Peripheral Th17/Treg imbalance in Chinese patients with untreated antisynthetase syndrome associated interstitial lung disease.

Interstitial lung disease (ILD) is a common and fatal manifestation of antisynthetase syndrome (ASS). The aim of this study was to provide new insight into investigate peripheral blood lymphocytes, CD4+ T cells, cytokine levels and their relation to the clinical profile of untreated patients with ASS-ILD. The retrospective study population included thirty patients diagnosed with ASS-ILD and 30 healthy controls (HCs). Baseline clinical and laboratory data were collected for all subjects, including peripheral blood lymphocyte, CD4+ T cell subsets measured by flow cytometry, and serum cytokine levels measured by multiple microsphere flow immunofluorescence. Their correlations with clinical and laboratory findings were analyzed by Pearson's or Spearman's correlation analysis. In addition, the Benjamini-Hochberg method was used for multiple correction to adjust the p-values. Patients with ASS-ILD had lower CD8+ T cells, higher proportion of Th17 cells and Th17/Treg ratio than HCs. Serum cytokine levels (IL-1ß, IL-6, IL-12, IL-17, IL-8, IL-2, IL-4, IL-10, TNF-α and IFN-γ) were higher in patients with ASS-ILD than HCs. Moreover, Th17/Treg ratio was negatively correlated with diffusing capacity of carbon monoxide (DLCO)%. Our study demonstrated abnormalities of immune disturbances in patients with ASS-ILD, characterized by decreased CD8+ T cells and an increased Th17/Treg ratio, due to an increase in the Th17 cells. These abnormalities may be the immunological mechanism underlying the development of ILD in ASS.

Associations of Digital Ulcers in Patients with Systemic Sclerosis: An 8-Year Retrospective Study.

INTRODUCTION: This study aimed to investigate the associations of digital ulcers (DUs) in patients with systemic sclerosis (SSc). METHODS: This retrospective study investigated the demographic characteristics, specific autoantibodies, organ involvement, and laboratory tests in patients with SSc from our hospital. RESULTS: This study enrolled 144 patients with SSc. The DU+ group consisted of 15 (10.4%) patients. Patients with SSc having DUs have longer disease duration, higher fibrinogen, higher fibrin degradation product, and lower cholesterol. None of the patients used cholesterol-lowering drugs before onset of DUs. The study also demonstrated a higher prevalence of anti-dsDNA and anti-histone antibodies in patients with SSc with DUs. Anti-dsDNA antibody is a specific antibody for SLE with a specificity of 96-99%. A total of 86.1% (124/144) of patients suffered from diffuse cutaneous SSc, and 28.5% (41/144) of patients suffered from overlap syndrome. CONCLUSION: Our study indicated that patients with SSc with fibrinogen of >2.895 g/L (p = 0.043) and cholesterol of <3.340 mmol/L (p = 0.036), which is equal to 129.258 mg/dL, are at high risk of developing DUs.

Xylan derived fluorescence carbon dots composite with cotton cellulose paper as 'turn-off' fluorescence platform for sensitive and selective detection Cu2+ in real samples.

The pollution of heavy metals such as Cu2+ is still serious and the discharge of sewage of Cu2+ will cause damage to soil environment and human health. Herein, a biomass-based solid-state fluorescence detection platform (CPU-CDs) was developed as fluorescent sensor for detection Cu2+ via fluorescence and colorimetric dual-model methods in real time. CPU-CDs was composed of xylan-derived CDs (U-CDs) and cotton cellulose paper, which exhibiting good reusability, non-toxicity, excellent fluorescence characteristics and high biocompatibility. Further, CPU-CDs displayed high effectiveness and sensitivity for Cu2+ with the detection limit as low as 0.14 µM, which was well below U.S. EPA safety levels (20 µM). Practical application indicated that CPU-CDs could achieve precision response of Cu2+ change in real environment water samples with good recovery range of 90 %-119 %. This strategy demonstrated a promising biomass solid-state fluorescence sensor for Cu2+ detection for water treatment research, which is of great significance in dealing with water pollution caused by heavy metal ions.

Identifying Key Genes and Functionally Enriched Pathways in Osteoporotic Patients by Weighted Gene Co-Expression Network Analysis.

Osteoporosis is a systemic bone disease characterized by low bone mineral density and bone microstructure damage, resulting in increased bone fragility and fracture risk. The present study aimed to identify key genes and functionally enriched pathways in osteoporotic patients. Weighted Gene Co-expression Network Analysis (WGCNA) was applied to microarray datasets of blood samples of osteoporotic patients from the Sao Paulo Ageing & Health [SPAH] study (26 osteoporotic samples and 31 normal samples) to construct co-expression networks and identify hub gene. The results showed that HDGF, AP2M1, DNAJC6, TMEM183B, MFSD2B, IGKV1-5, IGKV1-8, IGKV3-7, IGKV3D-11, and IGKV1D-42 are genes which were associated with the disease status of osteoporosis. Differentially expressed genes are enriched in proteasomal protein catabolic process, ubiquitin ligase complex, and ubiquitin-like protein transferase activity. Functional enrichment analysis demonstrated that genes in the tan module were enriched in immune-related functions, indicating that the immune system plays a critical role in osteoporosis. Validation assay demonstrated that the HDGF, AP2M1, TMEM183B, and MFSD2B levels were decreased in osteoporosis samples compared with healthy controls, while the levels of IGKV1-5, IGKV1-8, and IGKV1D-42 were increased in osteoporosis samples compared with healthy controls. In conclusion, our data identified and validated the association of HDGF, AP2M1, TMEM183B, MFSD2B, IGKV1-5, IGKV1-8, and IGKV1D-42 with osteoporosis in elderly women. These results suggest that these transcripts have potential clinical significance and may help to explain the molecular mechanisms and biological functions of osteoporosis.

Decreased expression of TIGIT on CD14 + monocytes correlates with clinical features and laboratory parameters of patients with primary Sjögren's syndrome.

OBJECTIVES: The purpose of this study was to investigate the expression of T-cell immunoglobulin and ITIM domain (TIGIT) in peripheral circulation of primary Sjögren's syndrome (pSS) and its role in the development of pSS. METHODS: The expression of TIGIT on T cells, B cells, natural killer (NK) cells, and CD14 + monocytes was detected by flow cytometry in pSS and healthy control (HC). The correlations between expression of TIGIT and clinical features and laboratory parameters of pSS were analyzed. Meanwhile, we analyzed the change in expression of TIGIT before and after treatment, and its role in the prognosis of pSS treatment was evaluated. RESULTS: The expression of TIGIT on CD3 + , CD4 + , and CD8 + T cells increased and decreased on CD14 + monocytes in pSS compared to HC; however, there was no significance of B lymphocytes and NK cells. The correlation analysis between the expression of TIGIT on T lymphocytes and CD14 + monocytes and clinical features of pSS showed that the decrease in TIGIT expression on CD14 + monocytes was more closely related to pSS. The expression of TIGIT + CD14 + monocytes negatively correlated with the disease activity of pSS. The expression of TIGIT + CD14 + monocytes of pSS with arthralgia, fatigue, decayed tooth, xerostomia, interstitial lung disease, anti-Ro52 positive, and high IgG decreased compared to that in negative patients. Furthermore, it was significantly lower in active patients than in nonactive patients. After treatment, the expression of TIGIT + CD14 + monocytes tended to increase. CONCLUSION: Our study suggested that decreased TIGIT expression on CD14 + monocytes was associated with the clinical manifestations, disease activity, and prognosis of pSS patients. TIGIT + CD14 + monocytes may present as a potential target and a biomarker of the prognosis for immunomodulatory therapy in pSS. Key Points • The expression of TIGIT+CD14+ monocytes significantly decreased in pSS patients compared to HC. • There was a negative correlation between TIGIT+CD14+ monocytes and the disease activity of pSS. • TIGIT+CD14+ monocyte expression was associated with the clinical manifestations, autoantibodies, IgG, and prognosis of pSS patients.

[Analysis of genes related to female bone peak and osteoporosis based on bioinformatics].

OBJECTIVE: To explore and verify the genes related to female peak bone mass(PBM) and osteoporosis (OP) based on bioinformatics. METHODS: Using GEO data, DNA microarray technology to conduct genome-wide analysis of adult female monocytes with high and low PBM. Cluster analysis, GO enrichment and KEGG analysis were used to analyze the differential genes, and the interaction network of differential genes was further analyzed. OP rat model was established and femur neck tissue staining was performed to further verify the expression of differential genes. RESULTS: A total of 283 genes were obtained by differential gene screening. Compared with the high PBM samples, 135 genes were up-regulated and 148 genes were down-regulated in the low PBM samples. A total of 7 pathways and 12 differential genes were enriched, and there were differences in the expression of several genes involved in mineral absorption and transport, cellular immunity and other aspects. Among them, voltage-gated Ca2+ channel 1.3(CaV1.3) encoded by CACNA1D gene was significantly enhanced in the femoral neck tissue of OP rat model. CONCLUSION: The above results suggest that the difference in the expression level of CaV1.3 gene may lead to the occurrence of OP in women with low PBM, which provides us with a potential target for the prevention and treatment of OP.

Biomass-based indole derived fluorescence sensor composited with cellulose paper: Detection of picric acid in food and environment samples.

Picric acid (PA) is highly water-soluble, the fact makes it stand out as the most hazardous environment pollutant. Therefore, accurate determination of PA is of great significance for human health and environmental protection. Herein, a novel indole-based fluorescent sensor (H1) with good water solubility and fluorescence stability was reported. H1 exhibited 'turn-off' fluorescence response for PA with fast reaction rate (<30 s), unique specificity and excellent selectivity and high sensitivity (limit of detection = 34 nM). Further, H1 was successfully applied to detect PA in real samples (tap water, Yangtze River, Xuanwu Lake, soil, food, fish and shrimp) with satisfactory recoveries at three spiking levels ranging from 98.0 to 112.0 %. In addition, H1 displayed high biocompatibility in mung beans and fresh blood. Moreover, aiming to attain portable analysis, H1 was composited with biomass cellulose paper (H1-FP) and integrated with smartphone for construction as a solid-state fluorescence platform to achieve fast and visual detection of PA in suit with significant stability, high sensitively and selectivity. The establishment of this sensing approach is expected to offer new insight into rapid, selective, and sensitive detection of major pollutants for food and environmental safety.

Tripartite motif­containing 14 may aggravate cardiac hypertrophy via the AKT signalling pathway in neonatal rat cardiomyocytes and transgenic mice.

Tripartite motif­containing 14 (TRIM14) is an E3 ubiquitin ligase that primarily participates in the natural immune response and in tumour development via ubiquitination. However, the role of TRIM14 in cardiac hypertrophy is not currently clear. The present study examined the role of TRIM14 in cardiac hypertrophy and its potential molecular mechanism. TRIM14 was overexpressed in neonatal rat cardiomyocytes using adenovirus and cardiomyocyte hypertrophy was induced using phenylephrine (PE). Cardiomyocyte hypertrophy was assessed by measuring cardiomyocyte surface area and markers of hypertrophy. In addition, TRIM14­transgenic (TRIM14­TG) mice were created and cardiac hypertrophy was induced using transverse aortic constriction (TAC). Cardiac function, heart weight­to­body weight ratio (HW/BW), cardiomyocyte cross­sectional area, cardiac fibrosis and hypertrophic markers were further examined. The expression of AKT signalling pathway­related proteins was detected. TRIM14 overexpression in cardiomyocytes promoted PE­induced increases in cardiomyocyte surface area and hypertrophic markers. TRIM14­TG mice developed worse cardiac function, greater HW/BW, cross­sectional area and cardiac fibrosis, and higher levels of hypertrophic markers in response to TAC. TRIM14 overexpression also increased the phosphorylation levels of AKT, GSK­3ß, mTOR and p70S6K in vivo and in vitro. To the best our knowledge, the present study was the first to reveal that overexpression of TRIM14 aggravated cardiac hypertrophy in vivo and in vitro, which may be related to activation of the AKT signalling pathway.

The proportion and phenotypic changes of CD4+CD25-Foxp3+ T cells in patients with untreated rheumatoid arthritis.

OBJECTIVE: CD4+CD25+Foxp3+ regulatory T (Treg) cell-mediated immunosuppression is an essential mechanism of rheumatoid arthritis (RA). However, little is known regarding the specific role of CD4+CD25-Foxp3+ Treg cells in RA. This study aimed to investigate the frequency of circulating CD4+CD25-Foxp3+ Treg cells and their role in RA. METHODS: Sixty-one untreated RA patients and 40 healthy controls (HCs) were enrolled in this study. The proportion of CD4+CD25-Foxp3+ T cells and CD4+CD25+Foxp3+ Tregs; the levels of CTLA4, GITR, Helios, and ICOS; and the production of IL-17A, IFN-γ, and IL-10 were assessed by flow cytometry. The correlation of CD4+CD25-Foxp3+ T cells and CD4+CD25+Foxp3+ Tregs with the clinical indicators was conducted by Spearman correlation analysis. RESULTS: The proportion of CD4+CD25-Foxp3+ T cells was elevated in RA and positively correlated with disease activity. CD4+CD25-Foxp3+ T cells expressed less Helios and produced more IFN-γ than conventional Tregs in RA. Additionally, the proportion of CD4+CD25-Foxp3+ T cells was positively correlated with DAS28 score, IgG titer, and anti-CCP titer. CONCLUSIONS: These data indicate that CD4+CD25-Foxp3+ T cells in RA exhibit several different functional properties from conventional Tregs and are correlated with RA disease activity.

Tripartite motif 27 promotes cardiac hypertrophy via PTEN/Akt/mTOR signal pathways.

Tripartite motif-containing 27 (Trim27) is highly expressed in tumor cells and regulates natural immunity and apoptosis. However, the effects of Trim27 in cardiac hypertrophy are not fully elucidated. In this study, we tried to explore the potential role of Trim27 in pressure overload-induced cardiac hypertrophy and the underlying mechanism. The results indicated that compared to sham operation (Sham) group, transverse aortic constriction (TAC) group showed significantly up-regulated Trim27 protein expression (P < 0.05). The neonatal rat cardiomyocytes (NRCMs) were isolated and stimulated with PBS, angiotensin (AngII) and phenylephrine (PE). NRCMs were collected to detect the protein expression of Trim27. The results were consistent with the results in vivo. Compared to PBS treatment, the expression of Trim27 protein in NRCMs was significantly increased after PE or AngII stimulation (P < 0.05, respectively). Knockout of Trim27 can reduce the size of cardiomyocytes and reduce the proteins expression of ANP, BNP, and ß-MHC, improve cardiac function, and reverse myocardial hypertrophy (P < 0.05). Trim27 may be involved in regulating the development of cardiac hypertrophy. Further results showed that Trim27 can increase the protein expression of phosphorylation of Akt, GSK3ß, mTOR, and P70s6k by interacting with PTEN (phosphatase tensin homolog). These findings revealed that Trim27 can promote cardiac hypertrophy by activating PTEN/Akt/GSK3ß/mTOR signaling pathway.

Biomarkers of joint metabolism and bone mineral density are associated with early knee osteoarthritis in premenopausal females.

INTRODUCTION: Biomarkers of bone and cartilage metabolism were proposed as early diagnosis indicators for knee osteoarthritis (OA), however, which were influenced by disease stage, age, and menopause state. Accurate diagnosis indicators are eagerly awaited. The current study aims to investigate associations of joint metabolism biomarkers and bone mineral density (BMD) with early knee OA in males and premenopausal females before age 50 years. METHOD: A total of 189 patients aged before 50 years with early knee OA and 152 healthy participants were enrolled. Levels of bone biomarkers (PINP, OC, and CTX-I) and cartilage biomarkers (PIIANP, COMP, CTX-II, and MMP-3) were assessed. BMD was measured at the lumbar, femoral neck, and hip. Multivariate regression analyses were performed to evaluate the relationship between biomarkers, BMD, and early knee OA. RESULTS: Serum COMP, urine CTX-II and BMD at femoral neck and hip were increased in premenopausal patients as compared to control; with serum PINP and OC reduced. Meanwhile, serum COMP, urine CTX-II, and BMD at femoral neck and hip showed positive associations with premenopausal early knee OA, while serum PINP had negative association. However, in male patients, only serum COMP was higher than control, and no association of biomarkers or BMD was found with early knee OA. CONCLUSIONS: The joint metabolism biomarkers and BMD showed multiple associations with early knee OA in premenopausal females, but not in males aged before 50 years. It was suggested that sex differences should be taken into account when evaluating cartilage and bone metabolism in early knee OA. Key Points • The joint metabolism biomarkers and BMD are associated with early knee OA in premenopausal females, but not in males aged before 50 years. • Sex differences should be taken into account when evaluating cartilage and bone metabolism in early knee OA.

Cav 1.3 damages the osteogenic differentiation in osteoporotic rats by negatively regulating Spred 2-mediated autophagy-induced cell senescence.

Cav 1.3 can affect the classical osteoclast differentiation pathway through calcium signalling pathway. Here, we performed cell transfection, real-time fluorescence quantitative PCR (qPCR), flow cytometry, SA-ß-Gal staining, Alizarin Red S staining, ALP activity test, immunofluorescence, Western blot and cell viability assay to analyse cell viability, cell cycle, osteogenesis differentiation and autophagy activities in vitro. Meanwhile, GST-pull down and CHIP experiments were conducted to explore the influence of Cav 1.3 and Sprouty-related EVH1 domain 2 (Spred 2) on bone marrow-derived mesenchymal stem cells (BMSCs). The results showed that OS lead to the decreased of bone mineral density and differentiation ability of BMSCs in rats. Cav 1.3 was up-regulated in OS rats. Overexpression of Cav 1.3 inhibited the activity of BMSCs, the expression of alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2) and osteocalcin (OCN), as well as promoted the cell cycle arrest and senescence. Furthermore, the negative correlation between Cav 1.3 and Spred 2 was found through GST-pull down and CHIP. Overexpression of Spred 2 increased the expressions of microtubule-associated protein 1 light chain 3 (LC3) and Beclin 1 of BMSCs, which ultimately promoted the cell activity of BMSCs and ALP, RUNX2, OCN expression. In conclusion, Cav 1.3 negatively regulates Spred 2-mediated autophagy and cell senescence, and damages the activity and osteogenic differentiation of BMSCs in OS rats.

Cav1.3 is upregulated in osteoporosis rat model and promotes osteoclast differentiation from preosteoclast cell line RAW264.7.

BACKGROUND: Osteoporosis (OP) is a systemic osteopathy with increased bone fragility and increased risk of fracture. Osteoclasts (OC) are the key target cells in the treatment of osteoporosis. We aimed to research the role of L-type calcium channel protein Cav1.3 in OC differentiation in this study. METHODS: OP rat model was established to detect the expression level of Cav1.3. Tartrate-resistant acid phosphatase assay was used to measure the differentiation of osteoclast during receptor activator of nuclear factor κ-Β ligand (RANKL)-induced osteoclasts formation. The expression of bone differentiation-related proteins were detected by western blot analysis. RESULTS: Cav1.3 is upregulated in OP rats. Knockdown of Cav1.3 inhibits the differentiation of RAW264.7. Cav1.3 regulates the cell differentiation and bone resorption of RAW264.7 during RANKL-induced osteoclasts formation, which is accompanied by upregulation of CaMK II, p-CERB, AP-1, NFATC1, and NF-κB. CONCLUSION: Cav1.3 plays an important role in osteoporosis and the differentiation of osteoclast, which might be involved with the bone differentiation-related proteins.

Association of Fcgamma receptor type 2A and 3A genotypes with rheumatoid arthritis in Chinese population.

AIM: SNPs of FcγRs were implicated in pathogenesis of rheumatoid arthritis (RA) and treatment efficacy of TNF inhibitors (TNFi). This study aims to investigate the associations of FcγRIIa and FcγRIIIa genotypes with autoantibody production and treatment response to TNFi in Chinese patients with RA. PATIENTS & METHODS: FcγRIIa and FcγRIIIa polymorphisms were genotyped in 158 RA patients. Response to TNFi was evaluated in 18 patients at 3 and 6 months after treatment. RESULTS: FcγRIIa-131H allele was significantly increased in autoantibody-negative RA patients. FcγRIIa-131H/H+H/R was closely associated with differences in 28-joint disease activity score in patients at months 3 and 6 of TNFi treatment. CONCLUSION: FcγRIIa-131H allele may have a protective role in autoantibody production and might be a biomarker for predicting good response to TNFi in Chinese RA patients.

Prevalence and possible risk factors of low bone mineral density in untreated female patients with systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by chronic inflammation. Different studies have shown decreased bone mineral density (BMD) in patients with SLE. The objective of this study was to investigate the prevalence and possible risk factors of low BMD in untreated female patients with SLE in Chinese population. A total of 119 untreated female patients with SLE were included. BMD was measured at lumbar spine and at total hip by dual-energy X-ray absorptiometry. The associations between decreased BMD and demographic variables, clinical variables, and bone metabolism variables were analyzed. These SLE patients had the following characteristics: mean age was 32.6 ± 11.9 years, mean disease duration was 22.1 ± 34.5 months, and mean SLEDAI was 11.4 ± 5.4. Osteopenia was present in 31.1% of the patients and osteoporosis in 8.5%. A significant negative association between low density lipoprotein cholesterol (LDL-c) and BMD at the lumbar spine (correlation coefficient = -0.242; P = 0.023) and total hip (correlation coefficient = -0.259; P = 0.019) was shown. These results seem to indicate that increased LDL-c may be an important risk factor for low BMD at lumbar spine and total hip in untreated female SLE patients.

[Role of glucocorticoid receptor in the pathogenesis of systemic lupus erythematosus].

OBJECTIVE: To investigate the expressions of GRα mRNA and GRß mRNA in the peripheral blood mononuclear cells (PBMCs) of systemic lupus erythematosus (SLE) patients, in order to reveal the role of GR mRNA in the pathogenesis of SLE and analyze the relationship between GR mRNA and SLEDAI score, dsDNA, cardiovascular involvement. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) technique was applied to semiquantitatively analyze GRα mRNA and GRß mRNA expressions in 104 SLE patients and 56 volunteers. RESULTS: The level of GRα mRNA was lower in the SLE group (the relative level was 1.24±0.97)than in the control group (the relative level was 2.31±1.42, P<0.05), and the level of GRß mRNA was higher in the SLE group(the relative level was 0.61±1.23) than in the control group(the relative level was 0.18±0.21, P<0.05). The level of GRα was lower in the active group (the relative level was 0.68±0.40) than in the inactive group(the relative level was 1.65±1.06, P<0.01), but the level of GRß was higher in the active group(the relative level was 0.88±1.56) than in the inactive group(the relative level was 0.24±0.23, P<0.01); GRα mRNA was related negatively to the SLEDAI score and dsDNA, but GRß mRNA was related positively to the SLEDAI score and dsDNA(P<0.01).The level of GRα mRNA was lower in the dsDNA positive group(the relative level was 0.89±0.66) than in the dsDNA negative group (the level was 1.54±1.10), the level of GRß mRNA was higher in the dsDNA positive group (the relative level was 0.95±1.60) than in the dsDNA negative group (the relative level was 0.22±0.21). The level of GRß mRNA and the value of GRß mRNA/ GRα mRNA was obviously higher in the SLE group with cardiac involvement (the relative level was 1.02 ±1.76, the valve of GRß / GRα was 1.10±2.02)than in the SLE group without cardiac involvement (the relative level was 0.28±0.31, the valve of GRß / GRα was 0.32±0.32, P<0.05), and the level of GRα mRNA wasn't significant in the two groups (P>0.05). CONCLUSION: The levels of GRα mRNA and GRß mRNA maybe play an important role in the pathogenesis of SLE. And the levels of GRα mRNA and GRß mRNA are related to the activity of SLE. The level of GRß mRNA and the value of GRß mRNA/ GRα mRNA are related with cardiovascular involvement in SLE.