Human insulin inhibits apoptosis and promotes proliferation of rat liver cell line BRL-3 A / 基础医学与临床

Objective_To study the effect of human insulin on cell cycle progression and apoptosis of rat liver cell line BRL-3A in vitro.Methods_MTT method was used to observe the effect of insulin on cell activity, and flow cytometry was used to detect cell apoptosis and cell cycle.qRT-PCR was used to evaluate the expression of related genes.Results_Human insulin induced the proliferation of BRL-3A cells in a dose-dependent manner ( P<0.05 or P<0.01);After 3 days treated by human insulin (500 nmol/L), the proportion of cells in G0/G1 phases re-markably decreased (P<0.05).Moreover, pro-apoptotic BAX was down-regulated (P<0.05), while cell prolif-eration-related gene CCNA2 was up-regulated (P<0.05).Conclusions_Human insulin may inhibit the apoptosis of BRL-3 A cell line and induce proliferation due to the down-regulated expression of BAX and up-regulated expres-sion of CCNA2 .

Balloon dilation versus Amplatz dilation during ultrasound-guided percutaneous nephrolithotomy for staghorn stones / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Amplatz dilation and balloon dilation are different methods in creating the accesses during percutaneous nephrolithotomy (PCNL). The aim of this study was to review the surgical experiences of managing staghorn calculi by Amplatz dilation and balloon dilation for 3 years.</p><p><b>METHODS</b>We retrospectively analyzed clinical data from 125 patients (129 kidneys) with staghorn kidney stones who underwent PCNL from January 2010 to December 2012, of whom 60 patients underwent Amplatz dilation (AD group) and 65 underwent balloon dilation (BD group) during PCNL.</p><p><b>RESULTS</b>The AD and BD groups were similar in age, male-female ratio, stone burden, stone type, hydronephrosis, and proportion of patients who had undergone extracorporeal lithotripsy. However, these two groups showed significant differences in terms of duration of percutaneous access (15.1 ± 3.6) minutes vs. (10.0 ± 3.3) minutes, one-attempt success rate of dilation via a single access 88.9% (72/81) vs. 97.8% (91/93), hemoglobin drop after surgery (3.5 ± 0.9) g/dl vs. (1.7 ± 0.9) g/dl, number of cases requiring intraoperative and postoperative blood transfusion 27.9% (n = 17) vs. 13.2% (n = 9), changes of central venous pressure before and after surgery (2.3 ± 1.2) cmH2O vs. (1.2 ± 0.7) cmH2O, number of patients who experienced postoperative fever >37.5°C 21 (34.4%) vs. 13 (19.1%) (all P < 0.05). No injury of adjacent organs, including pleura, liver, spleen, or bowel, was noted in patients.</p><p><b>CONCLUSIONS</b>During ultrasound-guided PCNL for staghorn stones, balloon dilation and Amplatz dilation are all effective and safe. Compared with Amplatz dilation, balloon dilation is a better choice, as it has a higher access creation success rate, shorter access creation time less blood loss, and lower proportions of circulatory overload and postoperative fever.</p>

Immune responses induced by DNA vaccine combined with recombinant adenovirus containing MOMP gene of Chlamydia trachomatis serovar E / 中华微生物学和免疫学杂志

Objective To investigate the specific immune responses induced by DNA vaccine com-bined with recombinant adenovirus carrying major outer membrane protein ( MOMP) gene of Chlamydia trachom-atis (Ct) serovar E.Methods Recombinant eukaryotic expression plasmid pVAX 1-MOMP and recombinant adenovirus Ad-MOMP were constructed and purified .Four immunization strategies were designed including DNA immunization (DNA group), recombinant adenovirus immunization (Ad group), DNA prime-recombinant ade-novirus boost regimen ( DNA/Ad group ) and recombinant adenovirus prime-DNA boost regimen ( Ad/DNA group) .Two weeks after the final immunization , vaginal wash specimens , blood samples and spleens were col-lected from all mice for the evaluation of humoral and cellular immune responses .Results Th1 responses and mucosal responses were not found in DNA group .Ad group induced both Th 1 responses and local mucosal re-sponses , and its IgG and IgA levels were significantly higher than those in DNA group .DNA/Ad group and Ad/DNA group induced stronger Th 1 responses and humoral responses than DNA and Ad group .Serum antibodies and mucosal antibodies in DNA/Ad group were higher than those in Ad/DNA group, but Th1 responses in DNA/Ad group were impaired than those in Ad/DNA group.Conclusion The recombinant adenovirus could induce Th1 responses and genital mucosal responses .Combined immunization of DNA/Ad or Ad/DNA showed enhanced specific immune responses in comparison with the single immunization with DNA or Ad .The differ-ences between the two combination methods were that DNA /Ad group stimulated higher levels of specific anti-body responses , while Ad/DNA group produced stronger Th 1 responses .The study provided a reference for the enhancement and development of vaccines against Chlamydia trachomatis in further studies .

Situation and Related Factors of Maintenance Medication Treatment in Schizophrenic Patients Re-hospitalized for Relapse / 中国康复理论与实践

@#Objective To survey the situation and related factors of maintenance medication treatment in schizophrenic patients re-hospitalized for relapse. Methods 362 patients re-admitted with schizophrenia and their relatives were interviewed with questionnaire which included 16 factors that might affect maintenance medication treatment. Results 63.8% patients stopped medication and 21.1% patients were on low-dose maintenance medication treatment by themselves, while 7.7% patients were on reasonable dose anti-psychotics in 2 years. The time of maintenance medication treatment related with insight (OR=2.2144, P=0.001), family care (OR=4.8842, P=0.025), outcomes of treatment (OR=2.2056, P=0.007) and negative life events (OR=0.4529, P=0.003). Conclusion Schizophrenic patients re-admitted with relapse often withdraw or reduce their medication by themselves, which risked with poor insight, poor therapeutic effect, poor care from their family, and more negative life events

Construction and identification of prostate-specific double gene expression vector pIRES-PSMAe/p-TKCx43 / 中华泌尿外科杂志

Objective To construct the prostate-specific double gene expression vector pIRESPSMAe/p-TK-Cx43 and establish the foundation for experimental prostate cancer gene therapy research. Methods Cx43 gene was amplified and cloned into pMD19-T Simple vector. HSV-TK gene was then synthesized and cloned into multiple clone site (MCS) A of the eukaryotie vector plRES. The new plasmid was named plRES-TK: PSMAe/p was obtained and cloned into plRES-TK by replacing CMV promoter. The new plasmid was named plRES-PSMAe/p-TK; Fourth, Cx43 gene was cloned into the MCS B of pIRES-PSMAe/p-TK and the new plasmid was named pIRES-PSMAe/p-TK-Cx43.This plasmid was identified by double digestion with Sal Ⅰ/Not Ⅰ and sequenced; Finally, LNCaP cells were transfected by the plasmid plRES-PSMAe/p-TK-Cx43 and the mRNAs expression of HSV-TK gene and Cx43 gene was tested by RT-PCR. Results The plasmids synthesized in this experiment were double digested respectively and the specific bands of the inserted genes were confirmed by RTPCR. plRES-PSMAe/p-TK-Cx43 was in line with the expected design by DNA sequencing. The mRNAs of TK gene and Cx43 gene were expressed and successfully confirmed by RT-PCR after LNCaP cells transfected with pIRES-PSMAe/p-TK-Cx43. Conclusion Double gene expression vector pIRES-PSMAe/p-TK-Cx43 containing HSV-TK gene and Cx43 gene is constructed successfully.

Meta-analysis of randomized trials of prostate specific antigen progression and death rate in patients with locally advanced prostate cancer / 中华泌尿外科杂志

Objective To verify the best treatment strategy in reducing prostate specific antigen (PSA) progression and death rate in patients with locally advanced prostate cancer by a meta-analysis. Methods The literature search strategy was followed according to the Collaborative Review Group search strategy. Published data of randomized clinical trials comparing radical prostatectomy (RP) plus adjuvant therapy to either RP alone or other treatment were analyzed. Both fixed effect model and randomized effect model were applied and odds ratio (OR) with its 95% confidence interval (95% CI) was also used as the effect size 'estimate. Results Eight clinical trials were chosen with total in-volved cases of 3826. There were 5 trials compared post radical prostatectomy plus adjuvant hormonal therapy with radical prostatectomy alone. PSA progression was used as the indicator of progression and the combined OR was 0.86 (95%CI 0.48-1.56). There were 3 trails compared the combination of radical prostateetomy with hormonal therapy and radical prostatectomy alone. Disease specific death rate was used as the evaluating criteria and the OR was 0.72(95%CI,0.51-1.02). Conclusion RP plus adjuvant hormonal therapy can reduce PSA progression of patients with locally advanced pros-tate cancer, but it has no significant effect on disease specific death rate.

Optimized codon usage enhances the expression and immunogenicity of DNA vaccine encoding the HPV 6b E7 gene / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To analyze the influence of optimal codon usage on the expression levels and immunogenicity of DNA vaccines, encoding the human papillomavirus type 6b (HPV 6b) E7 gene.</p><p><b>METHODS</b>The full length E7 gene of HPV 6b was modified to substitute human preferred codon for rarely used codon, and three mutations were introduced into the pRB binding site of HPV 6b E7 to eliminate its transformation potential. The codon optimized and mutated E7 gene (hu-mE7) were cloned into the Kpn I and EcoR I site of the pcDNA3 mammalian expression vector, the in vitro expression of the hu-mE7 gene and the immunogenicity of hu-mE7 DNA vaccine were compared with the wt-E7gene.</p><p><b>RESULTS</b>The in vitro expression of pcDNA3-hu-mE7 was much higher than the classical wt-E7 plasmid in monkey COS-1 cell line. Mice immunized intramuscularly with the pcDNA3-hu-mE7 showed that the codon modified E7 gene induced a stronger IFN-gamma ratios than the wt-E7 gene.</p><p><b>CONCLUSIONS</b>These results suggest that the optimized codon usage contributes to the enhancement of gene expression and immunogenicity of HPV 6b E7 gene.</p>

The diagnostic of six cases with gastric submucosal tumor / 中国癌症杂志

Purpose:To improve the diagnostic accuracy of gastric submucosal tumor whose endoscopy was negative. Methods:Six cases of gastric submucosal tumor with “negative” results of endoscopy were analyzed. Results:Before operation 3 were recieved by medicinal management, 5 were diagnosed as malignant tumor by CT, 2 were shown to be tumor by ultrasound. There were 3 gastric lymphoma, 1 gastric adenocarcinoma, 1 gastric Schwannoma and 1 gastric leiomyosarcoma confirmed by postoperational pathologic diagnosis. Conclusions:If suspicion remains in endoscopy diagnosis, procedure should involve careful physical examination, CT, B US, GI and so on to improve the diagnostic accuracy. [

Expression of calcium sensing receptor during ischemia/reperfusion myocardial damage and relationship between CaSR and injury of myocardium / 中国病理生理杂志

AIM: To investigate the expression of calcium sensing receptor(CaSR) during myocardial injury induced by ischemia/reperfusion and disclose the relationship between CaSR and myocardial ischemia/reperfusion. METHODS: The experimental model was established by the 30 min ligating and 1 h, 2 h, 4 h, 6 h reperfusing the left descending coronary artery (LAD) in rats. Wistar rats were randomly divided into 5 groups: sham group, ischemia/reperfusion 1 h, 2 h, 4 h, 6 h groups (I/R 1 h, 2 h, 4 h, 6 h group). CaSR mRNA expression was detected by RT-PCR. Left ventricular function was recorded. The levels of plasma lactate dehydrogenase (LDH), alondialdehyde (MDA) and superoxide dismutase (SOD) were measured. The change of ultrastructure in the ischemia/reperfusion myocardium of rats was observed by electron microscopy. RESULTS: LVSP,?dp/dtmax and SOD activity decreased gradually with the reperfusion time prolonged. LDH and MDA peaked at 2 h. The ultramicro-structural injury at the 1 h and 2 h was more serious than that at 4 h and 6 h. The expression of CaSR increased significantly after reperfusion of 1 h and 2 h, and decreased after 4 h and 6 h. CONCLUSION: The increased expression of CaSR mRNA and serious injure of myocardium were observed. CaSR may be associated with the myocardial ischemia/reperfusion injury.

Protective effect of COPP on hypoxia/reoxygenation injury of H9c2 myocytes / 中国药理学通报

Aim To study the effect and molecular mechanism of pretreatment with COPP on hypoxia/reoxygenation injury of H9c2 myocytes.Methods H9c2 myocytes model of hypoxia/reoxygenation injury was established and H9c2 myocytes were given COPP pretreatment before hypoxia/reoxygenation.Treatment with Znpp and all-trans retinoic acid(ATRA)inhibited HO-1 and Nrf2-ARE respectively.The level of LDH and CK in cell supernatants were measured.HO-1mRNA expression was analyzed by RT-PCR.HO-1 and Nrf2 protein expressions were analyzed by Western blot.Results Compared with hypoxia/reoxygenation group,the level of LDH and CK in COPP pretreatment groups decreased significantly and the level of HO-1mRNA,HO-1 protein expression and Nrf2 protein expression in the nucleus significantly increased.Znpp abolished protective effect of COPP pretreatment.ATRA blocked the nuclear accumulation of Nrf2 and decreased HO-1 protein expression that COPP pretreatment induced.Conclusions COPP can induce HO-1 overexpression which has protective effect on hypoxia/reoxygenation injury of H9c2 myocytes.Its mechanism is related to Nrf2-ARE signaling pathway.

Effect of quercetin on rat cardiomyocyte apoptosis induced by adriamycin in vitro / 中国药理学通报

Aim To observe the protective effect of quercetin on rat cardiomyocyte apoptosis induced by adriamycin and explore its possible mechanism.Methods Cultured neonatal rat cardiomyocytes were randomly divided into six groups:normal control group, adriamycin group,quercetin control group, adriamycin+quercetin(25,50,100 ?mol?L-1)groups. The activity of LDH was detected by chromatometry, the cardiomyocyte viability was measured by MTT, the ultrastructure of cardiomyocyte was observed by electron microscope, the expression of protein Bcl-2 and Bax was analyzed by immunocytochemical, and the mRNA and protein of caspase-3 were detected by RT-PCR and Western blot respectively.Results Compared with the control group, the activity of LDH was increased but the viability of cardiomyocyte was decreased; the expression of Bax and caspase-3 was up-regulated while Bcl-2 was down-regulated in ADR group.Compared with ADR group, the above changes were lightened in adriamycin+quercetin groups. But the quercetin control group, in which cultured myocardial cells only exposed to quercetin without ADR, had no obvious changes.Conclusions Quercetin significantly inhibits the apoptosis induced by ADR in the cultured myocardial cells. Its mechanism is involved in the apoptosis-related pathways, including caspase-3, Bax and Bcl-2.

CpG-oligodeoxynucleotides induce differentiation and apoptosis in HL60 cells / 中国病理生理杂志

AIM: To investigate the differentiative and apoptotic effect of CpG-oligodeoxynucleotides on HL60 cells and its mechanism. METHODS: After HL60 cells were exposed to synthetic CpG-oligodeoxynucleotides, non-CpG-oligodeoxynucleotides or ZpG-oligodeoxynucleotides for 72 hours, respectively, the inhibition of HL60 cells were detected using MTT method, NBT test was used and CD14 expression were determined. Apoptosis of HL60 cells were mensurated with flow cytometry and transmission electron microscope, and caspase 3, Bcl-2 and Bax expression of HL60 cells treated with oligodeoxynucletides were detected using immunohistochemistry. RESULTS: Treatment with CpG-oligodeoxynucleotides induced the differentiation and apoptosis in HL60 cells, but non-CpG-oligodeoxynu- cleotide and ZpG-oligodeoxynucleotide had no effect on HL60 cells. CONCLUSION: CpG-oligodeoxynucleotides can induce the differentiation and apoptosis in HL60 cells. It may provide a new approach for the immunological treatment of leukemia.