Objective To explore the role of type 1 plasminogen activator inhibitor (PAI-1 ) in mediating renal tubulointerstitial injury of patients with IgA nephropathy. Methods The mRNA and protein production of PAI-l in renal tubulointerstitium were defected using in stiu hybridization and immunohistochemistry. Detections of antigens of ?-smooth muscle actin(?-SMA) and proliferating cell nucleus antigen (PCNA) were also performed. Results PAI-l was normally expressed in the walls of vessels and distal tubules, but significantly increased in lesions of IgA nephropathy, including crescents, Bowman capsules and tubulointerstitial infiltrating cells. There was lithe expression of PAl-1 in the glomerular capillary tufts. The renal expression of PAI-l was significantly correlated with serum creatinine (P

Objective To observe the phenotypic changes between renal tubular epithelial cells and mesenchymal cells in rat tubulointerstitial fibrosis(TIF) .Methods The renal tubulointerstitial fibrosis models of Wistar rats were established by gavage with excessive adenine. The morphological changes of model kidney were observed by light microscopy, electron microscopy, polarizing microscopy. The?-smooth muscle actin(a-SMA), vimentin, and keratin were examined by immunohistochemistry staining. While the proliferating cell nuclear antigen (PCNA) was also assessed. Results There were many 2, 8-dioxyadenine crystals derived from the metabolite of adenine to deposit in the renal tubules, which induced the injury. Tubular epithelial cells degeneration and regeneration, loss of renal tubules, mononuclear cells infiltration in renal interstitium and the accumulation of extracellular matrix protein were observed. Both ?-SMA and vimentin were positive in some of renal tubules. And consecutive sections of immunohistochemical staining showed that the area of distribution of a-SMA positive cells was almost the same area as that of vimentin positive cells. Polarizing microscopy results showed that there was a mass of interstitial collagen ( Ⅲ and Ⅰ) accumulation. And electron microscopy examination proved that epithelial cells invaded into the renal interstitium. Conclusion There is tubular epithelial cells-myofibroblasts transdifferentiation in the process of TIF, and which plays a key role in the accumulation of extracellular matrix.

Objective To examine the expression of proliferating cell nuclear antigen (PCNA) of retinal pigment epithelial (RPE) cells, thus assessing the role of mechanism of contact inhibition playing in the process of experimental retinal detachment and reattachemnt. Methods Retinal detachment was produced in 72 cats by subretinal injection of 0.25% solution of healon through a micropipette three weeks after extracapsular lens extraction and vitrectomy. Some of the detached retinae were reattached 24 hours later. At different time, the cats were killed and eye globes were fixed and embeded in paraffin. Histologic sections were processed for immunohistochemistry examination using an antibody to detect PCNA protein. Labeled RPE cells were identified, and the proliferation was quantified in detached and un detached retinae of detachment group, and also in reattached retinae of reattachment group. The comparsion of PCNA labeled RPE cells in different groups were analyzed by ANOVA. Results In detached regions of detachment group, PCNA expression of RPE cells occured within 24 hours, and reached a maximum after 5 6 days, then gradually declined to barely detectable levels after 20 days. Similar tendency was found in reattached retinae, but the number of PCNA labeled RPE cells was obviously small. Fewer PCNA labeled RPE cells were found in regions of un detached retinae in detachment group. The difference of these three groups was significant. Conclusion Proliferation of RPE cells is induced when they lose contact with neural retina, but inhibited after neural retina reattached to RPE cells. It suggests that the mechanism of contact inhibition plays a role in the proliferative process after retinal detachment and reattachment.

Objective To study the dynamic change in plasma N-terminal pro-B-type natriuretic peptide (NT-proBNP) levels and its correlation with extravascular lung water index (EVLWI) in patients with septic shock. Methods Sixty-two patients with septic shock admitted to Department of Critical Care Medicine of Beijing Shijitan Hospital were enrolled. The patients were divided into survival group(39 cases)and non-survivors group (23 cases)according to 28-day prognosis. Venous blood was collected after intensive care unit(ICU)admission. The changes in plasma NT-proBNP and hemodynamics indexes levels were analyzed to evaluate their predictive value for clinical outcomes. Results Acute physiology and chronic health evaluation Ⅱ(APACHEⅡ)score(23.2±2.5 vs. 28.1±2.6),sequential organ failure assessment(SOFA score:7.74±2.80 vs. 12.43±3.00)and hemodynamics indexes including EVLWI〔ml/kg:7.0(6.0,8.0)vs. 9.0(7.0,12.0)〕,blood lactate(mmol/L):3.60±2.30 vs. 10.40±2.70)and NT-proBNP〔ng/L:945.0(228.0,1 246.0)vs. 5 471.0(3 308.0,11 174.0)〕in survivors were significantly lower than those in non-survivors,and cardiac index〔CI(L?min-1?m-2):4.23±0.85 vs. 3.31±0.74〕, global ejection fraction(GEF:0.205±0.054 vs. 0.149±0.054)were significantly higher than those in non-survivors (P<0.05 or P<0.01). Correlation analysis showed a positive correlation was found between NT-proBNP and EVLWI (r=0.277,P=0.010),and negative correlations were found between NT-proBNP and CI(r=-0.367,P=0.001), GEF(r=-0.259,P=0.017). No correlation was found between NT-proBNP and GEDVI,SVRI. Receiver operating characteristic curve(ROC curve)analysis showed that the area under the ROC curve(AUC)for plasma NT-proBNP predicting the outcome of septic shock patients was 0.869±0.042,95% confidence interval(95%CI)was 0.786-0.952,with the maximum sum of sensitivity and specificity 1.695 to determine NT-proBNP predicting septic shock patient's death cut-off value was 2 071 ng/L,under this cut-off value,the sensitivity was 81.4% and specificity,88.1%. The maximum AUC for EVLWI predicting the outcome of patients with septic shock was 0.690,cut-off value was 7.5 mL/kg,under this cut-off value,the sensitivity was 69.8% and specificity,66.7%. Maximum AUC for CI predicting the outcome of patients with septic shock was 0.785,cut-off value was 3.48 L?min-1?m-2,under this cut-off value,the sensitivity was 69.8%and specificity,66.7%. Maximum AUC for GEF predicting the outcome of septic shock patients was 0.794,cut-off value 0.175,under this cut-off value,the sensitivity was 76.2% and specificity, 81.4%. Multivariate analyses showed CI and NT-proBNP levels were independent predictors of the prognosis〔CI:P=0.001,odds ratio(OR)=9.183,95%CI 2.362-35.694;NT-proBNP:P=0.024,OR=1.000,95%CI 0.999-1.000〕. Conclusion The plasma NT-proBNP level which is correlated significantly to EVLWI can evaluate the severity of septic shock and can predict the prognosis of such patients.

Objective To investigate the value of inflammatory biomarkers such as procalcitonin (PCT),C-reactive protein (CRP),and endotoxin in early diagnosis of bacteriemia patients infected with gram-negative bacteria.Methods A cohort of 79 bacteriemia patients infected with gram-negative bacteria admitted from February 2011 to May 2013 were enrolled for retrospective study.Collected data for analysis included gender,age,disease severity (APACHE Ⅱ score),bacterial isolates from blood culture and other general information.The inflammatory biomarkers such as white blood cell (WBC),neutrophils (NEU),Creactive protein (CRP),procalcitonin (PCT),and endotoxin were assayed within 6 hours after admission.SPSS version 16.0 software was used for statistical analysis.The test of normality was used for analysis of continuous variables,t-test for inter-group comparison and non-parametric statistics for non-normal distribution variables.The AUC of ROC was calculated for determining the sensitivity and specificity of biomarkers for diagnosis of bacteriemia.Results (1) Statistically positive correlations were found among serum PCT,CRP,and endotoxin levels (PCT/CRP =0.916,PCT/endotoxin =0.496,Endotoxin/CRP =0.387),and between those and APACHE Ⅱ score were (PCT/APACHE Ⅱ =0.505,Endotoxin/APACHE Ⅱ =0.467,CRP/APACHE Ⅱ =0.278),respectively,in bacteriemia patients infected with gram-negative bacteria.(2) The receiver operating characteristic (ROC) curve indicated that AUC PCT =0.715 (sen 64.6％,spe 80.7％),AUC CRP =0.666 (sen 67.7％,spe 78.6％),AUC endotoxin =0.771 (sen 78.8％,spe 81.8％) in gram-negative bacteria bloodstream infection patients.(3) The AUC PCT =0.865 (sen 86.2％,spe 77.5％),AUC CRP =0.733 (sen 72.4％,spe 75.0％),AUCendotoxin =0.618 (sen 70.7％,spe 67.5％) in bacteriemia patients infected with gram-negative bacteria in severe sepsis and septic shock group.Conclusions The plasma PCT,CRP,and endotoxin have early predictive value in bacteriemia patients infected with Gram-negative bacteria.In sepsis stage,the level of serum endotoxin has the most significant value for diagnosis.In severe sepsis and septic shock stages,the PCT is the most value for diagnosis of bacteriemia.All biomarkers are positively correlated with severity of the disease.

Objective:To explore the predictive value of relevant hemodynemics indexes on cardiac shock prognosis by PiCCO technology.
Methods:A total of 54 consecutive patients with cardiac shock treated in our hospital from 2012-01 to 2013-05 were studied. The patients’ general information with hempdynemics indexes as cardiac index (CI), extra vascular lung water index (EVWI), general ejection fraction (GEF), general end diastolic index (GEDI), systolic vascular resistance index (SVRI) were monitored by PiCCO technology at before and 48 hours after treatment. Based on 28 days surviving condition, the patients were divided into 2 groups:Survival group, n=28 and Death group, n=26. The levels of above indexes were compared between 2 groups and their predictive values on cardiac shock prognosis were calculated by ROC curve analysis.
Results:Compared with Death group, Survival group presented much higher CI, GEF and much lower EVWI, SVRI at before and 48 hours after treatment, P<0.05-0.01, while GEDI was similar between 2 groups, P>0.05. With 48 hours treatment, the ROC analysis showed AUCEVWI=0.846 with cut-off point at 7.5ml/kg, AUCCI=0.884 with cut-off point at 3.46 L/(min·m2) and AUCGEF=0.853 with cut-off point at 16.5%;the sensitivity and speciifcity of EVWI, CI, GEF were 79.2%and 81.0%, 82.8%and 83.0%, 74.1%and 88.7%respectively. Multivariate Logistic regression analysis indicated that CI and EVWI were the independent predictors for cardiac shock prognosis.
Conclusion:PiCCO monitoring technology may guide the balance of relevant hempdynemics indexes in patients with cardiac shock. With 48 hours treatment, the levels of CI and EVWI had predictive value for cardiac shock prognosis.

Objective To observe the early changes of related indexes after high dose of 60Co γ-ray irradiation on rhesus monkey hematopoietic system.Methods A total of 33 rhesus monkeys were randomly divided into normal control and different irradiation control group,rhesus monkeys in irradiation control group were given different doses(4,8,12 Gy) irradiation to establish acute radiation sickness(ARS) models.XE-2100 automatic blood cell analyzer detected the peripheral blood before and after the irradiation of 3,6,9,12,24,48,80 h.The rhesus monkeys were sacrificed to have a observation of sternum pathological changes at 6,48 and 80 h after 4,8,12 Gy 60Co γ-ray irradiation.Results The number of white blood cell in peripheral blood of the rhesus monkeys after 4 and 8 Gy 60Co γ-ray irradiation were lower than that before irradiation at 3 h after irradiation,as was significant increased at 6 h after irradiation,the highest values were 136.04%.and 221.38% after 9 h(with before irradiation values was 100.00%,the same below),become obviously drooped from 12 h after irradiation,show clearly temporary peak.But the number of white blood cell after 12 Gy 60Co γ-ray irradiation was significant increased at 6 h after irradiation,at the highest of 9 h,become obviously drooped from 12 h after irradiation.Peripheral blood neutrophile count was significant increased at 6 h after irradiation,at the highest of 9 h,become obviously drooped from 12 h after irradiation.Peripheral blood lymphocyte count fell sharply after irradiation,3 h detection value was only 12.02%-25.04% of before irradiation.Sternal bone marrow nucleated cell number decreased sharply after irradiation,the more irradiation dose,the less residual hematopoietic cells.Conclusion In the early stage of BM-ARS,temporary peaktime node of the white blood cell and neutrophil count could be regarded as the best delivery time of hematopoietic cytokine therapy.

Objective To analyze the effects and influence factors of primary total hip arthroplasty in the treatment of senile femoral neck fractures,and to increase the excellent rate of hip joint function recovery.Methods Retrospective analysis was applied to study the clinical data of one hundred and ten senile femoral neck fractures patients who underwent primary total hip arthroplasty in the 210th Hospital of People's Liberation Army from January 2014 to March 2016,to explore the main factors affecting postoperative recovery of hip joint function.Univariate analysis was used to analyze the factors influencing the clinical efficacy of the patients in terms of gender,age,body mass index,cause of injury,type of fracture and postoperative rehabilitation time,and Logistic regression analysis was used for multivariate analysis.Results The 110 senile femoral neck fractures patients obtained primary total hip arthroplasty,36 cases have seen excellent hip joint function recovery,54 good,15 fair,and 5 poor,with an excellent rate of 81.8 %(90/110).Single factor analyses found that body mass index,preoperative concomitant diseases,surgical approach,operation time,total blood loss, postoperative analgesia,discrepancy in bilateral lower limb lengths,postoperative rehabilitation time and postoperative complications were significantly related to hip joint function recovery(χ2=8.528,12.742,9.912, 8.131,10.404,10.171,12.406,6.412,10.362,P<0.05).According to the multivariate analysis,body mass index,surgical approach,total blood loss,postoperative analgesia,discrepancy in bilateral lower limb lengths,and postoperative rehabilitation time were the main factors affecting hip joint function recovery(Wald χ2=6.213, 4.543,9.156,6.287,5.461,3.698,95%CI=1.17~6.74,1.43~9.15,1.03~5.82,1.29~9.35,0.05~0.62, 0.12~0.93,P<0.05).Conclusion Primary total hip arthroplasty may lead to excellent curative effects for elderly patients with femoral neck fracture.Smaller body mass index,minimally invasive approach,less discrepancy in bilateral lower limb lengths,postoperative analgesia,less total blood loss and longer postoperative rehabilitation time may facilitate postoperative recovery for hip joint function.

OBJECTIVETo determine whether increased expressions of gelatinases occur with aging in vivo in kidney tissue of autoimmune MRL/lpr mice by in situ zymography.

METHODSMRL/lpr mice at the age of 8 weeks, 16 weeks and 24 weeks were investigated. Kidney protein extracts were compared for activities of MMP-2/9 by gelatin zymography. Immunohistochemistry and SDS-PAGE gelatin zymography were used to determine the expressions and activities of gelatinase A (MMP-2) and gelatinase B (MMP-9). To determine the net gelatinase activities in murine lupus kidney, in situ zymography was used with autoradiographic emulsion as substrate.

RESULTSBoth gelatinase A and B were seldom detected in the kidney tissue in 8 week old mice, Increased expressions of both latent and activated form enzymes of MMP-2/9 were identified in kidney extraction by SDS-PAGE gelatin zymography and immunohistochemical staining showed both MMP-2 and MMP-9 were obviously up-regulated within glomerulus as well as tubular-interstitium in mice at the age of 16 and 24 weeks. In situ zymography showed markedly increased gelatinase activities in kidney tissue consistent with the results of immunohistochemical staining, it is mainly derived from MMPs and inhibited by EDTA but not by PMSF or aprotinin.

CONCLUSIONSThese in vivo results suggested that MMP-2/-9 expressions were significantly up-regulated with aging in murine lupus nephritis, which may play an important role in promoting the remodeling formation of ECM and thus contribute to the progression of renal damage in this model.

Aging ; physiology ; Animals ; Autoimmune Diseases ; enzymology ; immunology ; pathology ; Disease Models, Animal ; Electrophoresis, Polyacrylamide Gel ; Female ; Lupus Nephritis ; enzymology ; immunology ; pathology ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Inbred MRL lpr ; Up-Regulation

Objective: To study effects of cyclin D1 overexpression on the proliferation and differentiation of cervical squamous cell carcinoma SiHa cells and to investigate related signaling molecules. Methods: Primers were designed to amplify the full length of cyclin D1 gene and cyclin D1 gene was amplified by PCR for constructing pcDNA3.1 plasmid vector. The construct was then transfected into SiHa cells, and the cells with stable overexpression of cyclin D1 were established, cyclin D1 gene and protein expression were detected by RT-PCR and Western blot, respectively. Cell growth curve was documented by MTT assay. CK7, E-cadherin, vimentin, Snail gene and protein expression in transfected cells were detected by RT-PCR and Western blot. RT-PCR was used to detect the mRNA expression of proliferation and differentiation-related genes like CDK4, CDK2, p21, p27, cyclin E, Rb, E2F, E6/E7 and Ki-67. After synchronization of cells, RT-PCR was used to detect of cyclin D1 and p21 mRNA expression at different time points of the cell cycle. Results: The G-3 cells with cyclin D1 overexpression were successfully established. The growth curve and Ki-67 mRNA expression accelerated in G-3 cells.Vimentin and Snail expression significantly increased at both gene and protein levels, while E-cadherin, CK7 gene and protein expression significantly decreased, indicating epithelial mesenchymal transitionoccurred in G-3 cells.Meanwhile, mRNA expression of cyclin D1, CDK4, CDK2, p21, p27, cyclin E, E2F and Rb increased, while E6/E7 and p16 showed no significant change. The expression trends of p21 and cyclin D1 were almost identical with fluctuation at different time points in the cell cycle. Conclusions Overexpression of cyclin D1 induced by gene transfection promotes proliferation and epithelial mesenchymal transition in SiHa cells.The process is accompanied by up-regulation of CDK4, CDK2, p21, p27 and cyclin E genes.p21 expression increases synchronously with cyclin D1, suggesting a regulatory role in epithelial mesenchymal transition by affecting expression of vimentin in G-3 cells.