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| 1 | Reversal of multidrug resistance in drug-resistant human gastric cancer cell line SGC7901/VCR by antiprogestin drug mifepristone显示文摘AIM: To explore the reversal effect of mifepristone on multidrug resistance (MDR) in drug-resistant human gastric cancer cell line SGC7901/VCR and its mechanisms. METHODS: Expression of multidrug resistance-associated protein(MRP) was detected using reverse transcriptionpolymerase chain reaction(RT-PCR). Flow cytometry was used to assay the expression of P-glycoprotein(P-gp), Bcl-2,Bax, and the mean fluorescent intensity of intracellular rhodamine 123 in the cells. Meanwhile, the protein levels of Bcl-2 and Bax were also detected by Western blotting analysis. The sensitivity of cells to the anticancer agent, vincrimycin(VCR), and the intracellular [^3H]VCR accumulation were determined by tetrazolium blue (MTT) assay and a liquid scintillation counter, respectively. RESULTS: Expression of MRP and P-gp in SGC7901/VCR cells was 6.04-and 8.37-fold higher as compared with its parental SGC7901 cells, respectively. After treatment with 1, 5, 10, and 20 umol/L mifepristone, SGC7901/VCR cells showed a 1.34-, 2.29-, 3.11-, and 3.71-fold increase in the accumulation of intracellular VCR, a known substrate of MRP, and a 1.03-, 2.04-, 3.08-, and 3.68-fold increase in the retention of rhodamine 123, an indicator of P-gp function, respectively. MTT assay revealed that the resistance of SGC7901/VCR cells to VCR was 11.96-fold higher than that of its parental cells. The chemosensitivity of SGC7901/VCR cells to VCR was enhanced by 1.02-, 7.19-, 12.84-, and 21.17-fold after treatment with mifepristone at above-mentioned dose. After 96 h of incubation with mifepristone 10 umol/L, a concentration close to plasma concentrations achievable in human, the expression of Bcl-2 protein was decreased to (9.21+0.65)% from (25.32+1.44)%, whereas the expression of Bax protein was increased to (19.69+1.13)% from (1.24+0.78)% (P<0.01). Additionally, the effects of mifepristone on the expression of Bcl-2 and Bax proteins in SGC7901/VCR cells were further demonstrated by Western blotting analysis. CONCLUSION: Mifepristone has potent reversal effect on MDR in SGC7901/VCR via inhibiting the function of MRP and P-gp, modulating the expression of Bcl-2 and Bax proteins, and enhancing the sensitivity to anticancer agent VCR. | Da-QiangLi Zhi-BiaoWang JinBai JieZhao YuanWang KaiHu Yong-HongDu | 2004 | World Journal of Gastroenterology2004，10，12： | 29 |
| 2 | Identification of an epitope of SARS-coronavirus nucleocapsid protein显示文摘The nucleocapsid (N) protein of severe acute respiratory syndrome-coronavirus (SARS-CoV) is a majorvirion structural protein. In this study, two epitopes (N1 and N2) of the N protein of SARS-CoV werepredicted by bioinformatics analysis. After immunization with two peptides, the peptides-specific antibodieswere isolated from the immunized rabbits. The further experiments demonstrated that N1 peptide-inducedpolyclonal antibodies had a high affinity to bind to E. coli expressed N protein of SARS-CoV. Furthermore, itwas confirmed that N1 peptide-specific IgG antibodies were detectable in the sera of severe acute respiratorysyndrome (SARS) patients. The results indicated that an epitope of the N protein has been identified andN protein specific Abs were produced by peptide immunization, which will be useful for the study of SARS-CoV. | YINGLIN XuSHEN RUIFuYANG YIXUELI YONGYONGJI YouYuHE MuDESHI WEILU TIELIUSHI BINGSUN JINWANG HONGXIAWANG HUALIANGJIANG JIANHUASHEN YOUHUAXIE YUANWANG GANGPEI BBIFENSHEN JIARUIWU | 2003 | Cell Research2003，13，3： | 22 |
| 3 | A candidate DNA vaccine elicits HCV specific humoral and cellular immune responses显示文摘AIM: TO investigate the immunogenicity of candidate DNA vaccine against hepatitis C virus (HCV) delivered by two plasmids expressing HCV envelope protein 1 (El) and envelope protein 2 (E2) antigens respectively and to study the effect of CpG adjuvant on this candidate vaccine.METHODS: Recombinant plasmJds expressing HCV EI and E2 antigens respectively were used to simultaneously inoculate mice with or without CpG adjuvant. Antisera were then collected and tJters of antJ-HCV antibodies were analyzed by ELISA. One month after the last injection, animals were sacrificed to prepare single-cell suspension of splenocytes.These cells were subjected to HCV antigen specific proliferaion assays and cytokine secretion assays to evaluate the cellular immune responses of the vaccinated animals.RESULTS: Antibody responses to HCV EI and E2 antigens were detected in vaccinated animals. Animals receiving CpG adjuvant had slightly lower titers of anti-HCV antibodies in the sera, while the splenocytes from these animals showed higher HCV-antigen specific proliferation. Analysis of cytokine secretion from the splenocytes was consistent with the above results. While no antigen-specific IL-4 secretion was detected for all vaccinated animals, HCV antigen-specific INF-γ, secretion was detected for the splenocytes of vaccinated animals. CpG adjuvant enhanced the secretion of INF-γ, but did not change the profile of IL-4 secretion.CONCLUSION: Vaccination of mice with plasmids encoding HCV E1 and E2 antigens induces humoral and cellular immune responses. CpG adjuvant significantly enhances the cellular immune response. | Li-XinZhu JingLiu YeYe You-HuaXie Yu-YingKong Guang-DiLi YuanWang | 2004 | World Journal of Gastroenterology2004，10，17： | 12 |
| 4 | Effects of mifepristone on invasive and metastatic potential of human gastric adenocarcinoma cell line MKN-45 in vitro and in vivo显示文摘AIM: To investigate the effects of mifepristone on the invasive and metastatic potential of human gastric adenocarcinoma cell line MKN-45 and its mechanisms. METHODS: After incubation with various concentrations of mifepristone (5, 10, 20 umol/L), the adhesion to artificial basement membrane, Matrigel, and the migration of MKN-45 cells were assayed using MTT assay and Transwell cell culture chambers, respectively. Enzyme- linked immunoabsorbent assay (ELISA) and flow cytometry were used to determine the expression of vascular endothelial growth factor (VEGF) and integrin 133 in the cells. After subcutaneous transplantation of MKN-45 cells in nude mice, mifepristone (50 mg/kg.d) was administrated subcutaneously for 8 wk to assess its effects on tumor metastasis. Immunohistochemical analysis was used to detect the expression of VEGF and microvascular density (MVD) in xenografted tumors. RESULTS: Mifepristone dose-dependently inhibited the heterotypic adhesion to Matrigel of MKN-45 cells. The inhibition was accompanied by a significant down-regulation of integrin 133 expression in the cells. After incubation with 5, 10, 20 umol/L mifepristone, the number of migrated MKN-45 cells was 72+8, 50+6, 41+5 in experiment group, and 94+16 in control group (P<0.01). Meanwhile, secreted VEGF protein of MKN-45 cells in mifepristone-treated group (14.2+2.9, 8.9+3.1, 5.4+2.1 ng/g per liter) was significantly lower than that in control group (22.7+4.3 ng/g per liter, P<0.01). In vivo, mifepristone decreased the number of metastatic foci in lungs of nude mice and down-regulated the expression of VEGF and MVD in the xenograted tumors. CONCLUSION: Mifepristone can effectively inhibit the invasive and metastatic potential of human gastric adenocarcinoma cell line MKN-45 in vitro and in vivo through inhibition of heterotypic adhesion to basement membrane, cell migration and angiogenesis. | Da-QiangLi Zhi-BiaoWang JinBai JieZhao YuanWang KaiHu Yong-HongDu | 2004 | World Journal of Gastroenterology2004，10，12： | 10 |
| 5 | Gene expression profiling of rat livers with Yin-deficiency-heat syndrome显示文摘OBJECTIVE: To explore the nature of 'Yin internal heat caused byYin-deficiency,' in terms of the theory of Traditional Chinese Medicine, by studying energy metabolism in rats with Yin-deficiency-heat syndrome and analyzing the gene expression profile of their livers. METHODS: A Yin-deficiency-heat syndrome model was induced in rats using three Chinese medicinal herbs. Glycogen and triglycerides in blood plasma, and the enzyme activity of ATP in livers were measured colorimetrically. Triiodothyronine (T3 ), thyroxine (T4 ), and thyroid stimulating hormone levels in blood plasma were also measured with enzyme linked immunosorbent assay. The gene expression profile of livers was detected with gene chip analysis. Differentially expressed genes were screened out and classified according to Gene Ontology. The accuracy of results were examined with reversetranscription-polymerase chain reaction. RESULTS: Compared with the control group, body weight (P<0.05) and hepatic glycogen (P<0.05) were significantly lower in the Yin-deficiency-heat syndrome group. Moreover, toe temperature (P< 0.01) and triglyceride (P<0.05), Na+ -K+ -ATPase (P< 0.01), Mg2+ -ATPase (P<0.01), T3 (P<0.05), and T4 (P< 0.01) levels were significantly higher. There were 99 differentially expressed genes in livers from the Yin-deficiency-heat syndrome group. Genes were mainly related to sterol synthesis (Pc=0.0392), defense response (Pc=0.0448), and sterol metabolism (Pc=0.0533). CONCLUSION: Abnormal expression genes in rats with Yin-deficiency-heat syndrome prompted the synthesis and metabolism of cholesterol, increased energy consumption, and reduced defense response. This gene expression might be the molecular mechanism underlying 'internal heat caused by Yin-deficiency' in the rats with Yin-deficiency-heat syndrome. | Bingbing Han Shijun Wang Lin Li YuanWang Haijun Zhao | 2013 | Journal of Traditional Chinese Medicine2013，33，3： | 8 |
| 6 | Expression and purification of the complete PreS region of hepatitis B Virus显示文摘AIM: To express the complete PreS region of HBV in E.coli with good solubility and stability, and to establish an effective method for purification of the recombinant PreS protein.METHODS: The complete PreS region (PreS1 and PreS2) was fused into a series of tags including glutathione Stransferase (GST), dihydrofolate reductase (DHFR), maltose binding protein (MBP), 6x histidine, chitin binding domain (CBD), and thioredoxin, respectively. Expression of recombinant PreS fusion proteins was examined by SDS-PAGE analysis and confirmed by Western blot. Two fusion proteins, thio-PreS, and PreS-CBD, with desirable solubility and stability, were subjected to affinity purification and further characterization. RESULTS: Recombinant PreS fusion proteins could besynthesized with good yields in E.coli However, most of these proteins except for thio-PreS and PreS-CBD were vulnerable to degradation or insoluble as revealed by SDS PAGE and Western blot. Thio-PreS could be purified by affinity chromatography with nickel-chelating sepharose as the matrix. However, some impurities were also copurified. A simple freeze-thaw treatment yielded most of the thio-PreS proteins in solution while the impurities were in the precipitate. Purified thio-PreS protein was capable of inhibiting the binding of HBV virion to a specific monoclonal antibody against an epitope within the PreS1 domain. CONCLUSION: Increased solubility and stability of the complete PreS region synthesized in E.coli can be achievedby fusion with the thioredoxin or the CBD tag. A simple yet highly effective method has been established for the purification of the thio-PreS protein. Purified thio-PreS protein likely assumes a native conformation, which makes it an ideal candidate for studying the structure of the PreS region as well as for screening antivirals. | QiangDeng Yu-YingKong You-HuaXie YuanWang | 2005 | World Journal of Gastroenterology2005，11，20： | 8 |
| 7 | Identification of the immunogenic domains in HBsAg preS1 region using overlapping preS1 fragment fusion proteins显示文摘AIM: The incorporation of hepatitis B virus (HBV) preS1 region into epitope-based vaccines against HBV has been accepted widely, but the incorporate site and size of preS1 sequence is controversial. Therefore our purpose was to further investigate its immunogenic domains for the epitopebased hepatitis B vaccine design.METHODS: Eight GST fusion proteins containing overlapping preS1 fragments in preS1 (21-119) region were expressed in E.coli. Using these purified fusion proteins, the immunogenic domains in preS1 region were identified in detail in mice and humans by Western blot analysis and ELISA.RESULTS: The results in mice showed that the immunogenic domains mainly existed in preS1 (21-59) and preS1 (95-109). Similarly, these fragments had strong immunogenicity in humans; whereas the other parts except for preS1 (60-70) also had some immunogenicity.More importantly, a major immunogenic domain, preS1 (34-59), which has much stronger immunogenicity, was identified. Additionally, the antibodies against some preS1 fragments, especially preS1 (34-59), were speculated to be virus-neutralizing.CONCLUSION: Eight GST fusion proteins containing overlapping preS1 fragments were prepared successfully. They were used for the study on the immunogenic domains in preS1 (21-119) region. The preS1 (34-59) fragments were the major immunogenic domains in the preS1 region, and the antibodies against these fragments were speculated to be virus-neutralizing. Therefore, the incorporation of preS1 (34-59) fragments into epitopebased HBV vaccines may be efficient for enhancement of immune response. Additionally, the results also imply that there are more complex immune responses to preS1 region and more abundant immunogenic domains in humans. | Wei-GuoHu JunWei Heng-ChuanXia Xin-XiuYang FengLi Guang-DiLi YuanWang Zu-ChuanZhang | 2005 | World Journal of Gastroenterology2005，11，14： | 7 |
| 8 | LRH-1/hB1F and HNF1 synergistically up-regulate hepatitis B virus gene transcription and DNA replication显示文摘Enhancer Ⅱ(ENII)is one of the critical cis-elements in the Hepatitis B Virus(HBV)genome for the hepatic viral gene transcription and DNA replication.The liver-specific activity of ENII is regulated by multiple liver-enriched transcription factors,including LRH-1/hB 1 F,HNF 1,HNF3β,HNF4 and C/EBP.Knowledge on the interplay of these important factors is still limited.In this study,we demonstrate a functional synergism between the orphan nuclear receptor LRH-1/hB 1F and the homeoprotein HNF1 in up-regulating the liver-specific activity of ENII.This synergism is sufficient for initiating the viral gene transcription and DNA replication in non-hepatic cells.We have defined the activation domains in hB 1F and HNF 1 that contribute to the synergism.We further show that hB 1F and HNF1 can interact directly in vitro and have mapped the domains required for this interaction. | YANNINGCAI QINGZHOU YUYINGKONG MEILI BENOITVIOLLET YOUHUAXIE YUANWANG | 2003 | Cell Research2003，13，6： | 6 |
| 9 | Construction of prokaryotic expression system of ItB-ureB fusion gene and identification of the recombinant protein immunity and adjuvanticity显示文摘AIM: To construct ItB-ureBfusion gene and its prokaryotic expression system and identify immunity and adjuvanticity of the expressed recombinant protein.METHODS: The ureB gene from a clinical Helicobacter pylori (H pylon) strain Y06 and the ItB gene from Escherichia coli (E. coli) strain 44851 were linked into ItB-ureB fusion gene by PCR. The fusion gene sequence was analyzed after T-A cloning. A prokaryotic recombinant expression vector pET32a inserted with ltB-ureB fusion gene (pET32a-ltB-ureB) was constructed. Expression of the recombinant LTB-UreB protein (rLTB-UreB) in E. coliBL21DE3 induced by isopropylthio-β-D-galactoside (IPTG) at different concentrations was detected by SDS-PAGE. Western blot assays were used to examine the immunoreaction of rLTB-UreB by a commercial antibody against whole cell of Hpylori and a self-prepared rabbit anti-rUreB serum, respectively,and determine the antigenicity of the recombinant protein on inducing specific antibody in rabbits. GM1-ELISA was used to demonstrate the adjuvanticity of rLTB-UreB.Immunoreaction of rLTB-UreB to the UreB antibody positive sera from 125 gastric patients was determined by using ELISA.RESULTS: In comparison with the corresponding sequences of original genes, the nucleotide sequence homologies of the cloned ltB-ureB fusion gene were 100%. IPTG with different dosages of 0.1-1.0 mmol/L could efficiently induce pET32a-ltB-ureB-E, coli BL21DE3 to express the rLTB-UreB.The output of the target recombinant protein expressed by pET32a-ureB-E.coli BL21DE3 was approximately 35% of the total bacterial proteins, rLTB-UreB mainly presented in the form of inclusion body. Western blotting results demonstrated that rLTB-UreB could combine with the commercial antibody against whole cell of H pylori and anti-rUreB serum as well as induce rabbit to produce specific antibody. The strong ability of rLTB-UreB bindingbovine GM1 indicated the existence of adjuvanticity of the recombinant protein. All the UreB antibody positive sera from the patients (125/125) were positive for rLTB-UreB.CONCLUSION: A recombinant prokaryotic expression system with high expression efficiency of the target fusion gene ltB-ureB was successfully established. The expressed rLTB-UreB showed qualified immunogenicity, antigenicity and adjuvanticity. All the results mentioned above laid a firm foundation for further development of Hpylorigenetically engineered vaccine. | JieYan YuanWang Shi-HeShao Ya-FeiMao Hua-WenLi Yi-HuiLuo | 2004 | World Journal of Gastroenterology2004，10，18： | 6 |
| 10 | Expression of hepatitis C virus envelope protein 2 induces apoptosis in cultured mammalian cells显示文摘AIM: To explore the role of hepatitis C virus (HCV) envelope protein 2 (E2) in the induction of apoptosis. METHODS: A carboxyterminal truncated E2 (E2-661) was transiently expressed in several cultured mammalian cell lines or stably expressed in Chinese hamster ovary (CliO) cell line. Cell proliferation was assessed by ^3H thymidine uptake. Apoptosis was examined by Hoechst 33258 staining, flow cytometry and DNA fragmentation analysis. RESULTS: Reduced proliferation was readily observed in the E2-661 expressing cells. These cells manifested the typical features of apoptosis, including cell shrinkage, chromatin condensation and hypodiploid genomic DNA content. Similar apoptotic cell death was observed in an E2-661 stably expressing cell line. CONCLUSION: HCV E2 can induce apoptosis in cultured mammalian cells. | Li-XinZhu JingLiu You-HuaXie Yu-YingKong YeYe Chun-LinWang Guang-DiLi YuanWang | 2004 | World Journal of Gastroenterology2004，10，20： | 6 |
| 11 | Effects of mifepristone on proliferation of human gastric adenocarcinoma cell line SGC-7901 in vitro显示文摘AIM: To explore the effects of mifepristone, a progesterone receptor (PR) antagonist, on the proliferation of human gastric adenocarcinoma cell line SGC-7 901 in vitro and the possible mechanisms involved.METHODS: In situ hybridization was used to detect the expression of PR mRNA in SGC-7 901 cells. After treatment with various concentrations of mifepristone (2.5, 5, 10,20μmol/L) at various time intervals, the ultrastructural changes, cell proliferation, cell-cycle phase distribution, and the expression of caspase-3 and Bcl-XL were analyzed using transmission electron microscopy (TEM), tetrazolium blue (MTT) assay, ^3H-TdR incorporation, flow cytometry, and reverse transcription-polymerase chain reaction (RT-PCR).RESULTS: Mifepristone markedly induced apoptosis and inhibited cell proliferation of PR- positive SGC-7 901 cells revealed by TEM, MTT assay and ^3H-TdR incorporation, in a dose- and time-dependent manner. The inhibitory rate was increased from 8.98% to 51.29%. Flow cytometric analysis showed mifepristone dose-dependently decreased cells in S and G2/M phases, increased cells in G0/G1 phase,reduced the proliferative index from 57.75% to 22.83%.In addition, mifepristone up-regulated the expression of caspase-3, and down- regulated the Bcl-XL expression,dose-dependently.CONCLUSION: Mifepristone effectively inhibited the proliferation of PR-positive human gastric adenocarcinoma cell line SGC-7 901 in vilrothrough multiple mechanisms, and may be a beneficial agent against human adenocarcinoma. | Da-QiangLi Zhi-BiaoWang JinBai JieZhao YuanWang KaiHu Yong-HongDu | 2004 | World Journal of Gastroenterology2004，10，18： | 5 |
| 12 | The role of Arabidopsis 5PTase13 in root gravitropism through modulation of vesicle trafficking显示文摘肌醇 polyphosphate 5 磷酸酶(5PTases ) 是 phosphatidylinositol 新陈代谢的酶影响植物生长和开发的各种各样的方面。Arabidopsis 5PTase13 调整植物生长素动态平衡和荷尔蒙相关的子叶静脉开发,并且这里,我们证明它的猛烈变异的 5pt13 在根 gravitropic 回答提高了敏感到 gravistimulation。到 1-N-naphthylphthalamic 酸(一个植物生长素运输禁止者) 的 5pt13 异种的改变的回答显示 5PTase13 可能涉及植物生长素运输的规定。确实,植物生长素流出搬运人 PIN2 在 5PTase13 缺乏下面更广泛地被表示,并且膜选择的染料 FM4-64 的成为主观的观察在 5pt13 异种揭示改变的泡 trafficking。与相比野类型, 5pt13 异种幼苗对由泡骑车,幼苗生长,并且 PIN1 和 PIN2 蛋白质的细胞内部的骑车的 brefeldin A 的抑制不太敏感。进一步,在 gravitropic 刺激之上的植物生长素再分配在 5PTase13 缺乏下面被刺激。这些结果建议 5PTase13 可以由调整泡 trafficking 调制植物生长素运输并且从而在根 gravitropism 起一个作用。 | YuanWang Wen-Hui Lin Xu Chen Hong-Wei Xue | 2009 | Cell Research2009，19，10： | 5 |
| 13 | A note on overshoot estimation in pole placements显示文摘In this note we show that for a given controllable pair ( A, B ) and any λ> 0, a gain matrix K can be chosen so that the transition matrix e(A+BK)t of the system x = (A + BK)x decays at the exponential rate e-λt and the overshoot of the transition matrix can be bounded by MλL for some constants M and L that are independent ofλ. As a consequence, for any h >0, a gain matrix K can be chosen so that the magnitude of the transition matrix e(A+BK)t can be reduced by -1/2- (orby anygivenportion) over [0, h ] . An interesting applicafon of the result is in the stabilization of switched linear systems with any given switching rate ( see [ 1 ] ). | DaizhanCHENG LeiGUO YuandanLIN YuanWANG | 2004 | 控制理论与应用(英文版)2004，2，2： | 5 |
| 14 | Screening for PreS specific binding ligands with a phage displayed peptides library显示文摘AIM: To construct a random peptide phage display library and search for peptides that specifically bind to the PreS region of hepatitis B virus (HBV).METHODS: A phage display vector, pFuse8, based on the gene 8 product (pⅧ) of M13 phage was made and used to construct a random peptide library. E. coli derived thioredoxin-PreS was purified with Thio-bond beads, and exploited as the bait protein for library screening. Five rounds of bio-panning were performed. The PreS-binding specificities of enriched phages were characterized with phage ELISA assay.RESULTS: A phage display vector was successfully constructed as demonstrated to present a pⅧ fused HBV PreS1 epitope on the phage surface with a high efficiency.A cysteine confined random peptide library was constructed containing independent clones exceeding 5±108 clone forming unit (CFU). A pool of phages showing a PreS-binding specificity was obtained after the screening against thioPres with an enrichment of approximately 400 times. Five phages with high PreS-binding specificities were selected and characterized. Sequences of the peptides displayed on these phages were determined.CONCLUSION: A phage library has been constructed,with random peptides displaying as pⅧ-fusion proteins.Specific PreS-binding peptides have been obtained, which may be useful for developing antivirals against HBV infection. | QiangDeng MingZhuang Yu-YingKong You-HuaXie YuanWang | 2005 | World Journal of Gastroenterology2005，11，26： | 4 |
| 15 | Expression, Purification and Sublocalization of SARS-CoV Nucleocapsid Protein in Insect Cells显示文摘The causative agent of severe acute respiratory syndrome (SARS) is a previously unidentified coronavirus, SARS-CoV. The nucleocapsid (N) protein of SARS-CoV is a major viral protein recognized by acute and early convalescent sera from SARS patients. To facilitate the studies on the function and structure of the N protein, this report describe the expression and purification of recombinant SARS-CoV N protein using the baculovirus expression system. Recombinant hexa-histidine-tagged N protein with a molecularmass of 47 kD was produced in insect cells. Recombinant N protein was purified to near homogeneity by Ni^2+-NTA affinity chromatography. In addition, we examined the subcellular localization of the N protein by confocal microscopy in Trichoplusia ni BT1 Tn 5B1-4 cells infected with recombinant baculovirus. The N protein was found localized in the cytoplasm as well as in the nucleolus. The purified recombinant N protein can be used in further functional study of SARS-CoV. | Ai-XiaREN You-HuaXIE Yu-YingKONG Guan-ZhenYANG Yao-ZhouZHANG YuanWANG Xiang-FuWU | 2004 | Acta Biochimica et Biophysica Sinica2004，36，11： | 4 |
| 16 | Distribution and expression of non-muscle myosin light chain kinase in rabbit livers显示文摘AIM: To study the distribution and expression of non-muscle myosin light chain kinase (nmMLCK) in rabbit livers.METHODS: Human nmMLCK N-terminal cDNA was amplified by polymerase chain reaction (PCR) and was inserted into pBKcmv to construct expression vectors. The recombinant plasmid was transformed into XL1-blue. Expression protein was induced by IPTG and then purified by SDS-PAGE and electroelution, which was used to prepare the polycolonal antibody to detect the distribution and expression of nmMLCK in rabbit livers with immunofluorescene techniques.RESULTS: The polyclonal antibody was prepared, by which nmMLCK expression was detected and distributed mainly in peripheral hepatocytes.CONCLUSION: nmMLCK can express in hepatocytes peripherally, and may play certain roles in the regulation of hepatic functions. | Hua-QingZhu YuanWang Ruo-LeiHu BinRen QingZhou Zhi-KuiJiang Shu-YuGui | 2003 | World Journal of Gastroenterology2003，9，12： | 3 |
| 17 | Vibration Performance Analysis of a Mining Vehicle with Bounce and Pitch Tuned Hydraulically Interconnected Suspension显示文摘The current investigations primarily focus on using advanced suspensions to overcome the tradeo design of ride comfort and handling performance for mining vehicles. It is generally realized by adjusting spring sti ness or damping parameters through active control methods. However, some drawbacks regarding control complexity and uncertain reliability are inevitable for these advanced suspensions. Herein, a novel passive hydraulically interconnected suspension(HIS) system is proposed to achieve an improved ride-handling compromise of mining vehicles. A lumped-mass vehicle model involved with a mechanical–hydraulic coupled system is developed by applying the free-body diagram method. The transfer matrix method is used to derive the impedance of the hydraulic system, and the impedance is integrated to form the equation of motions for a mechanical–hydraulic coupled system. The modal analysis method is employed to obtain the free vibration transmissibilities and force vibration responses under di erent road excitations. A series of frequency characteristic analyses are presented to evaluate the isolation vibration performance between the mining vehicles with the proposed HIS and the conventional suspension. The analysis results prove that the proposed HIS system can e ectively suppress the pitch motion of sprung mass to guarantee the handling performance, and favorably provide soft bounce sti ness to improve the ride comfort. The distribution of dynamic forces between the front and rear wheels is more reasonable, and the vibration decay rate of sprung mass is increased e ectively. This research proposes a new suspension design method that can achieve the enhanced cooperative control of bounce and pitch motion modes to improve the ride comfort and handling performance of mining vehicles as an e ective passive suspension system. | Jie Zhang Yuanwang Deng Nong Zhang Bangji Zhang Hengmin Qi Minyi Zheng | 2019 | Chinese Journal of Mechanical Engineering2019，32，1： | 3 |
| 18 | Construction of prokaryotic expression system of 2 148-bp fragment from cagA gene and detection of cagA gene,CagA protein in Helicobacter pylori isolates and its antibody in sera of patients显示文摘To construct a prokaryotic expression system of a Helicobacter pylori ( H pylori) cagA gene fragment and establish enzyme-linked immunosorbent assays-(ELISA) for detectin.g Ca.gA.and its antibody, so as to understand the manner in which the infection of CagA-expressing Hpylori(CagA+ Hpylori) isolates cause diseases. | JieYan YuanWang Shi-HeShao Ya-FeiMao Hua-WenLi Yi-HuiLuo | 2004 | World Journal of Gastroenterology2004，10，8： | 3 |
| 19 | Loss of heterozygosity on chromosome 10q22-10q23 and 22q 11.2-22q 12.1 and p53gene in primary hepatocellular carcinoma显示文摘AIM: To analyze loss of heterozygosity (LOH) and homozygous deletion on p53 gene (exon2-3, 4 and 11), chromosome 10q22-10q23 and 22q11.2 -22q12.1 in human hepatocellular carcinoma (HCC).METHODS: PCR and PCR-based microsatellite polymorphism analysis techniques were used.RESULTS: LOH was observed at D10S579 (10q22-10q23) in 4 of 20 tumors (20%), at D22S421 (22q11.2-22q12.1) in 3 of 20(15%), at TP53.A (p53gene exon 2-3) in 4 of 20 (20%), at TP53.B (p53gene exon 4) in 6 of 20(30%), and at TP53.G (p53gene exon 11)in 0 of 20(0%). Homozygous deletion was detected at 10q22-10q23(8/20; 40%), 22q11.2-22q12.1(8/20; 40%), p53 gene exon 2-3(0/20;0%), p53gene exon 4(6/20; 30%), and p53gene exon 11(2/20; 10%).CONCLUSION: There might be unidentified tumor suppressor genes on chromosome 10q22-10q23 and 22q11.2-22q12.1 that contribute to the pathogenesis and development of HCC. | Guang-NengZhu LiZuo QingZhou Su-MeiZhang Hua-QingZhu Shu-YuGui YuanWang | 2004 | World Journal of Gastroenterology2004，10，13： | 3 |
| 20 | Establishment of transgenic mice carrying the gene of human nuclear receptor NR5A2 (hB1F)显示文摘AIM: Human hepatitis B virus enhancer II B1 binding factor (hBIF) was cloned and characterized as a novel member of the Ftz-F1 (NR5A) nuclear receptor subfamily. Although progresses have recently been made, its biological function remains largely unidentified. The aim of this study was to establish an hBIF transgenic mouse model to promote the functional study of hB1F.METHODS: Transgene fragments were microinjected into fertilized eggs of mice. The manipulated embryos were transferred into the oviducts of pseudopregnant female mice.The offsprings were identified by PCR and Southern blot analysis. Transgene expression was analyzed with RT-PCR and Western blot analysis. Transgenic founder mice were used to establish transgenic mouse lineages. The F1 and F2 mice were identified by PCR analysis.RESULTS: Seven mice were identified as carrying copies of transgene. RT-PCR and Western blotting results showed that the transgene was expressed in heart, liver, lung, kidney and stomach in one of the transgenic mouse lineages.Genetic analysis of the transgenic mice demonstrated that the transgene was integrated into the chromosome at a single site, and was transmitted stably.CONCLUSION: In this study we established an hB1F transgenic mouse model, which will facilitate the investigation of the biological function of hB1F in vivo. | Shui-LiangWang HuaYang You-HuaXie YuanWang Jian-ZhongLi LongWang Zhu-GangWang Ji-LiangFu | 2003 | World Journal of Gastroenterology2003，9，6： | 3 |
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