ABSTRACT
Objective:This work aims to assess the distribution of peripheral blood monocyte subsets, the expression level of the functional markers in rheumatoid arthritis (RA) patients, and analyze the correlation between the above indexes and the onset of RA.Methods:Peripheral blood mononuclear cells were collected and isolated from 62 RA patients, 52 healthy control (HC) and 12 disease control group′s patients via density centrifugation. The enrolled patients were attended or underwent physical examination in East Hospital, Tongji University from June 2020 to December 2021. Monocytes could be classified into classical (CM), intermediate (IM) and non-classical (NCM). Then, the flow cytometry was performed to examine the distribution of monocyte subsets and the measure the expression level of human leukocyte antigen DR (HLA-DR), intracellular tumor necrosis factor α (TNF-α) in peripheral blood monocytes. The statistical methods in this study mainly include: Kruskal-Wallis H test, Chi-Square test, Mann-Whitney U test, Wilcoxon matched-pairs signed ranks test, Spearman correlation coefficient test and Logistic regression analysis. The diagnostic value of IM proportion in RA was analyzed by ROC curve. Results:The monocytes number and monocytes proportion in white blood cells were much higher in RA [0.40 (0.40, 0.50), 7.60% (5.97%, 8.53%)] and disease control [0.40 (0.40, 0.68), 8.20% (5.85%, 10.28%)] compared with HC [0.30 (0.30, 0.40), 5.80% (5.03%, 6.38%)] ( H=24.733, P<0.001; H=27.469, P<0.001). A statistic-significant difference was detected among the proportion of CM[85.49%(76.91%,89.21%),88.94%(86.36%,91.72%),90.26%(80.25%, 92.56%)],IM[11.65%(8.47%,17.89%),7.89%(5.36%,10.75%), 5.56%(4.17%, 8.27%)], NCM[2.22%(1.39%, 3.74%), 2.49%(1.74%, 4.66%), 5.13%(3.39%, 9.85%)] in RA group, HC group and disease control group ( H=11.389, P=0.003; H=20.815, P<0.001; H=10.640, P=0.005). The proportion of CM was lower in RA and the IM proportion was increased in RA( P=0.003; P=0.003). The intracellular TNF-α level of monocytes in all three groups revealed the trend that IM>NCM>CM. The intracellular TNF-α in IM of RA was positively associated with serum TNF-α ( r=0.376, P=0.041). The HLA-DR expression in IM subsets were higher than CM and NCM subsets in all RA,HC and disease control groups. The expression of HLA-DR of IM in RA group and disease control was higher than HC group [8 611.50 (6201.3, 9890.8), 10 295.0 (7 899.0, 13632.0), 6 278.00(4 057.8, 9522.0), H=10.495, P=0.005]. There were no correlations between the proportion of peripheral blood IM and clinical characteristics CRP ( r=0.119, P=0.359), RF ( r=0.204, P=0.112) and ESR ( r=0.153, P=0.236). Logistic regression analysis showed that the proportion of IM ( OR=1.169, 95% CI 1.003-1.363, P=0.046), CRP ( OR=1.277, 95% CI 1.000-1.631, P=0.050), RF ( OR=1.179, 95% CI 1.080-1.287, P<0.001) are positively correlated with RA onset. The area under ROC curve for diagnosis of RA with IM proportion was 0.687, and the 95% confidence interval was 0.590-0.784, P<0.001. Conclusions:The distribution of monocyte subsets in peripheral blood of RA patients is abnormal. The increase in the proportion of IM, the enhanced antigen-presenting ability, and the increased level of TNF-α secretion in RA patients may play an important role in the pathogenesis of RA.
ABSTRACT
Objective:To collect the clinical materials of primary liver cancer patients treated by gamma knife and analyze the treatment methods as well as short-term efficacy,and to provide a reference for the clinical treatment of primary liver cancer patients.Methods:A total of 633 patients with inoperable primary liver cancer were treated by body gamma knife.According to the TNM staging method of Union for International Cancer Control(UICC), there were 351 cases with clear TNM staging.Among them, there were 251 cases (71.5%) at T3 stage and 57 cases(16.2%) at T4 stage.The prescription dose of 200-600 cGy each time to the 40%-85% dose line covering the planned target volume(PTV), this program was performed 5 times per week, and the number of treatment ranged from 2 to 13.The biochemical and imaging changes were observed 2-3 months after treatment to evaluate the short-term efficacy.Results:During the treatment, 229(36.2%)patients had adverse reactions,100 (15.8%) patients appeared the reduced white blood cells, and 137 (21.6%) patients appeared the reduced platelets.On discharge from the hospital, 601 patients were improved, 22 patients had no obvious change, 5 cases were worse, and 5 cases died.The proportion of improved patients who received the cumulative dose between 3 000 cGy to 4 000 cGy was higher than those who received the cumulative dose less than 3 000 cGy(P0.05).Conclusion:The proportion of adverse reactions in the primary liver cancer patients treated with body gamma knife is relatively low and the short-term efficacy is ideal.Body gamma knife treatment is a safe and effective treatment method for the primary liver cancer patients.
ABSTRACT
Objective To study the expression and function of TCRαβ+ CD3+ CD4-CD8-T (Double negative regulatory T cells,DN Tregs) in peripheral blood of Systemic lupus erythematosus (SLE) patients,and investigate their function in pathogenesis of SLE.Methods TCRαβ+ CD4-CD8 T cells were quantified as percentage of total CD3+ T cells in peripheral blood from 20 SLE patients and 20 healthy controls by flow cytometry.Used ELISA to test the cytokine levels (IFN-γ,TNF-α,IL-6,IL-17A) in the plasma of SLE patients.And analyzed the relationship between the percentage of DN Tregs and cytokines levels.Results The ratio of DN Tregs in TCRαβ+ T cells was significantly increased in SLE patients compared to healthy donors (t=3.54,P＜0.01).The levels of IFN-γ,TNF-α and IL-6 in plasma of SLE patients were higher than healthy donors (t=2.824,2.085,2.304,P＜0.05).The frequency of DN Tregs was found to correlate with IFN-γ (r=0.52,P=0.02) but not TNF-α (r=0.17,P=0.16) and IL-6 (r=0.47,P=0.49).Conclusion This study reveals frequency of DN Tregs in peripheral blood of SLE patients was higher than healthy controls,the frequency of DN Tregs was also found to correlate with IFN-γ levels,which means that DN Tregs may play an important role in pathogenesis of SLE.
ABSTRACT
Objective To analyze the species classification and chracteristics of drug resistance and virulence in CTX-M producing Escherichia coli isolated from urine culture.Methods Escherichia coli cultured by urine were collected from our hospital during 2014,the ring disk diffusion test was implemented to determine the bacterial susceptibility,the EBLs determination test was used to analyze the bacterial EBLs producing situation;the enterobactoer duplicated gene spacer consensus sequency PCR(ERIC-PCR) was adopted to perform the genetic relation analysis;PCR was used to amplify the CTX-M encoding genes and multiple virulence genes iutA,ompT,fyuA,fdeC,fimH,traT,cvaC,pap,kpsMT,pAI,usp,aer,hlyA,cnf and chuA;the multiple PCR was used to analyze the species calssification of CTX-M-producing Escherichia coli;these strains of bacteria were classified as the CTX-M-producing group and non-CTX-M-producing group according to the results of CTX-M coding gene detection,the differences in the antibacterial drug resistance and virulence genes between the two gorups were performed the contrastive analysis.Results One hundred and sixty-two strains of E.coli by urine culture had no genetic correlation,among 126 EBLs positive strains,91 strains produced CT-M,in which 57 strains of CT-M producing Escherichia coli belonged to type D,and 116 strains belong to Type B2.The statistical analysis found that the drug resistance rate in the CTX-M-producing group was significantly higher than that in the non-CT-M producing group (except for imipenem),the prevalence of virulence genes including iutA,chuA and traT in the CT-M producing bacteria group was significantly higher than that in the non-CTX-M-producing group(P=0.001,0.006,0.000)Conclusion CTX-M-producing E.coli is main pathogenic bacterium of urinary infection in our hospital,its majority belong to type D with increased drug resistance,moreover has close correlation with virulence genes iutA,chuA and traA and is a pertential threat in clinical treatment of urinary infection.
ABSTRACT
AIM: To evaluate the changes of microRNA (miRNA) in hepatocytes during hydrogen peroxide-induced oxidative stress injury, and to observe the alleviating effect of mesenchymal stem cell-conditioned medium (MSC-CM) in this progress.METHODS: The hepatocyte oxidative stress injury model was established using hydrogen peroxide and human normal liver cell line L02.MSC-CM was prepared using centrifugation and filter.The effects of MSC-CM on hepatocyte injury were evaluated by apoptosis analysis, cell viability detection, cell cycle, and mitochondrial membrane po-tential (MMP).Twenty-one differentially expressed miRNAs were selected by gene chip hybridization, in which miR-143, miR-145, miR-301a and let-7a were confirmed by RT-qPCR.Bioinformatics software was utilized to predict target proteins of these miRNAs, and then the proteins were verified by Western blot.RESULTS: MSC-CM markedly attenuated hydrogen peroxide-induced oxidative stress injury by reducing apoptosis, promoting cell viability and regulating cell cycle.The ex-pression of miR-143, miR-145, miR-301a and let-7a, indentified by RT-qPCR, increased under the condition of oxidative stress injury, while decreased after MSC-CM treatment.The expression of miR-143 predicted target proteins, HK2 and ADRB1, decreased under the hydrogen peroxide-exposure, while increased after MSC-CM treatment, which is consistent with the regulatory trend of miR-143.CONCLUSION: MSC-CM might attenuate hydrogen peroxide induced oxidative stress injury via inhibiting apoptosis and regulating some miRNA expression.
ABSTRACT
Objective To analyze the susceptibilities of Escherichia coli isolates collected from blood and the prevalence of ESBLs encoding genes.Methods A total of 121 Escherichia coli isolates collected from blood during 2012 were analyzed for antimicrobial susceptibilities by software of WHONET 5.6,the production of ESBLs was confirmed by confirmatory pheno-typic testing,PCR and DNA sequence were further implemented to analyze the ESBLs-encoding genes.Results 121 E.coli isolates displayed high resistance towards broad spectrum penicillin and 2nd or 3rd generation cephalosporins,levofloxacin and cotrimoxazole,with the resistance rates being more than 40%,susceptibilities to imipenem,piperacillin/tazobactam,ami-kacin were observed,with the resistance rates to be less than 12%,86(88.7%)out of 121 isolates were found to produce ESBLs.Among them,59.5% (72),38.8% (47)and 4.1% (5)were confirmed to carry blaCTX-M,blaTEM and blaSHV genes.Additionally,2(1.7%)isolates carried all the genes detected,30(24.8%)isolates carried both of blaCTX and bla-TEM,1(0.8%)isolate carried both of blaSHV andblaTEM.Conclusion Most of the E.coli isolates from the blood culture in Nanjing Gulou Hospital produce ESBLs,and displayed resistance towards most of the penicillins,cephalosporins and sin-gle amide antimicrobial agents should be chosen according to susceptibility results.