Bioinformatic identification and validation of autophagy-related genes in rheumatoid arthritis.

OBJECTIVES: Rheumatoid arthritis (RA) is a chronic systemic autoimmune disorder characterized by progressive synovial inflammation and joint destruction, with a largely unknown etiology. Studies have suggested that autophagy and its expression may be involved in the pathogenesis of RA; however, autophagy-related genes in RA are still largely unidentified. Therefore, in this study, we aimed to identify and validate autophagy-related genes in RA. METHODS: We identified differentially expressed autophagy-related genes between patients with RA and healthy individuals using gene expression profiles in the GSE55235 dataset and R software. Subsequently, correlation analysis, protein-protein interaction, gene ontology enrichment, and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were carried out using these differentially expressed autophagy-related genes. Finally, our results were validated by examining the expression of differentially expressed autophagy-related hub genes in clinical samples using qRT-PCR. RESULTS: We identified 52 potential autophagy-related genes in RA based on bioinformatic analyses. Ten hub genes, CASP8, CTSB, TNFSF10, FADD, BAX, MYC, FOS, CDKN1A, GABARAPL1, and BNIP3, were validated to be differentially expressed and may serve as valuable prognostic markers and new potential therapeutic targets for RA via the regulation of autophagy. CONCLUSIONS: Our results may help improve the understanding of RA pathogenesis. Autophagy-related genes in RA could be valuable biomarkers for diagnosis and prognosis and they might be exploited clinically as therapeutic targets in the future.

[Neonicotinoid Insecticides Threaten Surface Waters at the National Scale in China].

This study was conducted in response to the current situation in our country in which neonicotinoid pesticides (NNIs) are used in large quantities and their concentration in surface water is gradually increasing. Based on the species sensitivity distribution (SSD), the hazard quotient (HQ) and probabilistic risk assessment (PRA) were used to compare single and mixture risks of NNIs in the surface water in China. The target and recommended values of NNIs in China's surface water quality supervision were also presented. The results showed that:① in the single acute hazard assessment, imidacloprid (IMI) was the most harmful insecticide, and in the single chronic hazard assessment, imidacloprid (IMI) and acetamiprid (ACE) were more harmful. Furthermore, insects were the most sensitive creatures. ② Hainan province was the region with the highest single acute and chronic hazard in the study area. ③ Based on the joint probability curves of five neonicotinoids, the probability in which 5% of species would be affected by long-term exposure was approximately 91.12%. Thus, the combined ecological risk of these five neonicotinoids cannot be ignored. ④ Based on the toxicity reference value from SSD and the combined results of HQ and PRA, the regulatory values of surface water quality in China were as follows:acetamiprid (ACE) 0.04 µg·L-1, clothianidin (CLO) 0.22 µg·L-1, imidacloprid (IMI) 0.01 µg·L-1, thiacloprid (THI) 0.03 µg·L-1, and thiamethoxam (THIA) 0.24 µg·L-1. In short, the concentration of NNIs in the surface water in China has threatened the safety of aquatic organisms, and its supervision must be improved.

[Diversity and network features of fungal community in the soils of planted and natural Picea asperata forests.] / äººå·¥ä¸Žå¤©ç„¶äº‘æ‰æž—åœŸå£¤çœŸèŒç¾¤è½å¤šæ ·æ€§åŠèŒç¾¤ç½‘ç»œå ³ç³»ç‰¹å¾.

The diversity and interactions of soil fungal community are the key to maintain the diversity and stability of ecosystem. In this study, we examined the structure, diversity and co-occurrence networks of fungal community in rhizosphere and non-rhizosphere soils of planted and natural Picea asperata forests using high-throughput sequencing technique and bioinformatic methods. The results showed that Inocybaceae and Sebacinaceae were dominant family in soils of planted and natural forests, respectively. At the genus level, Inocybe was dominant one in soils of planted and natural forests. There were significant differences in ß-diversity of fungal communities between rhizosphere and non-rhizosphere soils in both planted and natural forests. There were no significant correlations between environmental variables and the relative abundance and α-diversity of fungal communities. Herb layer coverage, soil water content, total organic carbon concentration, and plant species richness played important roles in explaining the variations of ß-diversity of fungal communities. Results of the network analysis showed that the negative correlations were dominant among soil fungal communities in natural forest, suggesting that the competition of different groups in natural forest. Moreover, there were more negative correlations in non-rhizosphere soils than in rhizosphere soils, which indicated that fungal communities in non-rhizosphere soils comprised more competitive network structure than in the rhizosphere soils. Biomarker species were identified based on differential abundance analysis. Sebacinaceae was the single shared keystone species in the fungal network which had significant differences among rhizosphere and non-rhizosphere soils of planted and natural forests. Therefore, it is suggested that the variation of differential species in the soil fungal communities between the planted and natural forest might had limited influence on the stability of the community of planted and natural forests.

Molecular Signatures Related to the Virulence of Bacillus cereus Sensu Lato, a Leading Cause of Devastating Endophthalmitis.

Bacillus endophthalmitis is a devastating eye infection that causes rapid blindness through extracellular tissue-destructive exotoxins. Despite its importance, knowledge of the phylogenetic relationships and population structure of intraocular Bacillus spp. is lacking. In this study, we sequenced the whole genomes of eight Bacillus intraocular pathogens independently isolated from 8/52 patients with posttraumatic Bacillus endophthalmitis infections in the Eye Hospital of Wenzhou Medical University between January 2010 and December 2018. Phylogenetic analysis revealed that the pathogenic intraocular isolates belonged to Bacillus cereus, Bacillus thuringiensis and Bacillus toyonensis To determine the virulence of the ocular isolates, three representative strains were injected into mouse models, and severe endophthalmitis leading to blindness was observed. Through incorporating publicly available genomes for Bacillus spp., we found that the intraocular pathogens could be isolated independently but displayed a similar genetic context. In addition, our data provide genome-wide support for intraocular and gastrointestinal sources of Bacillus spp. belonging to different lineages. Importantly, we identified five molecular signatures of virulence and motility genes associated with intraocular infection, namely, plcA-2, InhA-3, InhA-4, hblA-5, and fliD using pangenome-wide association studies. The characterization of overrepresented genes in the intraocular isolates holds value to predict bacterial evolution and for the design of future intervention strategies in patients with endophthalmitis.IMPORTANCE In this study, we provided a detailed and comprehensive clinicopathological and pathogenic report of Bacillus endophthalmitis over the 8 years of the study period. We first reported the whole-genome sequence of Bacillus spp. causing devastating endophthalmitis and found that Bacillus toyonensis is able to cause endophthalmitis. Finally, we revealed significant endophthalmitis-associated virulence genes involved in hemolysis, immunity inhibition, and pathogenesis. Overall, as more sequencing data sets become available, these data will facilitate comparative research and will reveal the emergence of pathogenic "ocular bacteria."

Astragaloside IV protects rat gastric mucosa against aspirin-induced damage.

Aspirin (Asp) is commonly used as an anti-inflammatory drug, but the long-term usage of Asp can lead to severe gastrointestinal damage. Thus the co-administering of Asp with another drug that can suppress its side effect while having no impact on its anti-inflammatory activity would be ideal. Astragaloside IV (AST-IV) is a natural anti-inflammatory compound that has been shown to protect rat gastric mucosa from anhydrous ethanol-inflicted damage. In this study, we investigated whether AST-IV could protect rat gastric mucosa against Asp-induced gastric mucosal damage. Wistar rats administered 150mg/kg Asp showed significant damage to the gastric mucosa, as revealed by gastric damage score and histological evaluation. However, this was largely abolished by co-administering Asp and 25mg/kg or 50mg/kg AST-IV. The protective mechanism of AST-IV involved the suppression of Asp-induced inhibition of cycloxygenase-1 (COX-1) expression, prostaglandin E2 (PGE2) production, superoxide dismutase (SOD) activity and nitric oxide (NO) production. AST-IV blocked Asp-induced inhibition of SOD activity through preventing Asp from inhibiting the expression of SOD-1, both at the mRNA and protein levels. AST-IV did not appear to interfere with the anti-inflammatory activity of Asp since COX-2 level in model gastritis rats treated with Asp plus AST-IV was equally suppressed as in model gastritis rats treated with Asp alone. The results clearly showed that AST-IV could neutralize the toxicity of Asp while having no impact on its anti-inflammatory activity. AST-IV could therefore be considered as a potential drug for relieving the side effect associated with the long-term usage of Asp.

Anti-cancer activity and potential mechanism of a novel aspirin derivative.

Aspirin has been used in the treatment and chemoprevention of many malignant cancers. The mechanism of its anti-cancer activity mainly involves the inhibition of cyclooxygenase-2 (COX-2). However, the application of aspirin is limited by the serious gastric mucosal damage that accompanies its usage. We have previously reported the preparation of a novel aspirin derivative that we named Ca-Asp, and showed that it causes less damage to gastric mucosa of rat and inhibits the expression of COX-2 to higher degree than Asp. However, the anti-cancer effect and mechanism of Ca-Asp was not demonstrated. In this study, the anti-cancer effect of Ca-Asp was investigated and compared with those of Asp and Hydroxyapatite (Hap) at the cell level. The results showed that treatment of SGC-7901 cells (human gastric cancer cell line) with 200-400µg/ml Ca-Asp resulted in significant reduction in cell viability, compared to treatment with either Asp or Hap, and at a higher concentration (500µg/ml). Subsequent investigation into the possible underlying mechanism showed that Ca-Asp induced apoptosis and caused cell cycle arrest at the G1 phase. Ca-Asp also up-regulated the levels of caspase-3 and p53, but down regulated the level of cyclin D1, NF-κB, COX-2 and PGE2. Furthermore, simultaneous treatment of SGC-7901 cells with Ca-Asp and exogenous PGE2 reduced the anti-proliferative effect of Ca-Asp on the cells. Taken together, the results suggested that Ca-Asp might act as a potential anti-cancer drug, and that its suppression of PGE2 production might constitute an important part of its anti-cancer activity.

Inhibition of adjuvant-induced arthritis by nasal administration of novel synthetic peptides from heat shock protein 65.

BACKGROUND: Rheumatoid arthritis (RA) is a chronic systemic inflammatory disease mediated by T cells. The aim of the present study was to investigate the therapeutic efficacy of synthetic peptides (HP-R1, HP-R2 and HP-R3), derived from the sequence of 65-kD mycobacterial heat shock protein (HSP), in the treatment of RA using adjuvant-induced arthritis (AA) animal model. METHODS: AA was induced by a single intradermal injection Freund's complete adjuvant in male Lewis rats. At the first clinical sign of disease, rats were administered nasally by micropipette of peptides or phosphate buffer saline (PBS). Disease progression was monitored by measurement of body weight, arthritis score and paw swelling. The changes of histopathology were assessed by hematoxylin eosin staining. The serum levels of tumor necrosis factor (TNF) - alpha and interleukin (IL)-4 were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The peptides efficiently inhibited the footpad swelling and arthritic symptoms in AA rats. The synthetic peptides displayed significantly less inflammatory cellular infiltration and synovium hyperplasia than model controls. This effect was associated with a suppression of pro-inflammatory cytokine TNF-alpha production and an increase of anti-inflammatory cytokine IL-4 production after peptides treatment. CONCLUSIONS: These results suggest that the synthetic peptides derived from HSP65 induce highly effective protection against AA, which is mediated in part by down-regulation of inflammatory cytokines, and support the view that the synthetic peptides is a potential therapy for RA that may help to diminish both joint inflammation and destruction.

Synthetic peptides from heat-shock protein 65 inhibit proinflammatory cytokine secretion by peripheral blood mononuclear cells from rheumatoid arthritis patients.

1. Rheumatoid arthritis (RA) is a systemic autoimmune disease mediated by T cells. Proinflammatory cytokines plays a critical role in the pathogenesis of RA. The aim of the present study was to investigate the effects of synthetic peptides (HP-R1, HP-R2 and HP-R3), derived from the sequence of 65 kDa mycobacterial heat shock protein (HSP), on the proliferation of and cytokine secretion by peripheral blood mononuclear cells (PBMC) from RA patients. 2. The PBMC were obtained from RA patients and collected by Ficoll-Hypaque density centrifugation. Peripheral blood mononuclear cells were treated with one of the three synthetic peptides for 4 h, after which time proliferation and cytokine production were determined. The effects of the three peptides on the proliferation of PBMC were analysed by the colorimetric cell proliferation (CCK-8) assay. Cytokine production was measured in culture supernatants using specific ELISAs. 3. None of the three peptides had any significant effect on the proliferation of PBMC from healthy controls. However, the proliferation of PBMC from RA patients was inhibited by all three peptides. The production of tumour necrosis factor-α from RA patients was significantly inhibited by all three peptides. The secretion of interferon-γ was significantly suppressed by HP-R1 and HP-R2. Unlike the other two peptides, HP-R2 increased the secretion of interleukin (IL)-4. None of the peptides had any significant effect on the production of IL-10. 4. The results of the present study suggest that the synthetic peptides derived from HSP65 exhibit antiproliferative and anti-inflammatory activity, and support the potential use of synthetic peptides as therapeutic drugs in RA patients.