Reduced miR-99a-3p levels in systemic lupus erythematosus may promote B cell proliferation via NCAPG and the PI3K/AKT signaling pathway.

BACKGROUND: Systemic lupus erythematosus is an immunologically dysregulated disease characterized by the presence of multiple autoantibodies. In SLE, B lymphocytes contribute to the dysregulated production of autoantibodies and cytokines. Recently, we discovered that miR-99a-3p binds to both EIF4EBP1 and NCAPG mRNA and that lowering miR-99a-3p can promote B cell autophagy in SLE by increasing EIF4EBP1 expression. However, the functions of miR-99a-3p and NCAPG in SLE have not been extensively investigated. OBJECTIVE: This work aims to evaluate the levels of miR-99a-3p and NCAPG expression in SLE B cells and to determine whether the aberrant expression of miR-99a-3p and NCAPG contributes to the pathological mechanisms in SLE. METHODS: B lymphocytes were obtained through immunomagnetic negative selection. Using RT-qPCR, miR-99a-3p and NCAPG mRNA expressions in B lymphocytes and in the BALL-1 cell line were measured. To determine the relative abundance of NCAPG, PI3K, p-PI3K, AKT, and p-AKT, we normalize them to the level of ß-actin using Western blotting. Evaluation of miR-99a-3p and NCAPG's impact on cell proliferation was done utilizing CCK-8 assay. Using flow cytometry, the cell cycle and apoptosis were both measured. RESULTS: Comparing SLE B cells to healthy controls, miR-99a-3p expression was significantly downregulated. Additionally, it was observed that SLE B cells had significantly higher NCAPG mRNA expression. Blocking miR-99a-3p expression in BALL-1 cells with an antagomir elevated NCAPG expression, facilitated PI3K/AKT pathway activation, improved cell proliferation, raised the fraction of S-phase cells, and prevented cell apoptosis. The opposite effects of upregulated miR-99a-3p levels on BALL-1 cells were observed by using an agomir. Furthermore, the effect of decreased miR-99a-3p expression on cell proliferation was partially mediated by elevating NCAPG levels and activating the PI3K/AKT pathway. CONCLUSION: Our research indicates that lower miR-99a-3p expression in SLE B cells appears to boost B cell number via the NCAPG and PI3K/AKT pathways.

A comprehensive exploration of the heterogeneity of immune cells in Han and Zang systemic lupus erythematosus patients via single-cell RNA sequencing.

Systemic Lupus Erythematosus (SLE) is an autoimmune sickness with unclear pathogenesis. The goal of this research was to reveal the heterogeneity of immune cells in SLE patients of Han and Zang nationality by single-cell RNA sequencing (scRNA-seq) and bioinformatics profiling. METHODS: A total of 94,102 peripheral blood mononuclear cells (PBMCs) from six volunteers with SLE (3 Zang, 3 Han) and six healthy controls were first conducted through scRNA-seq analysis. The immune cell subsets in the pathogenesis of SLE were analyzed as well. Real-time quantitative PCR (RT-qPCR) was applied to confirm the results of sc-RNA seq analysis. RESULTS: For the Tibetan samples, the ratios of Naïve CD4 RPS4Y1 cells, Naïve CD4 cells, Memory BC CD24 and Memory BC differed significantly between the SLE and control samples, while that of CD8 CTL MAL cells was significantly different between the two groups in Han nationality samples. Variable differentiation states of CD8 CTL MAL cells, CD8 CTL GZMK cells, and Naïve CD4 cells were detected through pseudotime analysis. Moreover, T-cell receptor (TCR) abundance was notably higher in Tibetan SLE specimens than that in controls, while B-cell receptor (BCR) abundance in Tibetan and Han samples was higher than in control groups. CONCLUSIONS: In summary, the immune cellular heterogeneity of SLE patients both Han and Zang nationality was explored based on various bioinformatics approaches, providing new perspectives for immunological characteristics of SLE among different ethnic groups.

Early life history affects fish size mainly by indirectly regulating the growth during each stage: a case study in a demersal fish.

The complex life histories of demersal fishes are artificially separated into multiple stages along with changes in morphology and habitat. It is worth exploring whether the phenotypes expressed earlier and later during the life cycle are related or decoupled. The life stages of first year Pacific cod (Gadus macrocephalus) were tracked over different hatch years and regions to test whether the early life history had a long-lasting effect on subsequent growth. We further explored the contribution of growth in the early and subsequent life history stages to body size at the end of each stage. In addition to the accessory growth centre and the first annual ring, the other two checks on the otolith possibly related to settlement and entering deeper waters were identified in 75 Pacific cod individuals. The direct and indirect relationships among the life history stages was interpreted based on path analysis. The results showed that growth prior to the formation of the accessory growth centre had a significant effect on the absolute growth of the fish before and after settlement and migration to deep water. However, there was no or moderate evidence that early growth affected the body size at each stage, which was mainly regulated by growth during the stage. This study supports the lasting effect of early growth and clarifies that it affects size mainly by indirectly regulating staged growth. Quantifying the phenotype relationships and identifying the internal mechanisms form the basis for assessing population dynamics and understanding the processes behind the changes. Supplementary Information: The online version contains supplementary material available at 10.1007/s42995-022-00145-y.

Meta-analysis on the efficacy and safety of immunoadsorption for systemic lupus erythematosus among Chinese population.

To systematically evaluate the efficacy and safety of immunoadsorption (IAS) versus non-IAS for systemic lupus erythematosus (SLE) among Chinese population. A meta-analysis was performed by all the literatures germane to estimate the SLE patients treated with IAS and non-IAS from published randomized controlled trials (RCTs) from 1990 to February 2020. Mean differences (MDs), relative ratios (RRs), and 95% confidence intervals (CIs) were calculated, and the meta-analysis was conducted with Stata 12.0 software. A total of 18 RCTs involving 457 patients were included. The results of meta-analysis demonstrated that the IgG, Scr, Bun, ANA, 24-h urine protein, leptin, and TNF-α of IAS combined with a drug therapy group were lower than that of non-IAS, and the levels of C3 and C4 were higher than that of non-IAS after treatment in terms of laboratory parameters. In terms of adverse reactions, the incidence of fever or chills, low blood pressure, or bleeding risk was higher in the treatment group. However, there was no difference in the incidence of puncture point bleeding, thrombocytopenia, mild rash, death due to severe infection, tightness, palpitation, or chest tightness between the two groups. However, most of the adverse effects could be considered as tolerable after timely treatment. Our results indicate that IAS may be superior to non-IAS in treating SLE patients. However, due to the lower quality of included studies, high quality of multicenter, large sample size, randomized, and double-blind controlled trials are needed to validate the results.

MicroRNA-30a functions as tumor suppressor and inhibits the proliferation and invasion of prostate cancer cells by down-regulation of SIX1.

Increasing reports have demonstrated that aberrant expression of microRNAs (miRNAs) is found in multiple human cancers. Many studies have shown that down-regulated level of miR-30a is in a variety of cancers including prostate cancer (PCa). However, the precise mechanisms of miR-30a in PCa have not been well explored. In this study, we investigated the biological functions and molecular mechanism of miR-30a in PCa cell lines, discussing whether it could be a therapeutic biomarker of PCa in the future. We found that miR-30a is down-regulated in PCa tissues and cell lines. Moreover, the low level of miR-30a was associated with increased expression of SIX1 in PCa tissues and cell lines. Up-regulation of miR-30a significantly inhibited proliferation of PCa cells. In addition, invasion of PCa cells was suppressed by overexpression of miR-30a. However, down-regulation of miR-30a promoted cell growth and invasion of PCa cells. Bioinformatics analysis predicted that the SIX1 was a potential target gene of miR-30a. Next, luciferase reporter assay confirmed that miR-30a could directly target SIX1. Consistent with the effect of miR-30a, down-regulation of SIX1 by siRNA inhibited proliferation and invasion of PCa cells. Overexpression of SIX1 in PCa cells partially reversed the effect of miR-30a mimic. In conclusion, introduction of miR-30a dramatically inhibited proliferation and invasion of PCa cells by down-regulating SIX1 expression, and that down-regulation of SIX1 was essential for inhibition of cell growth and invasion of PCa cells by overexpression of miR-30a.