF-α,IL-1β,and IL-8 expression in intestinal epithelial cell post stress response.MethodsThe cultured intestinal epithelial cell line Lovo cells were divided into 5 groups,which included directly stimulated by lypopolysaccharide (LPS) (100 ng/ml) or H2O2 (200 μmol/L) for 3 hours groups,bifidobacterial sectory adhesin (30μg/ml) pre-incubation for 30 minutes and then treated by LPS (100 ng/ml) or H2O2 (200 μmol/L) for 3 hours groups and control group with no treatment.NF-κB DNA binding activity was evaluated by electrophoretic mobility shift assay (EMSA).The cytoplasm IκB and nuclei NF-κBp65 expression were determined by Western blot.The expressions of TNF-α,IL-1β and IL-8 at mRNA level were detected by semi-quantatitive assay of RT-PCR.ResultsThe expressions of NF-κB DNA binding activity [(6.20±0.35) times and (4.16 ± 0.52) times of that of non treated control group,respectively] and nuclei NF-κBp65 [ (0.64±0.05) and (0.67±0.06)] were higher in cells after LPS or H2()2 stimulated,however the cytoplasm IκB expression [ (0.28 ± 0.10) and (0.39 ± 0.12) respectively] was weaker.After bifidobacterial sectory adhesin pre-incubation,the expressions of NF-κB DNA binding activity and nuclei NF-κBp65decreased obviously,but cytoplasm IκB expression increased.After treated with LPS or H2O2,the mRNA expressions of TNF-α,IL-1β and IL-8 were significantly increased [LPS treated group2 (0.92±0.10)、(0.38±0.03)、(1.44±0.25),H2O2 treated group:(0.89±0.13)、(0.36±0.06)、(1.42±0.18)].After bifidobacterial sectory adhesin pre-incubation,their expressions decreased.The mRNA expressions of TNF-α,IL-1β and IL-8 have significantly positive correlation with NF-κB DNA binding activity.ConclusionsThere is a significant inhibition role of bifidobacterial sectory adhesin in LPS and H2O2 induced NF-κB DNA binding activity in intestinal epithelia cells.The activation of NFκB may associate with the regulation of TNF-α,IL-1β and IL-8 proinflammatory cytokines expression.
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<p><b>OBJECTIVE</b>To study the therapeutic effect adenovirus-mediated apoptin gene transfer combined with ADM and CDDP on hepatocellular carcinoma in mice.</p><p><b>METHODS</b>In c57BL/ 6 mice bearing hepatocellular carcinoma, the changes of tumor volume, histomorphology, tumor inhibition rate and the side effects were observed after intratumoral injection of adenovirus containing apoptin gene and ADM and CDDP.</p><p><b>RESULTS</b>Seven days after the treatment, the mean volume of the tumor in the mice receiving intratumoral apoptin-containing adenovirus injection combined with ADM and CDDP reduced significantly as compared with that in mice treated with adenovirus vehicle and control group. The tumor inhibition rate in the combined treatment group was 90.13%, significantly higher than that in the control group. No adverse effect of the treat was observed in the course of the experiment.</p><p><b>CONCLUSION</b>The adenovirus vectors containing apoptin gene combined with ADM and CDDP may serve a safe treatment of hepatocellular carcinoma.</p>
Subject(s)
Animals , Mice , Adenoviridae , Genetics , Metabolism , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Capsid Proteins , Genetics , Therapeutic Uses , Cisplatin , Combined Modality Therapy , Doxorubicin , Gene Transfer Techniques , Genetic Therapy , Liver Neoplasms, Experimental , Therapeutics , Mice, Inbred C57BL , Neoplasm Transplantation , Random AllocationABSTRACT
Objective To investigate the effects of silenced Racl on invasion and migration in LOVo cells.Methods The expression of Racl mRNA and protein in colorectal cancer cells(including LoVo SW480.SW620.SW1116,HT29)were detected by RT-PCR and Western blot,respectively.The changes of cytoskeleton were observed in LoVO cells after transfected with Racl-shRNA.then invasion and migration were recorded respectively in LoVo cells after transfected with Racl-N17 and Racl-L61.Results Racl mRNA and protein were overexpressed in all selected colorectal cancer cells.Deletion of Racl decreased the cross-linked actin network and pseudopodia,and inhibited the invasion and migration in LoVo cells.The migration experiment showed that the migrated cells were higher in Racl-shRNA[(75±5)cells].Racl-N17 [(93±5)cells]and Racl-L61[(267±7)cells]groups compared with control group[(214±8)cells,P＜0.01,＜O.01 and＜0.05,resprectively].The invasion experimental study revealed that the migrated cells were higher in Racl-shRNA[(35±5)cells],Racl-N17[(42±5)cells]and Racl-L61[(86±7)cells] groups compared with control group[(73±6)cells,P＜0.01,＜0.01 and＜0.05,resprectively].Condusion Deletoin of Racl can inhibit the invasion and migration in LDVo cells.
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Objective To investigate the expression changes of histidine decarboxylase(HDC)and H+,K+-ATPase in gastric mucosa during the healing of experimental gastric ulcer in rats.Methods The ulcers were caused by applying acetic acid to the serosal surface of the anterior face of the rat gastric body.At different time points during ulcer healing,HDC and H+,K+-ATPase mRNA and protein expressions were studied by using reverse transcription-polymerase chain reaction and Western blot respectively.Results An ulcer crater developed on the anterior face of the gastric body on day 1 after the induction of ulcers,and the ulcer area was biggest on day 3.On day 12,most of the gastric ulcers had healed.Compared with the control group,the HDC and H+,K+-ATPase mRNA expression in the gastric mucosa of ulcer rats showed a decrease on day 1,and increased back to initial level on day 9.The protein expression of HDC and H+,K+-ATPase in gastric mucosa of ulcer rats decreased immediately on day 1,more on day 6,and returned to the initial levels on day 12.Conclusion The mRNA and protein expressions of HDC and H+,K+-ATPase decrease in the healing process of gastric ulcers,resulting in accelerated ulcer healing through inhibiting gastric acid secretion.
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Objective To study H.pylori infection and the changes of muco sal inflammation， atrophy and intestinal metaplasia in the different histolog ical types of gastric polyps. Methods Two hundred and seventy－ eight cases of gastric polyps(12.6％ ) from 2 203 gastric mucosal biopsy cases were histologic ally classified and examined for the presence of Helicobacter pylori,for degree and type of inflammation,mucosal atrophy and intestinal metaplasia. Results 53. 9％ gastric polyps were infected with H.pylori,in which faveolar polyp was the highest with an infective rate of 73.1％ .The changes of active inflammation,atr ophy and metaplaisa in gastric mucosa were frequently accompanied with H.pylori positive faveolar polyps almost as same as those in adenoma.The fundic gland pol yps were usually with low rate of H.pylori infection,and the changes of mucosal active inflammation,atrophy and metaplasia were seldom observed. Conclusions Fav eolar polyps,which are different from fundic gland polyp,may caused by H.pylori infection and are usually with the changes of mucosal active inflammation,atroph y and metaplasia.
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Objective To study the effects of recombinant adenovirus vector containing vp3 gene (AdAFPvp3) on c57BL/6 murine hepatocellular carcinoma model. Methods The titers of adenovirus vectors AdAFPvp3 and AdCMV-eGFP were detected. c57BL/6 murine hepatocellular carcinoma models were reproduced by subcutaneous inoculation of murine Hepa1-6 cells, and tumors were monitored for their generation rates. Twenty-four mice were randomly divided into adenovirus vector AdAFPvp3 group, adenovirus vector AdCMV-eGFP control group and PBS control group (8 each) when the tumors grew into 5mm in diameter. AdAFPvp3 (5?108pfu/100?l), AdCMV-eGFP (5?108pfu/100?l) and PBS (100?l) was intratumorally injected, respectively, in the three groups every other day for 2 times. The volume of tumor and the presence of adverse effects were observed. Seven days after treatment, all mice were sacrificed for evaluation of antitumor effect. Tumors, livers and spleens were all harvested for routine pathological examination. At the same time, the way of tumor cell death induced by vp3 in vivo was identified by TUNEL. Results The titers of both adenovirus AdAFPvp3 and adenovirus AdCMV-eGFP were 5?109pfu/ml. The carcinogenesis rate of hepatocellular carcinoma was 100%. Compared with adenovirus AdCMV-eGFP control group and PBS control group, the volume of tumor was diminished evidently in AdAFPvp3 group after intratumoral injection (P
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Objectives In order to further investigate the biological characteristics and the developmental mechanisms of laterally spreading tumor, a cell line from the primary culture of human colon LST tissue was isolated. After 70 passages, we identified the cells by morphologic, genetic and immunohistochemistry analysis. Results (1) The origin of LST cells is epithelial, and the majority of which were multiform epithelial cells. It displayed an approximately similar growth characteristics of tumor cells. The nest corpus doubling time was 36 hours. (2) The number of chromosome is 42-66. Eighty-five percent of chromosomes were triploid, showing abnormal karyotypes. (3) Immunohistochemistry showed that ESA and CK20 were expressed. (4) The ultrastructure of LST was almost the same as that of a tumor cell. Conclusion A novel LST cell line has been established and named as LST-R1. It facilitates further study of the biological characteristics of LST cells and the development of colon cancer.
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Objective To investigate the relationship between early colonic cancer and the pit patterns of abnormal colonic mucosa. Methods We examined the pit patterns of 124 polypi in 139 patients by magnifying colonoscopy and stereomicroscopy, and compared the relations between the pit patterns and pathological diagnosis of polypi. Results 5 cases of LST(laterally spreading tumor)and 9 cases of advanced cancer were found out of 124 polypi in 139 patients. 1 among these 5 cases of LST showed Ⅲ L pit pattern, and 4 showed Ⅳ pit pattern with both magnifying colonoscopy and stereomicroscopy. Magnifying colonoscopy showed high coincidence rate with stereomicroscopy. Conclusion Pit patterns are found to be very important in disguishing tumorous lesions from non-tumorous lesions, and to discover early colonic cancer. Ⅴ pit pattern indicates that the polyp is carcinomatous.
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Objective To observe competitive inhibition of adherence of enterotoxigenic Escherichia coli (ETEC) and enteropathogenic Escherichia coli (EPEC) to intestinal epithelial cells by purified adhesin derived from bifidobacterium adolescentis 1027. Methods The number of bacteria adherent to intestinal epithelial cell line Lovo was counted and the results were analyzed by comparison. Results The purified adhesin in the concentration of 10?g/ml, 20?g/ml and 30?g/ml could significantly inhibit the adhesion of ETEC and EPEC to intestinal epithelial cell line Lovo. The higher the concentration of adhesin the higher inhibition rate was observed. Concentrations lower than 10?g/ml had no such effect. Conclusion The purified adhesin of bifidobacterium adolescentis 1027 could inhibit the adhesion of ETEC and EPEC to intestinal epithelial cell line Lovo, and the effect was dose-dependent.