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Comparative Study on the Immunogenicity between Recombinant MS-Sj26GST Vaccine and Recombinant BCG-Sj26GST Vaccine in Schistosoma japonicum

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2003年第3期23卷 213-215,218页

作者：戴五星高红黄海浪袁野胡佳杰皇甫永穆 department of Biochemistry and molecular biology

The BALB/c mice were immunized with rMS Sj26GST and rBCG Sj26GST vaccine in Schistosoma japonicum by subcutaneous injection After they were immunized for 8 weeks, the eyeballs were removed to get blood and macrophages of abdominal cavity and spleen cells were harvested The lymphocytic stimulating index (SI) was used to measure the cellular proliferating ability and NO release was used to measure the phagocytic activity of the macrophages By using ELISA kit, the levels of interleukin 2 (IL 2) and interferon γ (IFN γ) in serum and the splenic lymphocytic cultured supernatant were detected The results showed that after the mice were immunized with 10 6 CFU of rMS Sj26GST and rBCG Sj26GST vaccine separately by subcutaneous injection, proliferating ability of splenic lymphocytes in the mice showed no difference ( P >0 05), but both were significantly increased as compared with that in the control group( P <0 05); The contents of NO in the intraperitoneal macrophages of rMS Sj26GST vaccine group were significantly lower than in the control group ( P <0 001) and rBCG Sj26GST vaccine group ( P <0 01); The levels of serum IL 2 in the rMS Sj26GST vaccine group were significantly increased as compared with that in the control group ( P <0 001), vector group ( P <0 01) and rBCG Sj26GST vaccine group ( P <0 05); The contents of serum IFN γ in the rMS Sj26GST vaccine group were significantly increased as compared with that in the control group ( P <0 01) and rBCG Sj26GST vaccine group ( P <0 05) The contents of IFN γ in the cultured supernatant were significantly lower than those of rBCG Sj26GST vaccine group ( P <0 001), but were significantly increased as compared with that in the control group ( P <0 01) It was indicated that both vaccines could enhance the immune response of the mice, but rMS Sj26GST vaccine had stronger immunogenicity than rBCG Sj26GST vaccine

关键词： Schistosoma japonnicum Mycobacterium smegmatis mc 2155(MS) BCG Vaccine Immunogenicity

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Effect of Fragment Asp1961-Glu1978 in Fibronectin on the Expression of Triple-domain Polypeptide in E. coli

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Journal of Huazhong University of Science and Technology(Medical Sciences) 1998年第3期18卷 129-133页

作者：李东冯作化张桂梅张慧范曲 department of Medical molecular biology Tongji Medical University Wuhan

Two plasmids were constructed and used to express two triple-domain recombinant polypeptide of human fibronectin (FN). The cDNAs in plasmids code for two polypeptides, CH62 (Pro1239-Ser1515 of FN linked with Ala1690 -Val2049 through Met) and CH63 (CH62 without Ile1850-Glu1978). The expression level of CH62 in E. coli was very low, but that of CH63 was very high.The results suggests that Asp1961-Glu1978 in FN is a key sequence influencing the expression of triple-domain polypeptide in E. Coli. After being dissolved and renatured, CH63 can be purified by heparin-agarose affinity chromatography.Both of the cell-binding domains in the recombinant polypeptide were functional.The production of CH63 provides a fundamental basis for further study of recombinant products with better anti-metastasis function.

关键词： fibronectin recombinant polypeptide E. coli expression

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Comparative Study on the Inhibitory Effect of RecombinantFN Polypeptide CH50 and CH56 on the Metastasis ofMelanoma Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 1997年第3期17卷 129-131页

作者：张桂梅冯作化张慧李东范曲 department of Medical molecular biology Tongji Medical University Wuhan

On the basis of preparation of anti-metastatic recombinant FN polypeptides, CH50 and CH56, we further studied the function of these polypeptides.The capacity of CH50 binding with melanoma cells (ED50 30 mM) was higher than that of CH56 (ED50 45 mM). Both of the polypeptides could significantly suppress the binding of melanoma B16 cells to laminin. There was no significant difference in the inhibitory effect between two polypeptides. In the experimental metastasis of melanoma cells, both of CH50 and CH56 could significantly inhibit the metastasis of the tumor cells, and reduce the number of lung metastasis by about 80%. Our results suggest that Ⅲ-11 and ED-A repeats influenced, to some extent, the binding capacity of bifunctional-domain polypeptide to cells, but did not affect the inhibition of the polypeptide on the metastasis of melanoma cells. The presence and connection of cell Ⅰ and Hep Ⅱ domains are the elements which determine the ability of recoinbinant FN polypeptides to inhibit the metastasis of tumor cells.

关键词： recombinant FN polypeptide tumor metastasis melanoma laminin

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Involvement of insulin in early development of mouse one-cell stage embryos

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Science China(Life Sciences) 2008年第9期51卷 767-773页

作者： YU BingZhi, ZHANG Zhe, DENG Xin, XU XiaoYan, FENG Chen, LI YanXiao, CUI Cheng, SU WenHui, ZHAO HongMei & YU DaHai department of Biochemical and molecular biology, China Medical University, Shenyang 110001, China department of Biochemical and molecular biology China Medical University Shenyang China

Recent studies have suggested that growth factors and hormones play important roles in cell prolif-eration and differentiation during early embryonic development. In the present study, we examined the expression and localization of insulin in the mouse oocytes and one-cell stage embryos by quantitative ELISA, RT-PCR, Western blot and immunofluorescence. In the mouse oocytes and one-cell stage em-bryos, expression of insulin was uniformly distributed in the cytoplasm. We also examined the expres-sion, activity and localization of mTOR (mammalian target of rapamycin) and p70S6K. The expression of mTOR and p70S6K was not significantly different at the cell cycle of mouse one-cell stage embryos. mTOR and S6K were distributed evenly in the cytoplasm at G1, G2 and M phase phase, but at S phase, the distribution of mTOR and S6K was around the pronucleus. At different phases, the activity of mTOR fluctuated. We also used the PI3K specific inhibitor-Wortmannin to investigate the cleavage rate of eggs. The result showed that the rate obviously decreased. When the mTOR specific inhibitor Rapa-mycin was used, the first mitotic division of the mouse one-cell stage embryo was delayed. These re-sults suggested that insulin was expressed both in mouse oocytes and one-cell stage embryos, and may play functional roles in regulation of mouse early embryogenesis by activating the signal pathway of PI3K/PKB/mTOR/S6K.

关键词： mouse embryo insulin mTOR S6K

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Morphine enhances purine nucleotide catabolism in vivo and in vitro

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Acta Pharmacologica Sinica 2007年第8期28卷 1105-1115页

作者： Chang LIU Jian-kai LIU Mu-jie KAN Lin GAO Hai-ying FU Hang ZHOU Min HONG~1 department of Biochemistry and molecular biology,Basic Medical School,Jilin University,Jilin 130021,China department of Biochemistry and molecular biology Basic Medical School Jilin University Jilin China

Aim:To investigate the effect and mechanism of morphine on purine nucleotidecatabolism.Methods:The rat model of morphine dependence and withdrawal andrat C6 glioma cells in culture were used.Concentrations of uric acid in the plasmawere measured by the uricase-rap method,adenosine deaminase(ADA)and xan-thine oxidase(XO)in the plasma and tissues were measured by the ADA and XOtest kit.RT-PCR and RT-PCR-Southern blotting were used to examine the relativeamount of ADA and XO gene transcripts in tissues and C6 cells.Results:(i)theconcentration of plasma uric acid in the morphine-administered group was signifi-cantly higher(P<0.05)than the control group;(ii)during morphine administrationand withdrawal periods,the ADA and XO concentrations in the plasma increasedsignificantly(P<0.05);(iii)the amount of ADA and XO in the parietal lobe,liver,small intestine,and skeletal muscles of the morphine-administered groupsincreased,while the level of ADA and XO in those tissues of the withdrawalgroups decreased;(iv)the transcripts of the ADA and XO genes in the parietallobe,liver,small intestine,and skeletal muscles were higher in the morphine-administered group.The expression of the ADA and XO genes in those tissuesreturned to the control level during morphine withdrawal,with the exception of theskeletal muscles;and(v)the upregulation of the expression of the ADA and XOgenes induced by morphine treatment could be reversed by naloxone.Conclusion:The effects of morphine on purine nucleotide metabolism might be an important,new biochemical pharmacological mechanism of morphine action.

关键词： morphine uric acid purine nucleotide adenosine deaminase xanthine oxidase

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Down-regulation of survivin expression reversed multidrug resistance in adriamycin-resistant HL-60/ADR cell line

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Acta Pharmacologica Sinica 2003年第12期24卷 61-66页

作者： WANG Lei, ZHANG Gui-Mei, FENG Zuo-Hua department of Biochemistry and molecular biology, Tongji Medical College Huazhong University of Science and Technology, Wuhan 430030, China department of Biochemistry and molecular biology Tongji Medical College Huazhong University of Science and Technology Wuhan 430030

AIM: To investigate the ability of an antisense RNA eukaryotic expression plasmid pcDNA3.1/survivin in downregulating the expression level of survivin mRNA and survivin protein and reversed multidrug resistance (MDR) in adriamycin-resistant HL-60/ADR cell line. METHODS: The expression of survivin mRNA was measured by RTPCR and the expression of survivin protein was measured by Western blot. Caspase-3 activity was determined by Phar Mingen colorimetric assay kit. Apoptosis was assessed by flow cytometry. The chemosensitivity of HL-60/ADR cells to adriamycin (ADR) was measured by MTT assay. RESULTS: pcDNA3.1/survivin down-regulated the expression level of survivin mRNA and survivin protein obviously, and induced apoptosis of HL-60/ADR cells in a time-dependent manner during 12-48 h. After transient transfection with pcDNA3.1/survivin for 48 h, survivin mRNA decreased by 67％, survivin protein decreased by 57％, caspase-3 activity increased 4.37 times, and the apoptosis rate increased by 4.41％ compared wi

关键词： antisense RNA doxorubicin apoptosis survivin caspase-3 HL-60 cells multiple drug resistance

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Dynamic distribution of Ser-10 phosphorylated histone H3 in cytoplasm of MCF-7 and CHO cells during mitosis

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Cell Research 2005年第2期15卷 120-126页

作者： Deng Wen LI, Qin YANG, Jia Tong CHEN, Hao ZHOU, Ru Ming LIU, Xi Tai HUANG*department of Biochemistry and molecular biology, College of Life Sciences, Nankai University, Tianjin 300071, China department of Biochemistry and molecular biology College of Life Sciences Nankai University Tianjin China

The dynamic distribution of phosphorylated Histone H3 on Ser10 (phospho-H3) in cells was investigated to determineits function during mitosis. Human breast adenocarcinoma cells MCF-7, and Chinese hamster cells CHO were analyzedby indirect immunofluorescence staining with an antibody against phospho-H3. We found that the phosphorylationbegins at early prophase, and spreads throughout the chromosomes at late prophase. At metaphase, most of the phospho-H3 aggregates at the end of the condensed entity of chromosomes at equatorial plate. During anaphase and telophase,the fluorescent signal of phospho-H3 is detached from chromosomes into cytoplasm. At early anaphase, phospho-H3shows ladder bands between two sets of separated chromosome, and forms “sandwich-like structure” when the chro-mosomes condensed. With the cleavage progressing, the “ladders” of the histone contract into a bigger bright dot. Thenthe histone aggregates and some of compacted microtubules in the midbody region are composed into a “bar-like”complex to separate daughter cells. The daughter cells seal their plasma membrane along with the ends of the “bar”,inside which locates microtubules and modified histones, to finish the cytokinesis and keep the “bar complex” out of thecells. The specific distribution and kinetics of phospho-H3 in cytoplasm suggest that the modified histones may takepart in the formation of midbody and play a crucial role in cytokinesis.

关键词： mitosis cytokinesis midbody Ser10 phosphorylated H3 microtubule complex.

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Pyruvate kinase M in germ cells is essential for sperm motility and male fertility but not spermatogenesis

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Asian Journal of Andrology 2024年第2期26卷 212-219页

作者： Gao-Qing Qian Xiao-Chen Wang Xi Zhang Bin Shen Qiang Liu State Key Laboratory of Reproductive Medicine and Offspring Health Nanjing Medical UniversityNanjing 211166China Shanghai Key Laboratory of Reproductive Medicine Department of HistoembryologyGenetics and Developmental BiologyShanghai Jiao Tong University School of MedicineShanghai 200025China State Key Laboratory of molecular biology Shanghai Key Laboratory of Molecular AndrologyShanghai Institute of Biochemistry and Cell BiologyCenter for Excellence in Molecular Cell ScienceChinese Academy of SciencesUniversity of Chinese Academy of SciencesShanghai 200031China

Male germ cells employ specific metabolic pathways throughout their developmental stages.In a previous study,we discovered heightened expression of pyruvate kinase M(PKM),a pivotal glycolytic enzyme,in spermatogonia and spermatids.To gain deeper insights into PKM's roles in spermatogenesis,sperm function,and male fertility,we engineered a conditional-knockout mouse model(Pkm-vkO mice)to selectively disrupt the Pkm gene within germ cells.Despite maintaining regular testicular histology and sperm morphology,the male Pkm-vko mice were infertility,characterized by significant impairments in sperm motility and adenosine triphosphate(ATP)generation.In addition,Pkm-null spermatozoa exhibited similar deficits in protein tyrosine phosphorylation linked to capacitation,as well as compromised performance in in vitro fertilization experiments.To conclude,PKM's presence is not obligatory for the entirety of spermatogenesis in male germ cells;however,it emerges as a critical factor influencing sperm motility and overall male fertility.

关键词： glycolysis male fertility pyruvate kinase M sperm motility spermatogenesis

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Emerging role of cellular senescence in normal lung development and perinatal lung injury

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Chinese Medical Journal Pulmonary and Critical Care Medicine 2024年第1期2卷 10-16页

作者： Phyllis A.Dennery Hongwei Yao department of molecular biology Cell Biology&BiochemistryDivision of Biology and MedicineBrown UniversityProvidenceRI 02912USA department of Pediatrics Warren Alpert Medical School of Brown UniversityProvidenceRI 02903USA

Cellular senescence is a status of irreversible growth arrest,which can be triggered by the p53/p21cip1 and p16INK4/Rb pathways via intrinsic and external factors.Senescent cells are typically enlarged and flattened,and characterized by numerous molecular features.The latter consists of increased surfaceome,increased resid-ual lysosomal activity at pH 6.0(manifested by increased activity of senescence-associated beta-galactosidase[SA-𝛽-gal]),senescence-associated mitochondrial dysfunction,cytoplasmic chromatin fragment,nuclear lamin b1 exclusion,telomere-associated foci,and the senescence-associated secretory phenotype.These features vary depending on the stressor leading to senescence and the type of senescence.Cellular senescence plays pivotal roles in organismal aging and in the pathogenesis of aging-related diseases.Interestingly,senescence can also both promote and inhibit wound healing processes.We recently report that senescence as a programmed pro-cess contributes to normal lung development.Lung senescence is also observed in Down Syndrome,as well as in premature infants with bronchopulmonary dysplasia and in a hyperoxia-induced rodent model of this disease.Furthermore,this senescence results in neonatal lung injury.In this review,we briefly discuss the molecular features of senescence.We then focus on the emerging role of senescence in normal lung development and in the pathogenesis of bronchopulmonary dysplasia as well as putative signaling pathways driving senescence.Finally,we discuss potential therapeutic approaches targeting senescent cells to prevent perinatal lung diseases.

关键词： Senescence Normal lung development Bronchopulmonary dysplasia Down syndrome Senolytics

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Characterization of the infectivity of an Indonesian Zika virus strain in mammalian cell lines

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Asian Pacific Journal of Tropical Biomedicine 2024年第5期14卷 215-224页

作者： Justus Thomas Obiajulu Sievers Anom Bowolaksono R.Tedjo Sasmono Graduate School of biology Department of BiologyFaculty of Mathematics and Natural SciencesUniversitas IndonesiaDepok 16424Indonesia Eijkman Research Center for molecular biology National Research and Innovation AgencyCibinong 16911Indonesia Cellular and molecular Mechanisms in Biological System Research Group Department of BiologyFaculty of Mathematics and Natural SciencesUniversitas IndonesiaDepok 16424Indonesia

Objective:To characterize the infection patterns and growth characteristics of the Zika virus(ZIKV)strain JMB-185 from Indonesia in various mammalian cell lines.Methods:ZIKV was grown in human(A549,HEK293,HepG2,Huh7,Jurkat,and THP-1)and non-human mammalian(RAW264.7,Vero,and Vero76)cell lines.Viral replication kinetics were measured using plaque assay,while intra-and extracellular viral RNA concentrations were assessed using RT-PCR.Flow cytometry was used to quantify the infected cells and cell viability was measured using an MTT assay.The ability of ZIKV to infect cell lines was visualized using a fluorescence immunostaining assay.Results:This ZIKV strain preferentially infected the lung,kidney,and liver cell lines A549,HEK293,Huh7,Vero,and Vero76,but not the immune cells Jurkat,RAW264.7,and THP-1.By contrast,the ZIKV showed no sign of infection in HepG2 cells,while maintaining viral titer over 3 days post-infection,with no infection recorded in immunostaining,no increase in viral RNA,and no indication of cell deterioration.Conclusions:The Indonesian ZIKV strain has a similar infection profile as other strains,except for its poor infectivity on HepG2 cells.Information on the growth characteristics of Indonesia ZIKV will help expand our understanding of the biology of ZIKV which will be useful for various applications including antiviral discovery.

关键词： Zika virus Infectivity Replication Cell lines In vitro

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