Objective To explore the effects and mechanisms of urokinase-type plasminogen activator (uPA) on high glucose-induced rat mesangial cells proliferation and phenotype transformation. Methods Rat mesangial cells were cultured and incubated in media containing either 5 mmol/L D-glucose or 30 mmol/L D-glucose with or without addition of wortmannin, or uPA (105 U/L) for different time periods. At the end of the incubation period, mesangial cells proliferation was assessed by MTT assay and flow cytometric analysis. Cyclin-dependent kinase 2 (CDK2) and p27kip1 expression and activation of Akt were evaluated by Western blotting and Akt kinase assay respectively. Furthermore, the expression and distribution of α-SMA were detected with laser confocal microscopy. Results MTT assay and flow cytometric analysis demonstrated that high glucose induced mesangial cells proliferation (P＜0.05) and an incresed proportion of cells in G2/M+S stage after 24 h incubation (P＜0.01), which were attenuated by uPA or wortmannin (P＜0.01). High glucose induced the enhance of Akt activity after 3 h (P＜0.05), and the effect was inhibited by wortmannin or uPA (P＜0.01). High glucose did not alter CDK2 expression (P＞0.05),but significantly inhibited p27kip1 expression (P＜0.05), which was attenuated by wortmannin or uPA (P＜0.01). High glucose induced the up-regulation of α-SMA expression and perinucleus location in mesangial cells after 24 h (P＜0.01), which were alleviated by wortmannin or uPA (P＜0.01). Conclusion uPA up-regulates p27kip1 expression and counteracts high glucose-induced mesangial cells proliferation and phenotype transformation via blocking PI3K-Akt signaling pathway.

Detergents ; Immunohistochemistry ; instrumentation ; Oxalic Acid ; Potassium Permanganate ; Staining and Labeling ; instrumentation

Objective To investigate dexamethasone on airway inflammation and CD4+ CD25 + regulatory T cells (CD4+ CD25 +Tr) of asthmatic rats,and elucidate the possible mechanism of dexamethasone in treatment of asthma.Methods 30 Wistar rats were randomly divided into control group,asthma group and dexamethasone-treated group.Bronchoalveolar lavage fluid (BALF) was collected,and cytology study was conducted.The lung tissue was obtained and pathologic analysis was done through HE stain.Flow eytometry was used to detect the CD4+ CD25 +Tr ratio in PBMCs.Results Total cells number,the percentage of lymphocytes,neutrophils and eosinophils (Eos)in BALF of dexamethasone-treated group were lower than that of asthma group (P＜0.05,P＜0.01).Compared with the asthma group,less infiltration of inflammatory cells in lung tissues was observed in the dexamethasone-treated group.CD4+ CD25 + Tr of asthma group was lower than that of control and dexamethasone-treated group (P＜0.05).Conclusion Dexamethasone could suppress airway inflammation of asthmatic rats,which probably be due to increasing the number of CD4+ CD25 + Tr.

Objective To investigate the immunological mechanism of inhibitory effect of Danshen injection combined with dexamethasone(DXM) on asthmatic airway inflammation.Methods 50 Wistar rats were randomly divided into normal control(NC),asthma,Danshen,DXM and Danshen+DXM group.Cytology study of Bronchoalveolar lavage fluid(BALF) was conducted.Pathology of lung tissue was done through HE.Flow eytometry was used to detect CD4+CD25+ regulatory T Cells(CD4+CD25+ Treg) ratio in peripheral blood mononuclear cells(PBMCs).IL-4 and IL-5 levels in BALF were detected by ELISA.Results Total cells number,percentage of lymphocytes,neutrophils and eosinophils(Eos) in BALF of the three treated groups were lower than that in asthma group(P<0.05,P<0.01),particularly in Danshen+DXM group,which showed significant difference as compared with the other two treated groups(P<0.05).There was severe inflammation in lung tissue of asthma group,moderate inflammation in Danshen group and DXM group,and no inflammation of Danshen+DXM group.CD4+CD25+ Treg/CD4+ T ratio in the three treated groups were higher than that in asthma group,and the levels of IL-4 and IL-5 were lower than those in asthma group(P<0.05).In Dansben+DXM group,it showed significant difference on the change of CD4+CD25+ Treg,IL-4 and IL-5 as compared with other treated groups(P<0.05).Conclision Danshen injection combined with DXM could suppress airway inflammation in asthmatic rats,which may be through increasing the expression of CD4+CD25+ Treg,decreasing the levels of IL-4 and IL-5 and resuming the balance of Th1/Th2.

Objective To evaluate the effects of rehabilitation training on muscle strength and exercise toleranee in hemodialysis patients with muscle atrophy.Methods Nine hemodialysis patients with muscle atrophy because of end renal failure were recruited in this study. A structured exercise program(90 minutes a sedssion.3 sessions a week)was administered to all the subjects for 6 month.Immediately before and at the end of the exercise programme,the muscle strength of the lower limbs,the motor conduction velocity of the peroneal nerve and maximal oxygen consumption of the patients were examined. Results It was shown that all the patients had impaired exercise capacity,weakend muscle strength and slowed nerve conduction velocity before rehabilitation training.After the exercise programme,the patients' exercise capacity as reflected by the maximal oxygen consumption and exercise time was significantly increased.The muscle strength and the motor nerve conduction velocity were significantly increased.Conclusions Muscle atrophy in hemodialysis patients results in poor exercise tolerance, but rehabilitation exercise programme improves amyotrophy and therefore has beneficial effects on the patient's overall work performance.

Objective To observe the possible mechanism and inhibitory effects of curcumin on pulmonary fibrosis induced bleomycin in rats at the fibrosing stage. Methods 80 male Sprague-Dawley rats were random divided into 4 groups (20 rats in each group). Rats in the fibrosis model group, the prednisone group and the curcumin group were induced by instilled bleomycin through tracheal, rats in the control group with same volume normal saline. Since the 15th day after bleomycin administration, the curcumin group and prednisone group were given curcumin (300 mg/kg) or prednisone (5mg/kg) per day by intragastric administration, respectively. The normal control group and the model group were given 1% sodium carboxymethyl cellulose ( 10ml/kg). Six rats of each group were random sacrificed on the 21st, 28th, 42nd and 56th days after bleomycin administration. The histological changes of the pulmonary were evaluated by H. E and Masson dyeing. The expressions of transforming growth factor-β1 (TGF-β1), platelet-derived growth factor (PDGF) and hydroxyproline in the tissue of pulmonary were assessed by immunohistochemistry and digestion method. Results Pulmonary fibrosis and hydroxyproline level in the curcumin group were obviously reduced as compared with the model group on the 42nd and 56th day[42 d:1. 28 ±0. 61 vs 2. 28 ±0. 39,P <0. 01 ;(1.73 ±0. 22)mg/g vs (2.50 ±0. 37) mg/g, P <0.01;56 d:1.00 ±0.59 vs 1.73 ±0.36, P< 0. 05; ( 1.57 ± 0. 36) mg/g vs (2. 20 ± 0. 42) mg/g, P < 0. 01 ], and it was also lower than that in prednisone group on the 42nd day( P < 0. 05 ). The expression of TGF-β1 and PDGF in the curcumin group were obviously lower than that in the model group on the 28th, 42nd and 56th day[28 d:TGF-β1 :3642. 05 ±839. 31 vs 5067. 35 ±738. 39, P <0. 05 ;PDGF:2957. 55 ±739. 16 vs 4457. 75 ±568. 39, P <0. 05;42 d: TGF-β1: 2689. 73 ± 529.22 vs 4089. 50 ± 619. 37, P < 0. 01; PDGF: 2834. 46 ± 567. 16 vs 3239. 52 ±628. 26, P <0. 01 ;56 d:TGF-β1: 1968.57 ±408. 36 vs 2968.20 ±498.42, P <0. 01 ;PDGF: 1083.36 ±381.35 vs 2019. 40 ±412. 36, P <0. 01 ], which was lower than that in prednisone group on the 42nd and 56th day (42 d,TGF-β1 :3529. 07 ±981.35,PDGF:2618. 34 ±813. 34;56 d,TGF-β1 :2530. 83 ±439. 37,PDGF: 1738. 35 ±536. 62, Pall <0. 05 ) , and it had no obvious difference compared with control group on the 56th day ( P > 0. 05 ). Conclusion Curcumin could alleviate bleomycin-induced pulmonary fibrosis in rats at the fibrosing stage by inhibiting the expressions of TGF-β1 and PDGF.

Objective To analyze levofloxacin resistance in Helicobacter pylori (Hp) and the sequence difference of gyrA gene in levofloxacin resistance and sensitive Hp strains. To explore the function of gyrA gene mutation in the development of levofloxacin resistance Hp strain.Methods From July 2007 to December 2008 in Department of Gastroenterology, Jinhua People hospital of Zhejiang Province, the gastric mucosa from gastroscopy biopsy of chronic gastritis and peptic ulcer patients were cultured in Hp selective medium under microaerobic condition at 37 degrees for three to five days. The Hp strains were isolated and identified by oxidase test, catalase test,urease test and UreA gene detecting. The levofloxacin susceptibility was determined by E-test and then resistance and sensitive strains were screened. The genomic DNA of Hp strains was isolated. The gyrA gene was amplified by PCR and the sequences were analyzed. Results 38 clinical isolated Hp strains were passed the levofloxacin susceptibility E-test, among those the minimum inhibitory concentration (MIC) of 12 strains was over 1.0 μg/ml, the percentage of resistant strain was 31.58%, while the sensitive stains was 68.42%. The gyrA gene sequence result indicated 10 resistant strain with 261, 271 and 272 10 site mutation, 2 strains with C261A mutation, one strain with C261G mutation, two strains with G271A mutation, 2 strains with A272G mutation, 2 strains with C261A,G271T andA272G mutation, 1 strain with C261G and A272G mutation. However, no mutation sites were found in 26 sensitive strains. Conclusion The rate of levofloxacir-resistance in isolated clinical Hp strain was high. The drug-resistance was associated with 261,271 and 272. site mutation of gyrA gene.

Objective To observe the anticoagulant effect of low molecular weight heparin on induced hemodialysis in patients with acute renal failure.Method One hundred and eight patients with acute renal failure treated with induced hemodialysis were randomly divided into low molecular weight heparin(LMWH)group and unfractionated heparin(UFH)group.A bolus disc of UFH was given at first and then maintained by continuous infusion in UFH group,whereas a single bolus dose of LMWH with 2000AFXa IU to 4000AFXa IU in LMWH group.Results Anticoagulant effect between LMWH and UFH did not show significant discrepancy during induced hemodialysis.The bleeding from internal jugular vein catheter increased in the UFH group much more than that in the UFH group was significantly higher than that in the LMWH group.Anti-FXa blood levels were significantly higher in LMWH group than in UFH group.Conclusions LMWH has minor influence on aPTT and TT,while its anticoagulation effect approximates to that of UFH.LMWH represents a realistic alternative agent UFH in acute renal failure induced hemodialysis.

Objective: To evaluate the role of the MAPK subtypes (p38MAPK, ERK and JNK) in ANG Ⅱ induced apoptosis of cultured human podocytes. Methods: The cultured podocytes were incubated in media containing either vehicle, SB202190(5 ?mol/L, an inhibitor of p38MAPK), PD98059 (1 ?mol/L, an inhibitor of ERK), SP600125 (5 ?mol/L, an inhibitor of JNK), ANG Ⅱ (10 -8 mol/L) with or without SB202190、PD98059 and SP600125 for 18 hours; the cells were assayed for apoptosis by morphologic staining with H 33342 and propidium iodide and DNA fragmentation assays; the cell proteins were probed for phosphorylated MAPKs to determine the activation of specific MAPK subtypes. Results: ANG Ⅱ promoted podocyte apoptosis in a time and dose dependent manner; ANG Ⅱ stimulated p38MAPK, but inhibited JNK; SB202190 inhibited both ANG Ⅱ induced podocyte apoptosis and p38MAPK phosphorylation; Inhibition of ERK by PD98059 had no effect on ANG Ⅱ induced cell apoptosis. Conclusion: ANG Ⅱ induced apoptosis through stimulation of p38MAPK and inhibition of JNK in human podocytes.

Objective To understand the iodine deficiency disorders in Xunhua county Qinghai province, and to provide reference values for setting up prevention and control strategy. Methods According to historical data of Xunhua county, 1 township center elementary school and 1 village elementary school were sampled in Qingshui, Baizhuang, Hongqi, Jiezi and Galeng townships. A total of 9 elementary schools were investigated (Caleng has only 1 elementary school). Fourty children aged 8-10 years were checked thyroid size by palpation in every school. Twenty children were chosen to assay their urinary iodine and edible salt iodine in every school. Thirty 5-grade students were surveyed by questionnaire on health education in every school. Ten housewives were inquired prevention knowledge on iodine deficiency disorders and salt iodine level was tested in every village. Results Goiter rate was 7.9%(30/378) in children aged 8-10 years, with Baizhuang and Galeng the highest (11.9%,5/42), and Qingshui the lowest (2.4%, 1/42). The median was 196.5 μg/L in children aged 8-10 years, and urinary iodine level lower than 50 μg/L was accounted for 5.3% (11/208), but that value was 25.0% (5/20)in Galeng. The average qualified rate of using edible salt was 91.4%(159/174), and that rates were 79.0%(15/19), 83.3%(15/18) and 89.5%(17/19) in Chajia, Tuanjie and Minzhu villages, respectively. The average score were 2.8, 2.1 in health education survey of the 5-grade students and housewives. Conclusions After years effort in prevention and control of iodine deficiency disorders, remarkable progress has been made in Xunhua County. The monitoring of iodized salt and the knowledge spreading are still need to be strengthened.