Yisha Li

MicroRNA Expression Abnormalities in Limited Cutaneous Scleroderma and Diffuse Cutaneous Scleroderma

Scleroderma (systemic sclerosis, SSc) is a complex autoimmune disease caused by progressive fibrotic replacement of normal tissue architecture, a progressive and ultimately fatal process that currently has no cure. Although dysregulation of microRNAs (miRNAs) is known to be involved in a variety of pathophysiologic processes, the role of miRNAs in SSc is unclear. In comparison with the normal skin tissues, miRNAs were aberrantly expressed in limited cutaneous scleroderma and diffuse cutaneous scleroderma skin tissues. We also identified miRNAs whose expressions were correlated with SSc fibrosis: miR-21, miR-31, miR-146, miR-503, miR-145, and miR-29b were predicted to be involved. This study further confirmed that miR-21 was increased whereas miR-145 and miR-29b were decreased both in the skin tissues and fibroblasts. As predicted target genes, SMAD7, SAMD3, and COL1A1 were regulated by these miRNAs. After stimulation with transforming growth factor β, the expression of miR-21 was increased and that of SMAD7 mRNA was decreased. MiR-145 was upregulated whereas the mRNA level of SMAD3 was downregulated. The downregulation of miR-29b was correlated with the upregulation of COL1A1 mRNA. MiRNAs might play an important role in the pathogenesis of SSc and suggest a potential therapy.

MicroRNA-21 in Scleroderma Fibrosis and its Function in TGF-β- Regulated Fibrosis-Related Genes Expression

Uncontrolled fibrosis in multiple organs is the main cause of death in systemic sclerosis (SSc), and transforming growth factor-β (TGF-β) activation plays a fundamental role in the process. Our previous study demonstrated that miR-21 was significantly up-regulated in SSc fibroblasts. Here, we found that TGF-β regulated the expression of miR-21 and fibrosis-related genes, and decreased Smad7 expression. Over-expression of miR-21 in fibroblasts decreased the levels of Smad7, whereas knockdown of miR-21 increased its expression. Further study using a reporter gene assay demonstrated Smad7 was a direct target of miR-21. Similar to human SSc, the expression of miR-21 increased in the bleomycin induced skin fibrosis. Inhibition of fibrosis by treatment with anti-fibrosis drug bortezomib restored the levels of miR-21 and Smad7. MiR-21 may function in an amplifying circuit to enhance TGF-β signaling events in SSc fibrosis, and suggesting that miR-21 may act as a potential therapeutic target.Uncontrolled fibrosis in multiple organs is the main cause of death in systemic sclerosis (SSc), and transforming growth factor-β (TGF-β) activation plays a fundamental role in the process. Our previous study demonstrated that miR-21 was significantly up-regulated in SSc fibroblasts. Here, we found that TGF-β regulated the expression of miR-21 and fibrosis-related genes, and decreased Smad7 expression. Over-expression of miR-21 in fibroblasts decreased the levels of Smad7, whereas knockdown of miR-21 increased its expression. Further study using a reporter gene assay demonstrated Smad7 was a direct target of miR-21. Similar to human SSc, the expression of miR-21 increased in the bleomycin induced skin fibrosis. Inhibition of fibrosis by treatment with anti-fibrosis drug bortezomib restored the levels of miR-21 and Smad7. MiR-21 may function in an amplifying circuit to enhance TGF-β signaling events in SSc fibrosis, and suggesting that miR-21 may act as a potential therapeutic target.

MicroRNA-202-3p regulates scleroderma fibrosis by targeting matrix metalloproteinase 1

The leading cause of death in systemic sclerosis (SSc) is the uncontrolled fibrosis in multiple organs. The exact mechanism of fibrosis is not fully clear. Our previous studies using miRNA array analysis indicated that miR-202-3p was increased in SSc lesion skin tissues. Bioinformatics analysis suggested matrix metallopeptidase (MMP) 1 is the target gene of miR-202-3p. Here we confirmed that miR-202-3p was upregulated, and the mRNA and protein expression of MMP1 were significantly decreased in SSc skin tissues and primary fibroblast compared with normal skin. MMP1 expression was inversely correlated with the expression of miR-202-3p. Overexpression of miR-202-3p markedly increased collagen disposition in skin primary fibroblasts, while inhibitor of miR-202-3p decreased it. Furthermore, we demonstrated that MMP1 was a target of miR-202-3p detected by luciferase reporter assay, and played an essential role as a mediator of the biological effects of miR-202-3p in SSc fibrosis. Taken together, these findings suggest that miR-202-3p may function as a novel pro-fibrotic miRNA in SSc by inhibition the expression of MMP1.

Tumor necrosis factor antagonists in the treatment of multicentric reticulohistiocytosis: Current clinical evidence.

Multicentric reticulohistiocytosis (MRH) is a rare and debilitating systemic disorder characterized by cutaneous nodules and destructive polyarthritis. Due to its unknown etiology, the treatment of MRH varies with different rates of success, which causes treatment options to be rather independent and empirical. In the present study, a case of a 48-year-old woman with a 12-month history of polyarthralgia and skin nodules was reported. Biopsy samples, which were obtained from her skin eruption exhibited dermal infiltration with histiocytes and multinucleated giant cells. Immunohistochemical staining indicated positivity for CD68. The patient was diagnosed with MRH and treated with a combination therapy of infliximab, prednisolone and methotrexate. Her symptoms improved markedly within 2 weeks. Following the results of this case study, a systematic review of 17 cases of MRH treated with tumor necrosis factor (TNF) antagonists was performed, and the efficacy of anti-TNF treatment in MRH was analyzed.

Systematic approach to understanding the pathogenesis of systemic sclerosis

Systemic sclerosis (SSc) is a complex heterogeneous autoimmune disease. Progressive organ fibrosis is a major contributor to SSc mortality. Despite extensive efforts, the underlying mechanism of SSc remains unclear. Efforts to understand the pathogenesis of SSc have included genomics, epigenetics, transcriptomic, proteomic and metabolomic studies in the last decade.

Systemic lupus erythematosus with lung, brain, liver, and spleen tuberculosis.

To the Editor: We read the report of Tikly and Mia 1 in the Journal of Clinical Rheumatology with great interest, which emphasized an extremely rare site of tuberculous infection in a patient with lupus.Wewish to report a case of widespread (lung, brain, liver, and spleen) tuberculosis in a patient with systemic lupus erythematosus (SLE). A 22-year-old woman was admitted to the hospital in July 2009. She presented with bilateral joint swelling and pain in her knees, elbows, and peripheral interphalangeal joints since July 2008. She suffered from a headache since June 2009 and developed fever half a month ago, with the highest temperature reaching 39-C, and accompanied by cough. She was diagnosed with SLE and lupus nephritis based on arthritis; positive antinuclear antibody (1:320); positive anti-dsDNA antibody, nRNP, SSA, and antinucleosome antibody; and proteinuria (1.2 g/d). Additionally, she was diagnosed with lung tuberculosis based on a lung computed tomography (CT), which demonstrated lesions in the apical segment of bilateral upper lobes and the superior segment of the left lower lobe and tuberculous meningitis and encephalitis based on a decreased glucose and chloride level (0.65 and 113.3 mmol/L, respectively) and increased white blood cell count (60 cells/KL) in cerebrospinal fluid; and a right intracerebral lesionwas identified by a brain magnetic resonance imaging. She was treated with corticosteroids and antitubercular agents (isoniazid, rimactazid, ethambutol, and pyrazinamide). Her temperature fluctuated between 37-C and 38-C. Pyrazinamide was withdrawn because of hyperuricemia in November 2009. Her fever worsened after December 2009. Lung CT and brain magnetic resonance imaging showed improvement of previous lesions. Abdominal ultrasound and CT demonstrated emerging lesions in the liver and spleen (Figure). An ultrasound-guided liver biopsy was performed in March 2010. Pathologic examination revealed tuberculosis with caseous necrosis. Bacterial and mycobacterial cultures and acid-fast stain were negative. The patient was diagnosed with liver and spleen tuberculosis and treated with the former 4 antitubercular agents. Fever subsided in July 2010. As of the latest follow-up, in January 2011, fever had not relapsed, proteinuria was 1+, and the patient was still taking antitubercular agents and prednisone (10 mg/d). There is no literature discussing such widespread lung, brain, liver, and spleen tuberculosis in SLE patients. The incidence of liver and spleen tuberculosis is very low; it usually occurs in immunocompromised patients. Because of atypical symptoms and reliance on pathologic and pathogenic evidence, it is difficult to make an accurate diagnosis. Because of the immunologic disturbances intrinsic to SLE and the longterm administration of corticosteroids and immunosuppressive drugs, SLE patients are at increased risk of developing tuberculosis, especially extrapulmonary tuberculosis, and the incidence is as high as 17%. A high cumulative dose of corticosteroid and lupus nephritis are 2 important risk factors for the development of tuberculosis. Before the use of corticosteroid, the current patient had lung tuberculosis and tuberculous meningitis and encephalitis. During treatment with corticosteroids and antitubercular agents, she developed liver and spleen tuberculosis, even though the previous tuberculous lesions in the lung and brain were improved. In the case of tuberculosis in SLE patients, the course of antitubercular treatment should be prolonged. For the current patient, her fever was controlled after receiving antitubercular agents for 12months. If infective lesions in the liver and spleen are suspected, the pathologic and pathogenic evidence obtained should be as in-depth as possible.

Calcinosis and malignancy are rare in Chinese adult patients with myositis and nuclear matrix protein 2 antibodies identified by an unlabeled immunoprecipitation assay

Autoantibodies in patients with myositis may associate with specific clinical manifestations. This study aimed to identify a subset of patients with myositis carrying antinuclear matrix protein 2 (anti-NXP-2) antibodies using an unlabeled immunoprecipitation (IP) assay, and clarify the features of these patients in a Chinese cohort. We developed novel methods for unlabeled protein IP and immunoblotting of Myc-tagged truncated NXP-2 fragments for anti-NXP-2 detection. The sera of 120 Chinese adult patients with myositis were screened for anti-NXP-2 by IP and immunoblot. Anti-NXP-2 antibodies were detected in 10 of the 120 patients (8.3%) using the established unlabeled protein IP and immunoblotting, with 70% (7/10) exhibiting either heliotrope rash or Gottron’s papules. All 10 anti-NXP-2-positive patients exhibited myopathy and 60% complained of dysphagia. Severe diffuse calcinosis (10%) and nasopharyngeal carcinoma (10%) were each only present in single anti-NXP-2-positive patients with myositis. Antibodies against Ro-52 were found in four living but not in three deceased anti-NXP-2-positive patients. A comprehensive review of 13 anti-NXP-2 studies demonstrated markedly lower anti-NXP-2 prevalence among adult patients with myositis and lower association of anti-NXP-2 with calcinosis in Japan, China, and Hungary than in the USA and Italy. Anti-NXP-2 antibody association with internal malignancy in adult patients varied from 0 to 50% across different studies. A novel IP assay was developed to detect patients with myositis expressing anti-NXP-2. Calcinosis and malignancy are rare in Chinese adult patients with myositis positive for anti-NXP-2. Literature review indicated highest anti-NXP-2 prevalence and association of anti-NXP-2 with calcinosis in US and Italian myositis cohorts.

The role of IFI35 in lupus nephritis and related mechanisms

Abstract Objectives: It’s reported that multiple genes in the IFN-γ/STAT1 pathway were hypomethylated and associated with the pathogenesis of lupus nephritis (LN). Our previous study using microarray analysis suggested that interferon induced 35-kDa protein (IFI35) was hypomethylated and increased in LN. However, the role of IFI35 in LN and related mechanism remains to be elucidate. Methods: The expressions of IFNγR, STAT1, IFI35 and MBD2 in the human kidneys tissues was detected by real-time PCR and Western blot. The protein levels of IFI35 in the human kidney tissues were detected by immunohistochemistry. The methylation status of IFNγR, STAT1 and IFI35 were detected by methylation specific PCR. Cell proliferation assay was evaluated using cell counting kit 8; pcDNA-IFI35 (pcDNA-MBD2) or IFI35 RNAi (MBD2 RNAi) was used to upregulated or downregulated the expression of the IFI35 and MBD2. Results: The expressions of IFNγR, STAT1 and IFI35 in the LN kidneys were significantly higher than controls. IFI35 was expressed in mesangial cells, and positively correlated with the proliferation of mesangial cells. IFNγR, STAT1and IFI35 was hypomethylated and MBD2 was increased in LN kidneys. In vitro data confirmed those findings: after stimulating with the serum from LN patients, the proliferation of human renal mesangial cells (HRMCs) was increased. The expressions of the three members of IFNγ signal pathway were hypomethylated and upregulated. However, this effect was reversed by MBD2 knockdown. IFI35 promoted the proliferation of HRMCs and was regulated by MBD2. Conclusion: Our results demonstrated that IFI35 enhances the proliferation of mesangial cells and was regulated by MBD2 in LN.

408 Analysis of influencing factors on quality of life of patients with sle

Background and aims To investigate the influencing factors of quality of life (QOL) of patients with systemic lupus erythematosus (SLE). Methods QOL of 104 SLE patients were investigated by SF-36 scale (Chinese version). Depression, anxiety, social support, sleep quality and fatigue were evaluated by PHQ-9, GAD-7, social support rating scale (SSRS), Pittsburgh sleep quality index (PSQI) and VAS respectively. The demographic and clinical data were also recorded. SLE disease activity was assessed by SLEDAI. Results In SF-36 scale, scores of SLE patients were lower than normal people in global score and in all dimensionalities (p<0.05). SLEDAI, PHQ-9, GAD-7, PQSI and fatigue correlated negatively with SF-36 scores (p<0.01). SSRS correlated positively with SF-36 scores (p<0.01). In binary logistic regression analyses, disease activity, anxiety, social support, sleep quality and fatigue were the independent determinants of QOL in SLE (R2 =0.860, p<0.01). Conclusions QOL of SLE patients are lower than normal people. Disease activity, anxiety, social support, sleep quality and fatigue are the major influencing factors of QOL in SLE.

Investigation into the cause of mortality in 49 cases of idiopathic inflammatory myopathy: A single center study.

Idiopathic inflammatory myopathy (IIM) is an autoimmune disease characterized by chronic muscle weakness and myositis with unknown etiology. IIM may affect the function of multiple organs and has a poor prognosis. In the present study, the causes of mortality in patients with IIM admitted to the Xiangya Hospital during the last 14 years were investigated. The investigation included an analysis of frequent causes of IIM, and of infections and associated complications. A cohort study was conducted on 676 patients with IIM that were admitted to Xiangya Hospital from January, 2001 to January, 2015. There were 49 patient mortalities (7.2% of the total cases), of which 34 mortalities were infection-associated and 15 were not infection-associated. The proportion of infection-associated IIM mortalities had increased since 2001. Of the 34 infection-associated mortalities, 31 cases (63.3%) were of fungal and bacterial infections, most frequently infecting the lungs and the blood. Klebsiella pneumoniae and Acinetobacter baumannii were the most commonly isolated pathogens, and co-infection with the two pathogens was observed in the majority of cases. In the IIM mortalities not associated with infection, there were 2 acute myocardial infarction cases, 2 acute interstitial lung disease cases, 4 malignancies and 1 case of each of the following: Arrhythmia, pneumothorax, ventilator weakness, pulmonary artery hypertension, gastrointestinal bleeding, liver failure and renal failure. Three mortalities were secondary to viral hepatitis in the present study. Pathogenic infection was the most frequent cause of mortality in patients with IIM. The remaining causes of mortality included secondary to heart failure, lung dysfunction and malignancy. Following the ubiquitous application of glucocorticoids and immunosuppressants, the proportion of infection-associated mortalities increased in patients with IIM. Thus, in addition to focusing on the primary disease, infection should receive increased attention during clinical practice.