ObjectiveTo establish an analytical method with high sensitivity and wide range in biotin-avidin and time-resolved fluoroimmunoassay system for the quantitative detection of antinuclear antibody (ANA)-Ig (GAM).MethodsANA in standard preparation or sample was combined with solid nuclear antigen,biotinylation antibody and the europium(Eu3+)-labelled avidin to form the compounds of solid nuclear antigen-antibody-biotinylation antibody-SA-Eu3+.The fluorescence enhancement fluid was added to dis-sociate the Eu3+,the content of ANA was directly proportional to fluorescence intensity,the BAS-TRFLA was established for the quantitative detection of ANA-Ig (GAM).The sensitivity,specificity,reliability and range of detection were evaluated.Semm of 50 blood donor,105 patients with systemic lupus erythematosus (SLE),109 patients with rheumatoid arthritis(RA),25 patients with PBC,29 patients with Sj(o)gren's syndrome (SS),23 patients with scleroderma,25 patients with MCTD were included in this study.The positive rate was calculated.ResultsThe inner-group difference between high,medium and low densities mixture serum was 3.13％,3.74％ and 5.76％ and the inter-group precision rate was 5.31％,6.25％ and 7.46％ in BAS-TRFLA.The sensitivity of TRFIA was better than that of the ELISA method.The low detection limit was 2.24 U/ml.The mean recovery rate was 100.74％.The results measured by the TRFIA and ELISA methods were closely correlated(R2=0.978).The positive rate of blood donor,and patient with SLE,RA,PBC,SS,scleroderma and MCTD were 0,100％,23％,96％,38％,91％,92％ respectively.When compared with ELISA,the detection range of TRFIA was wider,and stability was better.ConclusionBAS-TRFIA is a stable method for detection of ANA with high sensitive and wide range of detection.It is important for the early diagnosis of autoimmune disease and monitoring the treatment efficacy of autoimmune disease.And this method may be widely used in clinical laboratories in the future.

Objective To study the effect of berberine on extracellular release of high mobility group box 1 (HMGB1) and its mechanism. Methods After LPS was activated by different concentration of berberine, the release of HMGB 1 and TNF-a, and expression of HMGB 1 mRNA in cultured mouse peritoneal macrophages was investigated. Results Berberine markedly suppressed the release of HMGB 1 and expression of HMGB 1 mRNA from LPS-stimulated macrophages (P<0.05). While the release ofTNF-a was't significantly affected. Conclusion Berberine suppressed extracellular release of HMGB 1 by decreasing expression of HMGB 1 mRNA.

Objective To investigate the significance of determination for binding ratio of L-PHA(leukoagglutinating phytohemagglutinin) with serum alkaline phosphatase in the diagnosis of primary hepatic cancer.Methods The binding ratios of L-PHA with serum alkaline phosphatase in 50 healthy individuals,103 cases of primary hepatic cancer,22 cases of hepatocirrhosis,36 cases of chronic hepatitis,18 cases of acute hepatitis and 13 cases of digestive tract tumor were determined by lectin affinity chromatography.Results The binding ratio of L-PHA with serum alkaline phosphatase in primary hepatic cancer(16.01% 5.42%) were significantly increased as compared with the healthy control(7.50% 2.53%)(P 0.01).The binding ratios were positively correlated with the tumor size and AFP level.Conclusion The binding ratio of L-PHA with serum alkaline phosphatase is a potential parameter for the laboratory diagnosis of primary hepatic cancer.

Objective To study the relationship between the expression of E-cadherin and the development, invasion and metastasis of esophageal carcinoma. Methods The expressions of E-cadherin in 58 cases with esophgeal carcinoma and 122 other cases with esophageal epithelial disease were examined by immunohistochemical technique. The relation between the expression of E-cadherin in esophageal carcinoma and the clinical pathological characteristics of esophageal carcinoma were analyzed. Results The expression rate of E-cadherin gene protein in esophageal squamous carcinoma and adenocarcinoma was 22.0%(11/50), 25.0%(2/8), respectively, compared with the normal group (100%, 20/20) and simple atypical hyperplastic group (92.9%, 26/28). The difference was significant (P

Objective To develop a method for detecting anti-cyclic citrullinated peptide (anti-CCP)in the serum of rheumatoid arthritis patients. Methods The range of lineage correlation, precision and detection range of time-resolved fluoroimmunoassay (TRFIA) was analyzed. Thirty-two positive serum of antiCCP, and the sera from 50 health blood donors, 32 SLE patients, 26 patients with SS, 10 patients with scleroderma, 20 patients with MCTD and 18 patients with MS were tested in this study. The clinical specificity was assessed. The consistency between TRFIA and ELISA was analyzed by Mc Nemar test. Results Analyzed by SPSS 13.0 statistical software, the intra-batch precision (n=20) rate was 2%, 3% and 4% respectively, and the inter-batch precision (n=8) was 3%, 4% and 7% respectively for 3 different concentrations. The clinical specificity was 98%, 97%, 96%, 100%, 95% and 100% in the sera of healthy blood donors, SL,E, SS,scleroderma, MCTD and MS patients respectively. The correlation coefficient was 0.989.The average recovery rate was 101%, and the sensitivity was 1 U/ml. When compared with ELISA, the detection range of TRFIA was wider and also with betterstability. Conclusion TRFIA is a stable method with high sensitivity and wide detection range. It can be used to detect anti-CCP antibody. It is important for early diagnosis of RA and monitor the curative effect of RA. And this method has potential to be used in clinical diagnosis.

Objective:Methamphetamine(METH)is an illicit psychoactive substance that can damage various organs,with the urinary system being one of its significant targets.This study aims to explore the role of microtubule affinity-regulating kinase 4(MARK4)in METH-induced acute kidney injury(AKI). Methods:A total of 10 healthy adult male C57BL/6 mice were randomly divided into a control group and a METH group,5 mice in each group.The METH group was administered METH(20 mg/kg,intraperitoneally,once daily for 3 consecutive days),while the control group received an equal volume of physiological saline.The mice were executed 24 hours after the final injection,and the success of the AKI model was detected by blood serum creatinine,blood urea nitrogen,and renal HE staining.Proteins differentially expressed between kidney tissues with METH-induced AKI and normal kidney tissues were screened by proteomics techniques and subjected to gene ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)and bioinformatics analysis.The accuracy of proteomic data was validated using Western blotting,and the expression levels of MARK4 and cleaved caspase-3 in mouse kidneys were measured.We further explored the role of MARK4 in METH-induced AKI.Firstly,a METH toxicity model was established in BUMPT cells to screen the appropriate concentration and time of METH treatment;the viability of BUMPT cells after METH treatment and the expression of cleaved caspase-3 were detected by interfering with MARK4 expression through inhibitors. Results:The proteomic analysis of kidney tissues from METH and control groups screened for a total of 17 differentially expressed proteins,of which 11 were up-regulated and 6 were down-regulated(all P＜0.05).The expression levels of MARK4 and cleaved caspase-3 were elevated in the kidneys of METH-treated mice(both P＜0.05).The activity of BUMPT cells gradually decreased with increasing METH treatment concentration(all P＜0.05),where the viability of BUMPT cells decreased to about 60％after METH treatment at 4 mmol/L.Compared with the control group,expression levels of MARK4 and cleaved caspase-3 were increased with higher METH concentrations and longer exposure times in a concentration-and time-dependent manner(all P＜0.05).Inhibition of MARK4 expression improved METH-induced decrease in BUMPT cell activity,down-regulated the expression of cleaved caspase-3,and decreased the apoptosis of BUMPT cells induced by METH. Conclusion:MARK4 is highly expressed in a mouse model of METH-induced AKI,and MARK4 mediates METH-induced AKI by regulating cell apoptosis.

【Objective】 To understand the current situation of blood components distribution in domestic prefecture-level blood stations through analyzing the components distribution data of 24 prefecture-level blood stations in China. 【Methods】 The data of components distribution of 24 blood stations from 2017 to 2020 as well as the data of blood deployment of 24 blood stations from 2019 to 2020 were collected and analyzed. 【Results】 From 2017 to 2020, positive annual growth in red blood cells, plasma and cryoprecipitate was observed in 22, 19 and 15 out of the 24 blood stations, and the annual growth median rate of above three components was 5.24%, 3.80% and 3.25%, respectively. Among the 24 prefecture-level blood stations, 23 carried out the preparation of cryoprecipitate. 【Conclusion】 The distribution of red blood cells, cryoprecipitate and plasma in prefecture-level blood stations is increasing year by year. However, there is a overstock of plasma, and most blood stations need blood employment.