Objective To explore the role of serum neutrophil extracellular traps (NETs) in rheumatoid arthritis (RA).Methods The serum circulating free DNA (cf-DNA) / NETs in 45 RA patients and 18 healthy controls were detected by Pico Green dsDNA Quantitation Kits,and the association between serum NETs and ESR,DAS28 score,anti-CCP antibodies was analyzed.T test,rank sum test,Pearson or Spearman correlation analysis were ued for statistical analysis.Results ① The levels of serum dsDNA/NETs in RA patients [0.523 0(0.282 0,1.637 0) μg/ml] were significantly higher than those in healthy controls [0.410 5 (0.140 0,0.966 0)μg/ml] (Z=-2.419,P=0.016).② The level of serum dsDNA/NETs in RA was positively correlated with ESR and DAS28 score (r=0.357,P=0.016; r=0.325,P=0.029),but not with anti-CCP antibodies(r=0.146,P=0.434).③ The levels of serum dsDNA/NETs in RA treated by DMARDs were lower than those of untreated with DMARDs[0.516 5(0.282 0,1.637 0) μg/ml,0.523 0 (0.369 0,1.485 0) μg/ml,Z=-1.215,P=0.225],but the difference was not significant.Conclusion NETs maybe associated with disease activity and play an important role in RA pathogenesis.

Objective To explore the significance of aryl hydrocarbon receptor (AhR) in patients with rheumatoid arthritis (RA) through detecting the levels of AhR and its response gene cytochrome P4501 A1 (CYP1 A1) in peripheral blood mononuclear cells (PBMCs).Methods Peripheral blood samples were collected from 35 patients with RA and 20 healthy subjects.The expression of AhR and CYP1A1 at mRNA level were detected by real-time PCR.The percentages of AhR-positive cells in PBMCs were detected by flow cytometry (FCM).The effects of leflunomide (LEF) on the expression of AhR and CYP1A1 were analyzed.The detailed clinical data of RA patients were recorded.The disease activity score (DAS) was calculated.Correlation analysis between AhR/CYP1A1 level and clinical data was conducted.Results (1) Both the expression of AhR at mRNA level and the percentage of AhR-positive cells in PBMCs from RA patients without LEF treatment were significantly higher than those from healthy subjects [(3.61±1.65) vs.(2.00±1.27),P=0.002; (34.21±11.30)％ vs.(18.83±7.32)％,P＜0.01].There were no statistically significant differences in the expression of AhR at mRNA level and the percentages of AhR-positive cells between patients with or without LEF treatment [(3.83 ± 1.62) vs.(3.61 ± 1.65),P =0.670 ; (36.69±10.61)％ vs.(34.21±11.30)％,P=0.462].(2) Non-LEF treatment group showed a higher relative expression of CYP1 A1 at mRNA level than that from control group [1.33 (0.08,7.86) vs.(0.62 ±0.29),z=-3.922,P＜0.01],but there was no statistical difference between LEF treatment group and non-LEF treatment group [(2.62±2.08) vs.1.33(0.08,7.86) z=-0.133,P=0.894].(3) Neither the expression of AhR and CYP1A1 at mRNA level nor the percentages of AhR-positive cells showed significant correlations with clinical data.Conclusion AhR was highly expressed in PBMCs from patients with RA,which might participate in the progression of rheumatoid arthritis.But the high expression of AhR did not reflect disease activity.Moreover,the treatment of LEF showed no significant influences on the expression of AhR and CYP1A1 in PBMCs from patients with RA.

OBJECTIVE To investigate the hepatotoxicity mechanisms of ethanol extract of Radix Polygoni Multiflori (RPM) and Radix Polygoni Multiflori Praeparata (RPMP) by high-content screen assay.METHODS HepG2 cells were treated with RPM (10,25,50,100,200 and 300 mg·L-1) and RPMP (10,50,100,300,600 and 1200 mg· L-1) for 3-24 h,respectively.The cell viability was detected by a CellTiter-GloTM luminescent cell viability assay kit.Cell count,reactive oxygen species (ROS),mitochondrial membrane potential (MMP),glutathione (GSH),superoxide dismutase 2 (SOD2),activating transcription factor 4 (ATF4),apoptosis,and cell cycles were investigated by high-content screen assay.Besides,SOD2 and ATF4 levels were confirmed by Western blotting.RESULTS RPM 300 mg· L-1 showed nearly 48 ％ reduction in cell viability compared with cell control (P＜0.01),while RPMP had no significant effect at the same concentration.Both RPM and RPMP decreased the level of MMP (P＜0.05) but incresed levels of GSH,ROS,SOD2 and ATF4 significantly (P＜0.05).Besides,RPM 200 mg· L-1 significantly increased the expression of SOD2 (P＜0.05) at 3 h by high-content screen assay,and the enhanced expression of ATF4 was shown at 6 h (P＜0.05).RPMP 300 mg· L-1 markedly increased the expression of ATF4 at 6 h (P＜0.05),while the expression of SOD2 significantly increased at 24 h (P＜0.05).CONCLUSION Both RPM and RPMP have some cytotoxicity,and the cytotoxicity of RPM is stronger than that of RPMP.The hepatotoxicity mechanisms of RPM and RPMP may be related to cell apoptosis caused by long-term oxidative stress and endoplasmic reticulum stress.

OBJECTIVETo investigate the cell-killing effect of adenovirus-mediated TK-ganciclovir (GCV) gene therapy in combination with tumor necrosis factor-alpha (TNF-alpha) against murine bladder carcinoma cells in vitro.

METHODSMurine bladder carcinoma MB49 cells were transfected with the adenoviral vector containing TK gene and green fluorescent protein (GFP) gene. The transfection efficiency was observed and the TK gene expression in the transfected cells was detected by RT-PCR. The survival rate of MB49 cells in response to TNF-alpha treatment and that of the TK gene-transfected cells after treatment with GCV and GCV+TNF-alpha were determined by MTT assay. The apoptosis of the cells after the treatments was analyzed by flow cytometry.

RESULTSIn cells transfected with TK gene, the cell inhibition rate increased gradually with the increment of GCV and TNF-alpha concentration. GCV in combination with TNF-alpha resulted in significantly increased killing efficiency of the cells as compared with GCV or TNF-alpha treatment alone, and the effect of the combined treatment was enhanced as the TNF-alpha concentration increased. GCV treatment (50 microg/ml) alone produced a cell killing rate of (24.39-/+1.10)%, and when combined with 5 microg/ml TNF-alpha, the rate was increased to (40.05-/+0.97) %, and further to (65.47-/+0.67) % when TNF-alpha concentration increased to 20 microg/ml. Flow cytometry revealed obvious apoptosis of the cells 8 h after treatments with TK/GCV, TNF-alpha, or TK/GCV+TNF-alpha, and the combined treatment resulted in the highest cell apoptotic rate.

CONCLUSIONTK/GCV in combination with TNF-alpha can enhance the effect of suicide gene therapy against murine bladder carcinoma cells and effectively induce apoptosis of the cells.

Adenoviridae ; genetics ; Animals ; Antiviral Agents ; pharmacology ; Cell Line, Tumor ; Cell Survival ; drug effects ; Ganciclovir ; metabolism ; pharmacology ; Genetic Therapy ; methods ; Green Fluorescent Proteins ; genetics ; metabolism ; Mice ; Reverse Transcriptase Polymerase Chain Reaction ; Thymidine Kinase ; genetics ; metabolism ; Transfection ; Tumor Necrosis Factor-alpha ; pharmacology ; Urinary Bladder Neoplasms ; genetics ; metabolism ; pathology

OBJECTIVETo clarify the relationship between hepatitis B virus (HBV) infection and IgA nephropathy (IgAN).

METHODSHBV antigen (HBAg) in renal tissues of the patients with IgAN was detected by immunohistochemical technique, the carrier status and localization of HBV DNA in renal tissues were determined by Southern blot analysis and in situ hybridization.

RESULTSSerum HBsAg was detected in 18 of the 100 patients with IgAN (18%), HBAg was detected in 31 of 100 patients (31%) in their renal tissue and in 20 of 31 patients (65%) in their glomeruli, and both HBsAg and HBcAg were detected in 10 of 31 patients (32%), respectively. HBcAg was also found in tubular epithelia (45%, 14/31) and renal interstitium (6%, 2/31), respectively. Five of six cases were proved to be positive of integrated-form HBV DNA in their renal tissue by Southern blot analysis. In situ hybridization demonstrated that HBV DNA was 8/8 and 6/8 positive in their renal tubules and glomeruli of all eight specimens, localized in the nucleus of tubular epithelial cells, glomerular mesangial cells, as well as infiltrated interstitial lymphocytes.

CONCLUSIONHBV infection closely related with IgAN and HBV infection might be involved in pathogenesis of IgAN.

Adolescent ; Adult ; Female ; Glomerulonephritis, IGA ; complications ; virology ; Hepatitis B ; complications ; Hepatitis B Antigens ; analysis ; Hepatitis B virus ; isolation & purification ; Humans ; Male ; Middle Aged

Global Health ; History, 20th Century ; Humans ; Influenza A Virus, H2N2 Subtype ; genetics ; isolation & purification ; Influenza A Virus, H3N2 Subtype ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; history ; mortality ; virology

OBJECTIVETo investigate the expression of programmed cell death 5 (PDCD5) in tissues of normal human prostate (NP), benign prostatic hyperplasia (BPH), and prostate cancer (PCa) in order to assess the clinical role of PDCD5 in PCa.

METHODSPDCD5 expression was determined by EnVision immunohistochemical staining in formalin-fixed and paraffin-embedded specimens obtained from 12 subjects with NP, 22 with BPH, and 22 with PCa. In addition, PCa cases were classified as low/middle-risk (Gleason sum < or = 7) and high-risk (Gleason sum >7) on the basis of Gleason grade. Positive expression rates and intensity of PDCD5 protein were observed under light microscope and analyzed with computer imaging technique. Expression of PDCD5 was compared among different prostatic tissues.

RESULTSThe expression of PDCD5 was significantly lower in tissue of PCa than in tissues of NP and BPH (P<0.01). However, there was no significant difference in PDCD5 expression between tissues of NP and BPH. In addition, the expression of PDCD5 was further downregulated with the increase of Gleason sum in PCa.

CONCLUSIONSBy downregulating apoptosis, low PDCD5 expression may play an important role in the occurrence and development of PCa. PDCD5 is supposed to have a potential clinical value to be a new predictor of progression and target of gene therapy in PCa.

Aged ; Apoptosis ; Apoptosis Regulatory Proteins ; analysis ; physiology ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasm Proteins ; analysis ; physiology ; Prostate ; chemistry ; Prostatic Neoplasms ; chemistry ; etiology ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; analysis

Objective To analyze the causes and epidemiological characteristics of injury in Hunan Province, so as provide scientific basis for the development of intervention measures of injury. Methods The Excel 2007 software and SPSS 18.0 software were used to organize and analyze the injury surveillance system data. Results A total of 107 754 effective cases were collected in three years, and the male-female ratio was 1.65:1. The top three causes of injury were falls(36.0%), motor vehicle accident (20.6%) and sharp knife injury (11.8%). The locations of injuries were mainly roads (30.9%) and homes (28.3%). The top three activities at the time of injury were leisure activities (24.7%), driving (17.6%) and work (16.4%). 93.2% of injury was unintentional injury, and the degree of injury was mainly mild injury (74.3%). Conclusions The number of injuries is increasing year by year. The number of male injuries is more than that of females. Systematic research and intervention measures should focus on falls and motor vehicle accident .

Objective To review the existing literature and quantitatively evaluate the association of circulating vaspin levels and the risk of gestational diabetes mellitus ( GDM) ． Methods We systematically searched the PubMed，EMBASE，Web of Science，China National Knowledge Infrastructure，and WanfangData databases up to June 2019． Pooled standardized mean differences ( SMDs) with 95% confidence intervals ( CIs) were calculated using random－ or fixed－effects models based on the heterogeneity of studies． Subgroup analyses，Meta－regression，sensitivity and publication bias were assessed to analyze the heterogeneity and the robustness of the results． All statistical analyses were performed using STATA 12．0． Ｒesults Nine articles ( 11 comparisons) published from 2013 to 2019 were included in our final Meta－analysis，covering a total of 738 patients with GDM and 661 normal pregnant women． There was significant difference in the overall maternal circulating vaspin levels between GDM patients and healthy pregnant women ( SMD= 0．613，95% CI: 0．044－1．182，P= 0．035) ． Subgroup analyses stratified by trimester in which vaspin was measured and whether BMI was matched suggested the similar trend to the overall result． Subgroup analysis according to ethnicity found that circulating vaspin level might not be related to GDM in " European" subgroup; sensitivity analysis by excluding moderate－quality studies and BMI－unmatched studies found that circulating vaspin levels were still related to GDM risk． Conclusions Our Meta－analysis indicated that maternal circulating vaspin levels might be positively correlated with the risk of GDM in Asians．