1.The progress of copy number variation and its measurement methods in autoimmune diseases
Chinese Journal of Laboratory Medicine 2013;36(11):987-991
Copy number variation (CNV) is one kind of structural variations,which exists extensively in human genome.In addition,it is the crucial source of inter-individual gene differences.Recently,with the successful development of gene array and gene sequencing,molecular methods for CNV testing are more and more standardizing and automating.Therefore,ongoing attentions have been attached to the effects and mechanisms of CNV in complicated diseases like autoimmune diseases.Molecular methods for CNV testing can be divided into two categories:target screening and the whole genome scanning platforms.Currently,the studies about CNV and autoimmune diseases mainly focused on systemic lupus erythematosus and rheumatoid arthritis.Considering the limitations of measurement methods and testing costs,it is still in the infant.
2.Cerebrovascular reactivity by using arterial spin-labeling magnetic resonance imaging in subcortical ischemic vascular disease
Min SHU ; Junjian ZHANG ; Yongzhe GAO ; Hong ZHANG ; Guangyao WU
The Journal of Practical Medicine 2014;(17):2737-2739
Objective To assess regional cerebrovascular reactivity (CVR) and cognition impairment of subcortical ischemic vascular disease (SIVD). To show whether CVR affects cognitive impairment of SIVD patients. Methods Arterial spin-labeling (ASL) by MR image was applied in measuring regional cerebral blood flow (rCBF) of frontal lobe, temporal lobe, parietal lobe, occipital lobes after 5% CO2 inhalation, and the CVR was demonstrated by increase rate of rCBF in all the subject areas. Results (1)The patients with SIVD had reduced CVR in cortex of frontal lobes, white matter of frontal lobes, temporal lobes and occipital lobes(P < 0.05). (2) CVR in the SIVD patients with cognitive impairment decreased in frontal cortex and white matter when compared with the patients without cognitive impairment(P < 0.01). Conclusion These results showed that CVR decreased significantly in cortical gray matter and white matter in elderly patients with SIVD. Also the reduction of CVR in the frontal cortex and white matter was associated with cognitive impairment.
3.Effects of IL-24 gene combined with ionizing radiation on apoptosis in PC-3 cell line
Yongzhe LIU ; Congmei WU ; Guanying NI ; Shunzi JIN
Journal of Jilin University(Medicine Edition) 2006;0(02):-
0.05).The apoptotic percentage was increased significantly after X-rays irradiation with the dose of 6 Gy(P
4.Effects of whole-body irradiation with X-rays on apoptosis in mouse splenocytes and peritoneal macrophages
Ning WU ; Yongzhe LIU ; Ruiming XU ; Yang LIU ; Shunzi JIN
Journal of Jilin University(Medicine Edition) 2006;0(06):-
Objective To study the effects of whole-body irradiation(WBI)with different doses of X-rays on apoptosis in mouse splenocytes and peritoneal macrophages.Methods The apoptosis percentages of mouse splenocytes and peritoneal macrophages were detected with flow cytometry(FCM)at different time after the whole-body X-irradiation using the staining of Annexin-V and PI.Results As compared with the control,the percentage of apoptosis in mouse splenocytes began to increase gradually 24 h after WBI with 2 Gy X-rays(P
5.Cloning and expression of the human asialoglycoprotein receptor H1 subunit and preliminary application
Yan ZHANG ; Yongzhe LI ; Lin WU ; Guozhen LIU ; Shulan ZHANG ; Chaojun HU ; Dawei TONG
Chinese Journal of Laboratory Medicine 2009;32(6):659-663
Objective To clone and express the human asialoglycoprotein receptor(ASGPR) H1 subunit, purify and identify the immunoreactivity of the recombinant protein, and establish the enzyme linked immunosorbent assay (ELISA) to detect anti-ASGPR antibodies in diagnosis of autoimmune hepatitis. Methods The CRDHI cDNA (435 bp) was subcloned into eukaryotic vector PEGH, and the recombinant protein expression was induced by D (+)-Galactose. The recombinant CRDH1 was purified with Glutathione Sepharose 4B, and its immunoreactivity was identified by SDS-PAGE and western blot as well as MALDI-TOF. ELISA was established to detect the anti-ASGPR antibodies in serum samples of 45 patients with AIH, 30 patients with SLE, 30 patients with RA, 10 patients with SS and 30 normal controls. Results The sequencing of recombinant plasmid showed the CRDH1 gene was successfully inserted to the eukaryotic expression vector with correct sequence and open reading frame. The fusion protein showed a molecular weight of 42 500 Da on SDS-PAGE gel and confirmed to be the human ASGPR by MALDI-MS through peptide mass fingerprint analysis with Mascot in human protein database. It shared 98. 34% homology with ASGPR H1 subunit. Western blot analysis showed that the fusion protein had the same immunoreactivity as human ASGPR. The results of ELISA indicated that the positive rate of anti-ASGPR was 35.6% ( 16/45 ), but the ELISA was negative in other control. There was significant difference of positivity of the autoantibodies between AIH and non-AIH controls (χ2 = 31.85,P < 0. 01 ). Conclusions The human plasmid containing ASGPR is successfully clone into Saccharomyces cerevisiae Y258. The recombinant autoantigen owns good antigenicity and specificity. ELISA established with the purified protein shows good specificity for diagnosis of AIH.
6.Clinical significance of different antiphospholipid antibody isotypes in systemic lupus erythematosus
Shulan ZHANG ; Ziyan WU ; Si CHEN ; Yuan LI ; Wen ZHANG ; Yongzhe LI
Chinese Journal of Laboratory Medicine 2014;37(8):597-602
Objective To investigate the prevalence and clinical significance of different subtypes (IgG,IgM and IgA) of anticardiolipin antibodies (aCL) and anti-β2-glycoprotein Ⅰ antibodies (aβ 2GP1),as well as lupus anticoagulant (LA) in systemic lupus erythematosus (SLE).Methods IgG/IgM/IgA,IgG,IgM,IgA aCL and anti-β2GP1 were tested by enzyme-linked immunosorbent assay (ELISA) in 100 patients with SLE (42 patients were diagnosed as secondary antiphospholipid syndrome),44 healthy controls and 32 patients with other connective tissue diseases excluding SLE and antiphospholipid syndrome (APS).Meanwhile,LA was tested by modified Dilute Russell's viper venom time (dRVVT).The correlation between antiphospholipid antibodies and clinical manifestation was analyzed by Spearman correlation analysis.The postiverate of antiphospholipid antibodies in SLE patients,health controls and patients with other connective tissue diseases were compared by chi square test.The concentrations of antiphospholipid antibodies in different groups were compared using independent sample Kruskal Wallis test.The diagnostic efficacy of antiphospholipid antibodies in SLE patients was analyzed by crosstable using clinical diagnosis of APS as gold standard.P < 0.05 was considered statistically significant.Results The prevalence of IgG aCL (x2 =15.031,P < 0.001),IgA/G/M (x2 =11.678,P =0.003) and IgA (x2 =6.17,P =0.036) antiβ2GP1 were significantly higher in patients with SLE than in the other two groups.IgA/G/M (r =0.207,P=0.039),IgG (r=0.230,P=0.021) and IgA (r=0.217,P=0.030) aCL,IgA/G/M (r=0.218,P=0.029) and IgA (r =0.255,P =0.01) anti-β2GP1,as well as LA (r =0.233,P =0.02) were associated with thrombotic events.IgA/G/M anti-β2GP1 (r =0.22,P =0.029) and LA (r =0.254,P =0.011) were associated with pathological pregnancy.23.1% (6/26) aCL positive SLE patients were IgM and/or IgA aCL positive.53.6% (15/28) anti-β2GP1 positive SLE patients were IgM and/or IgA antiβ2GP1 positive.In SLE patients,the specificity and sensitivity of IgA/G/M aCL for APS were 98.3% and 26.2%,respectively.The specificity and sensitivity of IgA/G/M anti-β2GP1 were 84.5% and 40.5%,respectively.The specificity of at least two isotypes positive for APS (both aCL and anti-β2GP1 were 98.3%),was higher than IgG aCL (94.8%) or anti-β2GP1 (93.1%).The sensitivity of at least one isotype of aCL (47.6%) or anti-β2GP1 (42.9%) positive for APS were higher than IgG aCL (40.5%)and anti-β2GP1 (21.4%).Conclusions IgG and IgM aCL together would be better than IgA/G/M aCL for APS screening.IgA/G/M anti-β2GP1 would be better for APS screen due to higher sensitivity and strong association with thromboembolic events and pathologic pregnance.IgA aCL or anti-β2GP1 was associated with thromboembolic events.IgA aCL or anti-β2GP1 would be useful for APS diagnosis in IgG and IgM aCL or anti-β2GPl negative patients.
7.Risk factors for retained common bile duct stones of laparoscopic cholecystectomy
Hongli CUI ; Changyu ZHOU ; Jiandong LIU ; Hongchao AN ; Huazhi LI ; Hongzheng XU ; Yongzhe WU
International Journal of Surgery 2014;41(11):758-762
Objective To investigate the related risk factors caused the retained common bile duct (CBD)stones after laparoscopic cholecystectomy,to provide the evidence for preventing from retained common bile duct stones of laparoscopic cholecystectomy.Methods Selected 654 cases of laparoscopic cholecystectomy patients as the objects whom hospitalized in Beijing Chuiyongliu Hospital from January 2002 to June 2013.All cases were divided into the group of retained CBD stones (27cases) and the group of non-retained CBD stones (627 cases).Collected the potentially relevant factors of two groups with the retained CBD stones,including medical history of jaundice and pancreatitis,common bile duct internal diameter (B Ultrasound) ≥ 8 mm,acute cholecystitis,emergency surgery,gallbladder removal order,calot triangle adhesion,sludge calculus,stones neck incarcerated,fulltype stones,cystic duct thickening,cystic duct reserved≥ 1 cm,the minimum diameter stones ≤5 mm,the number of gallbladder stones ≥ 5,partial cholecystectomy,purulent bile.Statistics analyses was proceeded using the IBM SPSS 20.0.Result Through dichotomy logistic regression analysis to the univariate analysis results with statistical significance,sorted the results according to the influence degree,found the independent risk factors:common bile duct internal diameter (B Ultrasound) ≥8 mm,gallbladder removal order (retrograde removal),medical history of jaundice pancreatitis,cystic duct reserved ≥ 1 cm,sludge calculus,in total of 5 indexes,which caused the retained CBD stones after laparoscopic cholecystectomy.Conclusion There are several independent risk factors for retained CBD stoes after LC such as medical history of jaundice and pancreatitis,common bile duct internal diameter (B Ultrasound) ≥ 8 mm,the order of cholecystectomy (retrograde remoral),shudeg calculus and the remaining length of bile duct ≥ 1 cm.The surgeons should pay close attention to them and take appropriate measures in the preoperation and intraoperation of LC,which contribute to preventing the acurence of postoperative retaimed CBD stone.
8.A Maternal Health Care System Based on Mobile Health Care.
Xin DU ; Weijie ZENG ; Chengwei LI ; Junwei XUE ; Xiuyong WU ; Yinjia LIU ; Yuxin WAN ; Yiru ZHANG ; Yurong JI ; Lei WU ; Yongzhe YANG ; Yue ZHANG ; Bin ZHU ; Yueshan HUANG ; Kai WU
Journal of Biomedical Engineering 2016;33(1):2-7
Wearable devices are used in the new design of the maternal health care system to detect electrocardiogram and oxygen saturation signal while smart terminals are used to achieve assessments and input maternal clinical information. All the results combined with biochemical analysis from hospital are uploaded to cloud server by mobile Internet. Machine learning algorithms are used for data mining of all information of subjects. This system can achieve the assessment and care of maternal physical health as well as mental health. Moreover, the system can send the results and health guidance to smart terminals.
Algorithms
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Clothing
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Electrocardiography
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Equipment Design
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Female
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Humans
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Internet
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Machine Learning
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Maternal Health
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Monitoring, Ambulatory
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instrumentation
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Telemedicine
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instrumentation
9.Prozone effect on indirect immunofluorescence assay for anti-nuclear antibody testing
Qiongwen HU ; Chaojun HU ; Ping LI ; Chuiwen DENG ; Ziyan WU ; Xiaofeng ZENG ; Fengchun ZHANG ; Pu LIAO ; Yongzhe LI ; Shulan ZHANG
International Journal of Laboratory Medicine 2019;40(2):210-213
Objective To explore the influence of prozone effect on anti-nuclear antibodies (ANA) testing by indirect immunofluorescence assay (IIFA).Methods The samples with high titer of ANA (≥1∶1 000) were selected from 880fresh serum samples, and were subsequently diluted in 1∶100, 1∶1 000and 1∶10 000ratio.Prozone effect was defined as fluorescence intensity from 1∶1 000dilution was stronger than that from1∶100dilution.The samples with prozone effect were determined manually or by Sprinter XL and EUROPattern.The samples with prozone effect were further characterized by combinations of fluorescence patterns, fluorescence intensities and autoantibody specificities.Results A total of 880samples were tested.Importantly, 34samples displayed prozone effect (3.86%in total and 29.57%in samples with ANA≥1∶1 000).Interestingly, prozone effect was identified by manual detection as well as by Sprinter XL with similar fluorescence patterns and fluorescence intensities.Notably, EUROPattern can only select the central area for identification.Among all samples with prozone effect, 74.42%samples exhibited fluorescence intensities of≥1∶10 000.Speckled pattern was the most prevalent fluorescence patterns in samples with prozone effect (46.51%).In addition, anti-RNP antibodies (62.79%) were the most popular autoantibodies in samples with prozone effect, followed by anti-dsDNA antibodies (51.16%) and anti-SSA antibodies (51.16%).Conclusion Prozone effect was present in ANA testing, especially in samples with high titers, resulting in underestimating the titers.The study highlighted that special attention should be paid to the prozone effect in clinical practice.
10.Preliminary study on the clinical application of four cytokines in serum of autoimmune diseases
Wei LI ; Ziyan WU ; Leili MAO ; Xinyao ZHANG ; Songxin YAN ; Honglin XU ; Futai FENG ; Shulan ZHANG ; Yongzhe LI
Chinese Journal of Laboratory Medicine 2023;46(11):1173-1179
Objectives:the purpose of this study was to systematically evaluate the clinical value of cytokines in autoimmune diseases (AID). It was a kind of complex disease, and its pathogenesis involved cytokines, autoantibodies, immune cells and other immune factors. Especially some AID, such as Adult still′s disease (AOSD) and Takayasu arteritis(TA), had no specific biomarkers at present. This study was a retrospective case-control study.Methods:the data of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-8(IL-8) and interleukin-10(IL-10) in 834 AID patients from January 2019 to August 2022 were collected, and the serum levels of those cytokines in 30 healthy controls (HC) were detected at the same time. And AOSD, TA, systemic lupus erythematosus (SLE) and Behcet′s syndrome (BS) were divided into active group and inactive group. In addition, we also made a subgroup analysis of two important organs involved in SLE (kidney and nervous system). GraphPad Prism 9 and R 4.2.2 software were used. Nonparametric tests (Kruskal-Wallis H test, Mann-Whitney U test) were used to compare the differences among groups, and Dunn′s method was used to correct the false positive caused by multiple tests. Results:To compare the level of IL-6 in each group, except Behcet syndrome (BS) group and antiphospholipid syndrome (APS) group, the serum IL-6 level of AID group was higher than that of HC group, with antineutrophil cytoplasmic antibodies associated vasculitis(AAV) [3.85(2.00,8.55) pg/ml], idiopathic inflammatory myopathies(IIM) [7.80(2.50,6.50)pg/ml], IgG4-related disease(IgG4RD) [3.65(2.08,12.83) pg/ml], rheumatoid arthritis (RA) [5.50(2.20,16.10) pg/ml], SLE[4.70(2.75,16.55) pg/ml], Sj?gren syndrome(SS) [3.20(2.00,8.90) pg/ml], systemic sclerosis(SSc) [2.70(2.00,8.90) pg/ml], TA[3.40 (2.00,6.50) pg/ml], other AID diseases[4.40(2.00,11.10) pg/ml], especially AOSD [15.20(2.10, 39.20) pg/ml]. After correction, the differences were statistically significant ( P c<0.05). At the same time, the levels of serum TNF-α [7.40(5.60,10.95) pg/ml]and IL-10 [5.00(5.00, 7.58) pg/ml] in AOSD group were significantly higher than those in HC group[7.15(5.93,8.00) pg/ml,5.00(5.00,5.00) pg/ml] after correction ( P c<0.05). At the same time, the levels of serum TNF-α and IL-10 in AOSD group were higher than those in HC group. The serum levels of IL-6 and IL-8 in patients with active AOSD, BS, SLE and TA were significantly higher than those in patients without active disease (all P<0.05). In addition, the level of serum IL-8 in lupus nephritis group was significantly higher than that in non-lupus nephritis group ( P<0.05). At the same time, the serum levels of IL-6, IL-8 and TNF-α in neuropsychiatric lupus erythematosus group were significantly higher than those in non-neuropsychiatric lupus erythematosus group ( P<0.05), but there was no significant difference in IL-10 between neuropsychiatric lupus group and non-neuropsychiatric lupus erythematosus group. Conclusions:there was a close relationship between AID and cytokines. At present, the change of serum IL-6 level was the most classic one in clinical routine.