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Objective:To explore the clinical teaching effect of case-based learning combined with diagnosis and treatment guidelines in standardized residency training in the department of rheumatology and immunology.Methods:Forty standardized trainees were randomized into two groups. One was the observation group, which adopted case-based learning combined with diagnosis and treatment guidelines. The other one was the control group, which performed the traditional clinical teaching mode. After 4 weeks, the assessment and satisfaction evaluation were carried out among the two groups. SPSS 19.0 was used for t test and Mann-Whitney test. Results:Students in observation group showed significantly better ability of physical examination and case analysis than those in the control group. The satisfaction degree of the students in the observation group was significantly higher in terms of diagnosis and treatment standard, clinical thinking, problem solving ability, self-learning ability and personal profession benefits than that in the control group.Conclusion:Case-based learning with diagnosis and treatment guidelines is an ideal teaching method combining theory with practice perfectly, which can significantly improve the effect of clinical teaching of standardized residency training in the department of rheumatology and immunology.
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Objective To explore the effects of ring finger protein 43 (RNF43) on fibroblast-like synoviocytes (FLS) from patients with rheumatoid arthritis (RA).Methods Synovial tissues from patients with RA treated by knee arthroplasty were used to isolate FLSs by type 2 collagenase.RNF43 lentivirus overexpressing plasmid was constructed and transfected in to RA-FLS.After successful transfection,RNA and super natant of RA-FLS were extracted.QRT-polymerase chain reaction (PCR) and enzyme linked immunosorbent assay (ELISA) were used to detect the mRNA and protein expression levels of matrix metalloproteinase (MMP)-1,MMP-3 and MMP-13.Data were analyzed with Student's t test.Results Transfection efficiency could meet the test requirements when the multiplicity of infection was 40 and was in conjunction with appropriate concentration of polybrene.The mRNA of RNF43 increased for 26158-fold than the control group.In vitro,compared with the control group,RNF43 could significantly inhibit the mRNA of MMP-1,MMP-3 and MMP-13 and MMP-13 [(0.19±0.06),t=28.314,P＜0.05;(0.28±0.07),t=23.413,P＜0.05;(0.21±0.09),t=18.365,P＜0.05]and the protein of MMP-1,MMP-3 and MMP-13 and MMP-13 [(31.0±9.4) pg/ml,(17.1±2.1) pg/ml,t=3.198,P=0.029],MMP-3 [(38.7±8.1) pg/ml,(24.9±3.5) pg/ml,t=3.514,P=0.015],MMP-13 [(35.9±5.4) pg/ml,(20.6±2.9) pg/ml,t=5.632,P=0.001].Conclusion The results of study suggest that RNF43 could inhibit the secretion of MMPs in RA-FLS by suppressing the activity of Wnt signal pathway.
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Objective To explore the molecular pathological mechanism of gout, and to explore the mechanism of how interleukin (IL)-37 influencing PDZK1 protein in gout through nuclear factor κB (NF-κB) pathway. Methods HK-2 cells were stimulated with inflammatory signal IL-37. The expression of PDZK1 and its subcellular localization were detected by real-time fluorescence quantitative polymerase chain reaction (real-time PCR) at different concentrations of IL-37 (defined as group A), PDTC+IL-37 (defined as group B), Wortmannin+IL-37 (defined as group C), respectively.The changes of PDZK1 protein expression in HK-2 cells were detected by adding inhibitor PDTC (NF-κB pathway inhibitor) or Wortmannin (PI3K pathway inhibitor) and inflammatory signal stimulating protein imprinting method. The comparative t test was used for statistical analysis between groups A, B and A and C. Results The average levels of PDZK1 mRNA were as follows:(group A: 28.71 ±0.35, 28.57 ±0.31, 28.78 ±0.28, 28.63 ±0.29, 28.62 ±0.19; group B: 28.71 ±0.31, 28.83 ±0.27, 28.58±0.26, 28.73±0.36, 28.68±0.35;group C:28.81±0.32, 28.91±0.29, 28.72±0.24, 28.59±0.18, 28.58±0.22). There was no significant change in PDZK1 mRNA level in group A and B. The same IL-37 was found in group A and B. The values of T were 5.73, 4.72, 4.69, 5.86 and 6.79, respectively. The P values were all greater than 0.05, and there was no significant difference between the two groups. The values of T were 6.78, 7.13, 7.47, 6.38 and 5.98 in the same IL-37 concentration groups of A and C, respectively, and the P values were all greater than 0.05, with no significant difference. The levels of PDZK1 protein in the three groups by Western blot analysis were as follows: (group A: 0.200±0.082, 0.412±0.032, 0.723±0.063, 1.202±0.021, 1.222±0.023;group B: 0.124±0.064, 0.303±0.081, 0.325±0.062, 0.223±0.071, 0.343±0.052; group C: 0.017±0.022, 0.204± 0.015, 0.187±0.053, 0.302±0.051, 0.404±0.051), The levels of PDZK1 protein in group A treated with different concentrations of IL-37 followed by the concentration of IL-37. The level of PDZK1 protein in group B increased with the increase of IL-37 concentration, but the level of PDZK1 protein in group B was lower than that in the group treated with IL-37 only, and decreased when the concentration of IL-37 was 40 ng/ml.The level of PDZK1 protein in group C increased with the increase of IL-37 concentration, but the level of PDZK1 protein in group C was lower than that in the group treated with IL-37 only, and the same concentration of IL-37 in group A and B. The values of T were 1.83, 1.37, 1.64, 1.57 ,1.49, with P values greater than 0.05. There was no significant difference between the two groups. The values of T were 1.28, 1.37, 1.26, 1.42, 1.39 in the same concentration of IL-37 in group A and C, with P values greater than 0.05, with no significant difference.The results of immunofluorescence analysis showed that the trend of the three groups was basically consistent with that of Western-Blot. Conclusion The pathogenesis of gout induced by IL-37 through PDZK1 protein may not occur at the transcriptional level, but may occur at the level of protein translation.
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Objective To investigate the clinical characteristics of Macrophage activation syndrome (MAS) in patients with rheumatoid arthritis (RA), so as to reduce misdiagnosis. The objective of this paper was to improve the comprehensive and systematic understanding of rheumatoid arthritis complicated with macrophage activation syndrome. Methods The clinical data of one patient with macrophage activation syndrome secondary to RA were analyzed retrospectively, and the related literatures were reviewed. Results The patient was a 65 year old male with ahistory of RA for 14 years. The patient presented with symmetrical multi-joint pain aggravated with stiffness for 14 years and was admitted because of aggravation for 2 weeks. He failed many drugs for the treatment of rheumatoid arthritis was ineffective accompanied with intermittent leukocytopenia. After bone marrow aspiration and biopsy, the phenomenon of phagocytosis of macrophages was clearly diagnosed. He was treatment with high dose corticosteroid +CsA+ human immunoglobulin and his condition wasimproved. Literature was searched in PubMed, Wan Fang medical network database, RA+MAS. Finally, 12 related articles was yielded, and a total of 14 patients, including 8 males, 6 females. Four patients were adults and 10 were children. The shortest duration was 0.5 months, the longest was 24 months. Fever, skin rash, arthritis, enlargement of the liver or spleen, decreased of blood cells count, elevation of liver transaminase, increase of triglyceride, and a series of symptoms and laboratory parameters were observed in the course of the disease. Conclusion When rheumatoid arthritis patients show decreased blood leukocytes and can not be explained by other causes, the differential diagnosis should be carefully performed to rule out secondary macrophage activation syndrome. Always be awake of the risks and dangers of rheumatoid arthritis complicated with macrophage activation syndrome. Early diagnosis and timely are important to improve prognosis.
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Objective To investigate the clinical characteristics of Macrophage activation syndrome (MAS) in patients with rheumatoid arthritis (RA), so as to reduce misdiagnosis. The objective of this paper was to improve the comprehensive and systematic understanding of rheumatoid arthritis complicated with macrophage activation syndrome. Methods The clinical data of one patient with macrophage activation syndrome secondary to RA were analyzed retrospectively, and the related literatures were reviewed. Results The patient was a 65 year old male with ahistory of RA for 14 years. The patient presented with symmetrical multi-joint pain aggravated with stiffness for 14 years and was admitted because of aggravation for 2 weeks. He failed many drugs for the treatment of rheumatoid arthritis was ineffective accompanied with intermittent leukocytopenia. After bone marrow aspiration and biopsy, the phenomenon of phagocytosis of macrophages was clearly diagnosed. He was treatment with high dose corticosteroid +CsA+ human immunoglobulin and his condition wasimproved. Literature was searched in PubMed, Wan Fang medical network database, RA+MAS. Finally, 12 related articles was yielded, and a total of 14 patients, including 8 males, 6 females. Four patients were adults and 10 were children. The shortest duration was 0.5 months, the longest was 24 months. Fever, skin rash, arthritis, enlargement of the liver or spleen, decreased of blood cells count, elevation of liver transaminase, increase of triglyceride, and a series of symptoms and laboratory parameters were observed in the course of the disease. Conclusion When rheumatoid arthritis patients show decreased blood leukocytes and can not be explained by other causes, the differential diagnosis should be carefully performed to rule out secondary macrophage activation syndrome. Always be awake of the risks and dangers of rheumatoid arthritis complicated with macrophage activation syndrome. Early diagnosis and timely are important to improve prognosis.
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Objective To optimize the culture method for rheumatoid arthritis fibroblast-like synoviocytes in vitro,and observe the effect of Dishevelled (Dvl) 2 on vascular endothelial growth factor (VEGF) in rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS).Methods Synovium from RA patients who underwent knee arthroplasties were cut into small piece,and RA-FLS were isolated and cultured in vitro using tissue block method.Dvl 2 lentivirus overexpressing plasmid was constructed and transfected into RAFLS.Q-polymerase chain reaction (PCR) and enzyme linked immunosorbent assay (ELISA) were used to detect the mRNA and protein expression levels of VEGF.Then we used 10 ng/ml tumor necrosis factor (TNF)-α recombinant protein to stimulate the transfected RA-FLS.24 h after stimulation,mRNA and protein expression of VEGF were detected again.Student's t test was used for two group analyses.Results RA-FLS was successfully isolated and cultured in vitro.The multiplicity of infection was 30 and was in conjunction with appropriate concentration of polybrene to promote transfection.Transfection efficiency could meet the test requirements.The mRNA of Dvl 2 increased for 79-fold than the control group.Compared with the control group,Dvl 2 could mildly inhibit RA-FLS secretion of VEGF.After TNF-α stimulation,Dvl 2 could significantly inhibit the VEGF's mRNA (2.15±0.10,2.92±0.47 fold,t=-3.924,P=0.003) and protein [(285±100) pg/ml,(155±61) pg/ml,t=-2.714,P=0.022] expression compared with the control group.Conclusion Dvl 2 can inhibit the effect of TNF-α induced secretion of VEGF in RA-FLS.The specific mechanism needs further study.
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Objective Using an in vivo adeno-associated virus(AAV)-mediated gene transfer technique,this study was designed to evaluate the protective effects of human osteoprotegerin(OPG)transgene against joint destruction in collagen induced arthritis(CIA)model.MethodsAfter CIA was established in the Sprague-Dawley rats,the experimental animals were treated with PBS or rAAV-EGFP or rAAV-hOPG (100μl/d)intra-articular injection 25 days after arthritis induction for 10 days.Paraffin-embedded joints were then analyzed histologically.The joint destruction was evaluated by Larsen Score.The protein expression of OPG,IL-1,MMP-3 was identified by enzyme-linked immunosorbent assay(ELISA).Results Suecessful trans-gene expression was confirmed by the detection of OPG by ELISA and positive fluorescence of the frozen joint section. Image analysis revealed that the expression of OPG significantly protected against joint destruction by 30% compared with the CIA group. Conclusion OPG gene transfer mediated by rAAV effectively protects against bone destruction induced by CIA model. Those data suggest that gene transferring using rAAV-OPG may be a feasible and effective therapeutic approach to treat or prevent joint destruction in inflammatory arthritis.
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Objective This study was designed to investigate the expression changes of osteopro-tegerin (OPG), tartrate-resistant acid phosphatase (TRAP) and vascular endothelial growth factor (VEGF) mRNA in collagen induced arthritis(CIA) rats. Methods After CIA was induced in Sprague-Dawley rats, the experimental animals were treated with PBS or rAAV-EGFP or rAAV-hOPG (100 μl/day) intra-articular injection for 10 days. Messenger RNAs (mRNAs) were obtained from CIA synovium 40days after first immun-ization. Reverse transcriptase-polymerase chain reactions (RT-PCR) were carried out to detect the mRNA encoding OPG, TRAP, VEGF and β-actin, which acted as inner control. The genes detected clearly by RT-PCR were quantified using real-time PCR. Results The expression of all genes was confirmed by specific single bands in RT-PCR. Real-time PCR showed that the expression levels of TRAP and VEGF were increased, whereas those of OPG mRNA were decreased in CIA group compared with normal controls. The intra-articular gene transduction markedly increased the gene copies of OPG by 128.21% (P<0.01). The expression change of OPG in synovium also caused the decrease of the expression levels of TRAP and VEGF by 58.79% (P<0.01)and 17.85% (P>0.05) respectively, however, the expression change of VEGF was not statistically significant. Conclusion OPG gene mediated by rAAV can be successfully tranfered to knee joint synovium in vivo. The results of this study suggest that gene transfer using rAAV-OPG may be a feasible and effective therapeutic approach to treat or prevent joint destruction in inflammatory arthritis.