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Construction of a bivalent vaccine candidate against HAdV4/HAdV7 based on capsid-display strategy via Red-homologous recombination and counter-selection methodology

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Biosafety and Health 2024年第2期6卷 70-79页

作者： Peng Wang Yunting Shao Xichun Yang Wenning Zhang Jianguang Zhou Fang Huang Shuang Liu Jiping Zheng Chengjun Wu Shanhu Li department of Cell Engineering Beijing Institute of BiotechnologyBeijing 100850China School of Basic Medical Sciences Faculty of MedicineDalian University of TechnologyDalian 116024China Lab of microbiological Engineering School of Life and Health SciencesHainan UniversityHaikou 570228China

Human adenoviruses(HAdvs)are major respiratory ***,human adenovirus type 4(HAdV4)and human adenovirus type 7(HAdV7)are known for causing fever and pneumonia,with docu-mented cases of fatalities among the *** recent years,HAdV4/HAdv7 has been implicated in caus-ing substantial outbreaks,leading to increased morbidity in multiple *** HAdV4 and HAdV7 infections have been reported in North America,Asia,Europe,Africa,South America,Oceania,and the Middle *** fatalities occurred in North America(the United States)and Asia(China and Singapore).Engineered recombinant adenoviruses have played a crucial role as vaccine *** this study,we con-structed a recombinant adenovirus,Ad4ITRmut-Ad7E3,and evaluated it in vitro and in *** observed that the replication rate of Ad4ITRmut-Ad7E3 was lower than that of the RI-67 strain,indicating that the mutation of inverted terminal repeats(ITRs)weakened the replication ability of *** of BALB/c mice with the bivalent Ad4ITRmut-Ad7E3 vaccine strain,administered by intraperitoneal injection and oral gavage,resulted in the elicitation of neutralizing antibodies targeting HAdV4 and *** finding not only pro-vides a novel method and technique for the efficient construction of a polyvalent recombinant adenovirus vac-cine candidate against HAdV4 and HAdv7 but also against other prevalent adenovirus serotypes such as HAdV3,HAdV11,HAdV14,and HAdv55,from various regions.

关键词： Bivalent adenovirus vaccine Capsid-display strategy ITRs modification Hexon epitope replacement

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B lymphocytes regulate airway granulocytic inflammation and cytokine production in a murine model of fungal allergic asthma

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Cellular & Molecular Immunology 2015年第2期12卷 202-212页

作者： Sumit Ghosh Scott A Hoselton Scott V Asbach Breanne N Steffan Steve B Wanjara Glenn P Dorsam Jane M Schuh department of Veterinary and microbiological Sciences North Dakota State University Fargo ND USA

Sensitization to fungi often leads to a severe form of asthma that is particularly difficult to manage clinically, resulting in increased morbidity and hospitalizations in these patients. Although B lymphocytes might exacerbate asthma symptoms through the production of IgE, these cells might also be important in the protective response against inhaled fungi. Through cytokine release and T-cell interactions, these lymphocytes might also influence the development and maintenance of airway wall fibrosis. JH-/- mice lack the JHgene for the heavy chain component of antibodies, which is critical for B-cell function and survival. These animals have facilitated the elucidation of the role of B iymphocytes in a number of immune responses; however, JH-/- mice have not been used to study fungal allergy. In this study, we examined the role of B lymphocytes using an Aspergillus fumigatus murine fungal aeroallergen model that mimics human airway disease that is triggered by environmental fungal exposure. We compared disease progression in sensitized wild-type BALB/c and JH-/- mice that were exposed to repeated fungal exposure and found no differences in airway hyperresponsiveness, overall pulmonary inflammation or collagen deposition around the large airways. However, the levels of the Th2-type cytokines IL-4 and IL-13 were significantly attenuated in the airways of JH-/- mice relative to the BALB/c controls. By contrast, levels of the inflammatory cytokines IL-17A and IL-6 were significantly elevated in the JH-/- animals, and there was significantly more robust airway eosinophilia and neutrophilia than in control animals. Taken together, these findings demonstrate that B lymphocytes help to regulate granulocytic responses to fungal exDosure in the pulmonary compartment.

关键词： Aspergillus fumigatus B lymphocytes inflammation

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RELATIONSHIP BETWEEN THE EXPRESSIONS OF SURVIVIN AND THE PROGNOSTIC RELATED FACTORS IN BREAST CANCER

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Chinese Journal of Cancer Research 2005年第4期17卷 270-272页

作者：沈敬华王晓娟苏和巴特赵晓霞陶格斯 department of microbiological Immunology Inner Mongolia Medical College

Objective： To study the relationship between the Survivin expression and the histological grade, status of ER, expression of PS2 and the prognosis of patients with primary invasive breast cancer. Method： By using LSAB and SP immunohistochemical method, the expression of Survivin, PS2 and ER in 95 cases of invasive breast cancer were detected. Results： the positive rate of Survivin was 70.5% （67/95） and the expression of Survivin was positively related to the histological grade and status of PS2 and ER. The survival time after operation of patients without expression of Survivin was longer than those with positive Survivin. Conclusion： These data suggest that Survivin expression may be considered as a new unfavorable prognostic factor of breast cancer.

关键词： Breast cancer Survivin PS2 Estrogen receptor

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Evidence for a role of mitogen-activated protein kinases in the treatment of experimental acute pancreatitis

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World Journal of Gastroenterology 2014年第44期20卷 16535-16543页

作者： Natasha Irrera Alessandra Bitto Monica Interdonato Francesco Squadrito Domenica Altavilla department of Clinical and Experimental Medicine University of Messina department of Paediatric Gynaecological Microbiological and Biomedical Sciences University of Messina

Acute pancreatitis(AP) is an inflammatory disease characterized by acute inflammation and necrosis of the pancreatic parenchyma. AP is often associated with organ failure, sepsis, and high mortality. The pathogenesis of AP is still not well understood. In recent years several papers have highlighted the cellular and molecular events of acute pancreatitis. Pancreatitis is initiated by activation of digestive enzymes within the acinar cells that are involved in autodigestion of the gland, followed by a massive infiltration of neutrophils and macrophages and release of inflammatory mediators, responsible for the local and systemic inflammatory response. The hallmark of AP is parenchymal cell necrosis that represents the cause of the high morbidity and mortality, so that new potential therapeutic approaches are indispensable for the treatment of patients at high risk of complications. However, not all factors that determine the onset and course of the disease have been explained. Aim of this article is to review the role of mitogen-activated protein kinases in pathogenesis of acute pancreatitis.

关键词： Experimental acute pancreatitis Mitogen-activated protein kinases Mitogen-activated protein kinases inhibitors Cytokines Cholecystokinin Cerulein

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Anti-inflammatory activity of probiotic Bifidobacterium: Enhancement of IL-10 production in peripheral blood mononuclear cells from ulcerative colitis patients and inhibition of IL-8 secretion in HT-29 cells

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World Journal of Gastroenterology 2008年第16期14卷 2511-2516页

作者： Akemi Imaoka Tatsuichiro Shima Kimitoshi Kato Shigeaki Mizuno Toshiki Uehara Satoshi Matsumoto Hiromi Setoyama Taeko Hara Yoshinori Umesaki Yakult Central Institute for microbiological Research Yaho Kunitachi department of Gastroenterology and Hepatology Nihon University School of Medicine

AIM: To determine the anti-inflammatory activity of probiotic Bifidobacteria in Bifidobacteria-fermented milk (BFM) which is effective against active ulcerative colitis (UC) and exacerbations of UC, and to explore the immunoregulatory mechanisms. METHODS: Peripheral blood mononuclear cells (PBMNC) from UC patients or HT-29 cells were co-cultured with heat-killed probiotic bacteria or culture supernatant of Bifidobacterium breve strain Yakult (BbrY) or Bifidobacterium bifidum strain Yakult (BbiY) to estimate the amount of IL-10 or IL-8 secreted. RESULTS: Both strains of probiotic Bifidobacteria contained in the BFM induced IL-10 production in PBMNC from UC patients, though BbrY was more effective than BbiY. Conditioned medium (CM) and DNA of both strains inhibited IL-8 secretion in HT-29 cells stimulated with TNF-α, whereas no such effect was observed with heat- killed bacteria. The inhibitory effect of CM derived from BbiY was greater than that of CM derived from BbrY. DNAs of the two strains had a comparable inhibitory activity against the secretion of IL-8. CM of BbiY induced a repression of IL-8 gene expression with a higher expression of IκB-ζ mRNA 4 h after culture of HT-29 cells compared to that in the absence of ***: Probiotic Bifidobacterium strains in BFM enhance IL-10 production in PBMNC and inhibit IL-8 secretion in intestinal epithelial cells, suggesting that BFM has anti-inflammatory effects against ulcerative colitis.

关键词： Bifidobacterium Ulcerative colitis Anti-inflammatory Intedeukin-10 Interleukin-8

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ANTISENSE RNA OF SURVIVIN ENHANCES THE SENSITIVITY OF PANCREATIC CANCER CELL LINE PANC-1 TO DOXORUBICIN

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Chinese Journal of Cancer Research 2006年第2期18卷 138-141页

作者：沈敬华王晓娟王秀梅苏和巴特 department of microbiological and Immunology Inner Mongolia Medical College Huhehaote 010059

Objective： This paper attempts to discuss the effects of surviving antisense RNA on doxorubicin-induced apoptosis in pancreatic cancer cell line PANC-1. Methods： A surviving antisense eukaryotic vector pcDNA3-SV Vas prepared in previous study was delivered into PANC-1 by electroperforation. Cell survival fraction and MTT assay were used to investigate the sensibility of transfected cells to doxorubicin. Apoptosis was detected by DNA gel electrophoresis. Results： We obtained two positive cell clone PANC-1/SVVas and PANC-1/neo cells, the growth of PANC-1/SV Vas cells was significantly reduced （P〈0.05）. By MTT assay, the IC50 to doxorubicin of PANC-1/SV Vas, PANC-1/neo and PANC-1 cells were （0.285±0.012） μmol/L, （1.528±0.317） μmol/L and （1.540±0.253） μmol/L respectively, the difference was significant by statistic analysis （P〈0.01）. Agarose gel electrophoresis of genomic DNA from PANC/SV Vas showed typical DNA ladder, but DNA from PANC-1/neo and PANC-1 did not. Conclusion： Survivin antisense RNA could enhance doxorubicin-induced apoptosis in pancreatic cancer cell line PANC-I. This may lay an experimental foundation for further research of gene therapy in pancreatic cancer.

关键词： Survivin Pancreatic cancer Antisense RNA Doxorubicin

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Secretory expression and characterization of a recombinant-deleted variant of human hepatocyte growth factor in Pichia pastoris

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World Journal of Gastroenterology 2005年第45期11卷 7097-7103页

作者： Zhi-Min Liu Hong-Liang Zhao Chong Xue Bing-Bing Deng Wei Zhang Xiang-Hua Xiong Bing-Fen Yang xue-Qin Yao department of microbiological Engineering Beijing instituteof Biotechnology 20 Dongdajie Street Fengtai District Beijing100071 China

AIM： To study the secretory expression of human hepatocyte growth factor （hdHGF） gene in Pichia pastoris. METHODS： The full-length gene of human cDNA encoding the deleted variant of hdHGF was cloned by RT-PCR and overlapping-fragment PCR technique using mRNA of human placenta as a template. The cloned hdHGF cDNA was inserted into the Escherichia coilyeast shuttle vector of pPIC9. The constructed plasmid, pPIC9-hdHGF, was transformed into the GSl15 cells of the methylotrophic yeast, P pastoris, using a chemical method. The Hut ＋ transformants were screened to obtain high-expression strains by the test and analysis of expressed products of shake-flask culture. A secretory form of rhdHGF was made with the aid of the leader peptide sequence of Saccharomyces cerevisiae α-factor. RESULTS： The expressed products, which showed a band of molecular mass of about 80 ku, were observed on 15% SDS-PAGE and identified by Western blotting and N-terminal amino acid sequencing. In the high cell density culture of 5 L fermentor by fed-batch culture protocol, the cell biomass was reached at approximately 135 g （DCW）/L. The productivity of secreted total supernant protein concentration attained a high-level expression of more than 8.0 g/L and the ratio of rhdHGF band area was about 12.3% of the total band area scanned by SDS-PAGE analysis, which estimated that the product of rhdHGF was 500-900 mg/***： The P pastoris system represents an attractive tool of generating large quantities of hdHGF for both research and industrial purposes.

关键词： Hepatocyte growth factor Pichia pastoris Secretory expressing

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Assessment of effect of hydroalcoholic and decoction methods on extraction of antioxidants from selected Indian medicinal plants

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Asian Pacific Journal of Tropical Biomedicine 2012年第3期2卷 195-202页

作者： Mital Kaneria Bhavana Kanani Sumitra Chanda Phytochemical Pharmacological and Microbiological LaboratoryDepartment of BiosciencesSaurashtra University

Objective:To assess the effects of extraction methods on antioxidant activities of selected Indian medicinal ***:Different parts of plants were extracted by hydroalcoholic and decoction methods using water and various concentrations of methanol(ME) viz.75%, 50%and 25%*** antioxidant activity of all the different extracts was evaluated using two different antioxidant assays viz.2,2-diphenyl-1-picryl hydrazyl(DPPH) free radical scavenging assay and superoxide anion radical scavenging *** phenol and flavonoid content was also ***:The results showed that the extracting solvent significantly altered the antioxidant property estimations of screened *** correlations between phenolic compositions and antioxidant activities of extracts were *** levels of antioxidant activities were detected in Manilkara zapota(***) as compared with other screened plants. Conclusions:The results obtained appear to confirm the effect of different methods on extraction of antioxidants and antioxidant property of ***.

关键词： Antioxidant Hydroalcoholic extraction Total phenol content DPPH Superoxide Manilkara zapota

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Protective effect of Polyalthia longifolia *** leaves on ethanol and ethanol/HCl induced ulcer in rats and its antimicrobial potency

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Asian Pacific Journal of Tropical Medicine 2011年第9期4卷 673-679页

作者： Sumitra Chanda Yogesh Baravalia Mital Kaneria Phytochemical Pharmacological and Microbiological LaboratoryDepartment of BiosciencesSaurashtra UniversityRajkot-360 005GujaratIndia

Objective:To explore antiulcer and antimicrobial properties of methanolic extract of Polyalthia longifolia ***:Gastroprotective potential of Polyalthia longifolia was studied on ethanol and ethanol/HCl induced ulcers at 2 different doses(270 and 540 mg/kg/body weight). Antimicrobial efficacy of Polyalthia longifolia(25 mg/mL) was also studied against six gram positive,seven gram negative bacteria and five fungi by agar well diffusion *** inhibitory concentration was determined by agar well diffusion method in two fold serial dilution, in the range of 97-25 000μg/***:The reduction of ulcer index in Polyalthia longifolia treated animals was found to be statistically significant with respect to control *** Polyalthia longifolia exhibited ulcer protection activity in dose dependent manner and was also better than the *** antimicrobial activity,gram positive bacteria were more susceptible to Polyalthia longifolia than gram negative bacteria and fungal ***:Results obtained confirm the antiulcer and antimicrobial potential of the Polyalthia longifolia.

关键词： Polyalthia longifolia Antimicrobial activity Antiulcer activity Ulcer index Methanol extract

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Hepatoprotective effect of Woodfordia fruticosa Kurz flowers on diclofenac sodium induced liver toxicity in rats

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Asian Pacific Journal of Tropical Medicine 2011年第5期4卷 342-346页

作者： Yogesh Baravalia Yogeshkumar Vaghasiya Sumitra Chanda Phytochemical Pharmacological and Microbiological LaboratoryDepartment of BiosciencesSaurashtra UniversityRajkot-360 005GujaratIndia

Objective:To evaluate the protective effect of Woodfordia fruticosa Kurz flowers against experimentally induced liver toxicity in ***:Two different doses of methanol extract of Woodfordia fruticosa(WFM) were evaluated for the hepatoprotective activity against diclofenac sodium induced hepatotoxicity in *** biochemical parameters like alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),total protein(TP),albumin(ALB),blood urea nitrogen(BUN) from serum;total protein(TP),glutathione (GSH) levels,catalase(CAT) and glutathione peroxidase(GPx) activities from liver were studied; histopathologic changes of liver were also ***:WFM effectively reduced the elevated levels of serum ALT,AST,ALP and BUN,enhanced the reduced TP,ALB and hepatic GSH,CAT,GPx *** histopathological analysis suggested that WFM decreased the degree of liver fibrosis induced by ***:This study demonstrates the hepatoprotective activity of WFM and thus scientifically support the use of this plant in traditional medicine for the treatment of liver disorders.

关键词： Woodfordia fruticosa Diclofenac Hepatoprotective Biochemical analysis Histopathology

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