摘要：
The present study was to determine the targeting effect of M13 phage peptide ZL4 (MppZL4) on Schistosoma japonicum (S.j). Mice infected with S.j were injected with MppZL4. Real-time PCR was used to detect the distribution and metabolism of MppZL4 in the livers and lungs of mice. In vivo refusion test was performed to detect the targeting of MppZL4. Western blotting was employed to determine the expression of MppZL4. Live imaging was used to detect the distribution of oligopeptide MppZL4. Immunohistochemistry was employed to determine MppZL4 location on adult S.j body surface. Gomori method was employed to detect the influence of oligopeptide MppZL4 on alkaline phosphatase activity. The distribution and metabolism of MppZL4 and M13KE are not significantly different from each other at each time point. The abundance of MppZL4 is changed as S.j migrates in mice. The targeted binding effect of MppZL4 varies at different stages. ZL4 oligopeptide targets S.j in mice. The specific binding sites of MppZL4 on S.j body are mainly located in syncytial cells. The binding sites of MppZL4 on S.j body surface might be ALP or ALP-related proteins. MppZL4 had targeted binding effect on S.j with its binding site being associated with proteins related to S.j alkaline phosphatase. S.j tegument had a specifically binding site with exogenous peptides, offering new means to explore the interactions between hosts and parasites. Additionally, MppZL4 can possibly be used as targeting molecules in worm-resistant drugs or as tracing molecules in imaging diagnosis technologies.
摘要：
FBXW7 gene (F-box and WD-40 domain protein 7) is also named HCDC4 and is a significant tumor suppressor gene, which can regulate human cell cycle, proliferation and differentiation. In this study, we tend to investigate protein expression and related biological functions of FBXW7 gene. FBXW7 expression level in renal cell carcinoma (RCC) tissues is highly related to its clinical pathologic grade (P = 0.0094) and TNM phase (P = 0.0080) and is highly lower than in paracancerous normal tissues through immunohistochemistry study. FBXW7 high-expression patients have overall better prognosis than low-expression patients (P < 0.001). After transfected with FBXW7 plasmid, the RCC cell lines ACHN and A704 showed a depressed proliferation activity and high proportion of apoptosis through CCK8, colony formation and flow cytometry assay studies. By Western blot analysis, expression of cell proliferation-activating protein c-Myc and c-Jun is downregulated in FBXW7 high-expression RCC compared with negative control. These data suggested that FBXW7 is a significant tumor suppressor gene in RCC.
摘要：
Oxidized low-density lipoprotein (ox-LDL) is an independent risk factor of atherosclerosis. However, the mechanism underlying its pro-atherosclerosis roles has not yet been well explored. DNA demethylation modification, via DNA methyltransferases or ten-eleven-translocation (TET) family, is a crisis epigenetic regulation for various biological and pathological processes. This study aimed to investigate the effects of ox-LDL on macrophage autophagy and its potential epigenetic mechanism. Results showed that after treatment with 0, 10, 20, 40 or 80 mg/L ox-LDL for 24 h, the autophagy markers Beclin 1 and LC3 expression were obviously decreased at protein levels (P < 0.05). The mRNA and protein expression of TET2 was evidently decreased (P < 0.05). After pre-treatment with TET2 siRNA, the mRNA and protein levels of Beclin 1 and LC3 decreased compared with the 80 mg/L treatment group (P < 0.01). The mRNA and protein levels of Beclin 1 and LC3-II were up-regulated (P < 0.05) in the 5-aza-2′-deoxycytidine (a DNA methyltransferase inhibitor) of pretreatment group. Consistent with the Western blot results, cell immunofluorescence showed that the protein concentration of LC3-II decreased in the TET2 siRNA group and increased in the 5-aza-2′-deoxycytidine group. Taken together, these results showed that DNA demethylation modifications regulate ox-LDL-treated THP-1 macrophages autophagy and TET2 might be a novel regulator.
期刊：
Infection and Immunity,2014年82(12):5327-5335 ISSN：0019-9567
通讯作者：
Zhong, Guangming
作者机构：
[Liu, Yuanjun; Yang, Zhangsheng; Baseman, Joel; Huang, Yumeng; Gong, Siqi; Zhong, Guangming] Univ Texas Hlth Sci Ctr San Antonio, Dept Microbiol & Immunol, San Antonio, TX 78229 USA.;[Liu, Quanzhong; Hou, Shuping] Tianjin Med Univ Gen Hosp, Dept Dermatovenereol, Tianjin, Peoples R China.;[Sun, Yina] Tianjin Med Univ, Hosp & Tianjin Inst Endocrinol, Key Lab Hormones & Dev Minist Health, Metab Dis, Tianjin, Peoples R China.;[Li, Zhongyu; Wu, Yimou; Chen, Chaoqun] Univ South China, Dept Microbiol & Immunol, Hengyang, Hunan, Peoples R China.
通讯机构：
[Zhong, Guangming] U;Univ Texas Hlth Sci Ctr San Antonio, Dept Microbiol & Immunol, San Antonio, TX 78229 USA.
摘要：
Hydrosalpinx induction in mice by Chlamydia muridarum infection, a model that has been used to study C. trachomatis pathogenesis in women, is known to depend on the cryptic plasmid that encodes eight genes designated pgp1 to pgp8. To identify the plasmid-encoded pathogenic determinants, we evaluated C. muridarum transformants deficient in the plasmid-borne gene pgp3, -4, or -7 for induction of hydrosalpinx. C. muridarum transformants with an in-frame deletion of either pgp3 or -4 but not -7 failed to induce hydrosalpinx. The deletion mutant phenotype was reproduced by using transformants with premature termination codon insertions in the corresponding pgp genes (to minimize polar effects inherent in the deletion mutants). Pgp4 is known to regulate pgp3 expression, while lack of Pgp3 does not significantly affect Pgp4 function. Thus, we conclude that Pgp3 is an effector virulence factor and that lack of Pgp3 may be responsible for the attenuation in C. muridarum pathogenicity described above. This attenuated pathogenicity was further correlated with a rapid decrease in chlamydial survival in the lower genital tract and reduced ascension to the upper genital tract in mice infected with C. muridarum deficient in Pgp3 but not Pgp7. The Pgp3-deficient C. muridarum organisms were also less invasive when delivered directly to the oviduct on day 7 after inoculation. These observations demonstrate that plasmid-encoded Pgp3 is required for C. muridarum survival in the mouse genital tract and represents a major virulence factor in C. muridarum pathogenesis in mice.
摘要：
High levels of aldehyde dehydrogenase (ALDH) activity have been regarded as a specific feature of progenitor cells and stem cells. Hence, as an indicator of ALDH activity, aldefluor fluorescence has been widely used for the identification and isolation of stem and progenitor cells. ALDH activity was recently detected in embryonic mouse pancreas, and specifically and exclusively in adult centroacinar and terminal duct cells, suggesting that these duct cells may harbor cells of endocrine and exocrine differentiation potential in the adult pancreas. Here, we report the presence of aldefluor+ beta-cells in a beta-cell proliferation model, partial pancreatectomy. The aldefluor+ beta-cells are essentially all positive for Ki-67 and expressed high levels of cell-cycle activators such as CyclinD1, CyclinD2, and CDK4, suggesting that they are mitotic cells. Our data thus reveal a potential change in ALDH activity of proliferating beta-cells, which provides a novel method for the isolation and analysis of proliferating beta-cells. Moreover, our data also suggest that aldefluor lineage-tracing is not a proper method for analyzing progenitor or stem activity in the adult pancreas.
作者机构：
[Yu, M. J.; Tang, S. Y.; Liu, A. Y.; Liu, Y.; Liu, L. Z.; Wan, Y. P.; Zhang, Y.] Univ South China, Pathogen Biol Inst, Hengyang 421001, Peoples R China.;[Li, L.] Univ South China, Sch Publ Hlth, Hengyang 421001, Peoples R China.
通讯机构：
[Tang, S. Y.] U;Univ South China, Pathogen Biol Inst, Hengyang 421001, Peoples R China.
关键词：
human papillomavirus type16 (HPV16);E2;Daxx;interaction
摘要：
The aim of the study was to explore the interactions of human papilloma virus 16 (HPV16) E2 protein and Daxx. The localization or co-localization of PML and E2 with Daxx in Caski cells was observed by indirect immunofluorescence. The interaction of E2 and Daxx was analyzed by co-immunoprecipitation, Western-blot and yeast-two hybrid assays. In Caski cells the fluorescence of Daxx and PML was mainly distributed in the cytoplasm or nucleus, respectively, and in the align image their signals did not overlap. However, when the red signal of HPV16 E2 and the green signal of Daxx in the cytoplasm of Caski cells were merged, the yellow signal appeared. The yeast co-transformed with pGBKT7/Daxx and pGADT7/E2 or pGADT7/E2 TAD can grow on SD/-Trp-Leu-His and SD/-Trp-Leu-His-Ade plates. So Daxx is not colocated with PML but with HPV16 E2 mainly in the cytoplasm of Caski cells. On the base of the results one can propose that HPV16 E2, in particularly its transcription-activity domain (TAD), interacts with Daxx.
作者机构：
[Tianping Tan; Guangli Ou; Xiaoxing You; Yanhua Zeng; Minjun Yu; Cuiming Zhu] Institute of Pathogen Biology, University of South China, Hengyang City, Hunan Province, P.R.China;[Tianping Tan] The 415 affiliated hospital, University of South China, Hengyang City, Hunan Province, P.R.China;[Yan Liu] Shaoyang Medical College, Shaoyang City, Hunan Province, P.R.China
会议名称：
The 6th Meeting of the Asian Organization for Mycoplasmology (AOM)The 7th Meeting of the Chinese Society for Mycoplasmology (CSM)(第七届全国支原体学术会议暨第六届亚洲支原体学术会议)
会议时间：
2014-8-22
会议地点：
张家界
会议主办单位：
中华医学会微生物与免疫学分会;亚洲支原体学组织
会议论文集名称：
The 6th Meeting of the Asian Organization for Mycoplasmology (AOM)The 7th Meeting of the Chinese Society for Mycoplasmology (CSM)(第七届全国支原体学术会议暨第六届亚洲支原体学术会议)论文集
摘要：
Objectives This study was to investigate the expression of heme oxygenase 1 (Heme oxygenase, HO-1) induced by the lipid-associated membrane proteins(LAMPs) derived from Mycoplasma pneumoniae in the human monocyte cell line THP-1 cells.Methods The cytotoxicity of LAMPs to THP-1 cells was investigated by CCK-8 test.THP-1 cells were stimulated by different concentration of LAMPs for different time, and the expression of HO-1 protein and mRNA were detected by Western blot and qRTPCR respectively.
作者机构：
Institute of Cardiovascular Disease, Key Laboratory for Arteriosclerology of Hunan Province, University of South China, Hengyang 421001, China;School of Pharmacy, Wenzhou Medical College, 1210 University Town, Wenzhou, Zhejiang 325035, China
通讯机构：
Institute of Cardiovascular Disease, Key Laboratory for Arteriosclerology of Hunan Province, University of South China, China
期刊：
CANADIAN JOURNAL OF MICROBIOLOGY,2012年58(7):898-908 ISSN：0008-4166
通讯作者：
Wu, Yimou
作者机构：
[Liu, Yan; Wu, Yimou; Liu, Liangzhuan; Zhu, Cuiming; You, Xiaoxing; Zeng, Yanhua] Univ S China, Inst Pathogen Biol, Hengyang 421001, Peoples R China.;[He, Jun] Univ S China, Affiliated Nanhua Hosp, Hengyang 421000, Peoples R China.
通讯机构：
[Wu, Yimou] U;Univ S China, Inst Pathogen Biol, Hengyang 421001, Peoples R China.
关键词：
phage display peptide library;mimic epitope;Mycoplasma genitalium;adhesion protein;banque d’exposition de peptides sur phage;mimes fonctionnels d’épitopes;Mycoplasma genitalium;protéine d’adhésion
摘要：
Mycoplasma genitalium adhesion protein (MgPa) is the major adhesion protein of M. genitalium, and its Cterminal domain (amino acid 1075-1444) is the most immunogenic region. However, the exact epitopes of the adhesion protein of M. genitalium are still unclear. We used the purified polyclonal antibody against the recombinant adhesion protein to screen the mimic epitopes of MgPa using a random 12-peptide phage display library. Immunoscreening via the phage display peptide library revealed that 3 motifs (P-S-A-A/V-X-R-F/W-E/S-L-S-P, A-K-I/L-T/Q-X-T-L-X-L, and K-S-L-S-R-X-DX- I) may represent 3 different mimotopes of MgPa. Results of bioinformatics analysis by MIMOX demonstrated that the key consensus amino acid residues in the aligned mimotopes may be S, A, and F for cluster 1; A, K, I, T, and L for cluster 2; and K, S, L, R, D, and I for cluster 3. Three representative phages could recognize sera from M. genitalium-positive patients to varying degrees, whereas they could not recognize the sera from Mycoplasma pneumoniae-positive patients or the sera from healthy people. These findings will help to clarify the mimic epitopes of MgPa to facilitate diagnosis of the antigen and to understand the antigenic structure of MgPa.
摘要：
Inducing apoptosis is an attractive antitumor strategy. PERP is an apoptosis-associated target of p53, and its activation alone is sufficient to induce apoptotic pathway leading to cell death. We have previously demonstrated that overexpression of PERP in tumor cell lines with low intrinsic PERP activity suppressed cancer cell growth and enhanced sensitivity to chemotherapeutical agents. We further identified that PERP was present in surgical normal lung tissue, but absent in cancerous tissue of the same patient. Here, we sought to investigate the anti-tumor effects of PERP gene therapy in vivo. Then nude mice were transplanted with p53-mutanted Anip973 human lung cancer xenografts and treated with normal saline, pcDNA3.1 (vector) and pcDNA3.1-PERP, respectively. Successful transfection and robust expression of PERP was detected. Treatment with pcDNA3.1-PERP increased apoptosis and retarded growth in the xenografts, which contributed to a 55% decrease in tumor volume compared with controls. Furthermore, PERP gene therapy activated pro-apoptotic Caspase-3 cascade and upregulated the expression of the second mitochondria-derived activator of caspase (Smac) and human TNF-related apoptosis-inducing ligand (TRAIL), while suppressed vascular endothelial growth factor (VEGF) expression, indicating apoptosis and anti-angiogenesis are involved in the inhibitory effect of the PERP gene therapy. Taken together, our results suggest PERP gene therapy may supply an alternative strategy for lung adenocarcinoma management. Furthermore, Anip973 is a p53-mutanted cell line and the findings of this study provide reference value for other p53-mutanted cancers which is common among malignant tumors.
摘要：
The role of renal lipoprotein lipase (LPL) per se in kidney diseases is still controversial and obscure. The purpose of this study was to observe the preventive effects of Ibrolipim, a LPL activator, on lipid accumulation and LPL expression in the kidneys of minipigs fed a high-sucrose and high-fat diet (HSFD). Male Chinese Bama minipigs were fed a control diet or HSFD with or without 0.1 g/kg/day Ibrolipim for 5 months. Body weight, plasma glucose, insulin, lipids, LPL activity, and urinary microalbumin were measured. Renal tissue was obtained for detecting LPL activity and contents of triglyceride and cholesterol, observing the renal lipid accumulation by Oil Red O staining, and examining the mRNA and protein expression of LPL by real time PCR, Western Blot and immunohistochemistry. Feeding HSFD to minipigs caused weight gain, hyperglycemia, hyperinsulinemia, hyperlipidemia and microalbuminuria. HSFD increased plasma LPL activity while it decreased the mRNA and protein expression and activity of LPL in the kidney. The increases in renal triglyceride and cholesterol contents were associated with the decrease in renal LPL activity of HSFD-fed minipigs. In contrast, supplementing Ibrolipim into HSFD lowered body weight, plasma glucose, insulin, triglyceride and urinary albumin concentrations while it increased plasma total cholesterol and HDL-C. Ibrolipim suppressed the renal accumulation of triglyceride and cholesterol, and stimulated the diet-induced down-regulation of LPL expression and activity in the kidney. Ibrolipim exerts renoprotective and hypolipidemic effects via the increase in renal LPL activity and expression, and thus the increased expression and activity of renal LPL play a vital role in suppressing renal lipid accumulation and ameliorating proteinuria in diet-induced diabetic minipigs.
