Objective To investigate the effect of advanced glycation end products (AGEs) on the disruption of tight junctions in rat glomerular endothelial cells (rGEnCs) and the role of renin-angiotensin system (RAS) in this pathological procedure.Methods Primary cultured rGEnCs were incubated with AGEs at concentrations of 20 mg/L,40 mg/L and 80 mg/L,for 6 h,12 h and 24 h respectively.The cells were treated with captopril (1 mmol/L) or valsartan (10 μ mol/L)to block RAS.The endothelial permeability was investigated by transendothelial electrical resistance and the flux of fluorescein isothiocyanate-conjugated bovine serum albumin.The expression of AGEs receptor (RAGE),tight junction proteins [occludin,claudin-5,junctional adhesion molecules A (JAM-A) and zona occludens-1 (ZO-1)]and RAS components [angiotensinogen,renin and angiotensin Ⅱ type 1 receptor (AT1)]were detected by Western blotting.Immunofluorescence was used to demonstrate the disruptions of the tight junction proteins.The activity of angiotensin converting enzyme (ACE) was evaluated by UV spectrophotometry.Angiotensin Ⅱ (Ang Ⅱ ) was measured by enzyme immunoassay.Results The monolayer permeability,the expression of RAGE,the activity of ACE,the concentration of Ang Ⅱ and the expression of AT1 of rGEnCs were increased after induced by AGEs.Meanwhile,AGEs decreased the expression of occludin,claudin5 and JAM-A and induced disruption of tight junction proteins.Pretreatment with anti-RAGE antibody (100 mg/L),captopril or valsartan could attenuate the detrimental effect of AGEs.Conclusion The changes of permeability induced by AGEs in glomerular endothelial cells are partly mediated by RAS through RAGE.

Objective To explore the efficacy and safety of percutaneous transhepatic portal vein or transjugular intrahepatic portosystemie shunt (TIPS)to implant the portal vein metallic stent in treatment of cavernous transformation of portal vein (CTPV).Methods Clinical and imaging data of 8 patients with CTPV were retrospectively analyzed who were treated in our hospital.All patients were treated with metallic stent implantation in portal vein including 3 patients by TIPS and 5 by percutaneous transhepatic portal vein.Results All patients were successful in the stent implantation without any occurrence of serious complications such as intra-abdominal hemorrhage and so on.Intraoperative angiography showed blood circulated freely in these stents.1 day-2 weeks later,the patients symptoms of abdominal pain and gastrointestinal bleeding were obviously relieved or disappeared.Follow up 1 month-3 years,1 patient with stent occlusion after one year of operation,the blood flow recovery after stent reimplantation,and the remaining patients,color doppler ultrasound reflected patency of blood flows in their stents.No one suffered from gastrointestinal bleeding or abdominal pain again.Conclusion Implantation of portal vein metallic stent via percutaneous transhepatic portal vein or via TIPS in treatment of cavernous transformation of portal vein is safe and effective.

AIM: To investigate the effects of adenovirus-mediated human tissue kallikerin (Ad-hKLK1) gene delivery on the proliferation, migration of VSMC_(SHR) induced by platelet derived growth factor-BB (PDGF-BB). METHODS: The VSMC_(SHR) proliferation induced by PDGF-BB was accessed by cell counting and methyl thiazolyl tetrazoliuin (MTT). The migration was assessed by modified Boyden chamber assay. Western blotting was used to determine the expressions of the cycle-independent kinase inhibitors p27~(Kip1) and p21~(Cip1).RESULTS: Proliferation of VSMC_(SHR) induced by PDGF-BB was inhibited after transfection of Ad-hKLK1 (20-100 MOI) in a MOI-dependent manner. The peak inhibition titer of Ad-hKLK1 fell on 100 MOI, with the peak inhibition rate of 39.3% (cell counting, n=3, P<0.01), 30.2% (MTT, n=3, P<0.01), peak stunning rate of cell-cycle in phase G0/G1 at 36.4%. The inhibitory effects of proliferation and cell-cycle caused by hKLK1 gene delivery were significantly abolished by Hoe140, a bradykinin B2 receptor antagonist. Migration of VSMC_(SHR) induced by PDGF-BB was inhibited after hKLK1 gene delivery, with the peak inhibitory rate of 34.6% (n=6, P<0.01). However the inhibitory effects of migration were not blocked by Hoe140. The protein expression of p27~(Kip1) and p21~(Cip1) increased significantly after the hKLK1 gene delivery, whereas Hoe140 nearly completely blocked these effects (n=3, P<0.01, respectively).CONCLUSION: The hKLK1 gene delivery may inhibit the proliferation and migration of VSMC_(SHR) induced by PDGF-BB. Bradykinin B2 receptor probably mediates the up-regulating expression of p27~(Kip1) and p21~(Cip1) that contributes to the inhibitory effects of proliferation of hKLK1. However, the inhibitory effects of migration by hKLK1 gene delivery may not be mediated by bradykinin B2 receptor.

Objective:To explore changes of central retinal vascular caliber and fractal dimension (Df) and their cor‐relation with blood pressure in hypertensive population .Methods :A total of 2169 subjects>30 years old were en‐rolled in this cross‐sectional study .They were divided into hypertension group (n=819) and non‐hypertension group (n=1350) .Fundus photos were collected in all subjects ,and semi‐automatic software was used to quantitatively ana‐lyze central retinal vascular caliber and Df ,and they were compared between two groups .Results:Compared with non- hypertension group ,there were significant reductions in central retinal arteriolar equivalent [CRAE ,(135.2 ± 10.72) μm vs .(132.25 ± 11.56) μm] ,central retinal venular equivalent [CRVE ,(184.95 ± 16.29) μm vs . (182.52 ± 17.07)μm] and Df [ (1.38 ± 0.05) vs .(1.34 ± 0.05)] in hypertension group , P<0.01 all .After adjus‐ting for age and gender ,analysis of covariance indicated that CRAE and Df of hypertension group were still signifi‐cantly lower than those of non - hypertension group (P<0.01 both) .Linear correlation analysis indicated that sys‐tolic blood pressure (SBP) ,diastolic blood pressure (DBP) and pulse pressure (PP) were inversely correlated with CRAE and Df ( r= -0.340~ -0.174 , P<0.01 all) .After adjusting for cardiovascular risk factors ,multi‐factor linear regression analysis indicated that CRAE and Df were still inversely correlated with SBP ,DBP and PP (stand‐ardizedβ= -0.190~ -0.134 ,P<0.01 all) .Df of hypertension course >5 years group was significantly lower than that of ≤5 years group [ (1.33 ± 0.05) vs .(1.35 ± 0.05)] , P<0.01. Conclusion:CARE ,Df are significantly in‐versely correlated to SBP ,DBP and PP in hypertensive population ,while correlation of Df is most .

Objective: To observe the effect of pyridoxamine and telmisartan on the indexes of myocardial remodeling in spontaneous hypertensive rats (SHR). Methods: A total of 48 male SHR at 22 weeks of age were randomly divided into 4 groups:①Hypertension (H) control group, the rats received distilled water 2 ml/d,②telmisartan (T) group, the rats received telmisartan 6 mg/(kg.d),③pyridoxamine (P) group, the rats received pyridoxamine 200 mg/(kg.d),④Combination (TP) group, the rats received both telmisartan and pyridoxamine. All animals were treated for 16 weeks and the systolic blood pressure (SBP) was measured before and after treatment. After treatment, the serum levels of nitric oxide (NO) and super oxide dismutase (SOD) were examined by chemiluminescent method, advanced glycation end-products (AGEs) was detected by ELISA, left ventricular weight index and collagen volume fraction (CVF) in myocardial tissue were calculated, the mRNA expression of myocardial receptor of advanced glycation end products (RAGE) was evaluated by real time RT-PCR. Results: Compared with H group, SBP levels were decreased in T and TP groups,P<0.01, while SBP in P group stayed similar,P>0.05. Compared with H group, the serum levels of NO and SOD were increased in T, P and TP groups,P<0.01; while compared with T group and P group, the levels of NO and SOD were further increased in TP group,P<0.05. Compared with H group, left ventricular weight index and CVF were decreased in T, P and TP groups,P<0.01; while compared with T group and P group, left ventricular weight index and CVF were further decreased in TP group,P<0.05. Pyridoxamine and telmisartan had the coordinative effect on left ventricular weight index and CVF, P<0.05. The serum levels of AGEs were lower in T group (5.99 ± 0.51) mg/L, P group (5.57 ± 0.91) mg/L and TP group (5.24 ± 0.63) mg/L than that in H group, (6.71 ± 0.50) mg/L,P<0.01; while compared with T group and P group, AGEs was further decreased in TP group,P<0.05. The mRNA expressions of RAGE were reduced in T group (0.035 ± 0.010), P group (0.036 ± 0.005) and TP group (0.024 ± 0.007) than that in H group (0.053 ± 0.010),P <0.01; while compared with T group and P group, the mRNA of RAGE was further reduced in TP group, P<0.05. Conclusion: Pyridoxamine could improve the oxidative stress and the indexes of myocardial remodeling independently from decreasing blood pressure in SHR. Combined intervention of P and T may break the coordination systems between AGEs-RAGE and renin-angiotensin, therefore further improve the indexes of myocardial remodeling in SHR.

Objective Sperm quality can be affected by many factors .The aim of this study was to observe the effects of di-etary n-6/n-3 polyunsaturated fatty acids ( PUFAs) on sperm concentration and motility . Methods Twenty healthy B6 male mice were equally randomized to a control and an experimental group to be fed with standard diet and standard diet +10%arachidonic acid (AA), respectively, both for 8 weeks.Then the testes and epididymal tails of the mice were collected for observation of the changes in the microstructure of the epididymal tissue and determination of sperm motility and concentration . Results Compared with the con-trol group, the experimental animals showed significantly decreased serum levels of α-linolenic acid ( ALA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), docosapentaenoic acid (DPA), and linoleic acid (LA) (P <0.01), but dramatically increased the levels of AA and n-6/n-3 PUFAs (P<0.01).The latter also exhibited remarkable reduction in sperm concentration (7 ×106/mL vs 80 ×106/mL, P<0.01), sperm motility (27%vs 86%, P<0.01). Conclusion High levels of dietary n-6/n-3 PUFAs ratios reduce sperm concentration and motility in mice .

Objective To investigate the pattern of antihypertensive medication prescribing in outpatients from the Second Hospital of Tianjin Medical University, and analyze the shortcoming and deficiency compared with 2010 Chinese guidelines for the management of hypertension. Methods A total of 154 262 electronic prescribing for outpatients with hy-pertension, from January-December 2012 in a Grade 3A hospital in Tianjin, were enrolled in this retrospective survey. Data of commonly used antihypertensive medication and combination therapy in patients were analyzed. The patient data collected were divided into different groups according to age, gender, high blood pressure level and the onset of the season. Results (1)The list of the drugs commonly used for treating hypertension in outpatients were calcium antagonist (52.3%), angiotensin receptor blockers (34.0%),βblockers (25.9%), angiotensin-converting enzyme inhibitors (12.1%), fixed-dose combination (11.0%) and diuretics (1.4%).(2)The fewer combination therapy was found in outpatients than that of monotherapy (43.9%vs 56.1%). Some prescriptions were not routinely recommended by the Guideline (4.6%).(3)The combination therapy used in patients with stage 3 hypertension was higher than that of patients with stage 1or stage 2 hypertension (44.5%vs 37.7%vs 37.7%, P<0.01). The rate of combination therapy was significantly higher in cardiology department than that of other clini-cal departments (P<0.01). The combination therapy tended to be used in the elderly patients than that of non-elderly pa-tients (P<0.01). The number of prescriptions was lower in summer than that of other seasons,but the rate of combination therapy was higher in summer than that of spring, autumn and winter (P<0.01). Conclusion The prescriptions of combina-tion therapy and diuretic were inadequate in outpatients with hypertension. These findings indicate the difference between clinical prescription and the guideline for the management of hypertension.

To understand the genetic origin and variation of porcine epidemic diarrhea virus (PEDV) prevailing in Henan province,a total of 22 PEDV positive samples from suckling piglets with severe diarrhea were collected from 16 pig farms and amplified by RT-PCR.ORF3 genes were then cloned into pMD18-T vector and sequenced.The result of sequencing showed that the length of ORF3 genes all were 675 bp which encoded 224 amino acids.The strains in this study had eight amino acid substitutions when compared with CV777 strain.The nucleotide homologies of the 22 strains were 95.9％-100％ and amino acid homologies were 96.4％-100％.By comparing with those of European strain CV777 and vaccine strain truncated CV777,the nucleotide homologies were 97.1％-97.9％ and 94.8％-95.4％,respectively.Based on the phylogenetic relationship of ORF3 genes,the PEDV field strains and PEDV reference strains could be divided into two groups,and all the field strains identified in this study belong to group 2.It suggested that prevalent PEDV strains in this study seem to be closely related to Henan isolates in previous years,domestic strains,Janpanese strains,American strains as well as Korean strains but differ genetically from European strains or vaccine strains used in China.This experiment analyzed the ORF3 gene sequence features in prevailing of Henan region in recent years,which provides a new support of epidemiology study and protein function research of PEDV ORF3 gene.

Objective To study the expression of mitochondrial tumor suppressor gene 1 (MTUS1) mR-NA in papillary thyroid carcinoma(PTC).Methods Reverse transcription polymerase chain reaction(RT-PCR) was used to detect MTUS1 mRNA level in 68 cases of human PTC and the adjacent non-cancerous epithelium (NCE) specimens.Results The relative expression value of MTUS1 mRNA in PTC was 0.31 ± 0.09,while it was 0.65 ±0.12 in NCE.The difference had statistical significance between the 2 groups(t =2.39,P ＜0.05).In PTC,mRNA expression of MTUS1 gene was related to the pathological grade,and lymph node metastasis(P ＜ 0.05)while it was not related to patients' age,sex,differentiation degree,tumor size or TNM stage(P ＞ 0.05).Conclusion mRNA expression of MTUS1 gene is lower in PTC than in NCE,which may play an important role in carcinogenesis and progression of PTC.

Aim To investigate the effect of MAPK signal transduction pathway on the expression of TNF-? in AM(alveolar macrophage) of CB(chronic bronchitis) rats,meanwhile,to initially discuss the possible regulated target of TAL(triterpene acids of loquat.leaf),to explain the mechanism about anti-inflammatory action of TAL on CB rats.Methods The depurated AM was incubated with the special inhibitors of MAPK signal transduction pathway respectively(ERK,p38 or JNK) and the expression of TNF-? was observed by RT-PCR and Western blot.Using factorial design,TNF-? level in AM culture supernatant was detected by ELISA.Results The special inhibitors of ERK(PD98059) and p38(SB203580) could change the expression of TNF-? in AM of CB rats apparently.Factorial analysis shows there is statistical significance between TAL,PD98059/SB203580 and mixture of them.Conclusions TAL reducing the expression of TNF-? may be related with the inhibition of ERK and p38 signal transduction pathway.It may be one of the mechanisms by which TAL has the preventive and therapeutic effects on chronic bronchitis.