A micro droplet generator based on V-shape linear ultrasonic motor was prepared to produce micro droplets with higher accuracy in the field of biochemistry.The device was composed of a micro droplet generator which was driven by the V-shaped linear ultrasonic motor, a three-dimensional displacement platform based on V-shaped linear ultrasonic motor, and a micro droplet separation unit based on the piezoelectric vibrator.The generating part consisted of an ultrasonic motor, a medical syringe, a silica flexible tube and a self-made micro nozzle based on glass.Utilizing the drive controller to drive the linear ultrasonic motor, the slipway pushes forward the syringe and the micro droplet was attached to the glass nozzle.The natural mode of the rod nozzle was excited by the piezoelectric vibrator.The attached droplet was separated from the tip of the nozzle after overcoming the viscous force.The separated droplet fell in a certain range.And the radius of the spherical droplet was calculated.In the experiment, distilled water was used as the initial liquid to investigate the characteristics of the micro droplets produced by the device.The experimental results showed that the droplet was attached to the tip of the micro nozzle which was formed by distilled water under the linear motor.By the vibration of the separation unit, the attached droplets formed the spherical droplets by overcoming the viscous forces in the tip of the nozzle.The radius of spherical droplets generated by this device was less than 40 μm by measuring the size.

The renin-angiotensin system plays an important role in the progression of chronic renal diesases. At present, the main approach for treating the chronic progressive renal disease is to block the role of renin-angiotensin system. Recently, angiotensin receptor blockers have being realized in width with its few side effect and good safety. A number of experimental and clinical resultsin recent 10 years have indicated that ARB can prevent or even reverse the progression of chronic renal diseases in many ways effectively and provided great evidences for current therapy of CPRD.

Objective To investigate the risk factors of chylous leakage after pancreatioduodenectomy so as to find effective measures to prevent this complication.Methods A retrospective analysis was conducted on 230 patients who underwent pancreatioduodenectomy at the First Affiliated Hospital of Zhejiang University from Jun.2012 to Jun.2014.Patients with chylous leakage were identified and a 1 ∶ 2 patients in the study and the control groups were selected.The parameters for matching included tumor volume,vascular invasion,and extent of lymph node dissection.A logistic analysis was performed to identify independent risk factors of chylous leakage.Results 15 (6.5％) patients developed chylous leakage after pancreatioduodenectomy.The average hospital stay after surgery of the study group was 20.8 days,compared to 13.5 days in the control-group (P =0.004).In the study group,chylous leakage rate increased in patients with 14th and 16th group of lymph nodes dissection (80％ vs 36.7％,P =0.006).Logistic analysis showed that 14th and 16th lymph nodes dissection was an independent risk factor of chylous leakage after pancreatioduodenectomy (P ＜ 0.05,OR =6.909,95％ CI 1.593 ～ 29.958).Conclusions Chylous leakage prolonged hospitalization after pancreatioduodenectomy.Dissection of the 14th and 16th lymph node groups was an independent risk factor of chylous leakage after pancreatioduodenectomy.Careful ligation of the gastrocolic vein near the lymphatic trunk and dissection of 14th and 16th group of lymph nodes were effective interventions to reduce postoperative chylous leakage.

Objective To survey the epidemiology of chronic kidney disease (CKD) among elderly people in two districts of Jiangsu province, China. Methods A total of 1404 residents aged 60 years or older in Huaian and Suzhou city in Jiangsu province were randomly recruited from the community population. All the people were screened for albuminuria (increased morning spot urine albumin-to-creatinine ratio, UACR) and reduced renal function (decreased eGFR by simplified MDRD equation). Urinary creatinine and albumin, serum creatinine, uric acid, cholesterol,triglyceride, low density lipoprotein-cholesterol, high-density lipoprotein cholesterol and fasting blood (1.73 m2) -1] and/or albuminuria (UACR ≥30 mg/g). The associations between healthy characteristics and indicators of kidney damage were examined. Results A total of 1316 (93.7%) elderly people completed the study. The prevalence of CKD was 32.3% and the awareness rate was only 9.6%. Albuminuria and reduced renal function were found in 30.2% and 3.2% of subjects respectively. The Logistic regression model showed that age, gender, hypertension,hypercholesterinemia and hyperuricaemia were independently associated with CKD. Systolic blood pressure (SBP) of 140-159 mm Hg exhibited a lower adjusted OR value (0.675) for CKD, while SBP of 160-179 mm Hg and of at least 180 mm Hg exhibited higher adjusted OR values (1.330 and 1.146). Similarly, FBG of 5.6-6.9 mmol/L exhibited a lower adjusted OR value for CKD as compared to FBG of at least 7.0 mmol/L (0.628 vs 1.941). Conclusions The prevalence of CKD in elderly people of Jiangsu province is quite high. Age, gender, hypertension,hypercholesterinemia and hyperuricaemia are independent risk factors for the development of CKD.

Objective To summarize the experiences in treatment of intertochanteric fractures with proximal femur nail antirotation (PFNA). Methods A retrospective study was done on 136 patients with intertrochanteric fractures treated with PFNA from March 2006 to September 2008. Postoperative reduction quality, long-term radiographic results and function of hip were evaluated separately. All the patients had closed fractures. The operation involved orthopedic traction bed, C-arm image intensifier,closed reduction or limited open reduction and locking technique. Results All patients were followed up for 10-18 months (average 14.5 months), which showed fracture healing in all patients. According to Harris criterion, the function of the hip joint obtained excellence in 129 patients ( 94.9% ). Conclusion PFNA has advantages of simple procedure, minimal invasion, firm fixation and early functional exercises and is an effective method for treatment of intertrochanteric fractures.

Objective To investigate the effects of low density lipoprotein receptor (LDLr) pathway on podocyte injury in diabetic nephropathy (DN) under inflammatory stress.Methods Male db/db mice and db/m mice were randomly divided into four groups (8 mice in each group):db/m group (control),casein injected db/m group (db/m + casein),db/db group (db/db),and casein injected db/db group (db/db + casein).An inflamed model of DN was established according to our previous study.24-hour urinary protein was measured every week.The plasma lipid profile was detected by clinical biochemistry assay.Podocyte changes were evaluated by electron microscope and immunofluorescent staining.Lipid accumulation in the kidney was evaluated by oil red O staining and intracellular cholesterol quantitative assay.The protein expression of Wilm's tumor-1 (WT-1),nephrin,α-smooth muscle actin (t-SMA),and molecules correlated with LDLr pathway were examined by immunohistochemical staining or Western blotting.The colocalized protein expression of LDLr with WT-1 was examined by immunofluorescent staining and laser confocal microscopy.Results There were no differences in plasma levels of LDL and HDL among four groups.Compared with db/db group,the db/db+ casein group showed markedly increased 24-hour urinary protein,more significant podocyte foot process effacement and podocyte damage,increased lipid droplet accumulation in kidneys,increased protein expressions of LDLr,SCAP and SREBP-2 in kidneys (all P ＜ 0.05).Interestingly,increased LDLr protein expression in kidneys of db/db mice was negatively correlated with decreased nephrin protein expression (r =-0.855,P ＜ 0.01) and positively correlated with increased α-SMA protein expression (r=0.768,P ＜ 0.01).Conclusions The disruption of LDLr pathway induced by inflammation contributes to podocyte injuries in diabetic nephropathy.

Objective To explore whether high glucose (HG)-induced endothelial-to-mesenchymal transition (EndMT) could be transitioned into mesenchymal stem cells (MSCs) and further differentiated into chondrocytes.Methods Human aortic endothelial cells (HAECs) were divided into three groups:normal glucose (NG,5.5 mmol/L glucose) group,HG (30 mmol/L glucose) group,and mannitol (5.5 mmol/L glucose + 24.5 mmol/L mannitol) group,and were cultured for 48 h.Immunofluorescence staining was performed to detect the co-expression of CD31 (endothelial markers),and fibroblast-specific protein 1 (FSP1,fibroblast markers).The expression of CD31 and FSP1 mRNA and protein was detected by real-time PCR and Western blotting.When endothelial-derived MSCs were grown in MSC medium for one week,the expression of the MSCs markers CD44,CD10 and the chondrocyte marker SOX9 was detected by Western blotting and RT-PCR.Chondrocyte expression was detected by alcian blue staining.Calcium deposit was analyzed by alizarin red staining.Pathological changes were investigated using electron microscopy.Results The expression of FSP1 mRNA and protein was significantly increased,but the expression of CD31 mRNA and protein was decreased (P ＜0.01),and the cells undergoing EndMT also significantly expressed CD10,CD44 and SOX9 in the HG group compared with those in normal glucose group (P ＜ 0.01).The incubation of HAECs exposed to HG resulted in a fibroblast-like phenotype,wherein increased microfilamentation and a roughened endoplasmic reticulum structure were observed in the cytoplasm.Double staining of the HAECs indicated a co-localization of CD31 and FSP1.After one week culture for chondrocyte medium,the expression of MSCs marker STRO-1 was significantly increased by immunofluorescence staining.Additionally,aleian blue staining in the HG group was positive compared to the NG group.Consistent with the elevation of SOX9 expression,calcium deposit also enhanced in the HG group.Conclusion HG can induce endothelial cells transdifferentiation into chondrocyte-like cells via the EndMT.

OBJECTIVE To investigate the molecular mechanisms of resveratrol( Res)in renal interstitial fibrosis(RlF)in rats with unilateral ureteral obstruction(UUO). METHODS Forty-eight Spra-gur-Dawley rats were randomly divided into UUO( normal saline,n = 16),UUO with Res treatment (Res,20 mg·kg-1 ,n=16),and sham-operation(sham,n=16)models. The kidneys were excised on the 7th and 14th day. The deposition of collagen fiber in the kidney was detected with HE and Masson staining. The levels of sonic hedgehog(SHH,an inducer of SHH pathway)in kidney tissues were deter-mined by ELlSA. lmmunohistochemical analysis was performed to evaluate the protein expression of SHH signaling-related molecules,including SHH,smoothened(Smo),patched-1(Ptch1),and Gli1, proliferating cell nuclear antigen(PCNA)and matrix component typeⅢ collagen. The mRNA expression levels of Smo,Ptch1 and Gli1 were detected by real-time RT-PCR. RESULTS The degree of RlF observed with HE and Masson staining was obviously increased in UUO kidneys,but decreased in Res-treated kidneys. Enhanced expression levels of typeⅢ collagen and PCNA in UUO rats were suppressed by Res treatment(P﹤0.05). Res administration decreased the expression levels of SHH,Smo,and Gli1 (P﹤0.05),but increased the expression of Ptch1(P﹤0.05),suggesting that Res inhibit the obstruction-induced activation of SHH signaling. CONCLUSION Res can attenuate RlF in UUO rats,and the possi-ble mechanism is that Res down-regulates the activity of SHH signaling and inhibits cellular proliferation, resulting in inhibition of matrix accumulation in renal interstitium of UUO rats.

AIM:To investigate the effect of cyclopamine on Hedgehog (HH) signaling, phenotypic transfor-mation and matrix accumulation induced by aristolochic acid (AA) in renal tubular epithelial cell NRK-52E.METHODS:NRK-52E cells were randomly divided into control group (treated with solvent only), AA group (treated with AA at con-centrations of 1, 5, 10 mg/L) and cyclopamine group (treated with AA at concentration of 10 mg/L plus cyclopamine at concentrations of 1, 5, 10μmol/L).After cultured for 24 h, the mRNA expression of Ptch1, Smo,α-SMA, E-cadherin, ZO-1, BMP-7, type I collagen and type III collagen was quantified by real-time PCR.The protein levels of Shh and TGF-β1 were detected by ELISA .Immunofluorescence staining was used to evaluate the expression of Ptch 1, Smo,α-SMA, E-cadherin and type III collagen in the NRK-52E cells.RESULTS: AA increased the expression of TGF-β1, α-SMA and type III collagen, decreased the expression of E-cadherin and ZO-1 protein, and down-regulated the expression of Ptch1, Shh and Smo mRNA in the NRK-52E cells, indicating that AA activated HH signaling , and phenotypic transformation and matrix accumulation occurred in AA-treated NRK-52E cells.Treatment with cyclopamine inhibited HH signaling by decrea-sing Smo expression and increasing Ptch 1 expression.Moreover, cyclopamine also down-regulated the expression of TGF-β1,α-SMA, type I collagen and III collagen , and up-regulated the expression of BMP-7, ZO-1 and E-cadherin.CON-CLUSION:AA induces phenotypic transformation and matrix accumulation in renal tubular epithelial cells , which can be inhibited by cyclopamine treatment .The possible mechanism is that cyclopamine suppresses the activation of HH signaling , resulting in the reduction of epithelial-to-mesenchymal transition and matrix deposition .

Objective To explore the effect of irbesartan on cardiac endothelial-mesenchymal transition (EndMT) in diabetic rats.Methods The model of diabetic rat was induced by intraperitoneal injection with streptozotocin (STZ,35 mg/kg) in spontaneous hypertensive rats (SHR).Diabetic rats were divided into diabetic group and the Irbesartan treated group.The pathological changes were investigated by fluorescence microscope and electron microscope.The EndMT was studied in human aortic endothelial cells (HAEC) exposure to high glucose.The concentration of angiotensin Ⅱ in the supernatant was detected by radioimmunoassay.Immunofluorescence staining was performed to detect the co-localization of CD31 and FSP1.Results The significant myocardial fibrosis was presented in the diabetic group.Endothelial protrusions were prominent feature in myocardial microvascular of diabetic rat compared with the control group rats.Double staining of HAEC showed co-localization of CD31 and FSP1,which was decreased by the treatment of Irbesartan (P ＜ 0.05).When HAEC was exposed to high glucose,it showed some cells acquired spindle-shaped morphology and lost CD31 staining,and FSP1 and α-SMA protein expression levels were markedly upregulated,which attenuated by the treatment of Irbesartan.Conclusion Irbesartan might prevent diabetes from myocardial fibrosis via inhibition of EndMT in diabetic rats.