In order to construct a recombinant attenuated Salmonella typhimurium expressing Helicobacter pylori (H. pylori) urease subunit B(UreB), UreB gene fragment amplified by PCR was cloned into the prokaryotic expression vector pTC01 after sequencing, then transformed into attenuated Salmonella typhimurium SL3261 to acquire SL3261/pTC01-UreB. The expression of H. pylori UreB in SL3261 was detected by Western blot. Tewelve weeks after oral immunization of mice with this vaccine, anti-UreB IgA antibodies in mouse intestinal fluid and IgG antibodies in serum were determined by ELISA. IFN-? and IL-10 contents in the supernatant of spleen cell culture were also assessed by ELISA. The results showed that 61kD protein was expressed in SL3261/pTC01-UreB that could be recognized by anti-H. pylori UreB antiserum by Western-blot. The multiple oral immunizations with SL3261/pTC01-UreB could induce significantly H. pylori-specific mucosal IgA response as well as serum IgG responses. Moreover, there was significant increase of IFN-? and IL-10 contents in the supernatant of spleen cell culture in SL3261/pTC01-UreB group. These results suggested that the attenuated Salmonella typhimurium expressing H. pylori UreB may be used as oral vaccine against H. pylori infection. Its effect against H. pylori infection also needs to be further evaluated in animal models.

Aim To observe the effects of cytokine signaling inhibition protein-3(SOCS3) on the liver fibrosis progression and reverse.Methods C57BL/6 mouse model was established by subcutaneous injection of carbon tetrachloride(CCl4).After a successful model of fibrosis, one-month normal diet was given to induce the reverse fibrosis model, while normal mice of the same gender and weight were as control group.Mice were sacrificed at 1, 2, 3, 4, 5, 6, 7, 8 weeks, respectively, then the liver tissue was harvested for the observation of its injury by hematoxylin and eosin(HE) staining.Then Masson staining was applied for the detection of changes in collagen, and the immunohistochemistry(IHC) for the observation of type Ⅰ Collagen(Colla-1), alpha smooth muscle actin(α-SMA), transforming growth factor beta 1(TGF-β1) and SOCS3 protein expression.In vitro formation of fibrosis was induced by TGF-β1 stimulating HSC-T6 cell lines, which was then reversed by MDI medium, with co-incubation of HSC-T6 cells with plasmid in the process of the reverse.Western blot was employed to detect SOCS3, Colla-1, α-SMA, TGF-β1 expression.Results The expression of SOCS3 and TGF-β1 increased in mouse model of fibrosis with the worsening fibrosis process and decreased in the reverse process.Over-expression SOCS3 in the reverse process reduced the development of liver fibrosis;meanwhile, the expression of TGF-β1 was also reduced accordingly.Conclusion SOCS3 may influence the development of the liver fibrosis and its reverse via regulating the expression of TGF-β1.

OBJECTIVETo establish attenuated Salmonella typhimurium producing Helicobacter pylori (H. pylori) urease subunit B (UreB) and determine whether it could be used as an oral vaccine against H. pylori.

METHODSH. pylori (SS1 strain) UreB gene fragment amplified by PCR was cloned into the prokaryotic expression vector pTC01 after sequencing, and then transformed into attenuated Salmonella typhimurium SL3261 to acquire SL3261/pTC01-UreB. The expression of H. pylori UreB in SL3261 was detected by Western blot. Twelve weeks after oral immunization of mice, antibody responses were evaluated using serum and intestinal fluid by ELISA assay. Interferon-gamma (IFN-gamma) and interleukin-10 (IL-10) in the supernatant of spleen cells culture were also assessed by ELISA. In vitro stability of pTC01-UreB plasmid in SL3261 was confirmed by growing in Luria Broth (LB) medium to 80 generations.

RESULTSThe UreB gene fragment amplified by PCR was consistent with the sequence of the H. pylori UreB as evidenced by sequence analysis. Enzyme digestion revealed that the correct pTC01-UreB was obtained. Western blot showed that a 61kDa protein was expressed in SL3261/pTC01-UreB, which could be recognized by anti-H. pylori UreB antiserum. After 80 generations of continuous culture, the recombinant plasmid pTC01-UreB was stable in SL3261 and had no obvious toxicity. Multiple oral immunizations with SL3261/pTC01-UreB could significantly induce H. pylori-specific mucosal IgA response as well as serum IgG response. Moreover, there were significant increases of IFN-gamma and IL-10 in the SL3261/pTC01-UreB group. Finally, no obvious side effects for mice and no change in gastric inflammation were observed.

CONCLUSIONAttenuated Salmonella typhimurium expressing H. pylori UreB may be used as oral vaccine against H. pylori infection.

Administration, Oral ; Animals ; Antibodies, Bacterial ; blood ; Bacterial Vaccines ; blood ; immunology ; Female ; Helicobacter Infections ; prevention & control ; Helicobacter pylori ; immunology ; Immunization ; Interferon-gamma ; biosynthesis ; Interleukin-10 ; biosynthesis ; Mice ; Mice, Inbred BALB C ; Plasmids ; Protein Subunits ; Salmonella typhimurium ; genetics ; Urease ; immunology ; Vaccines, Attenuated ; immunology ; Vaccines, Synthetic ; immunology

Colorectal cancer (CRC) belongs to multiple diseases of traditional Chinese medicine (TCM).CRC can exhibit sophisticated clinical manifestations and various TCM syndromes (Zheng).Modem studies have revealed that syndromes (Zheng) of CRC are related to pathological characteristics,cell types,staging and prognosis.Surgery,chemotherapy and other modem treatments can influence the manifestations of syndromes (Zheng) of CRC.Modern TCM doctors have extensively studied the biomedical basis of syndromes (Zheng) of CRC,including accumulation of damp-heat (Shi-Re-Yun-Jie),blood stasis (Yu-Xue-Nei-Zu),spleen (Pi) deficiency (Pi-Qi-Kui-Xu),deficiency of qi and blood (Qi-Xue-Liang-Xu),deficiency of yin of liver (Gan) and kidney (Shen) (Gan-Shen-Yin-Xu) and other syndromes (Zheng).These investigations may contribute to the objective and standardization study of syndromes (Zheng) in CRC.

Gastric cancer is one of the most common malignancies worldwide; however, the molecular mechanism in tumorigenesis still needs exploration. BCL2L11 belongs to the BCL-2 family, and acts as a central regulator of the intrinsic apoptotic cascade and mediates cell apoptosis. Although miRNAs have been reported to be involved in each stage of cancer development, the role of miR-24 in GC has not been reported yet. In the present study, miR-24 was found to be up-regulated while the expression of BCL2L11 was inhibited in tumor tissues of GC. Studies from both in vitro and in vivo shown that miR-24 regulates BCL2L11 expression by directly binding with 3'UTR of mRNA, thus promoting cell growth, migration while inhibiting cell apoptosis. Therefore, miR-24 is a novel onco-miRNA that can be potential drug targets for future clinical use.
Animals ; Apoptosis ; genetics ; Apoptosis Regulatory Proteins ; deficiency ; genetics ; Base Sequence ; Bcl-2-Like Protein 11 ; Cell Line, Tumor ; Cell Movement ; genetics ; Cell Proliferation ; genetics ; Down-Regulation ; genetics ; Gene Silencing ; Male ; Membrane Proteins ; deficiency ; genetics ; Mice ; MicroRNAs ; genetics ; Proto-Oncogene Proteins ; deficiency ; genetics ; Rats ; Stomach Neoplasms ; genetics ; pathology