Immunogenicity of multi-epitopes gene of major outer membrane protein of Chlamydia trachomatis / 中华预防医学杂志

<p><b>OBJECTIVE</b>To construct a recombinant eukaryotic expression plasmid pcDNA3.1-Ct MOMP168 including Ct MOMP multi-epitopes gene, and evaluate the Ct MOMP-specific humoral and cellular immune response induced by pcDNA3.1-Ct MOMP168 in BALB/c mice.</p><p><b>METHODS</b>Recombinant plasmid pcDNA3.1-Ct MOMP168 including Ct MOMP multi-epitopes gene was constructed. Then, BALB/c mice were randomly assigned to receive (intramuscular injection) either pcDNA3.1-Ct MOMP168 or pcDNA3.1 or PBS (n = 12, 100 microg/time per mouse), and the same immunization schedule was repeated for the third time at 2 week intervals. The titers of anti-Ct MOMP antibody and its antibody subtypes in sera, the cytotoxicity of Ct MOMP-specific cytotoxic T lymphocyte (CTL) in spleen, and the level of cytokine (IFN-gamma, IL-4, IL-10)-producing CD3(+) T cells in spleen were detected by ELISA, LDH release assays and intracellular cytokine staining-fluorescence activated cell sorter (ICS-FACS), respectively.</p><p><b>RESULTS</b>The recombinant plasmid pcDNA3.1-Ct MOMP168 was able to induce Ct-specific antibody response (A(490) = 0.973 +/- 0.136; serum titer was 1:1000) as compared with pcDNA3.1 (A(490) = 0.180 +/- 0.025) and PBS (A(490) = 0.110 +/- 0.015), and the major antibody subtype was IgG2a with statistical significance (F = 106.884, P < 0.05). When the ratio of effector cells and target cells reached to 50:1, the activity of cytotoxic T-lymphocyte in pcDNA3.1-Ct MOMP168 immunized mice (41.71% +/- 8.34%) was significantly higher (F = 22.315, P < 0.05) than that in pcDNA3.1 immunized mice (18.40% +/- 3.45%) and PBS immunized mice (14.50% +/- 2.42%). The levels of CD3(+) IFN-gamma(+) T cells in pcDNA3.1-Ct MOMP168 immunized mice (1.15% +/- 0.16%) were significantly higher (F = 99.638, P < 0.05) than that in pcDNA3.1 immunized mice (0.12% +/- 0.08%) and PBS immunized mice (0.09% +/- 0.03%), while the significant difference in the levels of IL-4(+) CD3(+) T cells and IL-10(+) CD3(+) T cells was not observed (F = 0.886 and 1.112, P > 0.05) between pcDNA3.1-Ct MOMP168 immunized mice (0.13% +/- 0.08% and 0.14% +/- 0.08%) and pcDNA3.1 (0.07% +/- 0.05% and 0.13% +/- 0.06%) or PBS immunized mice (0.08% +/- 0.04% and 0.07% +/- 0.04%).</p><p><b>CONCLUSION</b>In BALB/c mice, the recombinant plasmid pcDNA3.1-Ct MOMP168 might induce not only the generation of Ct-specific antibody, but also the high level of Ct MOMP-specific CD3(+) IFN-gamma(+) T cells.</p>

An effective combined therapy for simple premature ejaculation / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the clinical effect of a combined therapy in the treatment of simple premature ejaculation.</p><p><b>METHODS</b>A total number of 110 patients with simple premature ejaculation were divided into a control group (n = 50), given oral hydrochloric acid sertraline only, and a combined therapy group (n = 60), treated by oral administration of hydrochloric acid sertraline, local inunction of a traditional Chinese medicine and guidance in sexual psychology and knowledge. At the end of a 4-week treatment and 4 weeks after the drug withdrawal, the therapeutic effects were evaluated by ejaculation latency and satisfaction with sexual life.</p><p><b>RESULTS</b>The total effectiveness rates at the end of the 4-week treatment were 91.6% and 76% in the combined therapy and the control groups, while those 4 weeks after the drug withdrawal were 68.3% and 42% respectively, both with significant differences in between (P < 0.05 and P < 0.01).</p><p><b>CONCLUSION</b>The combined therapy has a satisfactory clinical effect and stability in the treatment of simple premature ejaculation.</p>

Anticancer drug resistance of HeLa cells transfected with rat glutathione S-transferase pi gene / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To establish a cytologic expressing system of rat glutathione S-transferase pi (GST-pi) cDNA for detecting the resistance of HeLa cells to anticancer drugs.</p><p><b>METHODS</b>The assessment was made with various anticancer drugs (adriamycin, mitomycin, cisplatinum and vincristine) that showed different cytotoxicities in transfectant HeLa cells with pSV-GT containing rat GST-pi cDNA (HeLa/pSV-GT) or control pSV-neo (HeLa/pSV-neo). Expression levels of GST-pi mRNA in HeLa/pSV-GT and HeLa/pSV-neo were measured by in situ hybridization using Digoxin-labelled cDNA probe.</p><p><b>RESULTS</b>HeLa/pSV-GT expressed significantly high degree of GST-pi mRNA, whereas both HeLa/pSV-neo and HeLa cells had very low expression. Cytotoxicities of HeLa/pSV-GT and HeLa/pSV-neo with 4 anticancer drugs were measured by MTT assay. Drug concentrations for yielding 50% inhibition (IC50) in HeLa/pSV-GT by adriamycin, mitomycin and cisplatinum were 70.13 microg/mL, 10.95 microg/mL and 16.52 microg/mL, respectively. In contrast, IC50 in HeLa/pSV-neo was 10.34 microg/mL, 7.48 microg/mL and 13.70 microg/mL, respectively. The cytotoxicities of vincristine on both HeLa/pSV-GT and HeLa/pSV-neo were not significantly different.</p><p><b>CONCLUSIONS</b>Our findings suggest that HeLa/pSV-GT containing rat GST-pi cDNA is resistant to some anticancer drugs due to overexpression of GST-pi. Also, HeLa/pSV-GT cell line could serve as a useful cytogenetic model for further research.</p>

Expression of human papillomavirus 11 type E7 protein in E.coli and its initial evaluation in diagnosis of condyloma acuminatum / 中华检验医学杂志

Objective To investigate expression of Human papillomavirus (HPV) 11 type E7 protein antigen in prokaryotic cells and its potential use for the serodiagnosis of condyloma acuminatum (CA).Methods The full-length gene encoding for HPV11 E7 protein was amplified by PCR,and cloned into vector pET32a(+) to form recombinant pET32a(+)/HPVll E7 plasmid.The fusion His-E7 protein was expressed and analyzed by using SDS-PAGE and Western blotting.Using ELISA assay,HPV11 E7 fusion protein were also used to screen human serum IgG antibody from 93 patients with CA,43 patients with cervix cancer and 58 healthy control subjects.Results Highly expressed fusion His-E7 protein was obtained,and purified protein served as a special diagnostic antigen to screen human serum antibody for CA serodiagnosis.It showed that CA group,cervix cancer group and healthy control human serum IgG antibody average value were 1.545?0.131,0.586?0.155 and 0.674?0.150 respectively,positive rate were 76.3% (71/93),11.6% (5/43) and 5.2% (3/58).There was significantly difference between the CA group to compare cervix cancer group and healthy control (P