[Medical visit status and clinical features in patients with IgG4 related disease].

OBJECTIVE: To understand the medical treatment and clinical characteristics of patients with IgG4-related disease (IgG4-RD) with complex clinical manifestations and easy to be misdiagnosed and missed, and to improve the recognition of this disease among doctors from relevant medical departments. METHODS: A retrospective analysis was conducted on the medical records of patients diagnosed with IgG4-RD who were hospitalized and discharged from Peking University Third Hospital from January 1, 2012 to December 31, 2022. The patient' s medical visit status, clinical manifestations, laboratory examinations, diagnosis, and treatment information were summarized. RESULTS: A total of 116 patients diagnosed with IgG4-RD were included in this study, with a male to female ratio of 2. 52∶ 1 and an average age of (61.83±10.80) years. The departments for initial visits were gastroenterology, general surgery, and ophthalmology. While the departments responsible for definitive diagnosis were gastroenterology, rheumatology and immunology, and respiratory medicine. Twenty-one patients (18. 10%) required consultation and treatment from three or more departments before receiving a definitive diagnosis. The median time from symptom onset to the initial clinic visit was 2 (1, 7) months, and the median time from symptom onset to diagnosis was 1 (1, 12) month. Twenty-four patients (20.69%) underwent surgical resection of the affected sites before diagnosis. According to the classification criteria of IgG4-RD, sixty-eight (58.62%) cases were diagnosed definitively, eight (6.9%) cases were likely to be diagnosed, and 40 (34.48%) cases were suspected to be diagnosed. In the 68 definitively diagnosed patients, the most commonly affected organs were submandibular gland, the pancreas, biliary tract, parotid in sequence. The median serum IgG4 (IgG4, immunoglobulin G4) level was 6.16 (3. 61, 12. 30) g/L. Fifty-seven patients (83.82%) were treated with glucocorticoids, and 14 patients (20.59%) were treated with immunosuppressants. The use of immunosuppressants was mainly in the rheumatology and immunology department (78. 57%). CONCLUSION: IgG4-RD is more common in elderly males, with submandibular gland, the pancreas, biliary tract, and parotid being most commonly affected. The distribution of initial visit departments in patients is wide. The proportion of definitive diagnosis based on pathology is relatively low. In terms of treatment, the main approach is steroid treatment, while the use of immunosuppres-sants is not widespread.

[Comparison of clinical and immunological characteristics between primary Sjögren's syndrome patients with positive and negative anti-SSB antibody].

OBJECTIVE: To analyze the differences of clinical manifestations and laboratory features between primary Sjögren's syndrome (pSS) patients with positive and negative anti-Sjögren's syndrome type B (SSB) antibody. METHODS: The clinical data of pSS patients hospitalized in Department of Rheumato-logy and Immunology, Peking University Third Hospital were retrospectively analyzed to investigate the differences of clinical and laboratory features between anti-SSB positive and negative groups. The t test, Mann-Whitney U test, Chi-square test and Fisher's exact probability were used for analysis. RESULTS: A total of 142 pSS patients were enrolled in this study, including 137 females and 5 males with a mean age of (54.8±13.3) years. The anti-SSB positive group included 44 patients accounting for 31.0% of the pSS patients. The anti-SSB positive pSS patients were younger at disease onset and at visit [age at visit: (50.9±14.5) years vs. (56.5±12.4) years; age at onset: (42.2±14.8) years vs. (49.5±15.3) years, P < 0.05]. The patients with anti-SSB positive more frequently presented with rash (29.5% vs. 14.3%, P < 0.05), enlargement of parotid glands (27.3% vs. 8.2%, P < 0.05), renal tubular acidosis (15.9% vs. 4.2%, P < 0.05), immune thrombocytopenia (9.1% vs. 1.0%, P < 0.05), rheumatoid factor (RF) positive (85.0% vs. 49.4%, P < 0.05), higher RF and antinuclear antibody (ANA) titers (median: 89.8 IU/mL vs. 20.5 IU/mL; median: 320 vs. 160, P < 0.05), anti-Sjögren's syndrome type A (SSA) antibody positive (97.7% vs. 64.3%, P < 0.05), elevation of γ globulin (71.4% vs. 38.5%, P < 0.05), higher levels of IgG (median: 21.0 g/L vs. 15.6 g/L, P < 0.05), higher proportions of CD3-CD19+ cells [(21.0±11.9)% vs. (13.7±9.6)%, P < 0.05] and lower proportions of CD3+ cells [(67.2±14.4)% vs. (76.6%±13.1)%, P < 0.05] than those negative. However, the anti-SSB positive group was less likely to show anti-mitochondrial antibodies (AMA)-M2 positivity (10.5% vs. 35.6%, P < 0.05). Glucocorticoids (90.9% vs. 73.5%, P < 0.05) and immunosuppressants (54.5% vs. 36.7%, P < 0.05) were more frequently used in anti-SSB positive pSS patients than those negative. CONCLUSION: The anti-SSB positive pSS patients were younger at disease onset while more frequently presenting with various symptoms, higher levels of other antibodies and activation of B cells than those negative. Glucocorticoids and immunosuppressants were more frequently used, indicating that anti-SSB positive group presented with a more severe clinal phenotype.

[Correlation analysis between body mass index and clinical characteristics of rheumatoid arthritis].

OBJECTIVE: To analyze the clinical features of overweight and obese rheumatoid arthritis (RA)patients, and the relationship between body mass index (BMI) and disease characteristics. METHODS: The demographic data, extra-articular manifestations, comorbidities, and disease activity of RA patients admitted to the Rheumatology and Immunology Department of Peking University Third Hospital from January 2015 to December 2020 were collected, and the above characteristics of overweight and obese RA patients were retrospectively analyzed. According to the WHO, BMI≥30 kg/m2 referred to obese individuals, 25≤BMI < 30 kg/m2 referred to overweight individuals, 18.5≤BMI < 25 kg/m2 referred to normal individuals, BMI < 18.5 kg/m2 referred to reduced body mass individuals. t test was used for the quantitative data in accordance with normal distribution. Wilcoxon rank sum test was used for the quantitative data of non-normal distribution. The qualitative data were analyzed by chi square test. But while 1≤theoretical frequency < 5, Chi square test of corrected four grid table was used. And Fisher exact probability method was used when theoretical frequency < 1. Analyzing whether overweight or obesity was associated with comorbidities using Logistic regression adjusted confounding factors. RESULTS: A total of 481 RA patients were included in this study, with an average BMI value of (23.28±3.75) kg/m2.Of the patients, 31 cases (6.5%) were with BMI < 18.5 kg/m2, 309 cases (64.2%) with 18.5≤ BMI < 25 kg/m2, amounting to 340 cases (70.7%). There were 119 overweight individuals (25≤ BMI < 30 kg/m2, 24.7%) and 22 obese individuals (BMI≥30 kg/m2, 4.6%), totaling 141 (29.3%).The proportion of the overweight and obese RA patients suffering from hypertension (57.4% vs. 39.1%, P < 0.001), diabetes (25.5% vs. 15.0%, P=0.006), hyperlipidemia (22.7% vs. 10.9%, P=0.001), fatty liver (28.4% vs. 7.4%, P < 0.001), osteoarthritis (39.0% vs. 29.4%, P=0.040) was significantly higher, and the proportion of the patients with osteoporosis(59.6% vs. 70.9%, P=0.016) and anemia (36.2% vs. 55.6%, P < 0.001) was significantly lower. However, there was no difference between the two groups in coronary heart disease (5.7% vs. 7.6%, P=0.442), cerebrovascular disease (6.4% vs. 8.8%, P=0.372) and peripheral atherosclerosis (9.2% vs. 7.6%, P=0.565).The median C-reactive protein (CRP, 1.52 mg/dL vs. 2.35 mg/dL, P=0.008), median erythrocyte sedimentation rate (ESR, 34.0 mm/h vs. 50.0 mm/h, P=0.003), pain visual simulation score (VAS) (3.66±3.08 vs. 4.40±2.85, P=0.011), and 28 joint disease activity scores (DAS-28, 5.05±1.60 vs. 5.45±1.52, P=0.010) in the overweight and obese RA group were all lower than those in the normal and reduced weight groups. Multivariate regression analysis showed that overweight and obesity was an independent risk factor for hypertension, diabetes, hyperlipidemia and fatty liver, and had protective effects on osteoporosis and anemia. CONCLUSION: In RA patients, RA disease activity is lower in overweight and obesity patients. Overweight and obesity is associated with hypertension, diabetes and hyperlipidemia, but not with cardiovascular and cerebrovascular diseases.

CXCL16 upregulates RANKL expression in rheumatoid arthritis synovial fibroblasts through the JAK2/STAT3 and p38/MAPK signaling pathway.

OBJECTIVE: To explore the influence of chemokine, CXCL16, on the expression of the receptor activator nuclear factor κB ligand (RANKL) in rheumatoid arthritis (RA) fibroblast-like synoviocytes (RA-FLS). METHODS: The expression of CXCL16/CXCR6 and RANKL in RA or osteoarthritis (OA) patient synovia was examined by Western blot and immunohistochemistry. The serum concentration of CXCL16 and RANKL was measured by enzyme-linked immunosorbent assay (ELISA). RA-FLS were treated with recombinant CXCL16, and RANKL mRNA and protein were measured using PCR, Western blot and ELISA. RESULTS: The synovial expression of CXCL16, CXCR6, and RANKL was higher in RA patients than in patients with OA. The serum CXCL16 and RANKL levels were higher in RA patients compared with OA patients and healthy controls. CXCL16 correlated with erythrocyte sedimentation rate, C reactive protein, disease activity, serum rheumatoid factor, and RANKL. RA-FLS treated with CXCL16 showed markedly increased expression of RANKL. When STAT3 or p38 activation was blocked by an inhibitor, CXCL16 failed to upregulate RANKL expression. In contrast, inhibiting the Akt or Erk pathway did not achieve the same effect. CONCLUSIONS: CXCL16 upregulates RANKL expression in RA-FLS and these effects are mainly mediated by the JAK2/STAT3 and p38/MAPK signaling pathways.

PTPN22 gene polymorphism (C1858T) is associated with susceptibility to type 1 diabetes: a meta-analysis of 19,495 cases and 25,341 controls.

The protein tyrosine phosphatase N22 (PTPN22) gene C1858T polymorphism has been reported to be associated with susceptibility to type 1 diabetes (T1D) in relatively small sample sizes. This study aimed at investigating the pooled association by carrying out a meta-analysis on the published studies. The Medline, EBSCO, and BIOSIS databases were searched to identify eligible studies published in English before June 2012. The association was assessed by odds ratio (OR) with 95% confidence intervals (CI). The presence of heterogeneity and publication bias was explored by using meta-regression analysis and Begg's test, respectively. A total of 28 studies were involved in this meta-analysis. Across all populations, significant associations were found between the PTPN22 C1858T polymorphism and susceptibility to T1D under genotypic (TT vs. CC [OR = 3.656, 95% CI: 3.139-4.257], CT vs. CC [OR = 1.968, 95% CI: 1.683-2.300]), recessive (OR = 3.147, 95% CI: 2.704-3.663), and dominant models (OR = 1.957, 95% CI: 1.817-2.108). In ethnicity- and sex-stratified analyses, similar associations were found among Caucasians and within Caucasian male and female strata. The meta-analysis results suggest that the PTPN22 C1858T polymorphism was associated with susceptibility to T1D among the Caucasian population, and males who carried the -1858T allele were more susceptible to T1D than females.

AG490 inhibits NFATc1 expression and STAT3 activation during RANKL induced osteoclastogenesis.

Commonly, JAK/STAT relays cytokine signals for cell activation and proliferation, and recent studies have shown that the elevated expression of JAK/STAT is associated with the immune rejection of allografts and the inflammatory processes of autoimmune disease. However, the role which JAK2/STAT3 signaling plays in the receptor activator of nuclear factor-κB ligand (RANKL)-mediated osteoclastogenesis is unknown. In this study, we investigated the effects of AG490, specific JAK2 inhibitor, on osteoclast differentiation in vitro. AG490 significantly inhibited osteoclastogenesis in murine osteoclast precursor cell line RAW264.7 induced by RANKL. AG490 suppressed cell proliferation and delayed the G1 to S cell cycle transition. Furthermore, AG490 also suppressed the expression of nuclear factor of activated T cells (NFAT) c1 but not c-Fos in RAW264.7. Subsequently, we investigated various intracellular signaling components associated with osteoclastogenesis. AG490 had no effects on RANKL-induced activation of Akt, ERK1/2. Interestingly, AG490 partly inhibited RANKL-induced phosphorylation of Ser(727) in STAT3. Additionally, down-regulation of STAT3 using siRNA resulted in suppression of TRAP, RANK and NFATc1 expression. In conclusion, we demonstrated that AG490 inhibited RANKL-induced osteoclastogenesis by suppressing NFATc1 production and cell proliferation via the STAT3 pathway. These results suggest that inhibition of JAK2 may be useful for the treatment of bone diseases characterized by excessive osteoclastogenesis.

[Comparison of ACR/EULAR remission criterion of rheumatoid arthritis published in 2011 year with the other common remission criteria].

OBJECTIVE: To evaluate and compare the advantage and utility of the 2011 ACR/EULAR criterion and the other remission criteria of rheumatoid arthritis. METHODS: The questionnaires for RA patients were used for the study. The remission rate and residual disease activity of RA patients were compared according to four criteria of remission, including 2011 ACR/EULAR remission criterion, DAS28, CDAI and ACR. RESULTS: Among the 310 cases, 254 effective questionnaires were obtained. The remission rates of ACR, CDAI,ACR/EULAR and DAS28 were 15.4%, 23.2%,25.2%,38.2%, respectively.ACR criteria is the most stringent criteria, the remission rate of ACR was significantly lower than the other three criteria (P<0.05). There was no residual disease activity of ACR criteria. DAS28 criteria is the laxest criteria,the remission rate and residual disease activity of DAS28 was significantly higher than the other three criteria (P<0.05).There was no difference between CDAI and ACR/EULAR, which were more suitable for clinical practice. CONCLUSION: Among the four criteria, ACR criteria is the most stringent criteria, DAS28 criteria is the laxest criteria, The CDAI and ACR/EULAR criteria were more suitable for clinical practice.

[Expression of Galectins in umbilical cord mesenchymal stem cells].

OBJECTIVE: To investigate the expression of Galectins in umbilical cord mesenchymal stem cells (UC-MSCs) from Wharton's jelly. METHODS: Umbilical cords were obtained sterilely from full term caesarean infants, then mesenchymal stem cells (MSCs) were isolated from Wharton's jelly of the umbilical cord via tissue cultivation. The morphology of UC-MSCs was observed under the optical microscope, and its immunophenotypes were analyzed by flow cytometry. The differentiation of UC-MSCs into the osteoblasts and adipocytes was determined utilizing von Kossa calcium node staining and oil red O staining, respectively. The expression of Galectins at mRNA level was measured by RT-PCR. The levels of secretory Galectin-3 in culture supernatants were detected by sandwich enzyme-linked immunosorbent assay. RESULTS: The UC-MSCs could be generated by tissue cultivation. Flow cytometry showed they highly expressed membrane molecules, such as CD29, CD44, CD73, CD90 and CD105, but did not express hematopoietic specific markers (CD14, CD34, and CD45) and immune rejection related molecule HLA-DR. UC-MSCs could differentiate into osteoblasts or adipocytes under appropriate experimental conditions. At the mRNA level, Galectin-1, 3, 4, 8 and 9 were detected in UC-MSCs. And they also could secrete soluble Galectin-3 in a cell number dependent manner. Statistical differences were obtained among the different cell number incubation groups (F=16.901,P=0.002). However, the secretory manner of Galectin-3 was not time dependent. CONCLUSION: UC-MSCs, derived from Wharton's jelly, were successfully cultured via tissue cultivation, and they could express secretory Galectin-3. All these data laid the foundation for further detecting the immunomodulatory mechanism of UC-MSCs.

The ethanol extract of Scutellaria baicalensis Georgi attenuates complete Freund's adjuvant (CFA)-induced inflammatory pain by suppression of P2X3 receptor.

ETHNOPHARMACOLOGICAL RELEVANCE: Scutellaria baicalensis Georgi (SBG) is a perennial herb with anti-inflammatory, antibacterial, and antioxidant activities, which is traditionally used to treat inflammation of respiratory tract and gastrointestinal tract, abdominal cramps, bacterial and viral infections. Clinically, it is often used to treat inflammatory-related diseases. Research has shown that the ethanol extract of Scutellaria baicalensis Georgi (SGE) has anti-inflammatory effect, and its main components baicalin and baicalein have analgesic effects. However, the mechanism of SGE in relieving inflammatory pain has not been deeply studied. AIM OF THE STUDY: This study aimed to evaluate the analgesic effect of SGE on complete Freund's adjuvant (CFA)-induced inflammatory pain rats, and to investigate whether its effect on relieving inflammatory pain is associated with regulation of P2X3 receptor. MATERIALS AND METHODS: The analgesic effects of SGE on CFA-induced inflammatory pain rats were evaluated by measuring mechanical pain threshold, thermal pain threshold, and motor coordination ability. The mechanisms of SGE in relieving inflammatory pain were explored by detecting inflammatory factors levels, NF-κB, COX-2 and P2X3 expression, and were further verified by addition of P2X3 receptor agonist (α, ß me-ATP). RESULTS: Our results revealed that SGE can notably increase the mechanical pain threshold and thermal pain threshold of CFA-induced inflammatory pain rats, and markedly alleviate the pathological damage in DRG. SGE could suppress the release of inflammatory factors including IL-1ß, IL-6, TNF-α and restrain the expression of NF-κB, COX-2 and P2X3. Moreover, α, ß me-ATP further exacerbated the inflammatory pain of CFA-induced rats, while SGE could markedly raise the pain thresholds and relieve inflammatory pain. SGE could attenuate the pathological damage, inhibit P2X3 expression, inhibit the elevation of inflammatory factors caused by α, ß me-ATP. SGE can also inhibit NF-κB and ERK1/2 activation caused by α, ß me-ATP, and inhibit the mRNA expression of P2X3, COX-2, NF-κB, IL-1ß, IL-6 and TNF-α in DRG of rats induced by CFA coupled with α, ß me-ATP. CONCLUSIONS: In summary, our research indicated that SGE could alleviate CFA-induced inflammatory pain by suppression of P2X3 receptor.

[A case report of IgG4-related sclerosing disease with lung involvement].

IgG4-related sclerosing disease(IgG4-RSD) is a kind of lymphoplasmacytic disease with multi-organ involvement and is characterized by serum IgG4 elevation and tissue IgG4 positive plasma cell infiltration. Autoimmune pancreatitis, sclerosing cholangitis, sclerosing sialadenitis, retroperitoneal fibrosis and lymphadenopathy make up its main clinical manifestations. This difficult case was a middle-aged female with onset as muiltiple lymph nodes and glands enlargement, including lacrimal gland, salivary glands and pancreas. Meanwhile, repeated examinations of auto-antibodies and serum IgG4 were all negative. The patient didn't respond well to glucocorticoid therapy, and further progressed to rare lung involvement presenting as lung nodule. This complex entity was eventually diagnosed as IgG4-RSD by the support of histopathology evidence of IgG4 immunohistochemistry stain. Though IgG4-RSD has been known for years, it is still underappreciated in China and case reports are scarce. The case report here with literature review is just to enhance the recognition of this disease regarding its pathogenesis, various clinical manifestations, diagnosis and therapy.

[Significance of the detection of anti-v-raf murine sarcoma viral oncogene homologue B1 antibodies in the diagnostic practice of rheumatoid arthritis].

OBJECTIVE: To clarify the clinical significance of the antibody against v-raf murine sarcoma viral oncogene homologue B1 (BRAF) in the diagnostic practice of rheumatoid arthritis (RA). METHODS: In the study, 112 patients with RA, 112 patients with other rheumatic diseases,and 73 healthy individuals were recruited . With recombinant human BRAF protein as antigen, we examined the level of anti-BRAF antibody in all the patients by enzyme-linked immunosorbent assay(ELISA), The clinical data of the RA patients were collected simultaneously, and analysed statistically by using SPSS 13.0. RESULTS: The positive rate of anti-BRAF antibody was 53.6% in the RA patients, which was significantly higher than that of the normal control group (4.1%,P<0.01)and other rheumatic diseases groups (P all<0.01)except osteoarthritis group. The titer of anti-BRAF antibody was also notably higher in the patients with RA than in other rheumatic diseases and normal control groups(P all <0.01).The diagnostic sensitivity and specificity of anti-BRAF antibody for RA were 53.6% and 84.3% respectively. The positive rate of anti-BRAF antibody in rheumatoid factor, anti-cyclic citrullinated peptide antibody, antikeratin antibody,antiperinuclear factor negative groups were 52.6%,38.2%, 30.3% and 31.0%respectively. It showed significant negative correlation between the titer of anti-BRAF antibody and patient's age, disease duration and the level of CRP. CONCLUSION: The anti-BRAF antibody contributes to the diagnosis of RA, and may act as a supplement of other autoantibodies.

Iguratimod inhibits osteoclastogenesis by modulating the RANKL and TNF-α signaling pathways.

BACKGROUND: Iguratimod, a small molecular drug, has been proven to have effective bone protection for treatment of patients with bone loss-related diseases, such as rheumatoid arthritis (RA). However, the exact bone protective mechanism of iguratimod remains to be determined. The purpose of this study was to better explore the underlying mechanism of bone protection of iguratimod. METHODS: Bone marrow monocytes from C57/BL6 mice were stimulated with either RANKL or TNF-α plus M-CSF. The effects of iguratimod on morphology and function of osteoclasts were confirmed by TRAP staining and bone resorption assay, respectively. The expression of osteoclast related genes was detected by RT-PCR and the activation of signal pathway was detected by Western blotting. We used rodent models of osteoporosis (ovariectomy) and of arthritis (modified TNF-α-induced osteoclastogenesis) to evaluate the osteoprotective effect of iguratimod in vivo. RESULTS: Iguratimod potently inhibited osteoclast formation in a dose-dependent manner at the early stage of RANKL-induced osteoclastogenesis, whereas iguratimod had no effect on M-CSF-induced proliferation and RANK expression in bone marrow monocytes. Bone resorption was significantly reduced by both early and late addition of iguratimod. Administration of iguratimod prevented bone loss in ovariectomized mice. The blockage of osteoclastogenesis elicited by iguratimod results from abrogation of the p38、ERK and NF-κB pathways induced by RANKL. Importantly, Iguratimod also dampened TNF-α-induced osteoclastogenesis in vitro and attenuated osteoclasts generation in vivo through disrupting NF-κB late nuclear translocation without interfering with IκBα degradation. CONCLUSIONS: Iguratimod not only suppresses osteoclastogenesis by interfering with RANKL and TNF-α signals, but also inhibits the bone resorption of mature osteoclasts. These results provided promising evidence for the therapeutic application of iguratimod as a unique treatment option against RA and especially in prevention of bone loss.

[Analysis for clinical features of cardiovascular complication of diffuse connective tissue disease].

OBJECTIVE: To analyze the clinical features and prognosis of cardiovascular complications of diffuse connective tissue disease(dCTD). METHODS: Clinical data of cardiovascular complications of 181 cases of dCTD were retrospectively analyzed. Clinical data of two subsets [rheumatoid arthritis (RA, 81) and systemic lupus erythematosus (SLE, 42)] were also analyzed. Follow-up was carried out for all the patients. RESULTS: RA and SLE were the most common disease complicated by cardiovascular disease needing hospitalization. The most frequent cardiovascular complications in dCTD patients were hypertension, hyperlipidemia and coronary artery disease. The most common echocardiographic abnormalities were valvular regurgitation, left ventricular diastolic dysfunction, enlargement of left atrium, pulmonary hypertension and pericardial effusion (57.8%, 50.6%, 33.7%, 21.7% and 19.3%, respectively). Compared with the subset of RA, patients were younger at onset of hypertension, coronary artery disease and hyperlipidemia in the subset of SLE [(40+/-11) vs (56+/-15), P<0.001; (53+/-12) vs (64+/-10), P=0.011; (44+/-16) vs (58+/-12), P=0.012, respectively]. Both pericardial effusion (P<0.001) and enlargement of left ventricle (P=0.03) were more frequent, and left ventricular diastolic dysfunction was less common (P<0.001). Median survival time of these dCTD patients was 9.8 years. CONCLUSION: RA and SLE are the most common diffuse connective tissue diseases complicated by cardiovascular disease needing hospitalization. The most frequent cardiovascular complications in dCTD patients are hypertension, hyperlipidemia and coronary artery disease. The prognosis of dCTD patients complicated with cardiovascular diseases is poor. SLE patients are younger at onset of cardiovascular diseases.

Clinical deep remission and related factors in a large cohort of patients with rheumatoid arthritis.

BACKGROUND: Clinical remission is the treatment target in rheumatoid arthritis (RA). This study aimed to investigate clinical remission and related factors in a large cohort of patients with RA. METHODS: This study composed of 342 patients with RA. Data were collected by face-to-face interview of 1049 patients with RA who visited the Department of Rheumatology of three teaching hospitals from September 2015 to May 2016. The patients with RA were clinically assessed by rheumatologists and a four-page questionnaire was completed on site. Subsequently, patients fulfilled remission criteria were further analyzed. The practicability of different definitions of remission of RA was rated by a panel of rheumatologists. Sustained intensive disease modifying anti-rheumatic drug (DMARD) treatment was defined as a combination treatment with two or more DMARDs for at least 6 months. RESULTS: In this cohort of 342 patients with RA, the proportions of patients achieving remission were 38.0%, 29.5%, 24.9%, 21.1%, 19.0%, 18.1%, and 17.0%, based on criteria of disease activity score in 28 joints (DAS28) using CRP (DAS28-CRP), DAS28 using ESR (DAS28-ESR), routine assessment of patient index data 3 (RAPID-3), Boolean, simplified disease activity index (SDAI), clinical disease activity index, and the newly described clinical deep remission (CliDR), respectively. Boolean and CliDR are the best in practicability scored by rheumatologists (7.5 and 8.0, respectively). Compared with the non-sustained intensive group, sustained intensive treatment with DMARDs yielded higher remission rates of 25.6%, 23.8%, and 21.3% in patients with RA based on Boolean (χ = 3.937, P = 0.047), SDAI (χ = 4.666, P = 0.031), and CliDR criteria (χ = 4.297, P = 0.038). The most commonly prescribed conventional synthesized DMARDs (csDMARDs) in patients with RA was leflunomide, followed by methotrexate, and hydroxychloroquine. Compared with the non-remission group, patients achieving remission had a longer median duration of DMARDs (45.0 [22.8-72.3] months, Z = -2.295, P = 0.022). CONCLUSIONS: The findings in this study indicated that clinical deep remission is achievable in patients with RA. Sustained intensive DMARD treatment is needed to achieve a better outcome in RA.

[Mucosal administration of altered CII263-272 peptide inhibits collagen-induced arthritis].

OBJECTIVE: To evaluate the effect of mucosal administration of altered collagen II(CII)263-272 peptide (267Q-->A, 270K-->A and 271G-->A) on collagen induced arthritis (CIA), and to explore the mechanism of the inhibitory effect of the altered CII263-272 peptide on CIA. METHODS: CIA was induced in Lewis rats by immunization with bovine CII. Altered CII263-272 peptide was given intranasally beginning from the onset of arthritis (100 microg/dose, daily for 5 doses and continuing every other day for other 7 doses). Wild CII263-272 peptide (100 microg/dose) or PBS was administered as controls with the same procedure. Therapeutic effects were evaluated by arthritis scores, body weight change, and joint pathologic scores. The anti-CII antibody and its subtypes were measured with ELISA. The cytokines of IFN-gamma and IL-10 were measured with ELISA. The induction of regulatory T cells was assessed by FACS analysis of percentage of peripheral CD4(+)CD25(+) T cells, and by real-time PCR analysis of the expression of Foxp3 and TGF-beta mRNA. RESULTS: (1) Following treatment with the altered CII263-272 peptide, arthritis scores were reduced and body weight was increased. The mean arthritis scores of rats treated with altered peptide, wild peptide and PBS were 2.50 +/- 2.43, 4.50 +/- 2.23 and 6.33 +/- 2.73, respectively. The altered peptide could retard the histologic lesion of the joints. (2) The titers of anti-CII antibodies IgG and IgG1 in the three groups were similar, but the IgG2a in altered peptide-treated rats decreased markedly as compared with PBS-treated rats (0.56 +/- 0.19 vs 0.95 +/- 0.29, P<0.05). The serum IFN-gamma in rats treated with altered peptide, wild peptide and PBS were (185.33 +/- 29.77), (231.62 +/- 41.82) and (220.64 +/- 83.61) ng/L, respectively (P<0.05). No difference was found in the levels of serum IL-10 among the three groups. (3) There was no significant difference in the percentage of peripheral CD4(+)CD25(+) T cells and the expression level of Foxp3 and TGF-beta mRNA. CONCLUSION: Mucosal administration of altered CII263-272 peptide could effectively inhibit the progression of CIA. It can decrease the IgG2a subtype of anti-CII antibodies and IFN-gamma, and inhibit Th1 response in vivo. Altered C II263-272 peptide may be therapeutic for RA.

[Establishment of Sjögren's syndrome models by immunization with alpha-Fodrin: experiment with mice].

OBJECTIVE: To explore the possibility to establish Sjögren's syndrome models by immunizing mice with alpha-fodrin. METHODS: Twenty-four 4-week-old BALB/C mice were randomly divided into 4 equal groups to undergo subcutaneous injection of alpha-Fodrin, submaxillary gland homogenate and glutathione S-transferase (GST) or phosphate-buffered saline (PBS) (negative control groups) on days 0, 14, 35, and 56 respectively. The drinking amount of water was measured. Blood samples were collected every 2 - 3 weeks. Munofluorescence assays and ELISA were used to examine the presence of anti-Fodrin, anti-type 3 muscarinic acetylcholine receptor polypeptide (M3RP), anti-SSA, anti-SSB, rheumatoid factor (RF), and antinuclear antibody (ANA). Immunochemistry was used to detect the levels of interferon (IFN)-gamma, interleukin (IL)-2, and IL-10. One mouse was killed from each group every 2 - 3 weeks. The salivary glands were examined. RESULTS: (1) No auto-immune antibody was found in the serum samples of the mice before immunization. Antibodies against alpha-Fodrin and M2RP, and ANA were positive in the serum samples of the alpha-Fodrin and submaxillary gland homogenate groups since the 35th day after immunization, and were all negative in the 2 control groups. However, no antibodies against SSA, SSB and RF were found in all 4 groups. (2) Lymphocytic infiltration could be seen in the salivary glands of the immunized animals since 50th days after the first immunization of alpha-Fodrin and submaxillary gland homogenate. Immunohistochemistry showed alpha-Fodrin expression in the submaxillary glands of the alpha-Fodrin and submaxillary gland homogenate groups, but not in the PBS and GST controls. (3) The serum IFN-alpha levels of the alpha-Fodrin and submaxillary gland homogenate groups were (81.6 +/- 7.1) and (90.5 +/- 4.9) pg/ml respectively, both significantly higher than those of the GST and PBS groups [(30.1 +/- 5.9) and (19.3 +/- 6.4) pg/ml respectively, both P < 0.05]. The serum IL-2 levels of the alpha-Fodrin and submaxillary gland homogenate groups were (18.7 +/- 2.3) and (19.8 +/- 0.9) pg/ml respectively, both significantly higher than those of the GST and PBS groups [(4.9 +/- 1.1) and (3.5 +/- 1.6) pg/ml respectively, both P < 0.05]. No difference was found in the level of serum IL-10 among the 4 groups. (4) There was no significant difference in the volume of water volume drunk among the 4 groups. CONCLUSION: (1) It is possible to establish mouse mice SS models by immunization with alpha-Fodrin or submaxillary gland homogenate that are reminiscent of human SS. (2) The appearance of multiple antibodies may be related to the antigen epitope spreading. (3) The pathogenesis of SS may be related to Th1 type response.

[Mucosal administration of alpha-fodrin inhibits experimental Sjögren's syndrome autoimmunity].

OBJECTIVE: To explore the effect of mucosal administration of alpha-fodrin in inhibition of autoimmunity in Sjögren's syndrome (SS). METHODS: Thirty-four 4-week-old NOD mice were randomly divided into 4 equal groups: to be immunized by nasal administration of alpha-fodrin 1 microg/dose and 10 microg/dose respectively every two days (experimental groups), and phosphate-buffered saline (PBS) or glutathion2 S-tansferase4 (GST) (control groups). The weekly volume of water drinking was calculated. The salivary flow was maintained. Serum samples were obtained to detect the anti-SSA, anti-SSB, RF, ANA, anti- -fodrin and anti-type 3 muscarinic acetylcholine receptor polypeptide (M3RP) by immunofluorescence or ELISA. The cytokines of IFN-gamma and IL-10 were measured with ELISA. The salivary glands were examined by HE staining and immunohistochemical analysis. Flow cytometry was used to detect the proportion of Foxp3+ CD4+ CD25+ T cells. RESULTS: The titers of anti- -fodrin antibody and M3RP antibody of the mice immunized with-fodrin were lower than those of the 2 control groups (all P < 0.05), however, there was not significant differences between these two a-fodrin immunized groups. Five of the 8 mice in the GST group, 5 mice in the PBS group, 2 mice in the alpha-fodrin 1 microg/dose group, and 3 mice in the alpha-fodrin 10 microg/dose showed ANA positive. The serum IFN-gamma levels in the mice of alpha-fodrin 1 microg/dose and 10 microg/dose groups, PBS group, and GST group were (42 +/- 16), (37 +/- 15), (87 +/- 18), and (72 +/- 11) pg/ml respectively, those of the fodrin groups being significantly lower than those of the control groups (all P < 0.05). There were not significant differences in the level of serum IL-10 among these four groups. The numbers of Foxp3+ CD4 CD25+ regulatory T cells were higher in the fodrin groups than in the PBS and GST control groups (all P < 0.05). The lymphocytic infiltration and expression of alpha-fodrin were decreased in the alpha-fodrin administrated groups. The volume of water drinking of the alpha-fodrin 1 microg/dose group, alpha-fodrin 10 microg/dose group, PBS group, and GST group were (39.2 +/- 2.1), (40.4 +/- 2.5), (49.3 +/- 3.1), and (51.6 +/- 2.8) ml respectively. CONCLUSION: Mucosal administration of alpha-fodrin effectively inhibits the progression of experimental Sjögren's syndrome autoimmunity.

Stattic inhibits RANKL-mediated osteoclastogenesis by suppressing activation of STAT3 and NF-κB pathways.

Tofacitinib, a small molecule JAK inhibitor, has been widely used to reduce inflammation and inhibit progression of bone destruction in rheumatoid arthritis. STAT3, a downstream signaling molecule of JAK, plays a key role in the activation of signaling in response to inflammatory cytokines. Thus, targeting STAT3 may be an inspiring strategy for treating osteoclast-related diseases such as rheumatoid arthritis. In this study, we first investigated the effects of Stattic, a STAT3 inhibitor, on receptor activator of NF-κB ligand (RANKL)-mediated osteoclastogenesis. Stattic inhibited osteoclast differentiation and bone resorption in RANKL-induced RAW264.7 cells in a dose-dependent manner. Stattic also suppressed RANKL-induced upregulation of osteoclast-related genes tartrate-resistant acid phosphatase, matrix metalloproteinase 9, cathepsin K, RANK, tumor necrosis factor receptor-associated factor 6, and osteoclast-associated receptor in RAW264.7 cells. Moreover, Stattic exhibited an inhibitory effect on cell proliferation and cell cycle progression at higher dosages. At the molecular level, Stattic inhibited RANKL-induced activation of STAT3 and NF-κB pathways, without significantly affecting MAPK signaling. In addition, Stattic inhibited RANKL-induced expression of osteoclast-related transcription factors c-Fos and NFATc1. Importantly, Stattic also prevented bone loss caused by ovariectomy. Together, our data confirm that Stattic restricts osteoclastogenesis and bone loss by disturbing RANKL-induced STAT3 and NF-κB signaling. Thus, Stattic represents a novel type of osteoclast inhibitor that could be useful for conditions such as osteoporosis and rheumatoid arthritis.

Macrophage activation syndrome in systemic lupus erythematosus: a multicenter, case-control study in China.

The objective of this study was to describe the clinical and laboratory characteristics, precipitating factors, treatment, and outcome of macrophage activation syndrome (MAS) complicating systemic lupus erythematosus (SLE). A multicenter case-control study was performed across six tertiary hospitals from 1997 to 2014. A total of 32 patients with SLE-associated MAS were enrolled. Sixty-four age- and sex-matched SLE patients diagnosed in the same period without MAS episodes were selected as controls. The most frequent clinical feature was fever, followed by splenomegaly. Hyperferritinemia, hypoalbuminemia, and hyper-lactate dehydrogenase (LDH)-nemia were among the most common laboratory abnormalities. Compared with pre-MAS visit, patients at the onset of MAS had greater frequencies of renal involvement, liver dysfunction, and cytopenia. Receiver operating characteristic (ROC) analysis identified optimal cutoff values of ferritin (>662.5 ng/mL) and LDH (>359 U/mL) to predict the occurrence of MAS in SLE. SLE flare and infection were the common triggers of MAS in SLE. Abortion and parturition were recorded as well. The overall mortality rate was 12.5%. All patients received corticosteroids. Cyclosporine A, cyclophosphamide, and etoposide were the three most commonly used immunosuppressants. Rituximab was given to one patient. Intravenous immunoglobulin (IVIG) was added for 46.9% patients. MAS is a potentially fatal complication of SLE. Its occurrence is most frequently associated with active SLE disease or infection. The presentation of unexplained fever, cytopenia, or liver dysfunction, with high levels of ferritin and LDH, in patients with SLE should raise the suspicion of MAS. Corticosteroids with immunosuppressants and IVIG may be an appropriate treatment.

[Inhibition of cartilage destruction in collagen-induced arthritis by altered CII 263-272 peptide: experiment with rats].

OBJECTIVE: To study the inhibitory effect of altered collagen II (CII) 263-272 peptide (sub268-270) with three consecutive substitutions of TCR-contacting residues on joint inflammation and cartilage destruction in collagen-induced arthritis (CIA). METHODS: Thirty-two Lewis rats were injected intracutaneously with bovine collagen type II so as to establish models of arthritis and then were randomly divided into 4 equal groups to be injected intravenously with sub268-270 30 microg, 5 microg, or 1 microg and PBS twice a week for 3 weeks. The therapeutic effect of the altered peptide on arthritis was evaluated by arthritis score. After the treatment the rats were killed and their ankle joints were taken to undergo pathological examination to observe the existence of synovitis, pannus formation, cartilage damage, and bone erosion. Blood samples were collected to detect the serum cartilage oligomeric matrix protein (COMP). Cartilage proteoglycan-specific dye safranin O was used on the joint sections to observe the coloration of the dye in the cartilage. RESULTS: The arthritis score in rats treated by 30 microg altered CII peptide was (5.6 +/- 2.63), significantly lower than those of the 5 microg, 1 microg, and blank control groups [(9.67 +/- 5.61), (10.02 +/- 5.06), and (11.8 +/- 5.34) respectively, all P < 0.01]. The synovitis score of the 30 microg group was (1.11 +/- 0.43), significantly lower than those of the 5 microg, 1 microg, and blank control groups [(1.87 +/- 0.78), (2.11 +/- 0.83), and (2.25 +/- 0.73) respectively, all P < 0.01]. The pannus score of the 30 microg group was (1.11 +/- 0.43), significantly lower than those of the 5 microg, 1 microg, and blank control groups [(1.83 +/- 0.79), (2.07 +/- 0.91), and (2.27 +/- 0.71) respectively, all P < 0.01]. The cartilage damage score of the 30 microg group was 0.56 +/- 0.23), significantly lower than those of the 5 microg, 1 microg, and blank control groups [(1.91 +/- 0.83), (2.13 +/- 0.79), and (2.29 +/- 0.69) respectively, all P < 0.01]. The bone erosion score of the 30 microg group was (0.53 +/- 0.21), significantly lower than those of the 5 microg, 1 microg, and blank control groups [(1.71 +/- 0.67), (1.88 +/- 0.93), and (2.01 +/- 0.93) respectively, all P < 0.01]. The serum COMP of the 30 microg group was (2.21 +/- 0.76), significantly lower than those of the 5 microg, 1 microg, and blank control groups [(5.63 +/- 1.73), (6.04 +/- 1.76), and (7.00 +/- 1.46) respectively, all P < 0.01]. The content of safranin O (A value) in the joint section of the 30 microg group was (2.35 +/- 0.76), significantly higher than those of the 5 microg, 1 microg, and blank control groups [(1.57 +/- 0.63), (1.37 +/- 0.53), and (1.00 +/- 0.41) respectively, all P < 0.01]. CONCLUSION: The altered CII peptide sub268-270 effectively ameliorates CIA and inhibits the cartilage damage in CIA, and may modify the disease course of rheumatoid arthritis.