in vitro fertilization in future.

Objective To investigate the effect of high glucose and losartan on cell proliferation and cyelooxygenase-2 (COX2) expression in normal human mesangial cells (NHMCs), and to examine the effect of losartan on COX2 and transforming growth factor-betal (TGF-β1) expression in a model of diabetic nephropathy (DN). Methods NHMCs were cultured in vitro in high glucose media with or without losartan. NHMCs proliferation and COX2 expression were determined by WST-1, Western blot,and RT-PCR. The rat model of DN was produced by injections of streptozocin (STZ). After the treatment with losartan for 4 weeks, glomerular hypertrophy, urinary thromboxane B2(TXB2) and 24 h urinary pro-tein counts were measured,and COX2 and TGF-β1 expressions were investigated using immunohistochem-ical techniques and RT-PCR. Results Losartan dose-dependently inhibited the proliferation of NHMCs in response to high glucose. Losartan also decreased COX2 expression in NHMCs at high or low glucose concentrations. In vivo experiments found kidney weight/body weight (KW/BW), urinary TXB2 and 24 hurinary protein counts increased significantly in the DN group. Losartan reduced KW/BW, urinary TXB2,and 24 h urinary protein counts and significantly suppressed the over-expression of COX2 and TGF-β1.Conclusion Losartan reduces COX2 expression in NHMCs, especially at high glucose concentrations.Losartan could suppress the expression of COX2 and TGF-β1 in the kidney of DN rats and attenuate the renal lesions caused by DN.

Objective To explore the association between fibroblast growth factor 23 (FGF-23) and calcium (Ca)-phosphorus(P) metabolism and bone mineral density (BMD) in patients on peritoneal dialysis.Methods Fifty-nine patients undergoing continuous ambulatory peritoneal dialysis(CAPD) were enrolled in this study.These patients were divided into three groups as normal, osteopenic and osteoporotic, according to World Health Organization criteria based on bone mineral density T scores.Another 30 healthy people were also enrolled as control group.Levels of serum FGF-23 and 1,25 (OH)2VitD3 were measured by ELISA.Parathyroid horomone(PTH) was detected by immunoradiometric assay.Calcium and phosphorus were assessed with autobiochemistry machine.Bone density was studied by dual-energy X-ray absorptiometry (DEXA).Results The incidences of osteoporosis at the femur neck and lumbar spine in CAPD patients were 23.7％ and 35.6％, respectively.Among three groups of CADP patients, no significant differences were found in the levels of serum FGF-23, while the level of serum FGF-23 in CAPD group was higher than that in control gronp(P＜0.01).A positive correlation was found between log [FGF-23]and serum phosphorus(r=0.604, P＜0.01).However, a negative correlation was found between log[FGF-23]and 1,25(OH)2VitD3 and GFR (r=-0.401, P＜0.05; r=-0.651, P＜0.01).There were no correlations of log [FGF-23]with PTH, Ca, T scores and the duration of dialysis.Conclusions In CAPD patients, serum FGF-23 increases significantly.Serum phosphorus, renal function and 1,25(OH)2VitD3may play an important role in adjusting the level of serum FGF-23, while FGF-23 has no direct effect on bone mineralization in CAPD patients.

Objective To explore the effect of aldosterone on renal epithelialmesenchymal transition in streptozocin(STZ)-induced diabetic nephropathy rats. Methods Wistar rats were intraperitoneally injected with STZ(60 mg/kg)for the preparation of diabetic model.After 4 weeks,the rats with urinary protein＞30 mg/d were regarded as successful diabetic nephropathy(n=16),and were randomly divided into diabetic nephropathy(DN group,n=8)and spironolactone group(SP group,n=8).Then eight healthy rats were selected randomly as control group(N group,n=8).SP group rats were treated with spironolactone 40 mg·kg-1·d-1,and N group and DN group rats were given equal water.After 8 weeks,rats were sacrificed to collect urine,blood plasma,kidney tissue for detection of 24 h urinary protein,creatinine and renal pathological changes.Aldosterone concentration in plasma and kidney tissue was detected by mdioimmunoassay;E-cadherin,α-SMA protein expression by immunohistochemistry,Western blotting; E-cadherin,α-SMA mRNA expression by RT-PCR. Results Compared with N group,serum creatinine, urinary protein excretion in the DN rats were significantly higher (P＜0.01,respectively), E-cadhefin protein and mRNA were significantly reduced (P＜0.01, respectively),α-SMA protein and mRNA expression was up-regulated (P＜0.01, respectively). Aldosterone level of kidney tissue in DN rats was increased obviously [(24.71±5.30) ng/g vs (16.38±2.85) ng/g, P＜0.01], which was positively correlated with urinary protein excretion, serum creatinine and α-SMA protein (r=0.737, 0.574, 0.688, P＜0.01, respectively), and negatively correlated with E-cadherin protein (r=-0.659, P＜0.O1). While no significant difference was found in serum aldosterone among three groups. Compared with DN rats, urinary protein excretion, serum creatinine were reduced (P＜0.01, respectively), E-cadherin protein and mRNA were increased (P＜0.01, respectively), α-SMA protein and mRNA expression were decreased (P ＜0.01, respectively) in SP group rats.Conclusions Local aldosterone involves in renal epithelial-mesenchymal transition in diabetic nephropathy rat. Spironolactone can block the effect of aldosterone and play a role in renal protection.

Objective To explore the clinical significance of lymph node micrometastases in stage Ⅱ rectal cancer patients. Methods Forty-two patients with rectal cancer underwent total mesorectal exci-sion between January 2000 and August 2001 were included, 484 lymph nodes were studied in paraffin blocks that had previously been considered free by conventional histopathological examination. These lymph nodes were submitted to immunohistochemical analysis using cytokeratin 20 (CK20) monoclonal antibodies to identify micrometastases. Five-year follow-up information was obtained on these patients. Observed survival rates and assessed respectively in the patients with and without micrometastases. Results Micrometastases were detected in 33 lymph nodes (6.8% ,33/484) of 15 cases (35.7%, 15/42). The five-year survival rate was 40.0% in the patients with micrometastases, whereas in the patients without micrometastases, the survival rate was 92.6%(P = 0.000,by the Log-rank test). By multivariate Cox regression analysis, lymph node mi-cromctastases was closely correlated with post-operative recurrence or metastases, the value of RR was 11.435. Conclusions Detection of micrometastases is an important prognostic tool in stage Ⅱ rectal can-cer. In this study, lymph nodes micrometastases is an independent prognostic factor for overall survival. These patients maybe get benefit from adjuvant chemotherapy.

Objective To evaluate the clinical effects of memory alloy embracing fixators in fixation of the rib fractures and investigate the related surgical indications and surgical techniques.Methods Retrospectively review was conducted on the clinical data of 30 patients with rib fractures treated with memory alloy embracing fixators from October 2010 to April 2011 at General Hospital of Guangzhou Military Command.The number of memory alloy embracing fixators used in operation,the number of fixed positions,and operation time were recorded.The pain scores before and after operation were comparatively studied.Operation efficacy and complications were analyzed.Results Of the 30 patients,the total operation time,number of fixators,and number of fixed ribs were (111.9±48.0) minutes,4.3±2.1 and 3.5±1.3,respectively.Meanwhile,the difference between pre-operative and post-operative pain scores was significant (6.93±0.88) points vs (4.04±0.62) points,P＜0.05).The ambulation perlod was (4.6±1.9) days and length of hospital stay was (27.2±10.8) days.Incisional and thoracic wall hematoma was detected in three patients and pulmonary infection in six post-operatively but none presented intractable chest pain,foreign body rejection or wound infection.Conclusion Memory alloy embracing fixators for rib fractures is reliable,easy,and effective in alleviating pain,improving lung function,reducing the frequency of ventilator use and preventing complications like lung infection.

OBJECTIVE: To investigate the pathogens and their resistance in continuous ambulatory peritoneal dialysis (CAPD) related peritonitis. METHODS: A total of 78 cases with CAPD related peritonitis from Xiangya Hospital between January 2007 and January 2011 were reviewed. Pathogens, resistance and outcomes of the 78 cases CAPD related peritonitis were analyzed retrospectively. RESULTS: Among them, 53 cases cultured positive (66.67%), 3 of which were combined infection and 2 strains were cultured. A total of 55 strains were cultured, including 32 gram-positive strains (58.18%), 18 gram-negative strains (32.72%) and 5 fungi (9.09%). The most common pathogens were coagulase negative staphylococcus, staphylococcus aureus and Escherichia coli. Drug sensitivity test of the gram-positive strains showed that the three with lowest antibiotic resistance were linezolid (0), teicoplanin (3.13%) and vancomycin (4.0%). Drug sensitivity test of the gram-negative bacteria showed that antibiotics with the lowest resistance were amikacin and imipenem, followed by piperacillin/tazobactam and cepoperazon/sulbactam. Cephazolin had the highest resistance rate of 83.33%. Clinical outcomes: 63 cases cured (80.77%); 11 cases transferred to hemodialysis (14.1%); 4 cases died (5.13%), including 2 cases fungus infections, 1 gram-negative bacteria infection and 1 combined infection. CONCLUSION The most common pathogens causing peritoneal dialysis associated peritonitis is gram positive bacteria. In the empirical treatment, in addition to traditional treatment of Cefazolin combined with aminoglycosides, cefazolin combined with piperacillin/tazobactam or cepoperazon/sulbactam is preferable for CAPD associated peritonitis.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bacteria ; drug effects ; isolation & purification ; Child ; Drug Resistance, Bacterial ; Female ; Humans ; Male ; Microbial Sensitivity Tests ; Peritoneal Dialysis, Continuous Ambulatory ; adverse effects ; Peritonitis ; etiology ; microbiology ; Young Adult

OBJECTIVE: To investigate the effect of cordyceps sinensis (CS) extract and losartan (Los) on the expression of Klotho (Kl), P53, P21, and apoptosis in renal tubular epithelial cell NRK-52E induced by angiotensin II (Ang II), and to elucidate its therapeutical mechanism in Ang II induced renal tubular epithelial cell apoptosis. METHODS: NRK-52E cells were incubated with CS with or without Ang II for 24 hours. Experimental groups were divided according to the increasing concentrations of CS:0 (serving as controls), 5, 10, 20, 40, and 80 mg/L. The optimal concentration of CS was selected and cells were divided into 5 groups: controls, Ang II (1*10(-8) mol/L), Ang II (1*10(-8) mol/L)+CS (40 mg/L), Ang II (1*10(-8) mol/L)+Los (1*10(-5) mol/L), and Ang II (1*10(-8) mol/L)+CS (40 mg/L)+Los (1*10(-5) mol/L). After 24 hours, cell proliferation was evaluated by MTT assay. The mRNA and protein expression of Kl, P53 and P21 were measured by RT-PCR. Activity of caspase-3 was evaluated by caspase-3 activity assay Kit. Cell apoptosis was determined by Annexin V-FITC/PI double staining and flow cytometry. RESULTS: Certain concentrations of CS promoted the proliferation of NRK-52E cells and increased cells proliferation inhibited by Ang II (P<0.01 or P<0.05 ). Ang II significantly down-regulated the mRNA and protein expression of Kl, and up-regulated the levels of P53 and P21. Caspase-3 activity and apoptotic rates were decreased, too (all P values<0.01). CS or/and Los significantly increased the expression of Kl mRNA and protein down-regulated by Ang II, decreased P53 mRNA and protein expression, P21 mRNA and protein expression,and inhibited caspase-3 activity and apoptotic rates(all P values<0.05). No cooperative effects were observed in the two drugs (P>0.05). CONCLUSION CS can increase the expression of Kl down-regulated by Ang II, decrease P53 and P21 expression and caspase-3 activity, and reduce Ang II induced NRK-52E cell apoptosis, which may be part of its mechanism of the protective effects on hypertensive renal damage.
Angiotensin II ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Cells, Cultured ; Cordyceps ; chemistry ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; cytology ; metabolism ; Glucuronidase ; genetics ; metabolism ; Humans ; Kidney Tubules ; cytology ; metabolism ; Losartan ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism

Objective:To investigate the effects of Cordyceps sinensis (CS) on cellular apoptosis and Sirt1 expression in HK2 cells followed by ischemia-reperfusion (I/R).Methods:HK2 cells were incubated with different concentrations of CS (10,20,40,80,160,320 mg/L) for 24 hours,and the optimal concentration of CS was selected by measuring cell proliferation.The confluent HK2 cells were incubated with 0.01 μmol/L antimycin A for 2 hours to induce ischemia in vitro,and then the reperfusion was achieved by incubating cells with glucose-replete complete growth medium for 24 hours.HK2 cells were divided into 4 groups:a control group,an I/R group,an I/R+CS (160 mg/L) group,and an I/R+CS (160 mg/L)+Sirtinol (25 μtmol/L) group.Twenty-four hours later,total RNA and protein were collected.The cell proliferation was evaluated by MTT assay;the mRNA and protein expression of Sirtl and the cleaved caspase-3 were measured by qRT-PCR and Western blot,respectively.The cellular apoptosis rate was determined by Annexin V-FITC/PI double staining and flow cytometry.Results:Certain concentrations (10-160 mg/L) of CS did not show effect on the proliferation of HK2 cells (P＞0.05),while 320 mg/L of CS inhibited cell proliferation significantly (P＜0.01);compared with the control group,the mRNA and protein expressions of Sirtl and the cleaved caspase-3 in the I/R group were up-regulated (P＜0.01) and the apoptosis rate was extremely high;compared with the I/R group,CS significantly up-regulated Sirt1 mRNA and protein expression (P＜0.01) while down-regulated cleaved caspase-3 mRNA and protein levels (P＜0.01),and reduced apoptosis rate (P＜0.05).The effects of CS were blocked in the presence of sirtinol,an inhibitor of CS.Conclusion:CS protects HK2 cells from I/R injury through activation of Sirt 1 pathway.

Objective:To investigate the status of vascular access in hemodialysis patients in our center.Methods:The general information of hemodialysis patients and types and complications of vascular access at Xiangya Hospital of Central South University from April 2015 to April 2016,were retrospectively analyzed.Results:Among 258 prevalent patients,87.60％ of them had arteriovenous fistula (AVF),while 12.40％ showed tunneled cuffed catheter.Of the 61 incident patients,80.33％ of them initiated dialysis with a non-tunneled and non-cuffed catheter,8.19％ with an AVF,9.84％ with a tunneled cuffed catheter,and 1.64％ with needle puncture.The types of AVF access included 76.55％ of wrist radiocephalic fistula,7.08％ of mid-forearm cephalic fistula,11.06％ of elbow brachiocephalic fistula,and 5.31％ of antecubital fistula and transposed basilic fistula.Seventy-seven (34.07％) patients with AVF suffered complications and wherein aneurysms accounted for 24.34％.Conclusion:In maintenance hemodialysis patients,autologous AVF is the prevalent vascular access.In the beginners for dialysis,non-tunneled and non-cuffed catheter are their choice.Additional efforts and incentives may be necessary to improve vascular access during the initiation ofhemodialysis.