1.Pay attention to the detection standardization and rational application of autoimmune liver disease relevant autoantibodies
Chinese Journal of Laboratory Medicine 2014;37(2):81-83
The detection of autoimmune liver disease (AILD) relevant autoantibodies is of important value in the diagnosis and treatment of AILD and especially in autoimmune hepatitis and primary biliary cirrhosis.With the increasing of patients clinically diagnosed AILD,the detection of AILD relevant autoantibodies is gradually clinically concerned and appreciated.As the detection of AILD relevant autoantibodies affected by various factors,there are still many problems in the detection and clinical applications of AILD relevant autoantibodies.We should promote the universal clinical application of AILD relevant autoantibodies,emphasis on the quality management and improve the quality of detection constantly,attend to the standard detection and rational application of AILD relevant autoantibodies.
2.Quality management of autoantibodies detection and the current situation and problems of its clinical applications
Chinese Journal of Laboratory Medicine 2013;36(8):673-676
The detection of autoantibodies is of great value in the diagnosis and treatment of autoimmune diseases.The popular autoantibody screening promotes the rapid development of the clinical awareness,diagnosis and treatment of autoimmune diseases,which in turn results in the increasing demand of autoantibody detection and continuous improvement the detection quality.In order to make better use of autoantibodies results during the diagnosis and treatment of autoimmune diseases,the workers of autoantibodies detection should understand various affecting factors of the autoantibodies detection completely,emphasis on the autoantibodies quality management and improve the quality of detection constantly ; Aware of the complexity and specificity of autoantibodies fully and participate in the selection of autoantibodies detection and correct interpretation of the clinical significance of autoantibodies actively;Emphasis on the clinical application of autoantibodies and promote the universal clinical application of autoantibodies.
3.A pilot study of protein fingerprinting in brain-gut interaction model of irritable bowel syndrome
Ru ZHANG ; Hong Lü ; Jiaming QIAN ; Yongzhe LI ; Chaojun HU
Chinese Journal of Internal Medicine 2010;49(2):134-137
Objective Matrix-assisted laser desorption ionization-time of might-mass spectrometry (MALDI-TOF-MS) was utilized to analyze the protein fingerprint in brain-gut interaction of irritable bowel syndrome (IBS) model rats' colon, so as to find the clues for IBS. Methods Fourteen healthy male adult Wistar rats were selected and divided into a control and a chronic and acute stress ( CAS) group. Colon motility, visceral sensation and behavior changes of rats were detected to evaluate the model. MALDI-TOF-MS was used to observe the overall view of protein in colon so as to study whether there are abnormalities of protein levels in IBS. Results As compared with those in the control group, the number of fecal pellets [ (6. 00 ± 1. 69 ) pellets/1 h vs ( 1. 14 ± 0. 69 ) pellets/1 h, P < 0. 01 ] and frequency of abdominal contraction induced by colorectal distention (CRD) increased, while the amount of weight gain [ (298. 88 ± 18.61)gvs (348. 00±12. 44)g, P<0.01] and consumption of sucrose solutions [ (13. 63 ± 1. 69) ml/1 h vs (19.00±3.06) ml/1 h, P<0.05] decreased in the CAS group (P <0. 05). As far as protein/peptide quality different peak was concerned, CAS rats had 12 different peaks compared with the control rats. The different proteins could be divided into 4 types, which were related to iron secretion, protein synthesis, G protein system and immunity. The protein levels of the model group were higher than those in the control group (P < 0. 05). Conclusions The CAS rats integrate the major characteristics of IBS such as altered colon motility, higher visceral hypersensitivity and psychiatric disorder and can mimic the brain-gut interaction of IBS partly. The detection of differential proteins provides reference for the pathogenesis and treatment of IBS.
4.Association between CD4~+CD25~+ regulative T cells and systemic lupus erythematosus disease active stage
Chaojun HU ; Yongzhe LI ; Dinghua LIU ; Dawei TONG ; Shulan ZHANG
Chinese Journal of Immunology 1985;0(05):-
Objective: To investigate the ratios of peripheral blood CD4+CD25+,CD4+CD8+ regulative T cells of systemic lupus erythematosus(SLE) patients, and explore the association with disease active stage,nephropathy,serum anti-ds-DNA antibody,and both IgG and C3 levels. Methods: The percentage of CD4+CD25+T cells and CD4+CD8+T cells of peripheral blood from patients with systemic lupus erythematosus(SLE)(30 females and 7 males),30 rheumatism controls and 30 normal individuals were measured by flowcytometry. Results: Patients with active disease had statisitically lower levels of CD4+CD25+T cells than did normal controls(P
5.The study of CD4~+ CD8~+、CD4~+ CD25~+ regulative T cells in peripheral blood of patients with primary biliary cirrhosis and its association with the hepatic damnification
Yongzhe LI ; Chaojun HU ; Dinghua LIU ; Dawei TONG ; Shulan ZHANG
Chinese Journal of Laboratory Medicine 2003;0(12):-
Objectives To investigate the ratios of peripheral blood CD4+CD8+ and CD4+CD25+ regulative T cells, and explore the association with hepatic damnification and anti-AMA-M2 antibodies.Methods The percentage of CD4+CD8+T cells and CD4+CD25+T cells in peripheral blood from patients with primary biliary cirrhosis(PBC) (n=27)、26 patients with other hepatic desease、30 normal individuals were measured by flowcytometry.Results Patients with PBC had statistically higher levels of CD4+CD25+T cells than the patients with other hepatic disease (P
6.DeveIopment and EvaIuation of ReIiabiIity and VaIidity of the Chinese Version of HHIE-S
Guoqing WANG ; Chaojun LI ; Wenjun GUAN ; Junwei XIONG ; Shaojing KUANG ; Yaqin HU
Journal of Audiology and Speech Pathology 2014;(6):568-572
Objective The study aimed to study and translate the English edition of HHIE-S into Chinese, and verify the reliability and validity of the scale.Methods The Chinese scale was formed by translation,back-translation,revision and other steps.A total of 170 elder sujbects with normal hearing and presbycusis were sur-veyed.One to two weeks later,they were resurveyed.ResuIts The Chinese version of HHIE-S included 10 entries which were completely retained for their strong correlations.The scale had two subscales:emotional and situation-al,the Cronbach's αcoefficient of the scale were 0.889 and 0.924,respectively,the total coefficient was 0.935;the split-half reliability outcomes of the two subscales were 0.836 and 0.903.The total split -half reliability was 0.836 and the test-retest reliabilities of the two subscales and the ten items were between 0.749 and 0.921.The total scale was was 0.963.For the validity test:the correlation between the ten items and two subscales were 0.750 and 0.927,the correlation between the ten items and the total were 0.659 and 0.878,respectively.The varimax ro-tation factor analysis identified two principal factors:emotional subscale and situational subscale.The two factors would be used to explain 73.874% of the ten items.The factor loadings of 10 entries in the scale were all between 0.684 and 0.871 and the factor loadings of all entries on the corresponding factors were greater than 0.60,consist-ent with the source scale.The average hearing threshold was considered as the gold standard,the two subscales and the total scale were positively correlated to the pure-tone average.ConcIusion The Chinese version of HHIE-S conformed to the characteristics of Chinese culture,and easily accepted by elderly people.The scale had stable structure,satisfactory reliability and validity,provided the basis for a preliminary hearing screening of elder Chinese.
7.The detection of scleroderma-related autoantibodies and its clinical significance in 135 Chinese patients with systemic sclerosis
Yina BAI ; Qian WANG ; Chaojun HU ; Dong XU ; Yong HOU ; Mengtao LI ; Jiulang ZHAO ; Xiaofeng ZENG
Chinese Journal of Microbiology and Immunology 2011;31(5):452-455
Objective To detect the expression of scleroderma-related autoantibodies, such as anti-Scl-70, anli-centromere antibody ( ACA)and anti-RNA polymerase Ⅲ ( ARA) , and their relationship with clinical features in Chinese systemic sclerosis (SSc) patients. Methods One hundred and thirty-five Chinese SSc patients from the clinical database of the Scleroderma Trials and Research Group proposed by European League Against Rheumatism's Scheroderma Trial and Research Group( EUSTAR) were consecutively enrolled. The expression of ARA, anti-Scl-70 and ACA were detected through linear immunoblotting, double immunodiffusion and indirect irnmunofluorescence, respectively. The relevance between the existing of autoantibodies and clinical manifestations was analyzed statistically. Results Among the 135 Chinese SSc patients, the prevalence of anti-Scl-70, ACA, ARA were 49. 6% , 13.3 % and 8.9% respectively. Patients with anti-Scl-70 antibody had significantly shorter disease course [(71 ±59) month vs (90 ± 103) month, P = 0.041] , higher proportion of interstitial lung disease ( P = 0. 031) but lower of pulmonary arterial hypertension (P =0.042). Modified Rodnan's skin score (P=0.008) and prevalence of facial and cervical cutaneous sclerosis (P = 0. 002) , distal (to elbow/knee ) cutaneous sclerosis ( P = 0. 004 ) and digital pitting scarring/disappear of digital pad were all significantly higher in anti-Scl-70 positive group. Patients with AC A had longer disease course ( P = 0. 036) , lower IgM level ( P = 0. 045) and were less prevalent of interstitial lung disease ( P =0. 045). Patients with ARA had higher serum creatinine and urea nitrogen level ( P < 0.001) although otherwise features had unremarkable differences. Conclusion Scleroderma-related autoantibodies have relevance with different clinical manifestation and detection of these autoantibodies may be helpful to the diagnosis of SSc, organ involvement evaluation and predicting outcomes. The clinical relevances of autoantibodies in Chinese SSc patients may differ from other areas or races.
8.Screening serum markers of primary biliary cirrhosis with high-throughput protein chip encoded by the human genes
Chaojun HU ; Yongzhe LI ; Xi LI ; Shulan ZHANG ; Ping LI ; Lijun LI ; Fengchun ZHANG ; Xiaofeng ZENG
Chinese Journal of Laboratory Medicine 2010;33(11):1073-1078
Objective To screen serum markers in patients with PBC by high-throughput protein chips encoded by the human genome. Methods High-throughput protein chips (contains a total of 38 400protein spots, including 17 718 human genes encoding proteins) were used to screen sera from 21 PBC patients, 20 disease control patients and 10 normal controls. Bioinformatics software was used to analyze information and statistical software was used to analyze the data to confirm the serum markers of PBC. Results The detection rate of protein spots using anti-GST antibody on the chip was 97. 6%, and the signal intensity correlation coefficient of double protein spots was 0. 98. Four serum markers( PDHA1, DBT,DLAT and HK1 )were screened by high-throughput protein chips between PBC group and the control group with a statistically significant. The positive rate of the four markers in the three groups was 66. 67% ( 14/21), 5.00% (1/20) and 0(0/10); 57. 14% ( 12/21 ), 5.00% (1/20) and 0(0/10); 52. 38% ( 11/21 ),0(0/20) and 0(0/10); 52. 38% ( 11/21 ), 0(0/20) and 0(0/10) respectively. All the four markers were different in the three groups with statistically significant (PDHA1 :x2 = 16. 79, P <0. 01 ;Fisher exact test,P=0. 000; DBT:x2 =12.86, P<0. 01;Fisher exact test, P=0. 004; DLAT and HK1:Fisher exact test,P <0. 01 or 0. 05). Of those markers, antibodies to PDHA1, DBT and DLAT were the component of AMAM2 which had been used as the marker of PBC. Antibody to HK1 was identified as new marker of PBC,whose sensitivity to PBC was 52. 38% and specificity was 100. 00%. There were no serum marker were screened between the AMA-M2 positive and negative PBC patients. Only antibody to CENPB was identified to be significantly expressed between the ACA positive and negative PBC patients ( Fisher exact test, P =0. 000). Conclusions High-throughput protein chip encoded by the human gene is a technology for quick and comprehensive screening of new markers of PBC. Antibodies to HK1 could be used as new marker for PBC with highly sensitivity and specificity. No serum marker is found between the AMA-M2 positive and negative PBC patients whereas only antibody to CENPB is identified as marker between the ACA positive and negative PBC patients.
9.Cloning and expression of histidyl-tRNA synthetase autoanfigen gene and its clinical application
Shanshan LI ; Yongzhe LI ; Zhixian ZHAO ; Dawei TONG ; Shulan ZHANG ; Chaojun HU ; Weiping YANG
Chinese Journal of Laboratory Medicine 2008;31(2):138-142
Objective To clone and construct the recombinant plasmid containing Jo-1 of HepG2 cells,then purify the protein and identify the immunoreactivity of the recombinant protein.and establish the enzyme linked immunosorbent assay(ELSA)to detect Jo-1 autoantigen correlative antibodies in diagnosis of polymyositis/dermatomyositis.Methods The constructed plasmid was transformed into E.coli.DH5α and BL21(DE3).This fusion protein was purified by Ni-NTA chromatography and its immunnoreactivity was identified by SDS-PAGE and Western blot.ELISA with the fusion protein was established to detect the Jo-1 autoantigen correlative antibodies in sernm samples of 75 patient with PM/DM,30 patients with SLE.30 patients with RA,10 patients with SS and 30 normal controls.Results The sequence of Jo-1 autoantigen gene Was the same as the sequence reported on the literatures.SDS-PAGE gel analysis showed the molecular weisat of fusion protein was approximately 55 000 Da. Western blotting analysis showed that the fusion protein had the same immunoreactivity as human Jo-1 autoantigen.The results of ELISA indicated that the positive rate of anti-Jo-1 antibody was 28%.but the antibody was negative in other controls.There was significant difierence of positivity of the autoantibody between PM/DM and disease controls or normal controls (x2=31.84,P<0.01).Conclusions The plasmid containing Jo-1 is successfully cloned into E.coli.DH5α and BL21 (DE3).EUSA analysis shows its good antigenicity and specificity.
10.Expression of TβRⅡRH mRNA in PBMCs of patients with lupus nephritis
Chen ZHAO ; Yongzhe LI ; Dinghua LIU ; Chaojun HU ; Dawei TONG ; Shulan ZHANG ; Yang ZHAO
Chinese Journal of Rheumatology 2008;12(6):395-397
Objective To investigate the association of TGF-β receptor typeⅡ(TβRⅡ)mRNA with lupus nephritis (LN) and disease activity by testing its expression levelin peripheral blood mononuclear cells (PBMCs).Methotis Forty-four patients with LN were included in this study.They were all had active LN.Twepty-eight LN patients were taking glueocorticoids and/or immunosuppressive agents and sixteen had never taken steroids or immunosuppressive agents.The expression levels of T13R H mRNA were semi-quantitativelydetermined by reverse transcription-polymerase chain reaction(RT-PCR).Resuits The expression levels of TβRⅡ mRNA in PBMCs from LN patients(1.7±1.0)were lower than those of non-lupus nephritis(4.0±3.1) and healthy subiects(4.1±2.5),(P<0.01).The difference of the expression levels between patients who took and had never taken glucocorticoids and/or immunosuppressive drugs was significantly statistically(P<0.05).The expression levels of TβRⅡ mRNA in PBMCs of patients with LN were correlated significantly with the systemic lupus erythematosus disease activity index (SLEDAI)scores(r-0.309.P<0.05),titers of anti-dsDNA antibody(r=-0.401,P<0.01)and serum complement C3 level(r=0.621,P<0.01).Conclusion This study suggests that TβRⅡ may be involved in the development of LN,and the TβRⅡ mRNA expression levels in PBMCs from patients with SLE are significantly correlated with LN activity.Glucocortieoids or immunosuppressive drugs can increase the expression levels of TβRⅡ mRNA and ameliorate renal damage.