Objective This study aimed to investigate the effects of angiotensin Ⅱ and Losartan pretreatment on regulating insulin secretion in human islet ? cells.Methods We measured changes in intracellular calcium by confocal laser scanning microscopy using Flou3-AM-loaded human islet cells.RT-PCR was used to measure changes in intracellular CaM.Dynamic insulin secretory responses were determined by chemiluminescence following perfusion of human islets.Results Exposure of the isolated islets to angiotensin Ⅱ induced glucose-stimulated insulin release coupled with intracellular calcium ascending in first phase and descending in second phase.Intracellular CaM concentration could not be affected by angiotensin Ⅱ.Conclusion The change of free Ca2+is induced by the combination of AngⅡ with ATI receptors of islet B cells,which results in the damage to islet B cells.Losartan pretreatment protects the islet B-cell function by inhibiting calcium overload.

Objective To examine the effects of mixed infusion of mesenchymal stem cells(MSCs) and bone marrow cells(BMCs) in the induction of chimerism and islet allograft tolerance.Methods BALB/C mouse was used as the recipient and C57BL/6 mouse was as the donor.BALB/C mice were rendered diabetic via injection of streptozotocin.The islet cells of donor mice were transplanted into the recipient mice under the capsule of kidney.Rat anti-mouse CD154 mAb was intraperitoneally injected to the recipient mice.All of recipient mice(n=25) were then randomly divided into five groups: A group(received nothing),B group(donor MSCs),C group(donor BMCs),D group(donor BMCs and MSCs) and E group(donor BMCs and the third strain-derived MSCs).The chimerism level of donor cells and the survival time of islet grafts were compared among these five groups on 7,30d and 60d after transplantation.Results On 30d and 60d after islet transplantation,the chimerism levels of donor cells in D and E groups,in which the recipient mice received the mixed infusion of MSCs and BMCs,were significantly higher than that in C group,in which the recipient mice received BMCs infusion only,and the survival time of islet graft prolonged from 53.0?16.4d to 77.0?7.7d and 61.0?2.2d,respectively(P

Objective To investigate the cause of refractory hiccups. Method 34 patients with refractory hiccups admitted to our hospital from January 1996 to December 2003 were examined with gastroscopy. The correlation of hiccups with reflux esophagitis and the efficacy of omeprazole treatment were studied. Results 10 patients were proven endoscopically to have esophagitis and duodenal ulcer, and the other 24 were suffering from esophagitis. According to Tokyo classification, 14 cases were having grade III esophagitis, and 20 were grade IV. All patients received omeprazole 20mg twice daily and cisapride 10mg thrice daily, and hiccups disappeared after 3 days. Conclusion Refractory hiccups may be the initial or primary symptom in moderate-severe esophagitis in aged people. Proton pump inhibitor therapy is effective in stopping refractory hiccup.

BACKGROUND:Percutaneous transforaminal endoscopic discectomy combined with radiofrequency annuloplasty is a minimaly invasive therapy for chronic discogenic low back pain, especialy suitable for lumbar disc herniation. OBJECTIVE:To evaluate the safety and short-term efficacy of percutaneous transforaminal endoscopic discectomy combined with radiofrequency annuloplasty for treatment of lumbar disc herniation. METHODS:Thirty-two patients with lumbar disc herniation were subjected to percutaneous transforaminal endoscopic discectomy combined with radiofrequency annuloplasty. Visual analogue scale and modified Macnab criteria were used to evaluate the relief of low back pain, and the Oswestry Disability Index was used for daily life evaluation of patients. Al the patients were folowed for 1-6 months, with an average of 3.5 months. RESULTS AND CONCLUSION:The operation was successfuly completed in al patients, and no complications such as damage to nerve and vascular systems occurred. Low back pain and living ability of the patients were improved obviously. Visual analog scale score and the Oswestry Disability Index were improved significantly after operation (P < 0.01); according to Macnnab's criteria, the results showed that at 1 week after the operation, the effective rate reached 94%; at 3 months after the operation, the effective rate was up to 97%. These findings indicate that percutaneous transforaminal endoscopic discectomy combined with radiofrequency annuloplasty in the treatment of lumbar disc herniation is confirmed in the short-term efficacy, and characterized as fewer complications and stable functional recovery.

Objective To investipate the cerebral autosomal dominat arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) through the clinical features , pathology, neuroimage, and molecular genetics as well as the family tree of a case as to exploring the diagnostic method of the disease .Methods Systematic study on the clinical manifestations, genetic site, neurologic and muscular biopsy was performed. Results The main clinical features including poor memory and history of stroke were found. Neuroimaging examination showed multiinfarct lesions and leukoencephalopathy. Biopsy indicated the arteriolar hyalinosis,GOM and osmiophilic granule. A family history was confirmed. A mutation on the fourth exon of notch 3 gene was revealed. And no risk factors of hypertension and arteriosclerosis were found. All these features are in conformity with those of CADASIL.Conclusion CADAIL is not rare and is possible to be defined in vivo by way of dermal biopsy,examination of gene and neuroimage characteristics.

Objective To investigate the effect of angiotenisn ⅡI (Ang Ⅱ) on RIN-m β-cell,and to explore the mechanism of β-cell function impairment caused by Ang Ⅱ.Methods RIN-m cells were cultured with various concentrations of AngⅡ (0.1,1,10,100 nmol/L).After incubation for 24 hours,the basal(3.3 mmol/L) and glucose-stimulated(16.7 mmol/L) insulin secretion(GSIS)were detected by radioimmunoassay,mRNA and protein expressions of uncoupling protein 2(UCP2)were determined by RT-PCR and Western blot,respectively.The intracellular ATP content was measured by luciferase bioluminescence.The mitochondrial membrane potential and cellular Ca~(2+) concentration were detected by flow cytometry.Results (1) Various concentrations of Ang Ⅱ had no significant influence on the basal insulin secrection of RIN-m cell(F=0.644,P = 0.634).Except for 0.1 nmol/L AngⅡ,the other concentrations of Ang Ⅱ markedly reduced GSIS of RIN-m cells(F= 118.528,P = 0.000).(2) Compared with the control group,Ang Ⅱ significantly increased mRNA and protein expression of UCP2(F= 1 370,P = 0.000;F=675.175,P = 0.000).(3)Except for 0.1 nmol/L Ang Ⅱ,the other concentrations of Ang Ⅱ significantly decreased the mitochondrial membrane potential,cellular ATP content,and cellular Ca2+ concentration of RIN-m cell(F=4.035,P=0.008;F=3.353,P = 0.013;F=5.867,P = 0.001).Conclusion Ang Ⅱ impairs GSIS of p-cell,the mechanism of impairment may be interpreted that Ang Ⅱ can increase the expression of UCP2,furthermore,it can reduce mitochondrial membrane potential,decrease the content of cellular ATP and the concentration of cellular Ca~(2+),can finally impair the function of β-cell.

[Objective]To study the mechanism of tubular epithelial hypertrophy induced by high glucose.[Methods]Human tubular epithelium line HK-2 cells were cultured in DMEM/F12 medium containing 1 g/L glucose(normal control group),4.5 g/L glacose(high slucose group),and 1 g/L glucose+3.5 g/L mannitol(iso-osmia control group),respectively.The cells of every group after cultured for 24 h,48 h.and 96 h were collected.The mRNA levels of CTGF and p27~(kip) were detected by real-time PCR.The protein levels of CTGF and p27~(kip) were detected by Western blot.The cellular proliferative activities were observed by MTT.The total cellular protein contents were detected by Bradford method.The cell cycle process was analyzed by flow cytometry.The cells of every group after being cultured for 30 win,1 h,24 h,and 48 h were collected.The levels of phospho-ERK1/2 were detected by Western blot.Every step was repeated for 3 times.[Results]High slucose could significantly up-regulate the mRNA and protein levels of CTGF and p27~(kip),activate ERK1/2 signal conductive pathway in HK-2 cells,make more cells arrest in G1 phase,decrease cellular proliferative activities,increase total cellular protein content reflecting cellular hypertrophy.[Conclusion]CTGF is an important mediator of tubular epithelial hypertrophy induced by high slucose.The mechanism is that CTGF up-regulates the p27~(kip) expression through activating ERK1/2 pathway.Up-regulated p27~(kip) lead proliferative cell to be arrested in G1 phase and cause cellular hypertrophy.

Objective To investigate the effects of injecting allogeneic mesenchymal stem cells(MSCs) on the cellular immunity in rat in vivo.Methods Bone marrow-derived MSCs were isolated from Wistar rats.The purity of MSCs was identified by morphological examination with microphotography,and the phenotypes were identified with flow cytometry.Twenty SD rats were randomly divided into 4 groups.Different concentrations of MSCs(5?106/ml for group A,5?105/ml for group B,and 5?104/ml for group C,respectively) and PBS(for group D) were given to allogeneic SD rats via intravenous infusions.The suppressive effect of MSCs on lymphocytes proliferation in recipient rat was analyzed using mixed lymphocyte cultures(MLR) 10 days after cultivation.At the same time,proportions of CD4+,CD8+ and CD4+CD25+/CD4+ T-lymphocytes in peripheral blood and spleen were analyzed with flow cytometry.Results Proliferation rate of splenic lymphocytes in group A(5?106/ml MSCs,8.58%?0.27%) was markedly lowered compared with that in group D(PBS,24.40%?5.21%,P

Objective To explore the changes in angiotensin Ⅱ(AngⅡ)produced by Langerhan islets of Syrian hamsters after blockage of renin-angiotensin system(RAS)by different inhibitors.Methods Islet cells from Syrian golden hamster were isolated and purified,and angiotensin Ⅰ was added.The experiment was then divided into six groups:control group(PBS was added),captopril group,chymostatin group,aprotinin group,?-antitrypsin group,and captopril+chymostatin group(according to inhibitors added).The content of AngⅡ in supernatant was detected by enzyme-linked immunosorbent assay(ELISA).Results Compared with control group,the AngⅡ content decreased by 42.50% and 50.94% in captopril group and chymostatin group,respectively(P

Objective To investigate the effect of angiotensin Ⅱ(AngⅡ) and losartan on ? cells,and its related molecular mechanisms.Methods Normally cultured RIN-m cells were divided into three groups:control group(cultured with RPMI 1640 medium),AngⅡ group(treated with 100 nmol/L AngⅡ),losartan group(treated with 1 ?mol/L losartan 15min before addition of 100nmol/L AngⅡ).After incubation for another 48h,the apoptosis rate of RIN-m cells was quantified by flow cytometry(FCM) with Annexin-V FITC/PI dual staining.The expressions of Bcl-2 and Bax mRNA and protein were determined by RT-PCR and Western blotting,and the activities of Caspase 3 and Caspase 9 were detected by spectrophotometry.Results The apoptosis rate of RIN-m cells was significantly higher in AngⅡgroup than in both control and losartan group(P0.05).In comparison to the control group,the expressions of Bcl-2 mRNA and protein significantly declined,while of Bax mRNA and protein increased obviously in AngⅡ group(P0.05),but the expressions of Bcl-2 mRNA and protein were significantly higher(P0.05).Conclusion AngⅡ may induce the apoptosis of ? cells through mitochondrial pathway,and pre-intervention with losartan,which partly reverses the effect of AngⅡ,may play a protective effect on ? cells.