Objective: To develop an HPLC method for the simultaneous determination of five active components ( 5-O-methyl-visammioside, prim-O-glucosycimifugin, hesperidin, rosmarinic acid and forsythin) in Sijiganmao tablets. Methods:The samples were separated on an Agilent C18 column (250 mm × 4. 6 mm,5 μm) by acetonitrile-0. 1% formic acid solution with gradient elution. The detection wavelength was 283nm,the flow rate was 1. 0μl·min-1 ,the column temperature was 30℃,the detection wavelength was 283 nm,and the sample size was 10μl. Results:The complete separation was obtained for the five active compounds. The five regression e-quations showed good linear relationships. The average recoveries of the compounds were between 95. 0% and 105. 0%. Conclusion:The established method is accurate, reliable, simple and effective, which can be used in the quality control of Sijiganmao tablets.

Primary biliary cholangitis (PBC) is an autoimmune liver disease with unclear pathogenesis.The amino acid composition and sequence in the complementarity-determining region 3 of T cell receptor (TCR) and B cell receptor (BCR) are highly diverse, which forms a large antigen recognition receptor repertoire, i.e., immune repertoire.In recent years, second-generation sequencing techniques combined with multiplex PCR or amplicon rescue multiplex PCR have been used to study the features of immune repertoire in PBC patients, and it has been found that PBC patients have clonal expansion of specific CD4+ T lymphocytes, clonal diversity of B lymphocytes, somatic hypermutation, and reduction in class switch, as well as increase in clonal diversity after treatment with ursodeoxycholic acid.These findings need to be confirmed by large-scale in vivo and in vitro studies and different immune repertoire research strategies.

Objective To investigate the significance of antibody to Ro-52 in patients with autoim-mune liver disease(AILD). Methods One hundred and fifteen patients with abnormal liver functions, who had anti-Ro-52 detection by immunological blotting, were reviewed retrospectively. According to types of AILD, the clinical features were compared between patients with and without anti-Ro-52, respectively, κ test of concordance was used to provide a chance-corrected valve for immune-serological results. Results The rates of anti-Ro-52 in autoimmune hepatitis( AIH), primary sclerosing cholangitis(PBC) and AIH/PBC o-verlap syndrome groups were 32.43%, 24.56% and 33.33%, respectively, there were no significant differ-enees among three groups ( x2 = 0. 949, P >0. 05). The rate of anti-soluble hver antigen/liver-pancreas ( an-ti-SLA/LP) in AIH patients with anti-Ro-52 (58.33%) was higher than AIH patients without anti-Ro-52 ( 16.00% ,P < 0.05 ). The rate of anti-SLA/LP in AIH/PBC overlap syndrome patients with anti-Ro-52 (85.71%) was also higher than that of control group (28.57% ,P <0.05). Anti-Ro-52 and anti-SLA/LP had concordance according to κ test( κ >0.40, P <0.05). The average level of IgG in AIH/PBC overlap syndrome patients with anti-Ro-52 was higher than patients without anti-Ro-52 ( t = 2. 508, P < 0.05 ). Conclusion The rates of anti-Ro-52 in AIH, PBC and AIH/PBC overlap syndrome were of no significant differences. Anti-Ro-52 may have correlation with anti-SLA/LP. AIH/PBC overlap syndrome patients with anti-Ro-52 shewed higher IgG level than patients without anti-Ro-52.

Objective To analyze the influence of the polymorphisms of human leucocyte antigen (HLA)-Ⅰ molecule and the effects on plasma viral load of human immunodeficiency virus (HIV) infected male homosexual population in Beijing.Methods The HLA-A,HLA-B,HLA-C allele were typed by sequence specific primer-polymerase chain reaction (SSP-PCR),and viral load was detected in 157 chronic HIV infected persons.Normally distributed measurement data were analyzed by one-way or multi-way analysis of variance,while data of abnormal distributions were analyzed by Mann-Whitney U test.Results Among 157 chronic HIV infected persons,the number of Bw4 motifs on HLA-B loci was associated with a lower level of viral load (F=3.01,P=0.045).In these HIV infected persons,the viral load in HLA-B carrying Bw4/4 homozygote was (4.19±0.76) lg IU/mL,in HLA-B carrying Bw6/6 homozygote was (4.63±0.74) lg IU/mL (t=2.27,P=0.010).The viral load of those who carried three,one or none Bw4 motifs on HLA-A and HLA-B loci were (3.92± 0.97),(4.54±0.88) and (4.60±0.72) lg IU/mL,respectively (three vs none:t=2.53,P=0.015; three vs one:t=2.11,P=0.039).HIV infected persons who carried homozygote on any loci of HLA-A,HLA-B,HLA-C had comparable levels of plasma viral load to those who carried heterozygote on HLA-A,HLA-B,HLA-C loci.Among the persons who carried heterozygote on HLA-A,HLA B,HLA-C loci,Bw4/4 homozygote on HLA-B had lower levels of viral load than Bw6/6 homozygote on HLA-B (median:4.09 lg IU/mL vs 4.55 lg IU/mL,U=210.50,P=0.041).HIV infected persons who carried A30/B13/C06 or A33/B58/C03 haplotype had comparable levels of plasma viral load to those without A30/B13/C06 or A33/B58/C03 haplotype (t=0.40,P=0.69; t=0.68,P=0.49,respectively).Conclusions Bw4/4 homozygote on HLA-B loci is associated with lower HIV viral load.Furthermore,the plasma viral load of HIV infected persons carrying heterozygote on HLA-A,HLA-B,HLA-C loci could be influenced by the Bw4/4 homozygote on HLA-B locus,with a lower viral load.

The core protein (CP) of the classical swine fever virus (CSFV) is one of its structural proteins. Apart from forming the nucleocapsid to protect internal viral genomic RNA, this protein is involved in transcriptional regulation. Also, during viral infection, the CP is involved in interactions with many host proteins. In this review, we combine study of this protein with its disorders, structural/functional characteristics, as well as its interactions with the non-structural proteins NS3, NS5B and host proteins such as SUMO-1, UBC9, OS9 and IQGAP1. We also summarize the important part played by the CP in CSFV pathogenicity, virulence and replication of genomic RNA. We also provide guidelines for further studies in the CP of the CSFV.
Animals ; Classical Swine Fever ; virology ; Classical swine fever virus ; genetics ; metabolism ; pathogenicity ; Genome, Viral ; Swine ; Viral Core Proteins ; chemistry ; genetics ; metabolism ; Virulence

Objective To establish the enzyme-linked immunosorbent assay (ELISA) for the detection of serum alcohol dehydrogenase (ADH) antibody and evaluate its role in its diagnosis of autoimmune hepatitis( AIH ). Methods The reactivity between yeast ADH and human anti-ADH serum antibody was tested by Western blot analysis. ELISA was established using yeast ADH. The method was applied in serums of 67 AIH patients,94 primary biliary cirrhosis(PBC) patients, 199 chronic hepatitis B (CHB) patients, 132 chronic hepatitis(CHC) patients, 24 alcohol hepatitis disease(ALD) patients, 99 connective tissue disease(CTD) patients and 31 healthy individuals. The positive rate of ADH antibody in the patients and healthy individuals was measured. The χ2 test was used to compare the positive rates. Results The ELISA method for detecting human anti-ADH serum antibody was established successfully and the optimum reaction conditions were defined. Western blot showed that yeast ADH has cross reactivity with human anti-ADH antibody. The positive rate of anti-ADH antibody in the AIH group [59. 7% ,40/67 ] was higher than that in the normal control group(0,χ2 = 31. 271 ,P <0. 05), PBC group (6. 4% ,χ2 =54. 492,P <0. 05), CHB group( 14. 1% ,χ2 =54. 848,P <0. 05) ,CHC group(21.2% ,χ2 = 29.269,P<0.05), ALl) group ( 25. 0% ,χ2 =8.512,P <0.05)and CTD group ( 43. 4% ,χ2 =4.229, P <0. 05). Conclusions Compared with the PBC, CHB, CHC, ALD and CTD group, the anti-ADH antibody positive rate in the serums of AIH was significantly increased. The antibody may be helpful to the diagnosis of autoimmune hepatitis.

OBJECTIVETo explore the differential characteristics of the AMA-M2 autoantibody in patients with primary biliary cirrhosis (PBC) and non-PBC patients.

METHODSPatients with abnormal liver function at the Capital Medical University affiliated to Beijing You-an Hospital were enrolled in this study between January 2011 and December 2013. Serum levels of ANA, AMA and AMA-M2 were detected by indirect fluorescence assay and enzyme-linked immunosorbent assay. The patients' clinical data was obtained for retrospective analysis. Statistical analyses were performed using the SPSS 16.0 software. Enumeration data have been presented as numbers and percentages, and were analyzed using the chi-square test and one-way ANOVA test.

RESULTSOf the 5315 patients with abnormal liver function, 15.3% (811/5315) were AMA-M2 positive patients; among those 811 patients, 78.4% (636) had PBC, 4.4% (36) had PBC overlapping with autoimmune hepatitis (AIH), 4.4% (36) had drug-induced liver injury, 6.5% (53) had hepatitis B, 3.3% (27) had hepatitis C, 0.6% (5) had hepatitis E, 0.9% (7) had alcoholic liver disease, 0.5% (4) had non-alcoholic fatty liver, 0.8% (6) had primary hepatic carcinoma, and 0.1% (1) had infectious mononucleosis. Serum AMA-M2 level was significantly higher in the PBC patients (vs. other groups, P less than 0.001) with the exception of the patients with PBC/AIH overlap syndrome. Among the 811 patients with AMA-M2 positivity, 88.5% (718) showed AMA positivity and 91.1% (739) showed ANA positivity. Serum alanine transferase (ALT) and aspartate transferase (AST) levels were significantly higher in the drag-induced liver injury patients (527.74+/-684.65 U/L, 490.60+/-716.89 U/L) and the hepatitis E patients (1015.94 ± 165.55 U/L, 665.4 ± 297.14 U/L) than in the PBC patients (96.02 ± 115.56 U/L, 94.82 ± 83.32 U/L) (ALT: F =8.041, P < 0.001, P < 0.001; AST: F =8.066, P < 0.001, P < 0.001). Serum alkaline phosphatase (ALP; 265.16 ± 179.08 U/L) and glutamyl transferase (GGT; 332.02 ± 279.29 U/L) were significantly higher in the PBC patients than in the hepatitis B patients (135.35 ± 123.17 U/L, 140.27 ± 229.24 U/L) and the hepatitis C patients (85.65 ± 27.77 U/L, 92.70 ± 125.72 U/L) (ALP: F=3.911, P =0.01, P=0.001; GGT: F=4.081, P <0.001, P < 0.001). The serum IgM level was significantly higher in the PBC patients (4.60 ± 2.67 g/L) than in the patients with drug-induced liver injury (1.76 ± 1.15 g/L), hepatitis B (2.02 ± 1.41 g/L), hepatitis C (1.48 ± 0.92 g/L), hepatitis E (1.40 ± 0.68 g/L), alcoholic liver disease (1.57 ± 1.07 g/L), non-alcoholic fatty liver (1.05 ± 0.72 g/L), and primary hepatic carcinoma (2.64 ± 2.26 g/L) (F=16.83, P < 0.001, P < 0.001, Probability value < 0.001, Probability value < 0.05, Probability value < 0.01, Probability value < 0.05 respectively).

CONCLUSIONAlthough detection of serum AMA-M2 is an important feature of PBC diagnostic testing,there is a high ratio of serum AMA-M2 detected in patients with drug-induced liver injury, hepatitis B, C and E, alcoholic liver disease, non-alcoholic fatty liver,and primary hepatic carcinoma. The AMA-M2 positive non-PBC patients still require close observation to watch for future development of PBC.

Autoantibodies ; Beijing ; Carcinoma, Hepatocellular ; Chemical and Drug Induced Liver Injury ; Enzyme-Linked Immunosorbent Assay ; Hepatitis B ; Hepatitis C ; Hepatitis, Autoimmune ; Humans ; Liver Cirrhosis, Biliary ; Liver Diseases, Alcoholic ; Liver Function Tests ; Liver Neoplasms ; Retrospective Studies

OBJECTIVETo analyze the HLA class I alleles and haplotypes in Chinese patients with primary biliary cirrhosis (PBC).

METHODSSequencing based typing-polymerase chain reaction (SBT-PCR) was used to investigate the HLA class I alleles of 146 PBC patients and 500 normal controls in northern China. The frequencies of alleles and haplotypes were calculated and compared for the two groups. The chi-square test and Fisher's exact test were used for statistical analyses.

RESULTSThere were 26, 51 and 21 alleles identified at the HLA-A, B and C loci respectively, and the frequencies of these alleles were not significantly different between the PBC and normal control groups.However, the frequencies of A *11:01-B*40:06 and A*02:01-B*l5:01 haplotypes were significantly higher in the PBC group than in the normal control group (7.53% vs. 1.40%, P<0.01, OR=5.38; 6.85% vs. 2.00%, P=0.003, OR=3.425).

CONCLUSIONThis study established the role of HLA class I haplotypes in determining PBC susceptibility in a Chinese population.

Alleles ; Asian Continental Ancestry Group ; China ; Gene Frequency ; Haplotypes ; Histocompatibility Antigens Class I ; Humans ; Liver Cirrhosis, Biliary ; Polymerase Chain Reaction

Objective: To understand the characteristics and clinical significance of anti-soluble liver antigen antibody (anti-SLA) in patients with liver diseases. Methods: Serum samples from seventy-seven patients with anti-SLA were collected from Beijing You'An Hospital during the period between January 2010 and December 2018. Anti-SLA, anti-liver cytosol type 1 antibody (anti-LC1), anti-glycoprotein 210 antibody(anti-gp210) and anti-nuclear body protein sp100 antibody(anti-sp100) were detected by immunoblotting; indirect immunofluorescence assay used for detecting anti-nuclear antibody (ANA), anti-mitochondrial antibody (AMA), anti-smooth muscle antibody (SMA), and anti-liver kidney microsome antibody (anti-LKM). One-way analysis of variance was used to compare the ages of different anti-SLA groups. The non-parametric rank sum test was used to compare the liver function indexes and immunoglobulins in different intensity groups of anti-SLA. P<0.05 was considered statistically significant. Further comparisons were made between the two groups, the correction level α′=0.008 3, P<0.008 3 was considered statistically significant. Results: The average age of 77 anti-SLA positive patients was (52.50±1.25) years old, 70 females (90.9%) and 7 males (9.1%). 80.5% of anti-SLA-positive patients (62/77 cases) were strongly positive at the time of initial diagnosis (+++ to ++++).The Alanine aminotransferase (ALT) level in the SLA++ group was higher than that in the SLA++++ group (232.7 U/L vs 65.6 U/L,χ²=7.751,P=0.005) and the immunoglobulin M(IgM) level in the SLA+++ group was lower than that in the SLA++++ group (1 270 mg/L vs 2 270 mg/L,χ²=8.337,P=0.004).There was no significant difference in age, other liver function and immunological indicators among the different groups.Seventy cases (90.9%) were both anti-SLA and ANA positive, 13 cases (16.9%) were positive with SMA, and none positive with anti-LKM and anti-LC1. Among anti-SLA positive patients, 58 cases were diagnosed with autoimmune hepatitis (AIH), 12 were AIH/primary biliary cholangitis (PBC) overlap syndrome (OS), 2 were drug-induced liver injury, 2 were chronic hepatitis B, and 3 were hepatitis A, hepatitis E and acquired immune deficiency syndrome (AIDS) with liver injury, respectively.Cases of AIH and AIH/PBC OS accounted for 90.9% (70/77 cases) of anti-SLA-positive patients, and 5 of 7 patients diagnosed with non-AIH (and OS) had elevated IgG, showing AIH feature.92.3% (12/13 cases) of anti-SLA with high titers of AMA were diagnosed as AIH/PBC overlap syndrome. Of the 77 anti-SLA-positive patients, 28 (36.4%) had advanced or end-stage liver disease, including decompensated cirrhosis (22 cases), chronic acute liver failure (4 cases), and liver transplantation (1 case) and death from liver failure (1 case). Conclusions Anti-SLA has high diagnostic specificity for AIH;anti-SLA positive in patients with PBC should be an important biomarker for the diagnosis of AIH/PBC overlap syndrome.

Objective To investigate the relation between course and effect of hyperbaric oxygen therapy on patients with spinal cord injuries. Methods The retrospective study of 147 patients with spinal cord injuries were reviewed and on the basis of comprehensive treatment the control group was not treated with hyperbaric oxygen (NHBO group,39 cases),whereas Hyperbaric oxygen treatment group(HBO group,108 cases)were treated with hyperbaric oxygen therapy. According to courses of HBO treatment ,from short term to long term ,the HBO group was divided into 3 subgroups,HBO1,HBO2 and HBO3 subgroup. Evaluation based on the result all selected patients assessed on the six months after the date of surgery. Results There was no significant difference in baseline data between the NHBO group and the HBO group and the overall effectiveness of the HBO group was better than the NHBO group(P<0.01). The efficacy of HBO2 and HBO3 subgroup was better than that of HBO1 subgroup. From the perspective of spinal cord function before and after treatment ,the efficacy of the HBO group was better than that of the NHBO group(P < 0.05).The efficacy of HBO2 and HBO3 subgroup was superior to that of HBO1 subgroup (P < 0.05). However there was no significant difference in total effective rate and spinal cord score between the HBO2 and the HBO3 subgroups. Conclusions Hyperbaric oxygen therapy is effective in the treatment of spinal cord injuries. The treatment for 3~4 courses play the best therapeutic effect.