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

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 com pare and explore the application value of diatom nitric acid digestion method and plankton 16S rDNA PCR method for drow ning identification. Methods Forty drow ning cases from 2010 to 2011 were collected from Department of Forensic Medicine of Wenzhou Medical University. Sam ples including lung, kidney, liver and field water fromeach case were tested with diatom nitric acid digestion method and plankton 16S rDNAPCR method, respectively. The Diatom nitric acid digestion method and plankton 16S rDNAPCR method required 20 gand 2g of each organ,and 15 mL and 1.5 mL of field water, respectively. The inspection time and detection rate were com pared between the two methods. Results Diatom nitric acid digestion method m ainly detected two species of diatom s, Centriae and Pennatae, while plankton 16S rDNA PCR method am plified a length of 162 bp band. The average inspection time of each case of the Diatom nitric acid digestion method was (95.30±2.78) min less than (325.33±14.18)min of plankton 16S rDNA PCR method (P<0.05).The detection rates of two methods for field water and lung were both 100% . For liver and kidney, the detection rate of plankton 16S rDNA PCR method was both 80% , higher than 40% and 30% of diatom nitric acid digestion method (P<0.05), respectively. Conclusion The laboratory testing method needs to be appropriately selected according to the specific circum stances in the forensic appraisal of drow ning. Com pared with diatom nitric acid digestion method, plankton 16S rDNA PCR method has practice values with such advantages as less quantity of sam ples, huge inform ation and high specificity.

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 explore the application value of plasma sHLA-G in diagnosis of CIN and cervical cancer. Methods The plasma sHLA-G levels were detected by ELISA in 102 cases with cervical cancer( FIGO Ⅰ stage 32 cases, Ⅱ stage 28 cases, Ⅲ stage 25 cases and Ⅳstage 17 cases; tumor size:＜4 cm 63 cases and ≥4 cm 39 cases; squamous cell carcinoma 78 cases and adenocarcinoma 24 cases;cell differentiation:well 57 cases, moderate 29 cases and poor 16 cases; lymph nodes metastasis negative64 cases and positive 38 cases ), 72 cases with CIN( Ⅰ grade 21 cases, Ⅱ grade 25 cases and Ⅲ grade26 cases ) and 20 cases of healthy controls. The diagnostic value of sHLA-G and its correlations with clinical parameters were analyzed. Results The plasma levels of sHLA-G were 193.6( 151.3-287.4 ) kU/L in cervical cancer group, 48.3( 34.6-57.2 ) kU/L in CIN Ⅰ group, 91.3( 68.2-118.6 ) kU/L in CIN Ⅱ group, 106.4( 73.8-165.7 ) kU/L in CIN Ⅲ group and 45.2( 38.0-55.5 ) kU/L in health control group.The level of sHLA-G was significantly higher in cervical cancer group than that in CIN Ⅰ group, CIN Ⅱ group, CIN Ⅲ group and healthy control group( U value of 8.832, 6.456, 4.017, 9.873, P ＜ 0.05,respectively ). The level of sHLA-G was significantly higher in CIN Ⅱ group and CIN Ⅲ group than that in CIN Ⅰ group and health control group( U value of 4.361,4.892, 5.139, 5.485, P ＜0.05, respectively ).The levels of SCC Ag in healthy control group, CIN Ⅰ group, CIN Ⅱ group, CIN Ⅲ group and cervical cancer group were 0.43( 0.38-0.69 )μg/L, 0.47( 0.35-0.72 )μg/L, 0.65( 0.53-0.81 )μg/L, 0.82( 0.54-1.03 )μg/L and 1.02( 0.62-1.87 )μg/L. The level of SCC-Ag was significantly higher in cervical cancer group than that in CIN Ⅰ group, CIN Ⅱ group and healthy control group( U value of 7.926, 4.877, 8.132,P ＜0.05, respectively ). The level of SCC-Ag was significantly higher in CIN Ⅲ group than that in CIN Ⅰ group and health control group( U value of 6.574, 6.763, P ＜0.05, respectively ). The levels of CA125 in healthy control group, CIN Ⅰ group, CIN Ⅱ group, CIN Ⅲ group and cervical cancer group were 14.38 ( 6.14-21.82 ) kU/L, 15.42( 6.25-23.53 ) kU/L, 21.34( 9.82-32.58 ) kU/L, 25.69( 14.47-38.71 )kU/L and 27.72( 14.29-43.87 ) kU/L. The level of CA125 was significantly higher in cervical cancer group than that in CIN Ⅰ group, CIN Ⅱ group and healthy control group( U value of 7.564, 4.522, 7.429, P ＜0.05, respectively ). The level of CA125 was significantly higher in CIN Ⅲ group than that in CIN Ⅰ group and health control group( U value of 5.871, 5.435, P ＜0.05, respectively ). ROC curve analysis showed AUC for sHLA-G was 0.828( 95% CI:0.768-0.879 ), which was high as compared with the AUC of SCC-Ag [ 0.727( 95% CI:0.658-0.788 );Z = 2.294, P ＜ 0.05 ] and the AUC of CA125 [ 0.705( 95% CI:0.636-0.769 );Z =2.842 ,P ＜0.05 ]. There was no significant difference of diagnostic efficiency between SCC and CA125( Z =0.672, P ＞ 0.05 ). When cutoff value of sHLA-G was 109.6 kU/L, the diagnostic sensitivity,specificity, positive predictive value, negative predictive value and accuracy rate were 86.3%, 76.1%,80.0%, 83.3%, and 78.4%, respectively. The levels of sHLA-G in cervical cancer patients were significantly correlated with FIGO stages and lymphoid node metastasis ( U value of 6.085, 4.451, P ＜0.05, respectively ), while there were no significant differences between the levels of sHLA-G and age,tumor size, histological type and cell differentiation( U value of 1.274, 1.956, 1.268, 2.719, P ＞0.05,respectively ). Conclusions sHLA-G can be used for the early screening of cervical cancer and its precancerous lesion. It could also be used as an index for judging progression and lymphoid node metastasis.

Objective To investigate the efficacy of platelet-rich plasma and hyaluronic acid intra-articular injection in patients with knee osteoarthritis. Method The following databases, including PubMed, Embase, the Cochrane Library and WANFANG, were used for collecting the randomized controlled trials, prospective control trials and experimental research about the therapy efficacies of platelet-rich plasma and hyaluronic acid on knee osteoarthritis. The meta-analysis was performed using RevMan 5.3 for Windows. Results Eleven publications, including six RCTs, one PCT and four ER, met the inclusion criteria. There were significant differences in WOMAC scores and IKDC scores between the PRP group and the HA group. but no significant differences were observed in Lequesne index between the PRP group and the HA groups at six month post treatment. Conclusion PRP intra-articular injection was better and more durable than HA in the treatment of knee osteoarthritis.

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 learn about the changes of the condition of coal-burning-borne endemic arsenism, the use of improved stoves and the formation of health-related behaviors in Shaanxi Province, and evaluate the effect of prevention and control measures.Methods:From 2015 to 2020, according to the "Implementation Plan for Monitoring Coal-burning-borne Endemic Arsenism in Shaanxi Province", regular field surveys were carried out in 4 natural villages, 2 counties in Ankang City and Hanzhong City, Shaanxi Province, to learn about the basic prevention and control situation in the monitoring village. Using the simple random sampling method, 10 families in each village were selected to investigate the use of stoves and the formation of health-related behaviors, and 5 of these families were selected to collect coal samples for arsenic content determination in accordance with the "Determination of Arsenic in Coal" (GB/T 3058-2008). According to the "Diagnosis of Endemic Arsenicosis" (WS/T 211-2015), the condition of the people exposed to high arsenic coal in the monitoring village was investigated. Urine samples of 30 adult patients (half males and half females) with arsenic poisoning were collected, the content of arsenic in urine was determined by the "Urine-Determination of Arsenic-Silver Diethyldithiocarbamate-Triethanolamine-Spectrophotometric Method" (WS/T 28-1996).Results:A total of 240 households were monitored in the past 6 years, and the quality conformance and correct utilization rates of improved stoves in the households monitored in the disease areas remained at 100.00% in 2018 - 2020. The utilization rate of clean energy increased from 75.00% (30/40) in 2015 to 100.00% (40/40) in 2018 - 2020 (χ 2trend = 25.5, P < 0.001). The 73.75% (177/240) of households using a variety of clean energy. From 2018 to 2020, the correct drying, storage and pre-processing washing rates of corn and pepper continued to reach 100.00% (40/40). The arsenic content of coal ( n = 120) in the disease areas was (118.09 ± 57.91) mg/kg, ranging from 16.70 to 280.94 mg/kg. The detection rate of arsenic poisoning decreased from 6.34% (231/3 646) in 2015 to 2.90% (109/3 754) in 2020 (χ 2trend = 121.8, P < 0.001), and no skin cancer or Bowen's disease was detected. The geometric mean of arsenic content in urine ( n = 720) was 0.038 1 mg/L, ranging from 0.000 5 to 0.312 9 mg/L. Conclusions:The condition of coal-burning-borne endemic arsenism areas in Shaanxi Province has reached the national elimination standard. The quality and correct utilization rate of improved stoves and the rate of using clean energy have increased. The healthy-related behaviors the people in the endemic areas have been basically formed. The prevention and control work has achieved good results.

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*