Objective To investigate the role of scavenger receptor class B type 1 (SR-B1) signaling pathway in serum amyloid A (SAA)-induced angiogenesis in rheumatoid arthritis (RA).Methods The expression and location of SR-B1 in RA and osteoarthritis (OA) synovial tissues were observed by immunohistochemistry.And SR-B1 expression in the resting human umbilical vein endothelial cells (HUVECs) was detected by immunoflourescence.Wound repair assessement and tube formation assessement were employed to evaluate the effect on cell migration and tube formation stimulated by SAA and/or anti-SR-B1 antibody.The t-test and one-way analysis of variance (ANOVA) were used for statistical analysis.Results ① SR-B1 was significantly highly expressed in RA tissue samples (A=6 788±819) when compared to the minimal expression in OA (A =31 849±6 977,t=3.567,P＜0.01).Positive staining of SR-B1 was observed in RA synovial vascular endothelial cells and perivascular areas.② Strong staining for SR-B1 was observed in all HUVECs tested.③ Significant wound healing induced by SAA (MI=2.50±0.17) was found compared with the untreated controls (MI=1.00±0.09,q=14.38,P＜0.01),and the effects were inhibited in the presence of anti-SR-B1 antibody (MI=1.16±0.14,q=13.02,P＜0.01).④ Compared to the untreated group (branch point number:6.6±0.8),there was an enhanced formation of branched and capillary-hke tube structure followed by SAA stimulation (branch point number:19.0±1.1,q=25.04,P＜0.01) after culturing for 72 h,whereas,tube formation decreased markedly upon pre-treated with anti-SR-B1 antibody (branch point number:7.6±1.3,vs SAA,q =23.32,P＜0.01).Conclusion Our present study suggests that serum amyloid A may induce angiogenesis via SR-B1 signaling pathway in RA.

Objective To investigate the neutrophil extracellular traps (NETs) formation and their molecular mechanisms induced by serum amyloid A (SAA) in rheumatoid arthritis (RA).Methods Neutrophils were isolated from peripheral blood of RA and healthy volunteers.① Neutrophils were cultured in vitro,the formation of NETs was observed and their percentage was calculated.② Neutrophils were cultured in vitro,divided into six groups:control,SAA,[SAA+anti-Toll like receptro4 (TLR4)-Ab],LPS,(LPS+anti-TLR4-Ab) and anti-TLR4-Ab.Appropriate stimulation was conducted for each group.NETs formation and their percentages were investigated.The concentration of DNA in supernatant was detected by fluorescent staining.F test and t test were used for statistical analysis.Results ① The purification of isolated neutrophils was higher than 95％.The network which was collocated with the spreading neutrophils nucleus and neutrophil elastase under the microscope,was NETs.In the RA group,the formation of NETs induced by SAA was significantly more than control [(19.1±0.8) vs (7.4±0.5),t=12.30,P＜0.05].② However,after pretreated with anti-TLR4 antibody,NETs formation was significantly less than the SAA group [(5.7±0.4) vs (14.7±1.1),t=7.825,P＜0.05].Moreover,the fluorescence intensity of DNA in supernatant was significantly higher in SAA group than that of anti-TLR4-Ab pretreatment group [(18.7 ±0.7) vs (12.9±0.8),t=5.552,P＜0.05].The concentration of DNA in supernatant of SAA group was higher than that of anti-TLR4-Ab pretreatment group [(36.9±1.3) μg/ml and (16.3±0.6) μ,g/ml,t=14.41,P＜0.05].Conclusion SAA can induce the formation of NETs from neutrophils by binding to TLR4 in RA.

Objective To investigate the effect of neutrophil extracellular traps (NETs) on inflammation of rheumatoid arthritis (RA), especially angiogenesis. Methods The presence of NETs in synovial tissues of RA and osteoarthritis (OA) was observed by immunofluorescence assay. Neutrophils were isolated from peripheral blood of health volunteers. Neutrophils were cultured in vitro, the formation of NETs was observed. NETs were extracted as a stimulating agent. The effects of NETs on the proliferation of human umbilical vein endothelial cells (HUVECs) and synovial fibroblasts (RAFLS) were evaluated by MTT, and which were classified into two groups: HUVECs group and RAFLS group, with the following treatment: control and NETs (0.28 mg/L). Wound repair assay was employed to evaluate the effect on the cell migration stimulated with NETs. The experiment was divided into three groups:control, VEGF (40μg/L VEGF) and NETs (0.28 mg/L NETs). Results (1) Compared with OA, NETs were found more in the synovial tissue of RA. (2) NETs formation was induced by stimulator in vitro. The concentration of extracted NETs-DNA was 27.8 mg/L. (3) MTT assay showed that compared with the control groups, low concentration of NETs (0.28 mg/L) promoted the proliferation of HUVECs (0.499 ± 0.011 vs. 0.393 ± 0.009, P<0.05) and RAFLS (0.266 ± 0.007 vs. 0.192 ± 0.007, P<0.05). (4) It was showed that a significant wound closure induced by low concentration of NETs (0.28 mg/L) was found compared with control. Conclusion Our present study suggests that NETs are found more in the synovial tissue of RA, and low concentration of NETs can promote angiogenesis in RA.

Objective To explore whether serum amyloid A (SAA) can induce the formation of neutrophil extracellular traps(NETs)in neutrophils in vitro. Methods A stable method for inducing NETs formation in vitro was established, in-cluding isolation of peripheral blood neutrophils, cell culture, and NETs formation and observation. The neutrophils were iso-lated from peripheral blood of healthy volunteers. And cells were cultured in vitro and classified into three groups:negative control (NC) group, SAA group and lipopolysaccharide (LPS) group. Following the distinct stimulation in three groups, NETs formation was observed and its percentage was calculated. The concentration of hinstone (h) 3 in supernatant was detected by ELISA. Results The purification and vitality of isolated neutrophils were both more than 95%. The nuclei of neutrophils lost their shape and spread, NETs formation was found. More NETs formation was found in SAA group than that in NC group (P < 0.05). Moreover, the concentration of h3 in supernatant was significantly higher in SAA group than that in NC group (P<0.05). Conclusion SAA can induce the formation of NETs in vitro.

Objective Fibroblast-like synoviocytes (FLS) play important roles in the pathogenesis of rheumatoid arthritis (RA).The present study was undertaken to investigate the mechanism of calreticulin (CRT) to promote FLS survival in RA.Methods FLS were isolated by enzymatic digestion of synovial tissue specimens obtained from RA and osteoarthritis (OA) patients and cultured in vitro.The expression of Bcl-XL and Mcl-1 in FLS at mRNA and protein level was detected by quantitative-polymerase chain reaction (q-PCR),Western blotting and immunofluorescence respectively.RA and OA FLS were cultured with different concentrations of recombinant human CRT for 48-72 h,the expression of Bcl-XL and Mcl-1 was detected by q-PCR and Western blotting.The proliferation of RA FLS following CRT stimulation was determined by MTT assay.Results ① Compared with FLS from OA patients (1.00±0.39;1.00±0.46),the anti-apoptotic Bcl-XL and Mcl-l mRNA expression (14.51 ±2.20;12.82±1.80) was significantly higher in the FLS from RA patients (t=10.47,1 1.02;P＜0.01);Western blotting analysis also showed increased protein levels of Bcl-XL and Mcl-1 in RA FLS;Immunofluorescence results showed higher expression of Bcl-XL and Mcl-1 in RA at the single FLS level;② CRT up-regulated the expression of Bcl-XL and Mcl-1 in RA FLS:compared with the control group (0 ng/ml),CRT stimulation at 10 ng/ml and 50 ng/ml increased the levels of Bcl-XL mRNA (1.70±0.28 vs 1.00±0.20,q=4.58,P＜0.05;1.87±0.35 vs 1.00±0.20,q=5.69,P＜O.05) and Mcl-1 mRNA (1.85±0.36 vs 1.00±0.20,q=5.63,P＜0.05;1.72±0.26 vs 1.00±0.20,q=4.77,P＜0.05) in RA FLS,while no significant effects of CRT on Bcl-XL and Mcl-1 mRNA expression were observed in OA FLS (F=1.49,1.60;P＞0.05);Western blotting results showed elevated protein levels of both Bcl-XL and Mcl-1 in RA FLS after CRT treatment at a concentration dependent manner.However,neither Bcl-XL nor Mcl-1 expression was significantly changed in OA FLS.③ MTT assay showed that CRT had no significant effect on the proliferation of RA FLS (F=2.88,P＞ 0.05).Conclusion Our results indicate that CRT-mediated up-regulation of anti-apoptotic Bcl-XL and Mcl-1 may inhibit apoptosis and promote the survival of FLS from RA patients.

Allotransplantation of peripheral blood-derived hemopoietic stem cells is the best therapy for acute leukemia. With increases of only-child families, sibling donors are decreasing. Moreover, the probability is low and time consuming is long to search a matched hemopoietic stem cell donor from the Chinese Marrow Donor Program. Haploidentical stem cell transplantation would bring a hope for donor resource. However, difficult transplantation and high incidence of graft versus host disease are two risks. Based on modified regimen and cyclosporine A+short-term amethopterin, mycophenolic acid, interleukin-11, anti-lymphocyte immunoglobulin and hemopoietic stem cells mobilized by recombinant human granulocyte colony-stimulating factor can prevent graft versus host disease. This therapy succeeded in two cases with no severe graft versus host disease.

OBJECTIVE: To assess the association of polymorphisms of rs3819024 and rs8193037 loci in the promoter region of IL-17A gene with the risk of ischemic stroke (IS) among ethnic Han Chinese from Guangxi. METHODS: The polymorphisms of rs3819024 and rs8193037 loci were detected by a SNaPshot assay and DNA sequencing among 392 IS patients and 443 healthy controls with matched age and gender. RESULTS: The genotypes, dominant model, recessive model, and alleles of rs3819024 polymorphisms showed no significant difference between the two groups, with the P values calculated as 0.150, 0.227, 0.125, 0.594 and 0.202, respectively, and OR (95% CI) as 1.27(0.92-1.74), 1.28(0.86-1.91), 1.27(0.94-1.72), 1.10(0.78-1.54), and 1.13(0.94-1.38), respectively. The genotypes, dominant model, recessive model, and alleles of rs8193037 polymorphisms also showed no significant difference between the two groups, with the P values calculated as 0.722, 0.352, 0.863, 0.345 and 0.969, respectively, and OR (95% CI) as 0.94(0.65-1.35), 2.25(0.41-12.35), 0.97(0.68-1.38), 2.27(0.41-12.48), and 1.01(0.72-1.40), respectively. CONCLUSION Polymorphisms of the rs3819024 and rs8193037 loci of the IL-17A gene are not associated with the susceptibility to IS among ethnic Han Chinese from Guangxi.
Alleles ; Asian Continental Ancestry Group ; Brain Ischemia ; genetics ; Case-Control Studies ; China ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Interleukin-17 ; genetics ; Polymorphism, Single Nucleotide ; Stroke ; genetics

Objective:To explore the role of Technique for Order Preference by Similarity to Ideal Solution(TOPSIS)combined with Rank Sum Ration(RSR)comprehensive evaluation method in establishing the China Healthcare Security Diagnosis Related Groups(CHS-DRG)operational performance system under the CHS-DRG payment system.Methods:SPSSPRO statistical analysis software was used,and TOPSIS method was used to regularly evaluate and rank the CHS-DRG disease groups in the hospital based on five indicators:Case Mix Index(CMI),total weight,average cost per weight,average length of stay per weight,and group profit and loss.RSR method was used to classify the TOPSIS disease group ranking results,establish performance reward standards for different grades of disease groups,and summarize and restore the reward results for each patient in the disease group to each clinical department,ultimately forming a department performance reward plan.The changes in key operational indicators related to medical insurance patients in hospitals were observed to verify the effectiveness of implementing performance reward programs.Results:After the application of TOPSIS combined with RSR comprehensive evaluation method in the construction of CHS-DRG operational performance system,the number of hospital medical insurance patients were increased,CMI value were stabilized,average cost per visit were decreased,average length of stay were shortened,and DRG disease group surplus were increased.Conclusion:The TOPSIS combined with RSR comprehensive evaluation method has played a good role in the construction of the CHS-DRG operational performance system.As a method of hospital economic operation evaluation,it is practical and innovative.

Objective:To explore the role of Technique for Order Preference by Similarity to Ideal Solution(TOPSIS)combined with Rank Sum Ration(RSR)comprehensive evaluation method in establishing the China Healthcare Security Diagnosis Related Groups(CHS-DRG)operational performance system under the CHS-DRG payment system.Methods:SPSSPRO statistical analysis software was used,and TOPSIS method was used to regularly evaluate and rank the CHS-DRG disease groups in the hospital based on five indicators:Case Mix Index(CMI),total weight,average cost per weight,average length of stay per weight,and group profit and loss.RSR method was used to classify the TOPSIS disease group ranking results,establish performance reward standards for different grades of disease groups,and summarize and restore the reward results for each patient in the disease group to each clinical department,ultimately forming a department performance reward plan.The changes in key operational indicators related to medical insurance patients in hospitals were observed to verify the effectiveness of implementing performance reward programs.Results:After the application of TOPSIS combined with RSR comprehensive evaluation method in the construction of CHS-DRG operational performance system,the number of hospital medical insurance patients were increased,CMI value were stabilized,average cost per visit were decreased,average length of stay were shortened,and DRG disease group surplus were increased.Conclusion:The TOPSIS combined with RSR comprehensive evaluation method has played a good role in the construction of the CHS-DRG operational performance system.As a method of hospital economic operation evaluation,it is practical and innovative.

Objective:To explore the role of Technique for Order Preference by Similarity to Ideal Solution(TOPSIS)combined with Rank Sum Ration(RSR)comprehensive evaluation method in establishing the China Healthcare Security Diagnosis Related Groups(CHS-DRG)operational performance system under the CHS-DRG payment system.Methods:SPSSPRO statistical analysis software was used,and TOPSIS method was used to regularly evaluate and rank the CHS-DRG disease groups in the hospital based on five indicators:Case Mix Index(CMI),total weight,average cost per weight,average length of stay per weight,and group profit and loss.RSR method was used to classify the TOPSIS disease group ranking results,establish performance reward standards for different grades of disease groups,and summarize and restore the reward results for each patient in the disease group to each clinical department,ultimately forming a department performance reward plan.The changes in key operational indicators related to medical insurance patients in hospitals were observed to verify the effectiveness of implementing performance reward programs.Results:After the application of TOPSIS combined with RSR comprehensive evaluation method in the construction of CHS-DRG operational performance system,the number of hospital medical insurance patients were increased,CMI value were stabilized,average cost per visit were decreased,average length of stay were shortened,and DRG disease group surplus were increased.Conclusion:The TOPSIS combined with RSR comprehensive evaluation method has played a good role in the construction of the CHS-DRG operational performance system.As a method of hospital economic operation evaluation,it is practical and innovative.