Landscape of immune cells in systematic lupus erythematosus patients with Epstein-Barr virus infection: assessed by single-cell sequencing.

OBJECTIVES: To analyze the immune cell and B cell receptor (BCR) profiles of patients with systematic lupus erythematosus (SLE), with or without Epstein-Barr virus (EBV) infection, and identify the differences between them. METHODS: We included two patients with SLE and positive detection of EBV infections (SLE-EBV+), four with SLE with negative detection of EBV infections (SLE-EBV-), and two healthy controls (HC). Single-cell RNA sequencing (scRNA-seq) was used to investigate the heterogeneity of cell populations by combining the transcriptomic profiles and BCR repertoires. RESULTS: A total of 83 478 cells were obtained and divided into 31 subtypes. The proportion of CD8+ proliferation T cells were higher in the SLE-EBV+ group than in the SLE-EBV- group. The interferon-alpha/beta pathways were upregulated in most T cells, monocytes, and B-cells in the SLE-EBV+ group, compared with the SLE-EBV- group. Moreover, T cell trajectory indicated CD4+ regulatory T cells (Tregs) may play crucial roles in SLE combined with EBV infection. In the BCR heavy chain, the IGHV3 and IGHV4 gene families were frequently present in all groups. Additionally, Immunoglobin (Ig) M was the largest component of five Ig isotypes, but its proportion was significantly decreased in the SLE-EBV+ group. CONCLUSION: This study provides a comprehensive characterization of the immune cell profiles and BCR repertoires of patients with SLE, both with or without concurrent EBV infections, contributing to a better understanding of the mechanism underlying the immune response to EBV infection in patients with SLE.

Application of logistic regression, support vector machine and random forest on the effects of titanium dioxide nanoparticles using macroalgae in treatment of certain risk factors associated with kidney injuries.

The use of titanium dioxide (TiO2) nanoparticles in many biological and technical domains is on the rise. There hasn't been much research on the toxicity of titanium dioxide nanoparticles in biological systems, despite their ubiquitous usage. In the current investigation, samples were exposed to various dosages of TiO2 nanoparticles for 4 days, 1 month, and 2 months following treatment. ICP-AES was used to dose TiO2 into the tissues, and the results showed that the kidney had a significant TiO2 buildup. On the other hand, apoptosis of renal tubular cells is one of the most frequent cellular processes contributing to kidney disease (KD). Nevertheless, the impact of macroalgal seaweed extract on KD remains undetermined. In this work, machine learning (ML) approaches have been applied to develop prediction algorithms for acute kidney injury (AKI) by use of titanium dioxide and macroalgae in hospitalized patients. Fifty patients with (AKI) and 50 patients (non-AKI group) have been admitted and considered. Regarding demographic data, and laboratory test data as input parameters, support vector machine (SVM), and random forest (RF) are utilized to build models of AKI prediction and compared to the predictive performance of logistic regression (LR). Due to its strong antioxidant and anti-inflammatory powers, the current research ruled out the potential of using G. oblongata red macro algae as a source for a variety of products for medicinal uses. Despite a high and fast processing of algorithms, logistic regression showed lower overfitting in comparison to SVM, and Random Forest. The dataset is subjected to algorithms, and the categorization of potential risk variables yields the best results. AKI samples showed significant organ defects than non-AKI ones. Multivariate LR indicated that lymphocyte, and myoglobin (MB) ≥ 1000 ng/ml were independent risk parameters for AKI samples. Also, GCS score (95% CI 1.4-8.3 P = 0.014) were the risk parameters for 60-day mortality in samples with AKI. Also, 90-day mortality in AKI patients was significantly high (P < 0.0001). In compared to the control group, there were no appreciable changes in the kidney/body weight ratio or body weight increases. Total thiol levels in kidney homogenate significantly decreased, and histopathological analysis confirmed these biochemical alterations. According to the results, oral TiO2 NP treatment may cause kidney damage in experimental samples.

Increased interleukin-22 levels in lupus nephritis and its associated with disease severity: a study in both patients and lupus-like mice model.

OBJECTIVES: Interleukin-22 (IL-22) has been considered as an inflammatory cytokine. In the present study, we investigated the potential role of IL-22 in lupus nephritis (LN). METHODS: We examined the IL-22 levels of serum and kidney tissue from LN patients and MRL/lpr mice. An intraperitoneal injection of saline, isotype control antibody (IgG), prednisone (3mg/kg/mouse), or anti-IL-22 mAb (5µg/kg/mouse or 25µg/kg/mouse) was administered twice a week from 6 to 18 weeks of age. RESULTS: IL-22 levels in both serum and kidney were significantly higher in LN patients as compared with those in healthy controls. The serum and renal levels of IL-22 in MRL/lpr mice were significantly increased over time. After MRL/lpr mice were treated with anti-IL-22 monoclonal antibody (mAb) for 12 weeks, significantly less urine protein and lower serum levels of creatinine and urea nitrogen were found. In addition, less renal injury score and few number of inflammatory cells per glomerulus were observed in MRL/lpr mice treated with anti-IL-22 mAb as compared with control groups. CONCLUSIONS: Our results suggest that IL-22 as a pathogenic cytokine might be a potential target for treatment of lupus nephritis.

Altered Amplitude of Low-Frequency Fluctuations in Inactive Patients with Nonneuropsychiatric Systemic Lupus Erythematosus.

Objective: This study is aimed at investigating the characteristics of the spontaneous brain activity in inactive patients with nonneuropsychiatric systemic lupus erythematosus (non-NPSLE). Methods: Thirty-one female inactive patients with non-NPSLE and twenty healthy controls were examined by resting-state functional magnetic resonance imaging (RS-fMRI). Three amplitude methods including amplitude of low-frequency fluctuations (ALFF), fractional amplitude of low-frequency fluctuations (fALFF), and percent amplitude of fluctuation (PerAF) (with and without standardization) were applied to evaluate the spontaneous brain activity. The correlation was performed between low-frequency oscillations and clinical and neuropsychological factors in inactive patients with non-NPSLE. Results: Compared to healthy controls, patients with non-NPSLE showed increased standardized ALFF (mALFF) in the left inferior temporal gyrus and left putamen, decreased PerAF in the right postcentral gyrus and bilateral precentral gyrus, and increased standardized PerAF (mPerAF) in the left putamen and decreased mPerAF in the right postcentral gyrus and bilateral precentral gyrus. By standardized fALFF (mfALFF), no significant brain regions were found between the two groups. Correlation analysis revealed significantly positive correlations between glucocorticoid dose and PerAF in the right precentral gyrus and mPerAF in the left putamen, and Complement 3 (C3) and mPerAF in the right postcentral gyrus. There was a significant negative correlation between C3 and mALFF in the left putamen. Conclusion: Abnormal low-frequency oscillations in multiple brain regions were found in inactive patients with non-NPSLE, indicating that the alteration of mALFF, PerAF, and mPerAF in specific brain regions might be an imaging biomarker of brain dysfunction in inactive patients with non-NPSLE.

Long Non-Coding RNA AL928768.3 Promotes Rheumatoid Arthritis Fibroblast-Like Synoviocytes Proliferation, Invasion and Inflammation, While Inhibits Apoptosis Via Activating Lymphotoxin Beta Mediated NF-κB Signaling Pathway.

Our previous study using RNA sequencing and reverse transcription quantitative polymerase chain reaction (RT-qPCR) validation identified a long non-coding RNA (lnc), lnc-AL928768.3, correlating with risk and disease activity of rheumatoid arthritis (RA), then the present study was conducted to further investigate the interaction of lnc-AL928768.3 with lymphotoxin beta (LTB) and their impact on proliferation, migration, invasion, and inflammation in RA-fibroblast-like synoviocytes (RA-FLS). Human RA-FLS was obtained and transfected with lnc-AL928768.3 overexpression, negative control overexpression, lnc-AL928768.3 short hairpin RNA (shRNA) and negative control shRNA plasmids. Then cell functions and inflammatory cytokine expressions were detected. Afterward, rescue experiments were conducted via transfecting lnc-AL928768.3 shRNA with or without LTB overexpression plasmids in RA-FLS. Lnc-AL928768.3 enhanced proliferation and invasion, inhibited apoptosis, while had little impact on migration in RA-FLS. In addition, lnc-AL928768.3 positively modulated interleukin-1ß (IL-1ß), IL-6 and IL-8 expressions in RA-FLS supernatant; moreover, it also positively regulated LTB mRNA expression, LTB protein expression, p-NF-κB protein expression, and p-IKB-α protein expression in RA-FLS. Furthermore, following experiment showed that lnc-AL928768.3 positively regulated LTB expression while LTB did not impact on lnc-AL928768.3 expression in RA-FLS. Furthermore, in rescue experiments, LTB overexpression curtailed the effect of lnc-AL928768.3 knock-down on regulating proliferation, invasion, apoptosis and inflammatory cytokine expressions in RA-FLS. Lnc-AL928768.3 promotes proliferation, invasion, and inflammation while inhibits apoptosis of RA-FLS via activating LTB mediated NF-κB signaling.

Clinical characteristics and risk factors of Helicobacter pylori infection-associated Sjogren's syndrome.

OBJECTIVE: Although infectious pathogens are predominant factors for inducing and maintaining immune system disorders, there exist few reports establishing the significant correlation between Helicobacter pylori (H. pylori) infection and Sjogren's syndrome. This study aims to demonstrate the correlation between Sjogren's syndrome and H. pylori infection in patients, highlighting various clinical characteristics and risk factors. METHODS: A single-center retrospective observational study was conducted in patients (n = 224) admitted from January 1, 2012, to February 10, 2021, in the First Affiliated Hospital of Wenzhou Medical University (Wenzhou, China). All the recruited subjects with Sjogren's syndrome and H. pylori infection were only included by validating the available medical records online. RESULTS: In this study, a total of 224 patients from January 1, 2012, to February 10, 2021, were diagnosed with Sjogren's syndrome. Among them, 94 patients (41.96%) with Sjogren's syndrome were infected with H. pylori. Accordingly, the clinical manifestations, serological and immunological characteristics, as well as gastroscopic biopsy outcomes of the recruited patients with primary Sjogren's syndrome (pSS) were reported. The multivariable analysis of the dry syndrome patients infected with H. pylori displayed hypergammaglobulinemia (odds ratio [OR], 0.354; 95% confidence interval [CI], 0.189-0.663), total cholesterol (OR, 1.158; 95% CI, 0.856-1.550), hypertension (OR, 0.227; 95% CI, 0.114-0.455), Female sex (OR, 5.778; 95% CI, 1.458-22.9), anti-SSA/Ro60 positive (OR, 2.384; 95% CI, 233-4.645), γ-GT (OR, 0.99; 95% CI, 0.99-1.00) and alkaline phosphatase (ALP, OR, 1.00; 95% CI, 0.99-1.00) levels. CONCLUSION: Together, our findings demonstrated that hypergammaglobulinemia could be the independent risk factors of H. pylori infection in patients with Sjogren's syndrome, requiring the physician's advice in the future.

Slit2 is a potential biomarker for renal impairment in systemic lupus erythematosus.

Slit2 glycoprotein has been described to regulate the inflammatory response and be involved in autoimmune diseases. Here, we investigated the expression of Slit2 and its potential significance in systemic lupus erythematosus (SLE). A total of 103 patients with SLE participated in our study. The levels of serum Slit2 were measured by enzyme-linked immunosorbent assay, and the expression of Slit2 in renal tissue was detected by immunohistochemistry. Patients with active disease had higher levels of serum Slit2 than patients with inactive disease and controls. Patients with sole skin impairment or sole renal impairment or both skin and renal impairment had higher levels of serum Slit2 than patients with neither skin nor renal impairment. Patients with chronic kidney disease (CKD) had higher levels of serum Slit2 than patients with no CKD. Levels of serum Slit2 in patients with active disease were positively correlated with the SLE Disease Activity Index, complement C4, and anti-dsDNA antibody. Levels of serum Slit2 in patients with CKD were positively correlated with serum creatinine, urine protein, and glomerular filtration rate. The expression of Slit2 and its receptor Roundabout1 (Robo1) in the renal tissue of patients with lupus nephritis were higher than controls. Moreover, renal Slit2 was positively correlated with renal chronic index. Our data indicated that Slit2 may contribute to renal impairment and this may be a potential biomarker for SLE.

Interleukin-22 From Type 3 Innate Lymphoid Cells Aggravates Lupus Nephritis by Promoting Macrophage Infiltration in Lupus-Prone Mice.

Lupus nephritis (LN) is one of the most severe manifestations of systemic lupus erythematosus (SLE). Our previous studies demonstrated increased serum and renal Interleukin (IL)-22 in LN patients and MRL/lpr mice. This study investigated the role of IL-22 and its mechanism in LN. Here, we found that IL-22 was mainly produced by type 3 innate lymphoid cells (ILC3) in kidney of MRL/lpr mice. The systemic illness and local renal lesion were significantly alleviated in IL-22 or IL-22R gene knockout (KO) mice (IL-22 KO or IL-22R KO MRL/lpr mice) than control mice (MRL/lpr mice). IL-22 KO or IL-22R KO MRL/lpr mice had significantly slighter infiltration of macrophage in kidney than MRL/lpr mice. Consistently, by RNA-Seq, the expression of (CC motif) ligand 2 (CCL2) and (CXC motif) ligand 10 (CXCL10) was decreased in kidney of KO mice compared with control mice. By immunoblotting, significantly increased levels of STAT3 phosphorylation were found in the kidney of control mice compared to KO mice. In vitro, primary kidney epithelial cells from control mouse stimulated with recombinant IL-22 (rIL-22) expressed higher levels of CCL2, CXCL10, and phosphorylated STAT3. At the same time, when primary kidney epithelial cells were treated with rIL-22, transwell assay demonstrated their supernatant recruited more macrophages. In human kidney epithelial cell line (HK2) cells, when treated with rIL-22, we observed similar results with primary mouse kidney epithelial cells. Moreover, when cells were stimulated with rIL-22 following pre-treatment with STAT3 pathway inhibitor, the expression of CCL2 and CXCL10 were significantly reversed. Our findings demonstrate that IL-22 binding to IL-22R in kidney epithelial cells activated the STAT3 signaling pathway, enhanced the chemokine secretion and then promoted macrophage infiltration to the kidney of MRL/lpr mice, thus aggravated LN in lupus-prone mice. These findings indicate that IL-22 may play a pathogenic role in LN and may provide a promising novel therapeutic target for LN.

Altered Temporal Dynamics of Brain Activity in Multiple-Frequency Bands in Non-Neuropsychiatric Systemic Lupus Erythematosus Patients with Inactive Disease.

PURPOSE: In this study, we seek to investigate dynamic changes of brain activity in non-neuropsychiatric systemic lupus erythematosus (non-NPSLE) patients with inactive disease. PATIENTS AND METHODS: Thirty-one non-NPSLE patients with inactive disease and 20 matched healthy controls underwent the blood oxygenation level-dependent fMRI examination. Dynamic regional homogeneity (ReHo) and fractional amplitude of low-frequency fluctuations (fALFF) were used to analyze the brain activity in typical band (0.01-0.08 Hz), slow-4 (0.027-0.073 Hz) and slow-5 (0.01-0.027 Hz). Pearson's correlation analysis was performed to correlate dynamic regional homogeneity (dReHo) and dynamic fractional amplitude of low-frequency fluctuations (dfALFF) values for clusters of voxels where significant group differences were found with clinical variables in non-NPSLE patients with inactive disease. RESULTS: In typical band, non-NPSLE patients showed increased dReHo in left middle occipital gyrus (MOG) compared to healthy controls. Meanwhile, patients showed decreased dfALFF in right superior frontal gyrus (SFG) and bilateral middle frontal gyrus (MFG) in typical band. In slow-4, increased dReHo in left MOG was found in non-NPSLE patients. In slow-5, non-NPSLE patients showed increased dReHo in left MOG, left calcarine fissure and surrounding cortex, right precentral gyrus (PreCG) and left postcentral gyrus (PoCG). Meanwhile, non-NPSLE patients showed decreased dfALFF in left SFG, right MFG, and right PreCG in slow-5. Moreover, the glucocorticoid dose showed significantly negative correlations with dReHo values in right PreCG in slow-5, left PoCG in slow-5, and left MOG in typical band. CONCLUSION: dReHo and dfALFF abnormalities in different frequency bands may be the key characteristics in the pathogenesis mechanism of non-NPSLE.

[Effects of sediment resuspension on nitrogen and phosphate exchange at the sediment-water interface in East Chongming Tidal Flat].

Sampling sediment and water to mimic sediment resuspension and researching its effects on nitrogen and phosphate exchange at the sediment-water interface in east Chongming tidal flat. Through the experiment, it was found that sediment resuspension causes NO3(-)-N, NH4(+)-N, NO2(-)-N and DIP releases. The release of NO3(-)-N is most marked, the increasing concentration of NO3(-)-N, NH4(+)-N, NO2(-)-N and DIP reach in turns: 11.869 mumol.L-1, 2.1713 mumol.L-1, 0.2 mumol.L-1, 0.02 mumol.L-1. At the same time, resuspension also change nitrogen and phosphate exchange at the sediment-water interface compare to without resuspension to great extent. NH4(+)-N and NO3(-)-N have absolutely opposite diversification with and without resuspension; NO2(-)-N shows complicated transformation because of many effected elements. The result of the experiment display the changes of NH4(+)-N, NO2(-)-N and DIP concentration in water have good relation to SPM (mg.L-1), this relation have not appear in change of NO3(-)-N.