Objective To study the expression and significance of the p38 mitogen-activated protein kinase (MAPK) and cAMP-responsive element binding protein (CREB 1) in experimental diabetic mellitus rats, and their relationship with extracllular matrix (ECM) restructuration. Methods Sixty male Wistar rats, with their kidneys removed surgically, were divided in two groups: control group (n=30) and diabetic group (n=30). Diabetic model was reproduced by intraperitoneal injection of Streptozocin (STZ, 65mg/kg). Six rats of each group were sacrificed at 1, 2, 4, 8 and 12weeks after STZ injection. Immunohistochemistry was used to assess the expression of p-p38 MAPK and p-CREB 1 in both control and diabetic groups. Western blot was performed to determine the activity of p-p38 MAPK and p-CREB 1 in both groups. Semiquantitative competitive reverse transcription polymerase chain reaction (PCR) was performed to detect the expression of mRNA of p38 MAPK, CREB 1 and fibronectin (FN). Results Compared with the control group, expression of p-p38 MAPK in diabetic glomeruli was significantly enhanced at 1, 2, 4, and 8 weeks, then it lowered at 12 weeks. P-CREB 1 was increased at 2, 4, 8 weeks, also decreased at 12 weeks. Elevation of FN mRNA began after 4 weeks in diabetic rats. Conclusions Glomerular p38 MAPK activity was increased in early stage of diabetes. This activated p38 MAPK pathway in diabetic glomeruli may in part play a role in the pathogenesis of extracellular matrix restructuring.

AIM: To investigate the effect of activation of JAK/STAT signaling pathway on the transdifferentiation induced by high concentration of glucose in renal proximal tubular epithelial cells(HKCs).METHODS: Cultured HKCs cells were divided into four groups: low glucose group(LG),high glucose group(HG),high mannitol group(LG+M),and HG+AG490 group(AG).Immunoprecipitation and Western blotting analysis were used to determine the expression of tryosine phosphorylated Janus kinase 2(p-JAK2).The protein expressions of STAT1,STAT3,p-STAT1 and p-STAT3 and expressions of ?-SMA,E-cadherin were observed by Western blotting.The contents of TGF-?1 and type I collagen in the supernatants of the cultured HKCs were detected by enzyme-linked immunoadsorbent assay(ELISA).TGF-?1 mRNA were measured by reverse transcription and polymerase chain reaction(RT-PCR).RESULTS: Compared with low glucose control group,the expression of JAK2,p-STAT1,p-STAT3 and TGF-?1 mRNA was significantly increased in HG group at different stimulated time from 6 h to 72 h.Meanwhile,the contents of TGF-?1 and collagen I in the supernatants and the expression of ?-SMA were increased,the expression of E-cadherin were decreased.The expression of JAK2,p-STAT1,p-STAT3 and TGF-?1 mRNA and the levels of TGF-?1,collagen I in the supernatants in HG+AG490 group were obviously lower than those in HG group.The expressions of ?-SMA and E-cadherin were also decreased in HG+AG490 group.CONCLUSION: Activation of JAK/STAT signaling pathway may be involved in the high glucose-induced transdifferentiation and overproduction of TGF?1 and ECM proteins in HKCs.

Objective To investigate the elationship between the expression of Cathepsin D and Laminin receptors with the invasion and metastasis of gastric cancers.Methods Cathepsin D and Laminin receptors immunoreactivities were evaluated in 96 patients with gastric cancer using streptavaclin pemxidase method.Results Over expression of Cathepsin D in gastric cancer tissue was significantly related with the depth of invasion,lymphnode metastasis and grade of differentiation (P<0.01,P<0.01 and P<0.05, respectively).Moreover,the high expression of Lamninin receptors in gastric cancer had significant correlations with the invasion depth and grade of differentiation(P<0.05 and P<0.01);The positive expression of Laminin receptors were not significantly related with lymph node metastasis(P>0.05).By statistics analysis Cathepsin D had a significant positive correlation with Laminin receptors (P<0.05). Conclusion Cathepsin D and Laminin Receptors play all important role in the invasion and metastasis of gastric Cancers.

Objective To investigate the effect of high glucose on the expression of transforming growth factor-?1(TGF-?1),?-SMA,E-Cadherin in renal proximal tubular epithelial cells(HKCs)and role of AG490,an antagonist of Janus kinase.Methods Cultured HKCs cells were divided into four groups:low glucose group(LG),high glucose group(HG),high mannitol group(LG+M),and HG+AG490 group(AG).Immunoprecipitation and Western blot analysis were used to measure the expressions of tryosine phosphorylated Janus kinase 2(p-JAK2),STAT1,STAT3,p-STAT1,p-STAT3 and ?-SMA,E-Cadherin.The contents of TGF-?1 and type I collagen in the supernatants of the cultured HKCs were detected by ELISA.TGF-?1 mRNA were measured by RT-PCR.ResultsCompared with low glucose control group,the expressions of JAK2,p-STAT1,p-STAT3 and TGF-?1 mRNA were significantly increased in HG group from 6 to 72 h.Meanwhile,the contents of TGF-?1 and collagen I in the supernatants and the expression of ?-SMA were increased,the expression of E-Cadherin was decreased.The expression of JAK2,p-STAT1,p-STAT3 and TGF-?1 mRNA and the level of TGF-?1,collagen I in the supernatant s in HG+AG490 group were obviously lower than those of the HG group.The expression of ?-SMA and E-Cadherinwas also decreased in HG+AG490 group.Conclusion Activation of JAK/STAT signaling pathway seems to be involved in the high glucos induced overproduction of TGF-?1 and ECM proteins in HKCs.

Objective To explore and evaluate the predictive value of the coagulopathy in patients with community-acquired pneumonia (CAP).Methods A retrospective study was carried out for investigating the prothrombin time (PT),activated partial thromboplastin time (APTT),plasma fibrinogen (FIB),thrombin time (TT),plateslets (PLT),D-dimer in 385 patients with CAP and 146 patients without infection,tumor,trauma,thrombosis as controls.All the patients were admitted to the Respiratory Medical Department from June,2010 to May,2011.The results of the aforementioned biomarkers were analyzed and compared between two groups.The Pneumonia Severity Index (PSI) was calculated to evaluate the correlation between coagulopathy and PSI.Results The comparisons of the abnormal rates of PLT,PT,APTT,Fib,D-dimer between the patients with CAP and the controls were 92/385 vs.9/146,39/385 vs.1/146,108/385 vs.7/ 146,331/385 vs.47/146,348/385 vs.5/146,respectively.The differences were statistically significant (x2 =21.608,13.557,33.747,149.280,365.619,respectively,P ＜ 0.01),while difference in TT was not statistically significant (8/385 and 0/146,x2 =1.839,P ＞ 0.05).The differences in abnormal rate of PLT,PT,D-dimer between high-risk group of CAP and the low-risk group of CAP were 45/148 vs.47/237,26/148 vs.13/237,146/148vs.202/237,respectively,and the differences were statistically significant (x2 =5.602,14.609,23.442,respectively,P ＜0.05),while there were no differences in TT,APTT,FIB between two groups (6/148 vs.2/237,47/148 vs.61/237,123/148 vs.208/237,x2 =4.614,1.635,1.638,respectively,P ＞0.05).D-dimer in patients with CAP was (3.8 ±6.1) mg/L,compared with the controls (0.3 ±0.1) mg/L,and D-dimer in high-risk patients with CAP was (7.5 ±8.3) mg/L compared with the low-risk group (1.6 ±2.0) mg/L (P ＜ 0.001).Rank correlation existed between D-dimer and PSI (r =0.798,P ＜ 0.01),while there was no correlation between PLT and PSI (x2 =6.040,P ＞0.05).Conclusions The coagulopathy commonly occurs in patients with CAP.D-dimer was significantly higher in patients with CAP.D-dimer level is positively correlated with severity of CAP.D-dimer can be an ideal biomarker to assess the severity of patients with CAP.

Objective To investigate the possible effects on radiosensitivity in human umbilical vein endothelial cells after transfection of pcDNA3.1+Ape1 plasmid. Methods The expressing vector pcDNA3.1+ Ape1, the control vector pcDNA3.1+or non-transfection cells was irradiated by 2, 4, 6, and 8 Gy photon beam at 48 h post-transfection. The value of initial and residual Oliver tail moment (OTM) under the alkaline single cell gelelectrophoresis assay and the colony forming test were utilized as the markers for the evaluaton of cells intrinsic radiosensitivity. The effect on radiosensitivity in human umbilical vein endothelial cells after transfection of the expressing vector pcDNA3.1+Ape1 was analyzed according to the radio-dose, compared to the empty vecor control and non-transfection cells. Results The initial and residual OTM value of endothelial cells transfected by 3 μg pcDNA3.1+Ape1 plasmid was lower significantly than ones of endothelial cells untransfected at 2 Gy irradiation (P<0.01), but was no significant difference at 8 Gy (P>0.05), and SF2 was higher remarkably in transfected cells than one in untransfected cells (P<0.05), but SF4, SF6 and SF8 were no significant differences (all of P>0.05). Conclusions The transfection of pcDNA3.1+Ape1 plasmid could enhance radioresistance of endothelial cells to the low-dose irradiation.

Aim To observe the effect of valsartan on expression of Smads of HKCs stimulated by TGF-?1.Methods HKCs were divided into three groups:control group,TGF-?1 group and valsartan group.Total protein was firstly abstracted at 30 min after stimulation to detect p-Smad2/3 protein by Western blot.Total RNA and protein were abstracted at 48 hours after stimulation.The expressions of Smad2/3,Smad7 were measured by Western blot.Smad2mRNA and Smad7mRNA were measured by RT-PCR.Meanwhile, The protein synthesis of Col Ⅰ in the supernatants of the HKCs was detected by Western blot.MTT assay was used to investigate the effect of valsartan on proliferation of HKCs stimulated by 5 ?g?L-1 TGF-?1 at different time and different drug concentration.Results Western blot analysis indicated that TGF-?1 stimulation could increase the expression of Smad2/3 and activate phosphorylation of Smad2/3 at 48 hour.At the same time,Smad7 decreased.Valsartan could reduce the level of Smad2/3 and p-Smad2/3 but it did not have any effect on the expression of Smad7 protein.RT-PCR analysis also showed that the expression of Smad2mRNA or Smad7mRNA was higher after TGF-?1 stimulation than control group,while valsartan could down-regulate the expression of Smad2mRNA.There was no difference of the expression of Smad7mRNA between the TGF-?1 stimulation group and valsartan treatment group.Valsartan could also greatly ameliorate the secretion of Col Ⅰ in the supernatants of the HKCs stimulated by TGF-?1 detected by Western blot.Compared with control group,TGF-?1 supressed the proliferation of HKCs,While 1,10 and 100 ?mol?L-1 valsartan ameliorated it and the effect was dependent of dose.Conclusion Valsartan has some renal protective effects on renal interstitial fibrosis,partly through inhibiting TGF-?/Smads signaling pathway.

Aim To investigate the effects of Ang Ⅱ receptor antagonist irbesartan on the expressions of connective tissue growth factor(CTGF) and membrane-type 1 matrix metalloproteinase(MT1-MMP) in high glucose-cultured rat glomerular mesangial cells(GMCs).Methods High concentration glucose and irbesartan were used to stimulate the cultured rat GMCs in vitro.The mRNA and protein expressions of CTGF and MT1-MMP were detected with semi-quantitative RT-PCR and Western blot.The secreted collagen Ⅳ in the supernatants of the GMCs was detected by enzyme-linked immunoadsorbent assay(ELISA).Results Compared with control group,the expressions of CTGF were continuously increased in GMCs under high concentration glucose medium;otherwise the mRNA and protein levels of MT1-MMP in GMCs were decreased in a time-dependent manner at the same time.These changes were accompanied by increased secretion of collagen Ⅳ.Irbesartan could inhibit those changes induced by high glucose.Conclusions High glucosecould induce the expression of CTGF and inhibit the expression of MT1-MMP in GMCs.Irbesartan could inhibit the secretion of ECM in GMCs under high concentration glucose medium,partly by regulating the expressions of CTGF and MT1-MMP.

Aim To investigate the effects of valsartan on the expression of TGF-?1 mRNA and the activation of p38 mitogen-activated protein kinase(p38MAPK),mitogen activated protein kinase kinasse3/6(MKK3/6)and cAMP response element-binding protein1(CREB1) in glomerular mesangial cells under high concentration of glucose.Methods High concentration glucose and valsartan were used to stimulate the cultured rat GMCs in vitro.The protein expressions of p38 MAPK,CREB1,p-p38 MAPK,p-MKK3/6 and p-CREB1 were observed by Western blot analysis.TGF-?1 and fibronectin(FN) mRNA were measured by reverse transcription and polymerase chain reaction(RT-PCR).The protein synthesis of FN and type Ⅳ collagen in the supernatants of the GMCs was detected by radioimmunoassay.Results Compared with low glucose control group,the expressions of p-p38 MAPK,p-MKK3/6,p-CREB1 protein,TGF-?1 and FN mRNA,FN and type Ⅳ collagen in the supernatants were significantly increased in GMCs under high concentration glucose medium.The expression levels of p-p38 MAPK,p-MKK3/6 and p-CREB1 were significantly lower in the valsartan group than those in the high concentration glucose group.So did the mRNA of TGF-?1 and FN.The concentration of FN and type Ⅳ collagen in the supernatants in the valsartan group was lower than that in the high concentration glucose control group.Conclusion Valsartan can inhibit over production of TGF-?1 and ECM proteins in GMCs under high concentration of glucose,partly by regulating the phosphorylation of p38 MAPK,MKK3/6 and CREB1.

Objective To investigate the effects of rapamycin on p-JAK2,STAT3,p-STAT3 and PCNA in ranal tissue of IgAN rats.Methods The animal models of IgA nephropathy were devided into control group,IgAN model group,losartan group and rapamycin group.The clinical efficacy of rapamycin,and its influences on the protein and mRNA expressions of STAT3,p-STAT3,p-JAK2 and PCNA were determined by western blot,RT-PCR and immunohistochemical techniques respectively.Results The protein and mRNA expressions of p-STAT3,STAT3,p-JAK2 and PCNA were significantly increased in kidney of IgAN rats(P