Objective To investigate the correlation between the connective tissue growth factor(CTGF)in serum and the stages of liver fibrosis of patients with chronic liver disease,and explore new marker for evaluation of liver fibrosis.Methods The serum levels of CTGF in 313 patients with liver disease was detected by ELISA.Hyaluronic acid,type III procollagen,type IV collagen and laminin in serum were determined by RIA.Liver biopsy was performed in 45 patients with chronic liver disease.The quantitative relationship between the levels of CTGF and the stages of liver fibrosis was statistically analyzed by SPSS 11.5 software.Results A positive correlation between the serum levels of CTGF and the severity degree of chronic liver disease was found and the correlation coefficient was strong(r=0.634,P

Objective: To explore the feasibility of observing hemodynamics on ischemia/reperfusion(I/R) rat heart model in vivo by studying the changes of hemodynamics,myocardial zymogram and electrocardiogram. Methods: Twenty SD rats were randomly divided into 2 groups(n=10)(IR group and control group).The left anterior descending coronary artery was ligated or left untouched,a catheter was put into the left ventricle to trace the pressure change index and peak pressure of left ventricle,and to record ST segment changes of ECG.At the end of experiment,the levels of myocardial zymogram were determined.The results of two groups were compared. Results: Compared with control group,IR group showed reduced dp/dt max,declined LVSP,raised ST segment in ECG,and elevated myocardium zymogram. Conclusion: Our I/R rat heart model in vivo is feasible and stable to explore the cardiac hemodynamics.

Purpose To explore the relationship between connective growth tissue factor(CTGF) in serum and the severity of liver fibrosis,and to determine the clinical value of CTGF in the assessment of liver fibrosis.Methods Serums CTGF were tested utilizing enzyme linked immunosorbent assay(ELISA).The correlation between serum CTGF concentration and fibrosis stage was assessed.Results The diagnostic performance of CTGF was assessed by comparing the area under receiver characteristic curves(AUC) with a panel of fibrosis markers.Correlation coefficient was 0.689(P＜0.001) between the levels of serum CTGF and fibrosis stages and AUC of CTGF was 0.841(95% confidence interval,0.762-0.920) in distinguishing mild fibrosis from significant fibrosis.Conclusion The present data revealed that serum CTGF was significantly correlated with the stage of liver fibrosis,suggesting that serum CTGF was an indicator for the stage of liver fibrosis,and serum CTGF could be used as a valuable marker assessing liver fibrosis.

Objective To study the risk factors of severe hand-foot-mouth disease (HFMD) among children.Methods The clinical data of 1 570 children with HFMD at Linyi People's Hospital in Shandong Province in 2011 were collected,retrospectively.The data were analyzed using univariate and multivariate Logistic regression.Results The mean age of severe HFMD (including severe and critical HFMD) was (25.0± 14.0) months old,predominantely aged between 1 and 5 years old,while mild HFMD was (27.1±15.8) months (t'=-2.717,P=0.007).There were 61.0％ and 65.9％ boys in two groups,respectively (x2 =3.894,P=0.048).Fever,convulsion,tremor,nausea and vomiting were more frequently seen in severe HFMD.The neutrophil count and the level of creatine kinase in severe HFMD were both significantly higher than that in mild HFMD.Univariate analysis revealed that age (odds ratio [OR]=1.799,95％CI:0.984-1.997),girl sex (OR=1.234,95％CI:1.001-1.522),high fever (OR=2.110,95％CI:1.816-2.452),convulsion (OR=1.878,95％CI:1.578-2.236),nausea and vomiting (OR=1.760,95％CI:1.456-2.128),neutrophil count (OR=1.031,95％CI:1.025-1.037) and creatine kinase (OR=1.002,95％CI:1.001-1.003) were risk factors for severe HFMD.Multivariate Logistic regression showed that high fever (OR =1.751,95％ CI:1.487-2.062),convulsion (OR=1.451,95％CI:1.204-1.749),nausea and vomiting (OR=1.269,95％CI:1.027-1.568),neutrophil count (OR=1.028,95％CI:1.021-1.035) were independent risk factors.Conclusions Body temperature,neurological manifestations and trend of neutrophil counts should be carefully monitored in children with HFMD.Prevention of the development of severe HFMD mainly relies on the identification of risk factors and adoption of precautions in time.

Objective To investigate the usage of defluoridation stove and the formation of related behavior in the disease affected areas and the current situation of coal-burning-borne endemic fluorosis,to evaluate the control effect,so as to provide a theoretical basis for superior department to adjust the control strategy in time.Methods According to The Implementation Plan for Prevention and Treatment of Coal-burning-borne Endemic Fluorosis in Shaanxi Province,Ziyang and Langao in Ankang City were chosen for epidemiological survey.Using stratified sampling method,according to the degree of disease situation,each county was divided into 3 layers and then 5％ from each layer was extracted to carry out an investigation,respectively.The number of investigated village was at least 1,up to 30.The number in a non-endemic area was 1-3.Questionnaire survey of all residents was carried out to ask of the changes of cooking and heating fuel;the correct usage of the improved stove;the changes of life behavior related to the corn and pepper for human consumption and the development of prevention and treatment of post management.In endemic areas and in non-endemic areas,according to the Determination of Fluoride in Foods (GB/T 5009.18-2003),fluoride levels of corn (or rice) and chili which collected from 10 families were determined.Meanwhile,the criteria for Diagnosis of Dental Fluorosis (WS/T 208-2011) was used to diagnose dental fluorosis and the criteria of Urine-determination of Fluoride-ion Specific Electrode Method (WS/T 89-1996) was used to evaluate urine fluoride;dental fluorosis of every child aged 8-12 was examined and 10 copies of urine samples were randomly collected in each age group.Housewives and students over the age of 16 in endemic areas were quizzed of the knowledge of health and diagnosed serious skeletal fluorosis according to The Diagnostic Criteria of Endemic Fluorosis (WS 192-2008).Results The rates of correct usage of improved furnace and oven were 91.43％ (3 019/3 302) and 95.72％ (3 578/3 738).The autonomous maintenance rate of stoves was 0.61％ (49/7 998).In endemic areas,the correct drying rate and the correct storage rate of corn and pepper were all 100.0％ (300/300);the washing rate of corn and pepper before eating was 98.3％ (7 863/7 998).The fluoride median contents of corn and pepper was 0.800 and 2.863 mg/kg,respectively.In non-endemic areas,The fluoride median contents of corn and pepper was 0.443 and 4.065 mg/kg,respectively.The awareness rates of health knowledge were 81.27％ in housewives and 90.32％ in students.Dental fluorosis detection rate of 8-12 years old children was 12.52％ (87/695);the fluoride content was between 0.04-7.00 mg/L and the geometric mean value of fluoride content was 0.61 mg/L in endemic areas.There was no new case of serious skeletal fluorosis.Conclusions The prevention effect is obvious.Disease surveillance,health education and management of defluoridation stoves are keys to prevent coal-burning-borne endemic fluorosis.

Objective To assess the clinical value of pentraxin-3 (PTX-3) in diagnosis and survey of therapeutic effect for lung cancer.Methods The serum level of PTX-3,carcinoembryonic antigen (CEA),cytokeratin 19 fragment (CYFRA 21-1) were measured in 802 patients with lung cancer,462 with benign lung diseases and 522 healthy controls from multiple research centers,using ELISA and electrochemiluminescent assays.The clinical value of PTX-3 was assessed by comparing the area under receiver characteristic curves (AUC) with CEA and CYFRA21-1.The optimum cutoff value for diagnosis of lung cancer was investigated by maximizing the sum of sensitivity and specificity.By following-up,the serum level of PTX-3 was measured at 3 day,7 day,and 14 day in 61 lung cancer patients after surgical resection of lung cancer.Results In test group and validation,the serum levels of PTX-3 (g/L) are significantly higher in lung cancer group [9.21 (6.13-12.80),10.4(5.54-13.11)] than in benign lung diseases [5.28 (3.42-8.53),6.52 (3.84-7.89)] and in healthy controls [2.18 (0.54-5.44),2.44 (0.67-5.87)],[Z =8.161,14.118,(test group,all P ＜ 0.05) ;Z =9.832,17.595 (validation group,all P ＜0.05)].ROC curve showed the optimal cut-off values for PTX-3 was 8.03 g/L [AUC of 0.831,with a sensitivity of 76.1％ and specificity of 75.2％ in the test cohort; 0.828,71.3％,89.2％ in the validation cohort].Similar results were noted for early-stage lung cancer [0.764,79.1％,and 62.2％ in the test cohort; 0.744,71.3％,and 69.6％ in the validation].In the diagnosis of early-stage lung,the AUC and sensitivity and specificity of PTX-3 were 79.1％,0.764,71.3％ (test group),and 75.2％,89.2％,0.824 (validation group) significantly higher in these patients than CEA and CYFRA21-1.In small cell lung cancer,PTX-3 and NSE shared similar AUC differentiating LC from benign lung diseases and health controls.In following-up 61 lung cancer patients,PTX-3 levels before surgical resection of tumours [11.12(9.12-12.59)] was significant high than following 3 day after surgery(Z =4.32,P ＜0.01),and 14 day (5.12 ±2.54) vs.7 day (7.13 ±3.42) (t =2.143,P =0.023).The correlation between PTX-3 and CRP in LC,benign lung diseases,health control was 0.364,0.592,0.512 (all P ＜ 0.05).Conclusion Serum PTX-3 is a valuable biomarker of lung cancer and early-stage lung cancer with high sensitivity and specificity and improved identification of patients with lung cancer from those with non-malignant chronic lung diseases.

Objective To explore the protective effects of erythropoietin (EPO) on cardiomyocytes against hypoxia/reoxygenation (HR) injury. Method Isolated and cultured rat eardiomyocytes were subjected to 2-hour hypoxia followed by 1 hour reoxygenation to establish model of HR injury. Cardiomyocytes were randomly divided into 4 groups: sham group, HR group, HR + EPO-treated group (EPO 10 U/ml), and HR + EPO + UO126-treated group (U0126 10?mol/L). The concentration of lactate dehydrogenase (LDH) in culture medium was detected by automatic biochemical analyzer; viability of eardiomyocytes was measured by MTT assay; apoptosis ratio was determined by TUNEL technology and Annexin-V-FITC with flow cytometer (FCM); level of extracellular signal- regulated kinase 1/2 (ERK_(1/2)) and phospho-ERK_(1/2) were measured by Western-blot analysis. Results EPO significantly decreased the leakage of LDH, enhanced activity, reduced apoptosis ratio, and increased level of phospho-ERK_(1/2).However, the effects were blocked by U0126, an inhibitor of MAPK. Conclusions EPO has the protective effects on cardiomyocytes against HR injury possibly via the mechanism of activation of ERK_(1/2) and inhibition of apoptosis.

As main recipient cells for porcine reproductive and respiratory syndrome virus (PRRSV), porcine alveolar macrophage (PAM) are involved in the progress of several highly pathogenic virus infections. However, due to the fact that the PAM cells can only be obtained from primary tissues, research on PAM-based virus-host interactions remains challenging. The improvement of induced pluripotent stem cells (iPSCs) technology provides a new strategy to develop IPSCs-derived PAM cells. Since the CD163 is a macrophage-specific marker and a validated receptor essential for PRRSV infection, generation of stable porcine induced pluripotent stem cells lines containing CD163 reporter system play important roles in the investigation of IPSCs-PAM transition and PAM-based virus-host interaction. Based on the CRISPR/Cas9- mediated gene editing system, we designed a sgRNA targeting CD163 locus and constructed the corresponding donor vectors. To test whether this reporter system has the expected function, the reporter system was introduced into primary PAM cells to detect the expression of RFP. To validate the low effect on stem cell pluripotency, we generated porcine iPSC lines containing CD163 reporter and assessed the pluripotency through multiple assays such as alkaline phosphatase staining, immunofluorescent staining, and EdU staining. The red-fluorescent protein (RFP) expression was detected in CD163-edited PAM cells, suggesting that our reporter system indeed has the ability to reflect the expression of gene CD163. Compared with wild-type (WT) iPSCs, the CD163 reporter-iPSCs display similar pluripotency-associated transcription factors expression. Besides, cells with the reporter system showed consistent cell morphology and proliferation ability as compared to WT iPSCs, indicating that the edited-cells have no effect on stem cell pluripotency. In conclusion, we generated porcine iPSCs that contain a CD163 reporter system. Our results demonstrated that this reporter system was functional and safe. This study provides a platform to investigate the iPS-PAM development and virus-host interaction in PAM cells.
Swine ; Animals ; Induced Pluripotent Stem Cells/metabolism* ; Receptors, Cell Surface/genetics* ; Antigens, CD/metabolism* ; Porcine respiratory and reproductive syndrome virus/genetics*