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Objective: To explore the effect of Sirt1 on the function of endothelial progenitor cells (EPCs) in rats with chronic obstructive pulmonary disease (COPD). Methods: A rat COPD model was established via smoking and endotoxin administration for three months. The peripheral circulating EPCs were isolated by gradient centrifugation, and their functions, cell cycle distribution, apoptosis, and Sirt1 expression were examined. The function changes of EPCs in the presence or absence of Sirt1 agonist and inhibitor were estimated; meanwhile, the expressions of Sirt1, FOXO3a, NF-κB, and p53 were also evaluated. Results: The proliferation, adhesion, and migration of EPCs decreased while the apoptosis rate was increased in the COPD rats. The expression of Sirt1 protein in EPCs of the COPD group was significantly lower than that in the control group (P<0.01). The overexpression of the Sirt1 gene using a gene transfection technique or Sirt1 agonists (SRT1720) improved the proliferation, migration, and adhesion, and decreased the apoptosis of EPC. However, Sirt1 inhibitor (EX527) decreased EPC functions in the COPD group. The effect of Sirt1 expression on EPC function may be related to reduction of FOXO3a and increase of NF-κB and p53 activity. Conclusions: Increased expression of Sirt1 can improve the proliferation and migration of EPCs and reduce their apoptosis in COPD rats. This change may be related to FOXO3a, NF-κB, and p53 signaling pathways.
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Objective: To explore the effect of Sirt1 on the function of endothelial progenitor cells (EPCs) in rats with chronic obstructive pulmonary disease (COPD). Methods: A rat COPD model was established via smoking and endotoxin administration for three months. The peripheral circulating EPCs were isolated by gradient centrifugation, and their functions, cell cycle distribution, apoptosis, and Sirt1 expression were examined. The function changes of EPCs in the presence or absence of Sirt1 agonist and inhibitor were estimated; meanwhile, the expressions of Sirt1, FOXO3a, NF-κB, and p53 were also evaluated. Results: The proliferation, adhesion, and migration of EPCs decreased while the apoptosis rate was increased in the COPD rats. The expression of Sirt1 protein in EPCs of the COPD group was significantly lower than that in the control group (P<0.01). The overexpression of the Sirt1 gene using a gene transfection technique or Sirt1 agonists (SRT1720) improved the proliferation, migration, and adhesion, and decreased the apoptosis of EPC. However, Sirt1 inhibitor (EX527) decreased EPC functions in the COPD group. The effect of Sirt1 expression on EPC function may be related to reduction of FOXO3a and increase of NF-κB and p53 activity. Conclusions: Increased expression of Sirt1 can improve the proliferation and migration of EPCs and reduce their apoptosis in COPD rats. This change may be related to FOXO3a, NF-κB, and p53 signaling pathways.
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Objective@#To investigate the functional classification of differentially expressed genes in manganese-poisoned rats and related metabolic pathways, and to provide a reference for the study of the mechanism of manganese poisoning and gene regulation in the prevention and treatment of manganese poisoning.@*Methods@#Six healthy specific pathogen-free male Sprague-Dawley rats were randomly divided into control group and experimental group according to body weight, with 3 rats in each group. Rats in the experimental group were injected intraperitoneally with MnCl2·4H2O (25 mg/kg) at 0.2 ml/100 g once every 48 h, and the control group was injected with phosphate-buffered saline at the same dose. After one month of exposure, the rats were anesthetized and then sacrificed by cardiac puncture blood collection. The striatum was isolated on ice, and RNA was extracted to establish a DNA data library. Whole genome sequencing was used to identify the differentially expressed genes in the rats with manganese poisoning. Gene Ontology functional enrichment analysis and pathway enrichment analysis were performed to investigate the possible metabolic pathways in which the differentially expressed genes may participate.@*Results@#A total of 18439 genes were detected in the striatum of rats, and 17 differentially expressed genes were screened out. Among them, 10 genes were up-regulated, and 7 genes were down-regulated. According to gene function analysis, 164 functional branches and 26 metabolic pathways with high gene enrichment were screened out. The genes were enriched in synaptic signaling, signal transduction, etc., especially behavioral function. The metabolic pathways with high gene enrichment were endocytosis pathway, PI3K-Akt pathway, and neuroactive ligand-receptor interaction pathway, in which the PI3K-Akt pathway had enrichment of the same differentially expressed gene (29 517) as the FoxO signaling pathway and mTOR signaling pathway, and the neuroactive ligand-receptor interaction pathway had enrichment of the same differentially expressed gene (24 415) as the glutamatergic synaptic pathway.@*Conclusion@#The differentially expressed genes in manganese-poisoned rats may influence the susceptibility to manganese poisoning through the PI3K-Akt pathway, mTOR metabolic pathway, or FoxO metabolic pathway, and may be involved in behavioral changes.
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<p><b>OBJECTIVE</b>To summarize the experience of radical retropubic prostatectomy (RRP) and the multi-factors which influence on the prognosis and long life quality.</p><p><b>METHODS</b>From January 1993 to March 2005, 132 cases radical retropubic prostatectomy were performed. The patients were divided into 2 groups: the early group and recent group. Eleven items in peri-operative time and follow up results were analysed. The erection function of 78 cases were investigated with international index of erectile function 5 score. In these patients, nocturnal electrobioimpedance volumetric assessment (NEVA) were observe in 19 cases.</p><p><b>RESULTS</b>Comparing of the 2 groups, the index connected with operative skill changed to optimization. No one died of prostate cancer in 63 follow up patients. Nine cases showed biochemical failure with criterion as prostate specific antigen > 0.4 microg/L. Fifty patients passed urine normal post-operation in 6 months. Eight patients had stress incontinence and 5 had entire incontinence at 6 month. Four patients had vesical neck stricture. Another follow up result shows 33 (58.9%) erection function recovered in 55 bilateral nerve-sparing operation and 7 recovered in 22 of unilateral nerve-sparing operation. NEVA shows 14 cases with artery supply insufficient in whom 4 regained erection function and 5 cases vein leakage in whom no one recovered.</p><p><b>CONCLUSIONS</b>The radical retropubic prostatectomy remains the procedure of choice for the cure of localized prostatic cancer. The keys for the operation are anatomic dissection, preservation of the neurovascular bundle and good skill. These are also important for a good life quality for the patients.</p>
Subject(s)
Aged , Humans , Male , Middle Aged , Erectile Dysfunction , Follow-Up Studies , Penile Erection , Postoperative Complications , Prostatectomy , Methods , Prostatic Neoplasms , General Surgery , Quality of Life , Retrospective Studies , Treatment Outcome , Urinary IncontinenceABSTRACT
<p><b>OBJECTIVES</b>To investigate whether the human PC-3 cell infected with recombinant Ad-PTEN and Ad-p27Kip1 can steadily produce PTEN and p27Kip1 protein and change the biologic behaviors such as cell proliferation, cell cycle and apoptosis. The synergistic effect of PTEN and p27Kip1 on the therapy for prostate cancer has also been investigated.</p><p><b>METHODS</b>We constructed recombinant adenovirus vector of human tumor suppressor gene PTEN and p27Kip1. The viral titer was examined by plaque assay and the mRNA and protein expressions of PTEN and p27Kip1 in human prostate cancer cell line PC-3 infected with Ad-PTEN and Ad-p27Kip1 were determined by RT-PCR and Western blot respectively. MTT assay was used to determine the effect of PTEN and p27Kip1 on growth and proliferation of PC-3 cell; the change of cell cycle and apoptosis was examined by flow cytometry, and to compare between the combined therapy group and single gene therapy group.</p><p><b>RESULTS</b>The viral titers of Ad-PTEN and Ad-p27Kip1 were 1.8 x 10(7) pfu/ml and 1.2 x 10(9) pfu/ml respectively. After infected by adenovirus, it had been verified that the mRNA and protein expression of PTEN and p27Kip1 were steady in human PC-3 cell. Ad-PTEN and Ad-p27 Kip1 inhibited the growth and proliferation of PC-3 cells. The progression of cell cycle of PC-3 cell was arrested in G(0)-G(1) phase, meanwhile the apoptosis rate of PC-3 was also affected after Ad-PTEN or/and Ad-p27 Kip1 infected. There was significant difference between combined therapy group and single gene therapy group.</p><p><b>CONCLUSION</b>The recombinant Ad-PTEN and Ad-p27Kip1 vector were constructed successfully and the expression of specific PTEN and p27Kip1 was high, steadily in PC-3 cell line. These results suggested that combination of PTEN with p27Kip1 has an application value in treatment of prostate cancer in future.</p>