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.

Objective To investigate the effects of salvianolate on noradrenaline(NE), angiotension-Ⅱ(Ang-Ⅱ)and tumor necrosis factor-α(TNF-α)in rats with congestive heart failure. Methods Sixty male SD rats were randomly divided into 6 groups, the normal control group(NCG), the model group, the captopril group(CAG), the low dosage of salvianolate group(LSG), the high dosage of salvianolate group(HSG), the captopril and high doseage of salvianolate group(CSG). The rats of Congestive Heart Failure were established with peritoneal injection of adriamycin except the rats in normal control group. The rats in normal control group were injected with the equal volume of normal saline once per week for 6 weeks. The medication was started in every group at the same time. Eight weeks later, the Left Ventricular Systolic Pressure(LVSP), Left Ventricular End-diastolic Pressure(LVDP), +dp/dtmax, -dp/dtmax were measured. The serum levels of NE, Ang-Ⅱ and TNF-αwere measured. Protein expressions of Ang-Ⅱ and TNF-αin cardiac muscle were detected by Western blot. Results Compared with the model group[(90.77±14.75)mmHg, (22.52±2.58)mmHg, (3 290.16± 109.61)mmHg/s, (3 114.07±112.39)mmHg/s], HSG[(114.10±13.71)mmHg, (19.97±1.14)mmHg, (3 504.97 ± 163.94)mmHg/s, (3 303.02 ± 121.98)mmHg/s ] and CSG [ (141.18 ± 15.42)mmHg, (15.58 ± 1.46)mmHg, (3 766.56±159.93)mmHg/s, (3 566.70±154.57)mmHg/s]had significant difference in LVSP, LVDP, +dp/dtmax, -dp/dtmax(P<0.05 or 0.01). Compared with the model group[(85.06±9.07)ng/ml, (180.11±25.45)pg/ml, (205.80±15.73)pg/ml], the serum levels of NE, Ang-Ⅱand TNF-αwere decreased in groups of HSG[(75.33±8.60)ng/ml, (149.21±25.39)pg/ml, (188.84±13.79)pg/ml], CAG[(71.49± 6.21)ng/ml, (139.15 ± 24.83)pg/ml, (183.73 ± 10.99)pg/ml ] and CSG [ (60.02 ± 7.38)ng/m, (110.68 ± 28.63)pg/ml, (165.93±16.22)pg/ml]in different degree(P<0.05 or 0.01), those of CSG were significantly lower than CAG(P<0.05). Compared with the model group[(1.043±0.044), (1.167±0.048)], the expression of protein in Ang-Ⅱand TNF-α were decreased in groups of HSG[(0.981±0.024), (1.069±0.055)]CAG [(0.954±0.031), (1.046±0.053)]and CSG[(0.886±0.044), (0.955±0.038)]in different degree(P<0.05 or 0.01), those of CSG were significantly lower than CAG(P<0.05). Conclusion Salvianolate can reduce the serum levels of NE, Ang-Ⅱand TNF-αin rats with congestive heart failure, decrease the expression of Ang-Ⅱand TNF-αin its cardiac muscle, improve the cardiac function.

Objective To study the effect of the extract of total flavonoids of Chrysanthemum indicum(TFC) on adjuvant arthritis(AA) synoviocytes.Methods Totally 0.1ml of the complete Freund's adjuvant was subcutaneously injected into the right hind feet pads of 20 SD rats.24 days after immunity synoviocytes in the knee joint were treated with TFC.Cell morphology was examined with electron microscopy.Protein level of caspase-3 cleaved fragments was analyzed by Western blotting.The annexin V stain assay was applied to explore the effect of caspase-3 inhibitor on the amelioration in synovial cells apoptosis of AA rats.Results Typical morphology and biochemical feature of apoptosis in synovial cells of AA rats were observed with TFC.The protein level of caspase-3 cleaved fragments increased obviously and was related with the concentration of TFC in synovial cells of AA rats.The apoptotic cells positively stained with annexin V were markedly reduced by caspase-3 inhibitor.Conclusion TFC can induce apoptosis in AA rats synoviocytes,which may achieve therapeutical effects in AA.The activation of caspase-3 may be one of the main causations.

Adult ; Humans ; Lipids ; blood ; Male ; Occupational Exposure ; Vanadium ; toxicity

ObjectiveTo investigate the role of myocardial calcium-sensing receptor (CaSR) in a rat model of high-level spinal cord injury (SCI).MethodsEighteen healthy male SD rats weighing 250-300 g were randomly divided into 2 groups:sham operation group(group S,n ＝6) and SCI group(n ＝ 12).SCI model was induced by dropping a 10 g weight onto spinal cord (C7) in freely vertical falling along the hollow glass tube from 5 cm height.The blood samples were taken 12 and 24 h after SCI in group SCI and 12 h after SCI in group S,and serum activity of creatine kinase(CK) and MB isoenzyme of creatine kinsse(CK-MB) were measured.Then myocardium specimens were obtained for uhrastructure examination and determination of CaSR mRNA and protien expression by fluorescence quantitative RCR and Western blot.Results Serum activities of CK and CK-MB and CaSR mRNA and protein expression were higher in group SCI than in group S.Serum activity of CK and CaSR mRNA expression were higher,and serum activity of CK-MB was lower at 24 h after SCI than that at 12 h after SCI.There was no significantly difference in CaSR protein expression between the two time points in group SCI.The ultrastructure examination showed that myocardial injury was found in group SCI.ConclusionThe expression of CaSR is up-regulated after SCI in rats,which might be the mechanism of myocardial injury after SCI.

Objective:To prepare the production of TIGIT-Fc fusion protein using H22 cells stably integrated the gene by lentivirus vector , and to explore the immunoregulatory effect on macrophages by TIGIT-Fc.Methods: TIGIT-Fc fusion gene were constructed by molecular cloning.The fusion gene was then subcloned to plasmids contained the secretion signaling peptide .The secrected TIGIT-Fc fusion gene was inserted into the lentivirus backbone vector.The purified lentivirus vector was the used to infect the murine H22 cell line.TIGIT-Fc protein was purified by protein A column from the ascites of H 22-injected C57BL/6 mice.Macrophages stimulated by lipopolysaccharide ( LPS ) was challenged to TIGIT-Fc treatment or control.Cytokine levels was then detected by ELISA.Results: TIGIT-Fc protein was purified from the ascites of H 22-injected mice.PVR was upregulated in LPS-treated macrophages.IL-10 level was upregulated in TIGIT-Fc treated macrophages.Conclusion: TIGIT-Fc promotes the mature macrophages to secrete anti-inflammatory cytokine IL-10.

Objective To investigate the effect of bifidobacteria on dextran sulphate sodium(DSS)- induced acute ulcerative colitis in mice.Methods Thirty BALB/C mice were randomly divided into nor- mal control group (n=10),0501 strain group (n=10) and c122 strain group (n=10).Fifty BALB/ C mice received 5% dextran sulphate sodium(DSS) for 7 days to induce ulcerative colitis.The mice were then divided to model group,negative control group(perfused with 0.9 NaCl solution ),positive control group(perfused with SASP of 20 mg/ml),DSS + 0501 strain group(perfused with 1?10~9 CFU/ml bifidobacteria 0501 strain solution and DSS + c122 strain group (perfused with 1?10~9 CFU/ml bifidobacteria c122 strain solution).All mice were sacrificed 9 days later.The colon specimens were measure by histoehemical staining with H-E.The expressions of interleukin-10 (IL-10) and its protein were detected by RT-PCR and immunohistochemistry respectively.Results The degree of colon inflam- mation in mice both in DSS+ 0501 strain and DSS+ c122 strain groups were aggravated and expressions of IL-10 mRNA and protein were reduced compared to model group.No colon inflammation was found in 0501 strain and c122 strain groups.Conclusion Some strain of bifidobaeteria may aggravate colon in- flammation in mice when mucosal harrier is destroyed.

A strain of Cellulosimicrobium cellulans Ha8 was studied on its morphological, biological characteristics and its utilization of several kinds of benzoic compounds, the results showed this strain was Gram-positive, the long rod-shaped cells were changed into short rod-shape gradually. pH value from pH 6.0 to pH 9.0 and the temperature from 20 ℃ to 40 ℃ were good for its growth. It could not only hydrolyze protein and starch, use cellulose and pectin, decomposite chitin, liquify gelatin and fix nitrogen, but also use phenol, xylene, benzoic, cinnamic acids and diphenlamine as the sole carbon resource for its growth. It could tolerate 0 mmol/L~30 mmol/L, 0 mmol/L~8 mmol/L, 0 mmol/L~30 mmol/L, 0 mmol/L~15 mmol/L and 0 mmol/L ~ 40 mmol/L of benzoic acids, phenol, xylene, cinnamic acids and diphenlamine seperately, but could not use 2,4-dinitrophenol, o-Nitrophenol, 2-Methoxyphenol, aminobenzenesulfonic acid, catechol and o-Phenanthroline as its sole carbon resource.

Objective To establish acute peritonitis induced by monosodium urate (MSU) of in mice and observe the significance of mitochondrial deoxyribonucleic acid (mtDNA) expression in the inflammatory processes.Methods The mouse models of acute peritonitis were made by intraperitoneal injection of MSU.Sixty-four male C57BL16 mice were randomly divided into the MSU group which were treated with 0.2 ml of 15 mg/ml MSU solution by i.p.injection and the control group which were treated with 0.2 ml of PBS.Respectively four mice from MSU group and four mice from control group were killed 2 hours, 4 hours, 6 hours, 8 hours 12 hour, 16 hours, 20 hours and 24 hours later and whole blood, peritoneal lavage and peritoneum were collected respectively.Four the mice from the MSU group and four mice from the control group were killed and whole blood, peritoneal lavage and peritoneum were collected.Immunoflourescence study of peritoneum tissues was performed.The levels of interleukin (IL)-1β, IL-18 in plasma and peritoneal lavage were examined by enzyme linked immunosorbent assay (ELISA).DNA was extracted from blood and peritoneal lavage, and mtDNA level was detected by using real-time polymerase chain reaction (PCR).The data was analyszied by multivariate analysis of variance.Results As compared with those killed at other time points from the MSU groups and the control group, the levels of IL-1β [(27.0±2.0) pg/ml vs (26.8±2.1) pg/ml], IL-18 [(673±454) pg/ml vs(752±495) pg/ml] in plasma and peritoneal lavage were increased progressively in those which were killed after i.p.injection of 2 hours and 4 hours from in the MSU group (F=22.778, P＜0.05;F=6.660, P＜0.05).The mtDNA in plasma and peritoneal lavage of the mice began to be expressed 4 hours after i.p.injection 4 hours from in the MSU group.The peak level was detected in those i.p.injected MSU 6 hours later [(9.85±4.59)×106 copies, (7.81±3.43)×106 copies].Then 8 hours later the mtDNA began to slowly decreased.At these three time points, the mtDNA were all increased progressively than those at the other time points of the MSN group or at all time points of the control group (F=6.719, P＜0.05;F=11.181, P＜0.05).By immunoflourescence study, there were neutrophil extracellular traps (NETs) were formed 12 hours later in the MSU group and aggregated NETs were found 24 hours later.Conclusion In the inflammatory processes of acute peritonitis induced by MSU of in mice, with the expression of mtDNA increasing, the inflammation is relieved, and aggregated NETs are formed in the end.Expression of mtDNA may be one for the protective factors of the inflammation induced by MSU.