OBJECTIVE To investigate the effect ofγ-secretase inhibitor N-[N-(3,5-difluorophen?acetyl)-L-alanyl]-S-phenylglycine t-butyl ester(DAPT)on phenotypic transformation and matrix accu?mulation induced by aristolochic acid(AA) in renal tubular epithelial cells(NRK-52E)and explore the mechanism. METHODS NRK-52E cells were divided stochastically into normal cell control group,AA 10 mg·L-1 group and AA 10 mg·L-1+DAPT 1 and 10μmol·L-1 group. After 24 h,the mRNA expressions of Notch1,Jagged1,Numb,E-cadherin,transforming growth factor-β1(TGF-β1),α-smooth muscle actin(α-SMA),bone morphogenic protein 7 (Bmp7),typeⅠ a1 (Col1a1) and Ⅲ collagens a1 (Col3a1)were quantified by quantitative real-time RT-PCR. The protein expressions of Notch1,Jagged1,α-SMA,and Col3a1 in NRK-52E cel s were detected by immunofluorescence staining. RESULTS In NRK-52E cells,AA enhanced the expression of TGF-β1,α-SMA and Col3a1 mRNA(P<0.05),reduced the expression of E-cadherin mRNA(P<0.05),up-regulated the mRNA expression of Notch1 mRNA(P<0.01)and Jagged1(P<0.05),and down-regulated the mRNA expression of Numb mRNA(P<0.05) compared with normal cell control group,indicating that phenotypic transformation and matrix accumu?lation occurred in AA-treated NRK-52E cells,accompanied by activated Notch signaling. Treatment with DAPT inhibited Notch signaling by decreasing the expression of Notch1 and Jagged1 (P<0.05),and increasing the expression of Numb mRNA(P<0.05). Furthermore, DAPT also down-regulated the expression levels of TGF-β1,α-SMA,Col1a1 and Col3a1 mRNA(P<0.05), and up-regulated the expression level of Bmp7 and E-cadherin mRNA(P<0.05) compared with AA group,suggesting that DAPT inhibited phenotypic transformation and matrix accumulation in AA-treated NRK-52E cells. CONCLUSION AA induces phenotypic transformation and matrix accumulation in renal tubular epithelial cells,which is inhibited by DAPT treatment. The possible mechanism is that DAPT suppresses the activation of Notch signaling,resulting in the reduction of epithelial-to-mesenchymal transition and matrix deposition.

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 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.

Objective To investigate the expression of monocyte-macrophage-related factors and interstitial fibrosis in kidney tissues of rats with ureter obstruction and recanalization .Methods Forty-eight male Spragur-Dawley rats were divided randomly into the obstructive group:sham (n=6), unilateral ureteral obstruction(UUO)3 days (n=6), UUO 7 days (n=6), and UUO 14 days (n=6) and recanalization group:bilateral ureteral obstruction(RBUO)0 day (n=6), 3 days after RBUO (n=6), 7 days after RBUO (n=6), and 14 days after RBUO (n=6).The kidneys were excised on day 3, 7, and 14, and the deposition of collagen fibers in kidney was detected with HE and Masson staining . Immunohistochemical analysis was performed to evaluate the protein expressions of monocyte chemoattractant protein -1 (MCP-1), macrophage colony-stimulating factor (M-CSF) and activated-macrophage marker CD68.Real-time PCR was used to detect the mRNA expressions of MCP-1 and M-CSF.TGF-β1 levels were determined by ELISA .Results Fibrosis observed with HE and Masson staining was obviously increased in kidney tissue of UUO rats , and aggravated as time prolonged, but alleviated in rats with recanalization .TGF-β1 levels were increased obviously in the UUO group , but decreased in rats with recanalization compared with those in BUO rats .In UUO rats, mRNA and protein expression levels of MCP-1 and M-CSF were increased .MCP-1 and M-CSF expression was gradually decreased in rats with recanalization compared with those in BUO rats .The dynamic change in expression of MCP-1 and M-CSF in both UUO rats and recanalization rats was consistent with the change in expression of CD 68. Conclusion Dynamic change in expression of MCP-1 and M-CSF in kidney tissues reflects change of activated and accumulated monocyte -macrophages , which may be one of the major mechanisms contributing to fibrosis induced by ureter obstruction .Renal fibrosis is alleviated by down-regulated expression of monocyte-macrophages factors with recanalization operation .

Objective To study the calcium metabolism in tacrolimus(FK506)induced rats nephrotoxicity and the preventive effect of calcium channel blocker.Methods Twenty-four Spragueinduced or FK506-induced nephropathy model.Blood creatinine,blood electrolytes,renal tissue histopathology(HE stain)and the change of ultrastructural organization in renal cells by transmission electron microscope were observed.Results The blood creatinine levels of both CsA and FK506 groups [(36.00±2.61)and(34.17±4.54)μmol/L] were significantly higher than those of the FK506+Dilgroup and control group(all P＜0.05).The blood calcium levels of both CsA and FK506 groups (2.00±0.04 and 2.05±0.04 mmol/L) were significantly lower than those of the FK506+Dil group and control group(all P＜0.05).The blood creatinine and calcium levels of FK506+Dil group were not significantly different with those of control group(P＞O.05).Histopathology examination showed cloudy swelling and vacuolization of the renal tubular epithelial cells and intra-cellular mitochondria swelling and vacuolization in the CsA and FK506 groups.However,the pathological changes of the FK506+Dil group were remarkably milder in comparison with the CsA and FK506 groups.Concluum channel blocker,Dil,could prevent the FK506-induced nephrotoxicity.

Objective To investigate the relationship between benign prostatic hyperplasia (BPH) andchronic prostatitis(CP). Methods Three hundred BPH patients were studied, aged from 51 to 96 (aver-age 72). All patients were divided into 3 groups (Ⅰ°、Ⅱ°and Ⅲ°)according to result of digital rectal examina-tion, which include 85 cases , 139 cases and 76 cases respectively. The incidence of CP among 3 groups were compared and analyzed. Results Two hundreds and thirty-five of the 300 cases with BPH were accompa-nied with CP(77.7%). Among the 233 cases, 53 cases were in Ⅰ degree BPH group (53 / 85, 62.4% ), 113 cases were in Ⅱ degree BPH group (113/139, 81.3%), 67 cases were in Ⅲ degree BPH group (67/76, 88.2%). Conclusions Many BPH patients were accompanied by CP. The prostate size and the inflamma-tion of prostate were positive correlated. The effect of anti-inflammatory treatment in Ⅰ degree and Ⅱ degreeBPH patients was better than Ⅲ degree BPH patients.

Objective To analyze clinical and laboratory features, viral load and viral shedding period of patients with mild or severe H1N1 influenza A infection. Methods Seventy mild cases and 16 severe cases with concurrent pneumonia were included from Shcnzhen area for analysis.Nasopharyngeal-swab specimens of patients were collected and viral load was detected by real-time quantitative polymerase chain reaction (PCR) assay during their hospitalization. The viral load and viral shedding period were compared between patients over 14 years old and less than 14 years old, and between 70 mild cases without pneumonia and 16 severe cases with pneumonia. The statistic analysis was performed using t test and chi square test. Results The most common symptoms and signs of the patients were fever, cough and enlargement of tonsils. However, the severe cases suffered more frequently from cough, dyspnea and high fever compared with the mild cases (x2 = 10. 9 and 14.3, respectively, t=3.65; both P＜0.01 ). The levels of white blood cell (WBC) count and alanine arninotransferase (ALT) of severe patients were both significantly higher than those of mild patients(t= 3.2, 2.4,respectively; both P＜0.05). The chest radiology of the severe cases showed interstitial pneumonia,mostly with ground glass image. The viral load of patients under 14 years was significantly higher than those over 14 years [(4.86± 1.23) lg vs (4. 17±0.89) lg; t=2.3, P＜0.05], and the viral shedding period of patients under 14 years was significantly longer than those over 14 years [(5.33±0. 49) d vs(3. 63±0.28) d; t=3.4, P＜0.01]. The severe patients also displayed significantly higher viral load and prolonged viral shedding period than the mild patients [(6. 36±1. 44) lg vs (4. 35±0.99) lg, t=6.1,P＜0.01; (5.75±1.77) d vs (4. 24±1. 96) d, t=3.2, P＜0.01]. Conclusion Age anddisease severity of patients with H1N1 influenza A infection are significantly associated with viral load and viral shedding period.

Objective To explore the value of retrograde recanalization technique in treatment of TASCⅡ C/D femoropopliteal arteriosclerosis obliterans.Methods Totally 36 patients with TASCⅡ C/D femoropopliteal arteriosclerosis obliterans were retrospectively analyzed.Preoperative and postoperative ankle brachial index (ABI) and intermittent claudication were compared,as well as postoperative vascular patency rate and complications were analyzed.Results The retrograde recanalization technique were successfully performed in all 36 patients.The walking distance and ABI of 7 days,and 3,6,12 months postoperation were increased significantly compared with preoperation (all P＜0.05).The postoperative vascular patency rate of 3,6 and 12 months was 97.22％,91.67％ and 72.22％,respectively.There was no death nor amputation during the follow-up period,and the incidence of perioperative complications was 11.11 ％ (4/36).Conclusion Retrograde recanalization technique can be used to treat TASC Ⅱ C/D femoropopliteal arteriosclerosis obliterans when anterograde access failed.

Objective: To learn the population,virus status and viral types of hantavirus（HV）hosts in Tiantai County of Zhejiang Province from 2011 to 2018,and to provide evidence for hemorrhagic fever with renal syndrome（HFRS）control. Methods: Rodents in Tiantai County were captured by night trapping method. After the species and age of rodents were identified,the composition of rodent species,dominant species and density of rodents were analyzed. The lungs and blood of rodents were sampled to detect the antigen and antibody of HV by immunofluorescence method. The HV antigen-positive lung samples were detected by RT-PCR with specific primers of HV S fragment,then HV was isolated and identified by inoculating Vero-E6 cells. Results: The average rodent density in Tiantai County from 2011 to 2018 was 4.44%. The rodent density in the field and residential areas were 4.94% and 2.23%,respectively. Ten species of rodents were identified,with Apodemus agrarius dominant in the field and Rattus norvegicus in the residential areas. Sixty-seven lung samples were HV antigen positive（4.13%）,one from Rattus norvegicus and the other sixty-six from Apodemus agrarius. Seventy-nine blood samples were HV antibody positive（4.86%）,all from Apodemus agrarius. Thirty-four HV antigen-positive lung samples were positive（50.75%）after RT-PCR amplification. Twenty-two strains of virus were isolated and all of them were from Apodemus agrarius,including twenty-one strains of Hantaan type（HTN）and one strain of Seoul type（SEO）. Conclusion In Tiantai County,Apodemus agrarius is the main source of HFRS infection;the main epidemic type of HV is HTN and SEO is first found in Apodemus agrarius.

Objective To compare the hemostatic safety and efficacy of a new butterfly femoral artery compression device (FACD) with those of manual compression (MC) in patients undergoing percutaneous peripheral endovascular interventions via femoral artery. Methods A total of 283 patients, who received percutaneous endovascular interventions via femoral artery during the period from September 2016 to December 2017, were enrolled in this study. After endovascular intervention, 167 patients received FACD to make hemostasis (FACD group), and 116 patients received MC hemostasis (MC group) . The patient's comfortableness, time used for hemostasis (min), limb immobilization time (h), and the incidence of vascular complications in both groups were analyzed. Results All 283 patients were included in analysis, the results indicated that the hemostatic success rates in FACD group and MC group were 96.4％ (161/167) and 94.0％ (109/116) respectively, the difference between the two groups was not statistically significant (P>0.05) . The postoperative Kolcaba Comfort Scale score of FACD group was (85.0 ±11.2) points, which was remarkably higher than (58.4±11.7) points of MC group (P<0.05), the time used for hemostasis in FACD group was (9.2 ±2.2) min, which was strikingly shorter than (18.5 ±2.9) min in MC group (P <0.05) . The limb immobilization time in FACD group was (10.4±2.4) hours, which was obviously shorter than (23.1±4.1) hours in MC group (P <0.05) . The incidence of vascular complications in FACD group was 3.6％, which was dramatically lower than 9.5％ in MC group (χ2=4.206, P=0.04) . Conclusion The use of the new butterfly FACD can promptly, safely and effectively stop bleeding of femoral artery puncture site. The new butterfly FACD is superior to MC in shortening hemostatic time and limb immobilization time, in reducing incidence of vascular complications, as well as in improving patient's comfortableness degree.