1.The study on the expression of EBV antibody in patients with systemic lupus erythematosus
Yan LIU ; Jing HUANG ; Lingyun SUN ; Xuebing FENG
Chinese Journal of Rheumatology 2011;15(5):291-295
objective To study the role of anti-EBV antibody in the development of systemic lupus erythematosus(SLE).Methods Peripheral blood was obtained from 55 SLE patients and 29 normal controls,and anti-EBV-VCA IgG,IgA,IgM antibody was detected by enzyme-linked immunosorbent assay[ELISA).Totat RNA was extracted from peripheral blood of 33 SLE patients and 26 normal controls and then reverse transcribed into complementary DNA.Real-time PCR technique was used to determine gene expressions at the transcription level.Comparison of anti-EBV antibody level and interferon inducible gene expression was made between SLE Datients and normal controls by t-test.The associations between anti-EBV antibody level,lupus activitv and IFN inducible gene expression were analyzed by Spearman's correlation analysis.Results (1)The levels of anti-EBV antibodies in SLE patients were significantly increased as compared to those in the normal controls[IgG(41±6)vs(19±6)U/ml,IgA(15.4±1.8)vs(8.3±2.1)U/ml,IgM(7.8±1.0)vs(3.7±1.2)U/ml]cantly elevated in SLE Datients as compared to normal controls[IFN scores 11±13 vs 0±3](all P<0.05),but not correlated with the levels of anti-EBV antibody in patients(all P>0.05).Conclusion Anti-EBV antibody is overproduced in SLE patients but not associated with disease activity as well as the expression level of several major types of I IFN inducible genes,suggesting that EBV infection may not be a major factor mediating the activation of IFN pathway and consequently the exacerbation of SLE.
2.Role of Olf-1/EBF associated zinc finger protein gene in bone marrow mesenchymal stem cells of systemic lupus erythematosus patients
Yan LIU ; Xiaolei MA ; Jing HUANG ; Lingyun SUN ; Xuebing FENG
Chinese Journal of Rheumatology 2012;16(6):364-367
Objective To study the role of OIf-1/EBF associated zinc finger protein (OAZ),a transcription factor encoded by a positional systemic lupus erythematosus (SLE) candidate gene,in the function of mesenchymal stem cells (MSC) of SLE patients by silencing this gene.Methods OAZ mRNA levels of bone marrow MSC obtained from 5 SLE patients and 5 healthy controls were detected by real-time PCR.Bone marrow MSC obtained from 6 SLE patients were incubated with specific siRNAs for 3 days,then cells were harvested for OAZ measurement,Idl-3 and CCL2 mRNA levels were tested by real-time PCR,and levels of CCL2 were detected in culture supernatants using ELISA.Differences between groups were analyzed using t-test or MannWhitney test.Results ① OAZ mRNA levels of bone marrow MSC were significantly elevated in SLE patients (0.013±0.016) compared to healthy controls (0.001±0.000,P=0.009).② After OAZ silencing,the expression levels of OAZ,Id1,Id2 and ld3 mRNA were significantly decreased (△Ct 10.3±0.7,15.2±1.6,8.1±1.4,10.5±0.6 vs 8.7±0.7,14.1±1.2,7.1±1.5,9.8±0.6) (P all <0.05).③ Both the expression levels of CCL2 mRNA (△Ct 2.2±1.1 vs 3.0±1.1 ) and the levels of CCL2 protein in culture supernatants [(341±29) pg/ml vs (304±19) pg/ml] were significantly increased in OAZ silencing group comparing to those in the control group (P all <0.05).Conclusion OAZ gene expression is significantly elevated in bone marrow MSC of SLE patients.OAZ may affect autoantibody production in SLE patients by regulating CCL2 expression.
3.Evaluating type I interferon-inducible gene expression in patients with systemic lupus erythematosus
Jing HUANG ; Rongliang LI ; Lina ZHU ; Lingyun SUN ; Xuebing FENG
Chinese Journal of Internal Medicine 2010;49(1):45-48
Objective To explore the expression levels of interferon-inducible genes in patients with systemic lupus erythematosus ( SLE) , and to validate these gene expressions as potential biomarkers for the differentiation of disease flare and infection.Methods Peripheral blood was obtained from 48 SLE, 16 rheumatoid arthritis ( RA) patients and 26 normal controls, and total RNA was extracted and reverse transcribed into complementary DNA.Real-time PCR technique was used to determine the gene expressions of MX1, OASL,OAS1, ISG15 and LY6E at transcription level.Univariate logistic regression analysis and multivariate conditional logistic regression model were applied to analyze 5 related factors for infection or activity.Results ( 1 ) The expression levels of MX1, OASL, 0AS1, ISG15, and LY6E mRNA in SLE patients were significantly increased as compared with normal controls ( P all < 0.01 ) , while the expression levels of OASL,OAS1 ,ISG15 and LY6E mRNA in SLE patients were also higher than those in RA patients (P all <0.05 ).(2)There were no significant difference between male and female patients of the 5 gene expression in SLE patients.(3) By logistic regression analysis, ISG15 and LY6E were independent risk factors for active SLE patients (P <0.01) , OASL expression was an independent risk factor for SLE patients with infection ( P = 0.003 ).Conclusion All the 5 interferon inducible genes are highly expressed in SLE patients, in which ISG15 and LY6E are independently associated with disease flare, while OASL may be helpful for the evaluation of infection in SLE patients.
4.Adjusting effect of human chorionic gonadotrophin in the vascular endothelial growth factor in rabbit phallus with hypospadias
Xiaofeng YANG ; Debiao LI ; Xuebing HAN ; Dongwen RONG ; Jing LIU
Chinese Journal of Urology 2008;29(9):643-645
Objective To study the function of human chorionie gonadotrophin(HCG) in adjus-ting the vascular endothelial growth factor(VEGF) in rabbit phallus with hypospadias. Methods Rabbit hypospadias mode was made by Finasteride. After the spontaneous delivery, 35 rabbits were divided into 5 groups with 7 in each group. Four groups accepted HCG intramuscular injection for 7 consecutive days with dosages of 100,200,400 and 600 U, respectively. The control group accepted the same dosage of saline injection. Another 7 normal rabbits were used as normal controls without in-tervention. After 3 weeks, the rabbit phallus tissue was collected and the VEGF levels were detected by ELISA. Results Rabbits with hypospadias accepted HCG 100, 200, 400, 600 U injection had the phallus tissue VEGF level of 5.00±2.37,5.63±1.73, 10.35±2.34 and 16.91±2.34 pg/ml, respectively. While the rabbits accepted saline injection had VEGF levels of 3.99±1.19 pg/ml. The normal rabbit phallus tissue contained VEGF level of 14.82±3.32 pg/ml. There were significant differences between normal group and the rabbits accepted HCG 100, 200, 400 U injection (P< 0.05), but there was no difference with the rabbits accepted HCG 600 U injection (P>0.05). The VEGF level in rabbits accepted 400 U HCG injection had significant difference with rabbits accepted 100, 200, 600 U HCG (P<0.05). The VEGF levels in rabbits accepted 100, 200 U HCG injection had no difference with rabbits accepted saline injection(P>0.05). Conclusions The VEGF in rabbit phallus with hypospadias is decreased. HCG of certain dosage could increase VEGF level in rabbit phallus with hypospadias.
5.Increased interferon-inducible gene expression, an useful marker in the diagnosis and disease activity evaluation of systemic lupus erythematosus
Jiyun ZHANG ; Jing HUANG ; Rongliang LI ; Lingyun SUN ; Xuebing FENG
Chinese Journal of Rheumatology 2010;14(12):803-807
Objective To study the role of the expression levels of 5 type Ⅰ interferon (IFN)-inducible genes (LY6E, OAS1, OASL, MX1, and ISG15) in the diagnosis and disease activity evaluation of systemic lupus erythematosus (SLE). Methods Peripheral blood was obtained from 68 SLE patients, 50 patients with other connective tissue diseases and 26 normal controls, and total RNA was extracted and reverse transcribed into complementary DNA. Real-time PCR technique was used to determine gene expressions at transcription level. An IFN score for each individual was calculated according to the expression of 5 1FN genes. Comparisons of gene expression and IFN score were made among groups. The genes expression levels in patients with SLE were analyzed using receiver operative characteristic curve. The association between IFN scores and disease activity, as assessed by the SLEDAI scores and 24 h proteinuria, was analyzed using Spearman correlation analyses. Results ① The expression levels of MX1, OASL, OAS1, ISG15 and LY6E mRNA in SLE patients were significantly increased as compared with normal controls and disease controls (P all<0.01 ).② IFN scores in SLE patients (17.9±29.1) were significantly increased as compared with normal controls (0±3.3)and disease controls (3.0±8.1) (P all<0.01 ). ③ IFN scores area under the ROC curve (AUCROC) was 0.846. When The IFN scores reached 2.56, its sensitivity and specificity for the diagnosis of SLE were 93.1%and 78.3%, respectively. ④ Levels of IFN score was positively correlated with SLEDAI scores (r=0.256,P<0.05) and 24 h proteinuria (r=0.337, P<0.05). Conclusion The 5 IFN-inducible genes are highly expressed in SLE patients. IFN score level is valuable for the diagnosis of SLE and a high IFN score is usually associated with an elevated disease activity.
6.Effect of mesenchymal stem cells transplantation on OAZ signaling pathway in patients with systemic lupus erythematosus
Rongliang LI ; Jing HUANG ; Dandan WANG ; Lingyun SUN ; Xuebing FENG
Chinese Journal of Rheumatology 2010;14(8):516-520
Objective To investigate the in vivo effects of allogeneic mesenchymal stem cells transplantation (MSCT) on OAZ signaling pathway in patients with systemic lupus erythematosus (SLE).Methods Isolated and expand human MSCs from bone marrow cells or umbilical cord of healthy donors were infused into SLE patients. Peripheral blood cells were collected from 10 pre-MSCT patients as well as 1 week and 4 week post-MSCT, and RNA was extracted and reverse transcripted to cDNA. mRNA expression levels of OAZ and Id1-3 were measured by using real-time PCR. Serum levels of IL-10, IL-12, IL-21, CCL2 and ANA were tested by ELISA. Relationships of the gene expression levels with levels of cytokines and ANA were analyzed. Results mRNA expression levels of OAZ, Id1 and Id3 gene in patients with SLE were significantly decreased at week 1(△C:12.4±1.1, 9.7±1.9, 9.7±1.9, 2.1±1.0) and at week 4 (△Ct:13.3±1.2, 10.4±1.5,10.8±1.2, 2.1±1.2) after MSCT when compared to those of the pre-MSCT (△Ct:11.0±0.9, 7.4±2.1, 7.8±2.1, 0.1±1.5 respectively, P all<0.05). Levels of IL-10, IL-21 and ANA were significantly lower 4 week after MSCT than those before (P<0.05); while level of CCL2 was higher than pre-MSCT (P<0.05). Cytokines and ANA levels 1 week after MSCT were not differentially changed comparing to those of the pre-MSCT. Alteration of OAZ mRNA expression levels pre- and post-MSCT were negatively correlated with changes of ANA, IL-21levels and positively correlated with changes of IL-12/IL-10 and CCL2 levels. Conclusions The expression of genes involving in the OAZ signaling pathway is effectively reduced along with the alteration of several cytokines and ANA after allogeneic MSCT in SLE patients. OAZ signaling pathway may play an important role in MSCT treatment for SLE.
7.Effect of apoptosis and gp 130 expression by Cap preconditioning on rats of acute myocardial injured
Ming GU ; Xuebing JING ; Zhaomei CHE ; Tongku LIU ; Yan ZHOU
Chinese Journal of Biochemical Pharmaceutics 2014;(1):43-45
Objective To study the influence of Captopril preconditioning on apoptosis and the expression of gp 130 in rats of acute myocardial injured. Method Wistar rats were randomly divided into three groups:control group, Iso group and Cap preconditioning group. Cardiomyocytes apoptosis was detected with TUNEL method. The expressions of Bcl-2 and Bax proteins in myocardium were tested by immunohistochemistry. The expression of gp 130 was tested with Western blot method. Results The index of cardiomyocytes apoptosis was decreased, the expressions of Bax proteins and gp 130 were decreased and the expression of Bcl-2 proteins was increased in Cap preconditioning group compared with Iso group (P<0.01 or P<0.05). Conclusion The expression of gp 130 could be decreased through Cap preconditioning, which can reduce the cardiomyocytes apoptosis.
8.Correlation between the expression of periostin and Epithelial-mesenchymal transition markers in colorectal cancer
Leilei YAN ; Jing YAO ; Xuebing YAN ; Yu WANG
International Journal of Surgery 2014;41(11):750-754
Objective To investigate the correlation between the Periostin and Epithelial-Mesenchymal Transition(EMT) markers(E-cadheirn and N-cadheirn) in colorectal cancer(CRC),and analyze the relationship between the expression of Periostin and clinicopathological parameters.Methods The expression of Periostin,Ecadheirn and N-cadheirn in 106 cases with primary CRC tumors and corresponding normal tissues were detected by Immunohistoehemistry and the results were analyzed.Results The expression of Periostin,E-cadheirn and N-cad-heirn in tumors were significantly high expression than those in corresponding normal tissues(62/106 vs 21/106,x2 =34.027,P < 0.05 ;43/106 vs 89/106,x2 =42.480,P < 0.05 ; 66/106 vs 19/106,x2 =43.382,P < 0.05).The expression of Periostin in tumors was associated significantly with tumor differentiation,tumor invasion,lymph node metastasis and distance metastasis (x2 =7.752,P =0.007; x2 =5.008,P =0.031 ; x2 =10.227,P =0.002;x2 =8.001,P =0.006).It was negatively correlated with E-cadherin expression (r =-0.435,P < 0.001),but positively correlated with N-cadherin expression (r =0.213,P =0.028).Conclusion Periostin may promote the occurrence and development of CRC by participating in EMT program.
9.A novel mutation in the ABCD1 gene causes adrenomyeloneuropathy by impairing autophagy level
Mingming MA ; Yongkang CHEN ; Xiaoyi JIANG ; Jing YAO ; Xuebing DING ; Xuejing WANG
Chinese Journal of Neurology 2021;54(4):336-342
Objective:To report a Chinese family with a novel ABCD1 gene mutation at c.332T>G (p.V111G) site and discuss its clinical characteristics and molecular mechanism.Methods:The clinical data, laboratory examination, and imaging examination results were analyzed to make the clinical diagnosis of a middle-aged onset patient from the First Affiliated Hospital of Zhengzhou University in May 2017. High-throughput sequencing was used to discover a novel ABCD1 gene mutation. Sanger sequencing was used to find out whether other family members contain the same ABCD1 gene mutation. The pathogenicity of this mutation was explored by protein structure prediction and pathogenicity analysis. Adrenoleukodystrophy protein-green fluorescent protein (ALDP-GFP) and ALDP-GFP (V111G) plasmids were constructed and human embryonic kidney 293 cells were transfected, then immunofluorescence and Western blotting were used to explore the molecular mechanism of this mutation (completed in Henan Provincial People′s Hospital).Results:The proband (a 39-year-old male) was diagnosed as adrenomyeloneuropathy, a subset of X-linked adrenoleukodystrophy, with a novel heterozygous missense mutation in the ABCD1 gene at c.332T>G (p.V111G) site, and his mother and two daughters were all carriers. Protein structure prediction and pathogenicity results suggested that this mutation is pathogenic. Overexpression of ALDP-GFP (V111G) in the human embryonic kidney 293 cells resulted in a significant decrease in the expression levels of ALDP and the abnormal localization from the peroxisomal membrane to the cytoplasm, accompanied by significant down-regulation of LC3-Ⅱ/LC3-Ⅰ and beclin-1.Conclusion:c.332T>G (p.V111G) is a novel pathogenic mutation in the ABCD1 gene, which causes adrenomyeloneuropathy by impairing autophagy.
10.The Status research of ICU Nurses career situations of tertiary general hospital in Shangdong Province
Hong LIU ; Xuebing JING ; Xiaojie FANG ; Tingting GAO ; Cuiping LI ; Xiaoting FAN
Chinese Journal of Practical Nursing 2016;32(25):1978-1981
Objective To understand the status quo of ICU nurses career situations and its influencing factors in Shandong province. Methods A total of 247 ICU nurses were investigated by questionnaires on career condition. Results The career condition score was 79.19 ± 11.63. In addition, statistical significances were revealed by occupational career questionnaire of ICU nurses in different aspect of the scores with age, gender, working years in the ICU, access years to ICU nurses specialist, whether to participate in career training. The scores of career condition and career identity ICU nurses were different, the 40-years-older group was higher than 31-40 years group (P=0.007, P=0.005). The dimension scores of male nurses organizational commitment was higher than that of female nurses(P=0.002). The scores of career condition whose ICU working life more than 16 years were higher than those of within 5 years(P=0.013). The career condition scores of the ICU nurses with access years of specialist qualifications 7-9 years was higher than those of within 3 years(P=0.011). Conclusions The career condition score of ICU clinical nurse specialists was at a medium level, it still needs to be further improved, managers should pay attention to the different levels of nurses to provide career training, in order to promote the professional development of critical care nurses.