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1.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 265-269, 2014.
Article in English | WPRIM | ID: wpr-351085

ABSTRACT

There is controversy regarding the roles of Ureaplasma urealyticum (U. urealyticum) colonization in the development of bronchopulmonary dysplasia (BPD). This study explored the association between U. urealyticum and bronchopulmonary dysplasia at 36 weeks post-menstrual age (BPD36). Studies published before December 31, 2013 were searched from Medline, Embase, Ovid, Web of Science, and Cochrane databases, with the terms "Ureaplasma urealyticum", "chronic lung disease", or "BPD36" used, and English language as a limit. The association between U. urealyticum colonization and BPD36 was analyzed with RevMan 4.2.10 software, using the odds ratio (OR) and relative risk (RR) for dichotomous variables. Out of the enrolled 81 studies, 11 investigated the BPD36 in total 1193 infants. Pooled studies showed no association between U. urealyticum colonization and subsequent development of BPD36, with the OR and RR being 1.03 (95% CI=0.78-1.37; P=0.84) and 1.01 (95% CI= 0.88-1.16, P=0.84), respectively. These findings indicated no association between U. urealyticum colonization and the development of BPD36.


Subject(s)
Humans , Bronchopulmonary Dysplasia , Microbiology , Pathology , Ureaplasma Infections , Microbiology , Pathology , Ureaplasma urealyticum , Virulence
2.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 535-541, 2014.
Article in English | WPRIM | ID: wpr-351044

ABSTRACT

The effects of over-expression of testis-specific expressed gene 1 (TSEG-1) on the viability and apoptosis of cultured spermatogonial GC-1spg cells were investigated, and the immortal spermatogonial cell line GC-1spg (CRL-2053™) was obtained as the cell model in order to explore the function of TSEG-1. We transfected the eukaryotic vector of TSEG-1, named as pEGFP-TSEG-1 into cultured spermatogonial GC-1spg cells. Over-expression of TSEG-1 inhibited the proliferation of GC-1spg cells, and arrested cell cycle slightly at G0/G1 phase. Transfection of TSEG-1 attenuated the transcript levels of Ki-67, PCNA and cyclin D1. In addition, over-expression of TSEG-1 induced early and late apoptosis, and reduced the mitochondrial membrane potential of GC-1spg cells. Moreover, transfection of TSEG-1 significantly enhanced the ratio of Bax/Bcl-2 and transcript levels of caspase 9, and decreased the expression of Fas and caspase 8 in GC-1spg cells. These results indicated over-expression of TSEG-1 suppresses the proliferation and induces the apoptosis of GC-1spg cells, which establishes a basis for further study on the function of TSEG-1.


Subject(s)
Animals , Male , Mice , Caspase 8 , Genetics , Cell Line , Cyclin D1 , Genetics , G1 Phase , Physiology , Histones , Genetics , Metabolism , Ki-67 Antigen , Genetics , Proliferating Cell Nuclear Antigen , Genetics , Resting Phase, Cell Cycle , Physiology , Spermatogonia , Cell Biology , Metabolism , bcl-2-Associated X Protein , Genetics
3.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 535-41, 2014.
Article in English | WPRIM | ID: wpr-636719

ABSTRACT

The effects of over-expression of testis-specific expressed gene 1 (TSEG-1) on the viability and apoptosis of cultured spermatogonial GC-1spg cells were investigated, and the immortal spermatogonial cell line GC-1spg (CRL-2053™) was obtained as the cell model in order to explore the function of TSEG-1. We transfected the eukaryotic vector of TSEG-1, named as pEGFP-TSEG-1 into cultured spermatogonial GC-1spg cells. Over-expression of TSEG-1 inhibited the proliferation of GC-1spg cells, and arrested cell cycle slightly at G0/G1 phase. Transfection of TSEG-1 attenuated the transcript levels of Ki-67, PCNA and cyclin D1. In addition, over-expression of TSEG-1 induced early and late apoptosis, and reduced the mitochondrial membrane potential of GC-1spg cells. Moreover, transfection of TSEG-1 significantly enhanced the ratio of Bax/Bcl-2 and transcript levels of caspase 9, and decreased the expression of Fas and caspase 8 in GC-1spg cells. These results indicated over-expression of TSEG-1 suppresses the proliferation and induces the apoptosis of GC-1spg cells, which establishes a basis for further study on the function of TSEG-1.

4.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 265-9, 2014.
Article in English | WPRIM | ID: wpr-636685

ABSTRACT

There is controversy regarding the roles of Ureaplasma urealyticum (U. urealyticum) colonization in the development of bronchopulmonary dysplasia (BPD). This study explored the association between U. urealyticum and bronchopulmonary dysplasia at 36 weeks post-menstrual age (BPD36). Studies published before December 31, 2013 were searched from Medline, Embase, Ovid, Web of Science, and Cochrane databases, with the terms "Ureaplasma urealyticum", "chronic lung disease", or "BPD36" used, and English language as a limit. The association between U. urealyticum colonization and BPD36 was analyzed with RevMan 4.2.10 software, using the odds ratio (OR) and relative risk (RR) for dichotomous variables. Out of the enrolled 81 studies, 11 investigated the BPD36 in total 1193 infants. Pooled studies showed no association between U. urealyticum colonization and subsequent development of BPD36, with the OR and RR being 1.03 (95% CI=0.78-1.37; P=0.84) and 1.01 (95% CI= 0.88-1.16, P=0.84), respectively. These findings indicated no association between U. urealyticum colonization and the development of BPD36.

5.
Journal of Zhejiang University. Science. B ; (12): 237-241, 2009.
Article in English | WPRIM | ID: wpr-335375

ABSTRACT

Gastric carcinoma with osteoclast-like giant cells (OGCs) is an extremely rare tumor. So far, only six cases have been reported in the literature. Here we report an additional case of this tumor in a Chinese 78-year-old man presented with abdominal pain, vomiting, and hematemesis. Physical examination and gastroscopy revealed a tumor in the gastric antrum. The biopsy and pathological findings indicated a gastric adenocarcinoma with OGCs, which were present in both the tumor and the metastatic lymph nodes. Further immunohistochemical staining indicated that OGCs were reactive with CD68, CD45, and vimentin protein, but not with pancytokeratin, carcinoembryonic antigen, or epithelial membrane antigen, suggesting the monocytic/histiocytic derivation of these OGCs. In situ hybridization for Epstein-Barr virus showed no nuclear positivity in either adenocarcinoma or OGCs. Postoperative follow-up showed that the patient had survived for at least 6 months without recurrence. Further investigation is warranted to clearly define the prognostic significance of OGCs in gastric carcinoma.


Subject(s)
Aged , Humans , Male , Giant Cells , Metabolism , Pathology , Immunohistochemistry , In Situ Hybridization , Osteoclasts , Metabolism , Pathology , Stomach Neoplasms , Genetics , Metabolism , Pathology
6.
National Journal of Andrology ; (12): 99-105, 2009.
Article in Chinese | WPRIM | ID: wpr-292417

ABSTRACT

<p><b>OBJECTIVE</b>To clone the mouse testis specific gene TSEG-2 via a bioinformatic approach.</p><p><b>METHODS</b>The expressed sequence tags (EST) in the normal mouse testis were obtained from the online EST database ZooDDD. Their highly homologous EST sequences were retrieved through the dbEST database to construct contigs and spliced with the biomedical software Biolign. The corresponding exons and introns within the genome sequences were predicted with the software GeneScan. Primers were designed according to the open reading frame. RT-PCR was applied in cloning the cDNA of the novel gene from the mouse testis tissue and analyzing its expression patterns in the undescended testis and various organ tissues as well as in different developmental stages of the mouse testis. The sequencing results of TSEG-2 underwent bioinformatic analyses.</p><p><b>RESULTS</b>The novel mouse testis gene TSEG-2 was successfully cloned, with full-length sequence of 451 bp. The open reading frame was 267 bp, coding a protein of 88 amino acid residues, and demonstrated to be correct by RT-PCR. The expression of TSEG-2 was high in the mouse testis, regular in the testis cDNA samples of different postnatal days, and down-regulated in the cryptorchidism model. No obvious homology with other mouse cDNA was found for TSEG-2. The GenBank accession number EU079025 was achieved. Function prediction showed that mouse TSEG-2 was probably a soluble non-secretary protein located at chromosome 15qE3, or a nucleoprotein with 2 phosphorylation sites of protein kinase C (PKC) and 1 of casein kinase II (CK2).</p><p><b>CONCLUSION</b>A novel mouse testis specific gene TSEG-2 was successfully cloned, which could be down-regulated by cryptorchidism-inducible 17-beta estradiol. This has prepared the ground for further researches on the biological function and expression regulation of TSEG-2.</p>


Subject(s)
Animals , Female , Male , Mice , Pregnancy , Amino Acid Sequence , Base Sequence , Cloning, Molecular , Expressed Sequence Tags , Gene Expression , Mice, Inbred Strains , Molecular Sequence Data , Open Reading Frames , Proteins , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA , Testis , Metabolism
7.
Acta Pharmaceutica Sinica ; (12): 584-590, 2008.
Article in Chinese | WPRIM | ID: wpr-277792

ABSTRACT

This study is to explore the inhibitory effect of methyl jasmonate on cell proliferation and expression of XIAP and survivin of human neuroblastoma cell line BE(2)-C. After cultivation of 1 - 2 mmol x L(-1) jasmonates with BE (2) -C cells for 6 - 24 h, the growth inhibiting rates of BE (2) -C cells were studied by MTT colorimetry. Cell proliferation was detected by colony formation assay. Cell cycle phases were assayed by propidium iodide staining flow cytometery. Cell apoptosis was inspected by acridine orange-ethidium bromide fluorescent staining, Hoechst 33258 fluorescent staining, and Annexin V-FITC and propidium iodide staining flow cytometry. Expressions of cyclin D1, XIAP and survivin were determined by RT-PCR and real-time RT-PCR. Methyl jasmonate inhibited the growth of BE(2)-C cells in a dose- and time-dependent manner. After addition of 1, 1.5 and 2 mmol x L(-1) of methyl jasmonate for 24 h, the inhibiting rates of cell growth reached 20.6% - 85.5% (P < 0.01), and the IC50 was 1.35 mmol x L(-1). The cell cycles were arrested at S phase. A part of cells presented the characteristic morphological changes of apoptosis. The early apoptotic rates were 13.51%, 17.32%, 24.59% (P < 0.01) and the cell death rates were 29.36% , 54.73% , 75.52% (P < 0.01), respectively. The expression of XIAP and survivin mRNA were downregulated by 18.5% - 68.9% , 22.4% - 48.7% (P < 0.05), respectively, without change in that of cyclin D1. The results indicated that methyl jasmonate could significantly inhibit the growth of BE(2) -C cells through inducing cell cycle arrest and apoptosis, downregulating the expression of XIAP and survivin might be one of its molecular mechanisms of action.


Subject(s)
Humans , Acetates , Pharmacology , Antineoplastic Agents, Phytogenic , Pharmacology , Apoptosis , Cell Line, Tumor , Cell Proliferation , Cyclin D1 , Genetics , Cyclopentanes , Pharmacology , Dose-Response Relationship, Drug , Down-Regulation , Inhibitor of Apoptosis Proteins , Microtubule-Associated Proteins , Genetics , Neuroblastoma , Metabolism , Pathology , Oxylipins , Pharmacology , RNA, Messenger , Metabolism , S Phase , X-Linked Inhibitor of Apoptosis Protein , Genetics
8.
Chinese journal of integrative medicine ; (12): 126-131, 2006.
Article in English | WPRIM | ID: wpr-314073

ABSTRACT

<p><b>OBJECTIVE</b>To explore the growth inhibition effects and apoptosis inducing mechanisms of curcumin on human ovarian cancer cell line A2780.</p><p><b>METHODS</b>After treatment with 10 - 50 micromol/L curcumin for 6 - 24 h, the growth activity of A2780 cancer cells were studied by [4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) colorimetry. Cellular apoptosis was inspected by flow cytometery and acridine orange-ethidium bromide fluorescent staining methods. The fragmentation of cellular chromosome DNA was detected by DNA ladder, the ultrastructural change was observed under a transmission electron microscope, and the protein levels of nuclear factor-kappa B (NF-kappaB, P65) and cysteinyl aspartate specific protease-3 (Caspase-3) in ovarian cancer cells were measured by immunohistochemistry.</p><p><b>RESULTS</b>After treatment with various concentrations of curcumin, the growth inhibition rates of cancer cells reached 62.05% - 89.24%, with sub-G(1) peaks appearing on histogram. Part of the cancer cells showed characteristic morphological changes of apoptosis under fluorescence and electron microscopes, and the rate of apoptosis was 21.5% - 33.5%. The protein expression of NF-kappaB was decreased, while that of Caspase-3 was increased in a time-dependent manner.</p><p><b>CONCLUSION</b>Curcumin could significantly inhibit the growth of human ovarian cancer cells; inducing apoptosis through up-regulating Caspase-3 and down-regulating gene expression of NF-kappaB is probably one of its molecular mechanisms.</p>


Subject(s)
Female , Humans , Acridine Orange , Apoptosis , Caspase 3 , Cell Division , Cell Line, Tumor , Colorimetry , Curcumin , Pharmacology , DNA Fragmentation , Down-Regulation , Ethidium , Flow Cytometry , Immunohistochemistry , Microscopy, Electron, Transmission , NF-kappa B , Ovarian Neoplasms , Pathology , Up-Regulation
9.
Chinese Journal of Pathology ; (12): 92-96, 2005.
Article in Chinese | WPRIM | ID: wpr-265186

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of overexpression of second mitochondria-derived activator of caspases (Smac) gene on apoptosis of gastric cancer cells.</p><p><b>METHODS</b>Under the induction of liposome, MKN-45 cells were transfected by Smac gene and incubated with G418 for subclone selection. Reverse transcriptase-polymerase chain reaction and Western blot were used to determine cellular Smac gene expression. After induction of apoptosis by mitomycin (MMC), cell viabilities were analyzed using trypan blue stain. Apoptosis was measured by electronic microscopy, acridine orange-ethidium bromide fluorescent staining and in situ terminally labelled transferase technique (TUNEL). Western blot and colorimetry were used to assess cellular caspase-3 expression and its activity.</p><p><b>RESULTS</b>The Smac mRNA and protein levels in MKN-45/Smac subclone cells (subclone consistently expressing Smac gene) were significantly higher than those in MKN-45 (P < 0.01). When compared with those in MKN-45, cell viabilities of MKN-45/Smac were reduced by 10.0% to 30.8% (P < 0.01), after treatment with 10 microg/ml MMC for 6 to 24 hours. Some of the MKN-45/Smac cells showed characteristic morphologic changes of apoptosis, their apoptotic rate being increased by 21.2% (P < 0.01). After treatment with MMC, caspase-3 expression and its activity in MKN-45/Smac cells were significantly higher than those in MKN-45 (P < 0.01).</p><p><b>CONCLUSIONS</b>Overexpression of Smac in gastric cancer cell line significantly improves expression and activity levels of caspase-3 after induction by MMC. Such apoptosis-inducing effect establishes a novel strategy for regulating the apoptosis activity of gastric cancer.</p>


Subject(s)
Humans , Apoptosis , Caspase 3 , Metabolism , Cell Line, Tumor , Intracellular Signaling Peptides and Proteins , Genetics , Mitochondrial Proteins , Genetics , Mitomycin , Pharmacology , RNA, Messenger , Genetics , Stomach Neoplasms , Metabolism , Pathology , Transfection
10.
Chinese Medical Journal ; (24): 50-55, 2005.
Article in English | WPRIM | ID: wpr-257325

ABSTRACT

<p><b>BACKGROUND</b>Recently, arsenic trioxide (As2O3) was considered as a novel anti-tumor agent. However, it showed severe toxicity effect on normal tissue at the same time. To improve its therapeutic efficacy and decrease its toxicity,we prepared arsenic trioxide-loaded albuminutes immuno-nanospheres [As2O3-(HAS-NS)-BDI-1] targeted with monoclonal antibody (McAb) BDI-1 and tested its specific killing effect against bladder cancer cell.</p><p><b>METHODS</b>As2O3-HAS-NS was prepared by chemical cross-linking method. Monoclonal antibody BDI-1 was purified with ammonium sulphate saltingout and chromatography. Albuminutes microspheres were conjugated with McAb by SPDP cross-linking method. Concentration of As in As2O3-(HAS-NS)-BDI-1 and As2O3-HAS-NS was measured by atomic fluometry method. As2O3-(HAS-NS)-BDI-1 and its activity were detected by SDS-PAGE reduction electrophoresis, indirect immunofluorescence test, light microscope and scanning electron microscope observation. Acridine orange staining and tritiated thymidine (3H-TdR) incorporation tests were used to indicate specific killing activity of As2O3-(HAS-NS)-BDI-1 in vitro.</p><p><b>RESULTS</b>In As2O3-(HAS-NS)-BDI-1 groups, we saw two protein bands in SDS-PAGE reduction electrophoresis. Albuminutes immuno-nanospheres were rounded with clear green fluorescence by immunofluorescence test. Under microscope, we observed that BIU-87 cells were covered with the As2O3-(HAS-NS)-BDI-1 and that As2O3-(HAS-NS)-BDI-1 moved with the BIU-87 cells. The albuminutes immuno-nanospheres were tightly junctioned with the BIU-87 cells. Specific killing activity of As2O3-(HAS-NS)-BDI-1 on bladder tumor cells was observed by acridine orange staining and 3H-TdR incorporation assays.</p><p><b>CONCLUSIONS</b>As2O3-(HAS-NS)-BDI-1 might bind specifically against BIU-87 cells, thus leading to high activity of killing bladder tumor cells.</p>


Subject(s)
Animals , Mice , Antibodies, Monoclonal , Pharmacology , Antineoplastic Agents , Apoptosis , Arsenicals , Pharmacology , Cell Line, Tumor , Cell Survival , Drug Delivery Systems , Fluorescent Antibody Technique , Mice, Inbred BALB C , Nanotubes , Oxides , Pharmacology , Serum Albumin , Pharmacology , Urinary Bladder Neoplasms , Drug Therapy , Pathology
11.
Chinese Journal of Urology ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-676087

ABSTRACT

Objective To explore the investigate of X-chromosome-linked inhibitor of apoptosis pro- tein(XIAP)and its effect on chemotherapeutic sensitivity in bladder carcinoma.Methods Using immu- nohistochemistry methods,the expression of XIAP was evaluated in 47 bladder carcinomas and 6 normal bladder tissues.The XIAP gene was transfected into bladder cancer cell line T24 by liposome and the positive clone was screened by G418.Cellular XIAP mRNA level was detected by RT-PCR.The apoptosis of T24 cells was induced by low-dose of mitocycin C(0.005 mg/ml and 0.05 mg/ml,respectively).The in vitro cellular growth activities were assayed by MTT color imetry;and the apoptosis rate was assayed by TUNEL methods. Results The expression rate of XIAP was 78.7%(37/47)in bladder carcinoma samples,with no corre- lation with carcinoma stages and grades(P>0.05).XIAP mRNA level in transfected T24 ceils was signifi- cantly increased by 3.8 times.Treated with 0.005 mg/ml and 0.05 mg/ml of mitomycin C,the growth rates of XIAP transfected T24 cells were increased [(11.60?0.25)% and(16.51?0.87)% ,respectively,P<0.05];and the apoptosis rates were decreased [(10.1?0.2)% and( 11.9?0.2)% ,respectively,P<0.05]compared with those in control cells.Conclusions XIAP is highly expressed in humun bladder car- cinoma samples.Overexpression of XIAP in T24 cells results in decrease in bladder carcinoma cell apoptosis induced by MMC,which may decrease the chemotherapeutic sensitivity of T24 cells.

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