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Objective To observe the effect of RNAi targeting Livin gene on biology characteristics such as apoptosis and proliferation in human prostate cancer cells.Methods siRNA expression vector targeting Livin gene was constructed and transfected into human prostate cancer cell line PC3.The expressions of Livin mRNA and protein were detected by real-time PCR and Western-blot,cell apoptosis and cell cycle were assayed by flow cytometry,proliferation and colony formation were detected by MTT and colony formation assay,and the tumor growth in vivo was observed in nude mice.Results After transfection,downregulation of Livin mRNA and protein expression in PC3 cells was observed (P＜0.01).Compared with the control group,the proliferation of cancer cells was inhibited significantly (P＜0.01) and the apoptotic ratio was (26.5±3.3) ％ (P＜0.01).The Caspase3 activity increased obviously (P＜0.05),and the experimental group showed a decreased colony formation rate (P＜0.01).The tumor volume of xenografts in nude mouse in experimental and control group was (1.79± 0.07) and (4.40 ± 0.06) cm3 respectively (P ＜ 0.01).Conclusions The siRNA recombinant expression vector targeting Livin gene was constructed and can knockdown the expression of Livin mRNA and protein.It can inhibit PC3 cell proliferation,induce apoptosis and inhibit tumor growth in vivo.
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Objective To analyze relationship between elderly patients with acute cerebral infarction hyponatremia and their nutrition status.Methods One hundred and twenty patients were selected who hospitalized from January 2011 to December 2012.All the patiens' fasting venous blood samples were taken in the next day morning after hospitalied.and then performed the routine blood test,blood biochemical testing items,observed the level of serum sodium (Na),hemoglobin (Hb),total protein (TP),serum albumin (ALB),prealbumin (PA),C-reactive protein (CRP) and lymphatic cell counts (LY) were measured.According to the value of serum sodium,all patients were classified to lower sodium group (serum sodium ＜ 135 mmol/L,36cases),normal group (serum sodium range from 136 to 145 mmol/L,76 cases),and higher sodium group (serum sodium ＞ 145 mmol/L,8 cases).The results were statistically analyzed.Results The incidence of hyponatremia was 30％ (36/120).The levels of TP,ALB,PA,Hb and LY in lower sodium group were (50.35 ± 8.61) g/L,(28.35 ± 6.98) g/L,(89.96 ± 12.13) mg/L,(94.13 ± 25.36) g/L and (0.87 ±0.51) × 109 respectively,lower than that of normal group ((65.30 ± 5.48) g/L,(37.50 ± 3.63) g/L,(178.14 ± 18.61) mg/L,(124.87 ± 29.08) g/L,(1.67 ± 0.98) × 109 respectively,t =5.0897,7.1058,4.3216,3.8174,5.4237 respectively,P ＜ 0.01),but the level of CRP ((76.55 ± 49.95) mg/L) in lower sodium group was obviously higher than the normal serum sodium group ((21.33 ±35.04) mg/L,t =0.1287,P ＜ 0.01).The level of the serum sodium in ICU was related to the TP,ALB,PA,Hb and CRP(r =0.3176,0.4369,0.3695,0.2408,0.3612,0.0753 respectively,P ＜0.05).Conclusion The level of the merger of hyponatremia was correlated with malnutrition in the elderly acute cerebral infarction patients in ICU.We must strengthen the nutrition support treatment while correcting hyponatremia,in order to improve the clinical curative effect.
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Objective To investigate mobilization of the bone-marrow-derived stem cell (BMSC) into peripheral blood by granulocyte-colony stimulating factor (G-CSF) to accelerate the renal regeneration.Methods Six-week-old transgenic C57BL/6J mice labeled with green fluorescent protein (GFP) as bone marrow donors and C57BL/6 mice without fluorescence label as recipients ( n =20 ) of bone marrow transplantation were used.All recipients received lethal dose of 8.5 Gy total body γ-ray irradiation with 137 Cs before bone marrow transplantation,and the transplantation of bone marrow mononuclear cells 2 × 105 by retrobulbar injection was done two hours later after irradiation. Bone marrow reconstruction after transplantation was proved by flow cytometry five weeks after transplantation.Six weeks after the bone marrow reconstruction completed,left renal pedicles of all mice were cross-clasped for 30 minutes followed by reperfusion to establish the animal model of ischemia-reperfusion injury.Mice were divided into two groups:( 1 ) Saline control group ( n =10),saline 0.2 ml/day was injected subcutaneously into chimeric mice from 3 days before to 4 days after operation ; (2) G-CSF mobilization group (n =10),chimeric mice were injected subcutanously with recombinant human G-CSF,200μg/kg/day,once a day from three days before surgery for a week.On the 1st day after mobilization,the percentage of stem cell in non-erythroid cells of peripheral blood was detected by using flow cytometry.One week after ischemia,the homing of BMSC to kidney was identified by flow cytometory.Renal tissue sections were stained with Hemotoxylin and Eosin staining method for pathological study,and the degree of renal tubular injury was analyzed by semiquantitative method of Vyacheslav.Four weeks after ischemia,the differences in degree of renal regeneration between the two groups by analysis the numbers of vascular endothelial cells in the kidney.Results After G-CSF mobilization,the percentage of stem cells with Sca-1 +,c-Kit +,CD29 and CD34 + antigen in peripheral blood in G-CSF mobilization group were higher than those in control group.One week after ischemia,mice of mobilization group showed higher percentage of Sca-1 +,c-Kit + and CD34 + bone marrow derived stem cells in tbe kidney compared to control group (P ＜0.05).One week after ischemia,the tubular epithelial damage score of mobilization group was lower significantly than that of the control group (P ＜ 0.05 ) studied by Hemotoxylin and Eosin staining. Four weeks after ischemia,mice of G-CSF mobilization group showed more CD31 positive cells in the kidney compared to control group (P ＜ 0.05 ).Conclusions G-CSF can effectively mediate the mobilization of bone marrow derived stem cells to peripheral blood and homing to kidney.G-CSF mobilization can accelerate renal regeneration and alleviate the degree of renal histopathological changes after ischemia.
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Objective To evaluate the effects of urodynamic studies on diagnosis and treatment of female stress urinary incontinence (SUI). Methods Urodynamic studies including the determination of stress leakage spot pressure were performed in 20 cases of SUI.Of them 6 cases (30%) had detrusor instability.Based on the types of SUI pathogenesis,7 cases were of Type Ⅲ,10 cases of mixed type (Type Ⅲ/Type Ⅱ) and 3 cases of Type Ⅱ.Of the 20 cases,16 underwent operation,including modified Gittes suspension in 12,sling with polyester pieces in 3 and MMK plus posterior vaginal wall repair in 1. Results The 16 operatied patients were followed up for 8～40 months(mean,31 months).The patients undergoing MMK or Sling operation were cured.But 3 patients (1 of Type Ⅲ and 2 of Type Ⅲ/Ⅱ) undergoing modified Gittes suspension had relapse (recurrence rate,25%). Conclusions Urodynamic studies are important to the diagnosis and differential diagnosis of SUI,and are indicative for choice of operation approaches.
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Objective To study the quantitative changes of gap junction (GJ) in the detrusors of unstable bladder(USB) in rats,and to deduce the functional changes of GJ,which mediates intercellular communication,so as to demonstrate one of the mechanisms of USB. Methods Thirteen Wistar rats, which were identified to have USB by manometry of filling bladder after establishment of bladder outlet obstruction (BOO) model, served as study,ie,USB group.Another 10 healthy female Wistar rats served as control group.The content and distribution of connexin (Cx) 43 of the detrusors, which were taken from these rats,were quantitatively analyzed by Western blot and laser confocal microscopy with a double-label immunohistochemical technique. Results Cx43 protein was expressed in both control and USB group, the relative molecular weight was 43000.The mean gray levels of the detrusor protein bands in USB group and control group were 31.066 and 11.701,respectively;the difference of the values between the 2 groups was significant (P
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Objective To explore the method for detecting the connexin 43(Cx43) in detrusor of bladder in rats with detrusor instability.Methods The model of rats with detrusor instability was established.Ten rats in unstable bladder group were determined by filling cystonetry method controlled by 10 normal rats.The fluorescein isothiocyanate(FITC) was used to label the Cx43 and the tritin conjugated rhodamine was used to label the cell membrane of detrusor muscle.The Cx43 and the cell membrane of detrusor muscle were detected by laser scanning confocal microscopy(LSCM),and the images were analyzed by the image analysis program.Results The distinct images of Cx43 and cell membrane of detrusor muscle could be obtained by the LSCM.The expression of Cx43 and pixel density were increased significantly in rats of unstable bladder group than those of the control group(all P
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Objective To study the pathogenesis of unstable bladder with partial bladder outlet obstruction through the experiment based research on connexins in both unstable bladder group and cell hypoxia group.Methods Unstable bladder rat models were established and smooth muscle cells were cultured in vitro.Then cell anoxia was induced.RT-PCR and Western blot were used to quantify the connexins.Results The mRNA levels of Cx40,Cx43 and the Cx43 protein expressed in the smooth muscle cells from the unstable bladder group and cell hypoxia groups were all higher than those in the control groups(the sham-operation group and normoxia groups)(P