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Objective To clone human autoimmune antigen SSA/Ro60 and to purify its expression to provide the material basis for the assisted diagnosis of human autoimmune diseases .Methods The SSA/Ro60 gene was cloned by RT‐PCR and directionally inserted into expression vector pPICZ .The recombinant plasmid was transformed into Pichia SMD1168 .The obtained recombinant protein was identified by SDS‐PAGE and Western blotting .Results The amplified full‐length sequence was about 1 .5 kb in size . The pPICZ‐SSA positive clone produced a 60 × 103 recombinant protein which had natural immunogenicity of human autoimmune antigen SSA/Ro60 by SDS‐PAGE and Western blot .Conclusion Human autoimmune antigen SSA/Ro60 is successfully cloned and expressed ,which lays a foundation for diagnosing autoimmune diseases .
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Definiting the workflow and key link of the risk management in medical laboratory by FMEA.Identifying risk factors of the workflow and key link of blood coagulation test by the criteria for laboratory accreditation , such as ISO15189 recognition criteria and CAP laboratory accreditation inspection . Through the evaluation of the blood coagulation test , effective corrective actions and examining performance data periodically , the quality of the blood coagulation test can be improved continuously.
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Objective To investigate the diagnostic value and monitoring application of anti-phospholipase A2 receptor antibodies ( PLA2R ) in patients with idiopathic membranous nephropathy ( IMN) .Methods A retrospective study was used .48 patients were diagnosed as idiopathic membranous nephropathy by puncturing kidney from January of 2013 to June of 2014 in the Hospital of Liaoning Traditional Chinese Medical University and Shengjing Hospital of China Medical University were included. 43 patients were diagnosed as non-idiopathic membranous nephropathy ( NIMN ).35 healthy volunteers were selected as control groups.Serum PLA2R were detected by indirect immunofluorescence.The intensity of antibodies fluorescence and the level of urine protein in 24 hours(24 h PRO) ,serum UREA ,serum CYSC, serum UA,GFR were compared in each groups.Measurement data was shown as mean ±standard deviation;Count data was shown as frequency or composition, t test andχ2 test were used as statistical method.Results (1) Among 48 cases with IMN,37 cases showed positive PLA2R (positive rate 77.08%).All of cases were negative in NIMN group.The sensitivity and the specificity of serum PLA2R were 77.08%and 100%.(2) Compared with control groups, the levels of UREA, CYSC and UA were found statistically significant differences in patients with IMN ( P 0.05 ) . ( 3 ) Positive correlation were found between 24 h PRO(r=0.877,P=0.00),serum UA (r=0.766,P=0.00 ) with the level of PLA2R antibodies fluorescence intensity.( 4 ) There were no correlation between serum PLA2R antibody with IMN pathology aging (r=0.087,0.194,0.182;P=0.598,0.399,0.667).PLA2R positive rate(Ⅰstage:37.50%,Ⅱstage:84.85%, Ⅲstage 85.71%) may increases with the increasing of illness severity.Conclusion Serum PLA2R antibody would be a new laboratory diagnosis standard in patients with IMN.(Chin J Lab Med, 2015, 38:595-599 ) Objective To investigate the diagnostic value and monitoring application of anti-phospholipase A2 receptor antibodies ( PLA2R ) in patients with idiopathic membranous nephropathy ( IMN) .Methods A retrospective study was used .48 patients were diagnosed as idiopathic membranous nephropathy by puncturing kidney from January of 2013 to June of 2014 in the Hospital of Liaoning Traditional Chinese Medical University and Shengjing Hospital of China Medical University were included. 43 patients were diagnosed as non-idiopathic membranous nephropathy ( NIMN ).35 healthy volunteers were selected as control groups.Serum PLA2R were detected by indirect immunofluorescence.The intensity of antibodies fluorescence and the level of urine protein in 24 hours(24 h PRO) ,serum UREA ,serum CYSC, serum UA,GFR were compared in each groups.Measurement data was shown as mean ±standard deviation;Count data was shown as frequency or composition, t test andχ2 test were used as statistical method.Results (1) Among 48 cases with IMN,37 cases showed positive PLA2R (positive rate 77.08%).All of cases were negative in NIMN group.The sensitivity and the specificity of serum PLA2R were 77.08%and 100%.(2) Compared with control groups, the levels of UREA, CYSC and UA were found statistically significant differences in patients with IMN ( P 0.05 ) . ( 3 ) Positive correlation were found between 24 h PRO(r=0.877,P=0.00),serum UA (r=0.766,P=0.00 ) with the level of PLA2R antibodies fluorescence intensity.( 4 ) There were no correlation between serum PLA2R antibody with IMN pathology aging (r=0.087,0.194,0.182;P=0.598,0.399,0.667).PLA2R positive rate(Ⅰstage:37.50%,Ⅱstage:84.85%, Ⅲstage 85.71%) may increases with the increasing of illness severity.Conclusion Serum PLA2R antibody would be a new laboratory diagnosis standard in patients with IMN.
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This review summarizes the epidemiology, etiology, clinical manifestation, treatment, follow-up of neonatal lupus erythematosus with focus on new discoveries on the etiology of the disease in recent years including anti-SSA/Ro and anti-SSB/La antibodies, serotonin (5-hydroxytryptamine 4), apoptosis of cardiac cells, calcium channels, maternal micro-chimera, genetic variants, to improve clinician awareness of the disease.
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ObjectiveTo investigate the combined effects of one to one whole process companion delivery model on emotional status and postnatal recovery of parturients.Methods350 cases adopted one to one whole process companion delivery model were set as the observation group,another 350 cases who did not received companion delivery model were set as the control group.The emotional state and postpartum recovery (the amount of vaginal bleeding after childbirth,time of the uterus return to normal size,and the incidence of urinary retention 6 h after delivery,colostrum time and fasting time) were compared between the two groups.ResultsIn the observation group,312 cases were mild anxiety(89.1％),24 cases were with moderate anxiety (6.9％),14 cases were with severe anxiety (4.0％).In the control group,32 patients with mild anxiety (9.1％),237 cases were with moderate anxiety(67.7％),81 patients were with severe anxiety(23.2％),the difference was significant.In the observation group,postpartum vaginal bleeding was(48.6 ± 9.7) ml,time length of the size of the uterus return to normal was(4.5 ± 3.1 ) weeks,the incidence of postpartum urinary retention 6 hours after delivery was 1.4％,colostrum time (16.4 ± 5.3) h,and fasting time was (12.4 ± 2.1 ) h.In the control group,postpartum vaginal bleeding was (56.5 ± 11.2) ml,time length of the size of the uterus return to normal was(7.1 ± 4.2) weeks,the incidence of postpartum urinary retention 6 hours after delivery was 6.9％,colostrum time was (23.5 ± 4.7) h,and fasting time was (15.3 ± 2.6) h,the difference was significant.Conclusions One to one whole process companion delivery model helps reduce maternal anxiety,fears,reduce the incidence of postpartum unfavorable situation effectively and promote early resumption of maternal,it is worthy of clinical application.
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BACKGROUND: Matrix metalloproteinase (MMP) degrades extracellular matrix, which is a necessity of joint destruction in rheumatoid arthritis patients. MMP-2 and MMP-9 can evaluate rheumatoid arthritis and serve as an index to predict progressive destruction of the joint. OBJECTIVE: To observe heterogenous allogeneic umbilical cord blood stem cell (UBSC) transplantation on MMP-2 and MMP-9 expression in the spleen of mice with type Ⅱ collagen-induced arthritis. METHODS: Fetus cord blood was sterilely obtained and cord blood stem cells were separated. The C57BL/6(H-2b) mice were assigned to five groups (n=10). Except normal control group, models of collagen-induced arthritis were established using complete Freund's adjuvant + type Ⅱ collagen. Mice from the methopterin group were intragastrically administered methopterin suspension 0.017 5 g/kg, once every 5 days. Other groups used caudal vein injection. Mice from the model and normal control groups were injected with saline. Mice from the mono-UBSCs group and double-UBSCs group were injected with 2×106/kg UBSCs from one and two parents. At 42 days following injection, animals were sacrificed and the ankle joint was obtained for histopathological detection. MMP-2 and MMP-9 mRNA expression in the spleen was examined using reverse transcription-polymerase chain reaction. RESULTS AND CONCLUSION: Double-UBSC transplantation could significantly inhibit inflammatory cell infiltration in synovial tissue of mice with type Ⅱ collagen-induced arthritis, repaired impaired cartilage tissue. The repair effect was better than that in methopterin group and mono-UBSCs group. MMP-2 and MMP-9 mRNA expression in the spleen was significantly lower in the double-UBSCs group than the mono-UBSCs group (P < 0.01). These suggest that heterogenous allogeneic double-UBSCs transplantation participated in pathological changes in rheumatoid arthritis cartilage and in synthesis of cartilage extracellular matrix and effectively treated rheumatoid arthritis by regulating MMP-2 and MMP-9 mRNA expression.
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BACKGROUND: The occurrenceand development of rheumatoid arthritis were strongly associated with unbalance proliferation and apoptosis of synovial cells and lymphocytes. Some synovial cell apoptosis was abnormal. OBJECTIVE: To observe effects of heterogenic double umbilical cord blood stem cell transplantation on Bcl-2 and Bax expression in mice with type Ⅱcollagen-induced arthritis. METHODS: C57BL/6(H-2b) mice were induced with Freund's complete adjuvant and type Ⅱcollagen to establish mouse models of type Ⅱcollagen-induced arthritis.At 2days after secondary immunity incubation, mice were injected with saline via tail vein in the model and normal control groups. Umbilical cord blood hemopoietic stem cells were injected into mouse tail vein in the UBSCs transplantation groups (single dose:2×106/50 g; double dose: 1×106/50 g each, totally 2×106/50 g). Mice were intragastrically administrated methotrexate in the methotrexate positive control group, 0.017 5 g/kg once, once every 5 days, totally six times. RESULTS AND CONCLUSION: Joint tissue below knee and elbow was obtained at42 days following transplantation. Histopathology displayed that smooth and glossy articular surface, no inflammatory cell infiltration in the synovial layer and normal chondrocytes in the normal control group. Hyperplasia, a lot of inflammatory cell infiltration and damaged cartilage surface were visible in the model group. Slight hyperplasia and a little inflammatory cell infiltration were detectable in the methotrexate positive controlgroup and single UBSCs transplantation group. Double UBSCs transplantation group exhibited smooth and glossy articular cartilage surface, no damage, a little inflammatory cell infiltration. Immunohistochemistry demonstrated that Bcl-2 and Bax expression was lower in the double UBSCs transplantation group compared with single UBSCs transplantation group (P 0.05). Results suggest that double umbilical cord blood stem cells can induce synovial cell apoptosis in mice with type Ⅱcollagen-induced arthritis in a certain number and action time, and protect synovial membrane against damage.
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Objective To study the changes of serum levels of T-cell subgroups,cytokines and immunoglobulins on the pathogenesis of asthma in children,and explore the role of them in the pathogenesis of asthma. Methods Forty children with asthma (asthma attack group and asthma convalescence group) and 25 healthy children (control group) were enrolled in this study.The level of CD3+CD4+ and CD8+T-cell were determined using flow cytometry (FCM).The levels of TNF-α,IL-6,IL-8 and IgE were determined using ELISA, the level of IgG,IgA and IgM were detected by immunoturbidimetry.Results Compated with control group,there were higher levels of CD3+ and CD4+T-cell and CD4+/CD8+ in asthma attack group (P＜0.01),and higher levels of CD4+T-cell and CD4+/CD8+ in asthma convalescence group (P＜0.05),while no difference in the level of CD8+T-cell The level of CD4+T-cell and CD4+/CD8+ were higher in asthma convalescence group than those in control group (P＜0.05).The level so IL-6,IL-8 and TNF-αin asthma attack group were higher than those in asthma convalescence group and control group (P＜0.05 or＜0.01),the level of TNF-α was significantly higher in asthma convalescence than that in control group (P＜0.05).There were higher levels of IgE and IgG in astham attack group than those in asthma convalescence group and control group (P＜0.01 or＜0.05),the level of IgA was lower than than in conrtol group (P＜0.01).There was higher level of IgE in asthma convalescence group than that in control group (P＜0.01).Conclusions The immune imbalance exists in children with asthma at both attack and convalescence stages.A long-time anti-allergy trealment for childhood nsthma is necessary.
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Objective To investigate the effect of compound preparation of Astragalus mongho-licus on subsets of lymphocytes in patients with myasthenia gravis (MG), and to explore its regulatory mechanism. Methods 60 MG patients were randomly divided into test group (n=30, given compoundpreparation of Astragalus mongholicus for 12 wks) and control group (n=30, given prednisone tablets for 12 wks). Flow cytometry was applied to examining the distribution change of the subsets of lym-phocytes in patients before and after treatment. Results After treatment, the percentages of CD4+ T cells and the ratio of CD4+/CD8+ in both groups decreased compared with those before treatment.The ratio of CD4+/CD8+ in test group was significantly lower than that in control group (P<0.05).The percentage of CD8+ T cells in test group after treatment increased significantly (P<0.05) com-pared with that before treatment, but had no significant difference compared with that in control group(P>0. 05). Conclusion The result from the current study suggests that one of the mechanisms of the compound preparation of Astragalus monghoticus regulating immune response may be achieved through its modulating effect on the distribution of subsets of lymphoeytes and humoral immune func-tion.