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A method of headspace-solid-phase microextraction-gas chromatography/triple quadrupole mass spectrometry was established for simultaneous determination of 10 types of 51 odorous substances in water. The effects of extraction head, extraction temperature, extraction time, salt content, desorption time and split ratio on detection results were investigated with optimized gas chromatography-mass spectrometry parameters using the multiple reaction monitoring (MRM) mode for triple quadrupole mass spectrometry. The results showed that the optimized conditions were polydimethylsiloxane/divinylbenzene as extraction head, 55 ℃ as extraction temperature, 20 min as extraction time, 300 g/L NaCl as salt content, desorption time of 0.5 min and the split ratio of 10∶1. The detection limits of 51 odorous substances ranged from 0.4 to 183 ng/L, and the linear correlation coefficients were greater than 0.995. The detection limits of indicator substances, such as geosmin, 2-methylisothiol, anisole, dimethyl disulfide, and dimethyl trisulfide were 0.4, 0.9, 1.9, 3.5, 1.7 ng/L, respectively, which met the limit requirement of “Standard for Drinking Water Quality (GB 5749—2022)” of China. The spiked recoveries of the low, medium and high concentration levels were 76.2%-118%, 78.9%-119% and 82.0%-119%, and the relative standard deviations were 2.9%-19.4%, 1.8%-13.4% and 0.9%-13.2%, respectively. This method was used to detect the odorous substances in 29 surface waters, in which 12 odorous substances such as methyl mercaptan, dimethyl sulfide, and DL-sec-butyl acetate were detected with the concentration range of 2.2~1 651 ng/L. There were no apparent interferences in the MRM channels for the detected odorous substances. This method has the characteristics of simple pretreatment, good reproducibility and high accuracy, which is suitable for simultaneous and rapid screening of various types of odorous substances in water bodies.

Pulmonary fibrosis (PF) is an interstitial lung disease with complex pathological mechanism, and there is currently a lack of therapeutics that can heal it completely. Using gene therapy with drugs provides promising therapeutic strategies for synergistically reversing PF. However, improving the intracellular accumulation and transfection efficiency of therapeutic nucleic acids is still a critical issue that urgently needs to be addressed. Herein, we developed lipid nanoparticles (PEDPs) with high transfection efficiency coloaded with pDNA of nuclear factor erythroid 2-related factor 2 (pNrf2) and pirfenidone (PFD) for PF therapy. PEDPs can penetrate biological barriers, accumulate at the target, and exert therapeutic effects, eventually alleviating the oxidative stress imbalance in type II alveolar epithelial cells (AECs II) and inhibiting myofibroblast overactivation through the synergistic effects of Nrf2 combined with PFD, thus reversing PF. In addition, we systematically engineered various liposomes (LNPs), demonstrated that reducing the polyethylene glycol (PEG) proportion could significantly improve the uptake and transfection efficiency of the LNPs, and proposed a possible mechanism for this influence. This study clearly reveals that controlling the composition ratio of PEG in PEDPs can efficiently deliver therapeutics into AECs II, improve pNrf2 transfection, and synergize with PFD in a prospective strategy to reverse PF.

Based on the coupled communication of smart elderly care and traditional Chinese culture, this paper proposes four paths of coupled communication, including in-depth communication, innovative cultural forms of communication, precise communication, and promotion of coupled development. Two systematic index systems have been constructed to disseminate traditional Chinese culture and develop smart culture and pension. The data is standardized using principal component analysis to establish the correlation coefficient matrix. The coordination index between the two subsystems is calculated, and the coordination model of the two is created based on the concepts of coordination coefficient and coefficient of variation. The subject area, Province S, is in the third stage of the coupled coordinated development during 2010-2023, with traditional culture dissemination and smart elderly care services coming to a high level, and is in a good development level during 2016-2023, with a coupled coordination degree of 8 or more. Using the VAR model, the effect of coupling development on the quality of life of elderly individuals is analyzed. Impulse response analysis of LNCCD and LNQOE: after LNCCD receives a positive shock, LNQOE shows a sharp shock effect. At this time, the response value is 0.07021, which plays a large positive effect, and then enters into a smooth development. The traditional Chinese culture dissemination and the wisdom culture of old age are in the stage of coupled development.

Background: Studies have shown that fine particulate matter (PM2.5) remains a significant problem in developing countries and plays a critical role in the onset and progression of respiratory illnesses. Circular RNAs (circRNAs) are involved in many pathophysiological processes,but their relationship to PM2.5 pollution is largely unexplored. Objectives: To elucidate the functional role of hsa_circ_0000992 in PM2.5-induced inflammation in a human bronchial epithelial cell line (16HBE) and to clarify whether the competing endogenous RNA (ceRNA) mechanism is involved in the interrelationships between hsa_circ_0000992 and hsa-miR-936 and the inflammatory signaling pathways. Methods: Detection of inflammatory factors in 16HBE cells exposed to PM2.5 by RT-qPCR and ELISA.High throughput sequencing and bioinformatics analysis methods were used to screen circRNA.The bioinformatics analysis method western blotting and dual-luciferase reporter gene system were used to verify mechanisms associated with circRNA. Results: PM2.5 cause inflammation in the 16HBE cells. High throughput sequencing and RT-qPCR result revealed that the expression of hsa_circ_0000992 was markedly up-regulated in 16HBE exposed to PM2.5. The binding sites between hsa_circ_0000992 and hsa-miR-936 was confirmed by dual-luciferase reporter gene system.Western blotting and RT-qPCR showed that hsa_circ_0000992 can interact with hsa-miR-936 to regulate AKT serine/threonine kinase 3(AKT3),thereby activating the PI3K/AKT pathway and ultimately promoting the expression of interleukin (IL)− 1β and IL-8. Conclusion: PM2.5 can induce the inflammatory response in 16HBE cells by activating the PI3K/AKT pathway. The expression of hsa_circ_0000992 increased when PM2.5 stimulated 16HBE cells,and the circRNA could then regulate the inflammatory response.Hsa_circ_0000992 regulates the hsa-miR-936/AKT3 axis through the ceRNA mechanism,thereby activating the PI3K/AKT signaling pathway,increasing the expression of cellular inflammatory factors,and promoting PM2.5-induced respiratory inflammation.

BackgroundAutophagy plays a critical role in the progression of osteoarthritis (OA), mainly by regulating inflammatory and immune responses. However, the underlying mechanisms remain unclear. This study aimed to investigate the potential relevance of autophagy-related genes (ARGs) associated with infiltrating immune cells in OA.MethodsGSE114007, GSE169077, and ARGs were obtained from the Gene Expression Omnibus (GEO) database and the Human Autophagy database. R software was used to identify the differentially expressed autophagy-related genes (DEARGs) in OA. Functional enrichment and protein–protein interaction (PPI) analyses were performed to explore the role of DEARGs in OA cartilage, and then Cytoscape was utilized to screen hub ARGs. Single-sample gene set enrichment analysis (ssGSEA) was used to conduct immune infiltration analysis and evaluate the potential correlation of key ARGs and immune cell infiltration. Then, the expression levels of hub ARGs in OA were further verified by the GSE169077 and qRT-PCR. Finally, Western blotting and immunohistochemistry were used to validate the final hub ARGs.ResultsA total of 24 downregulated genes and five upregulated genes were identified, and these genes were enriched in autophagy, mitophagy, and inflammation-related pathways. The intersection results identified nine hub genes, namely, CDKN1A, DDIT3, FOS, VEGFA, RELA, MAP1LC3B, MYC, HSPA5, and HSPA8. GSE169077 and qRT-PCR validation results showed that only four genes, CDKN1A, DDT3, MAP1LC3B, and MYC, were consistent with the bioinformatics analysis results. Western blotting and immunohistochemical (IHC) showed that the expression of these four genes was significantly downregulated in the OA group, which is consistent with the qPCR results. Immune infiltration correlation analysis indicated that DDIT3 was negatively correlated with immature dendritic cells in OA, and FOS was positively correlated with eosinophils.ConclusionCDKN1A, DDIT3, MAP1LC3B, and MYC were identified as ARGs that were closely associated with immune infiltration in OA cartilage. Among them, DDIT3 showed a strong negative correlation with immature dendritic cells. This study found that the interaction between ARGs and immune cell infiltration may play a crucial role in the pathogenesis of OA; however, the specific interaction mechanism needs further research to be clarified. This study provides new insights to further understand the molecular mechanisms of immunity involved in the process of OA by autophagy.

The systemic immune response, including B- and T-cell reactions, plays a corresponding role in syphilis infections. The TP0136 protein is a target of the immune response in infected hosts and may mediate the immune response. Here, we developed a method that combining reverse vaccine approach with Pepscan/T-cell proliferation to screen and identify three B-cell and two T-cell epitopes of TP0136, and explore the role of the B- and T-cell epitopes in immunized-infected animals. The results showed that immunized with B-cell epitopes not only had no protective effect but also aggravated the syphilitic lesion development. While immunized with T-cell epitopes of TP0136 could induce a strong Th1-cellular immunity response, which could attenuate syphilitic lesion development to a certain extent. The variation in exacerbation or attenuation of skin lesions, induced by distinct B- and T-cell epitopes of Tp0136, within the host's defense against syphilis warrants in-depth exploration.

Summary: Tubulointerstitial abnormalities contribute to the progression of diabetic kidney disease (DKD). However, the underlying mechanism of the pathobiology of tubulointerstitial disease is largely unknown. Here, we showed that MYCT1 expression was downregulated in in vitro and in vivo DKD models. Adeno-associated virus (AAV)-Myct1 significantly attenuated renal dysfunction and tubulointerstitial fibrosis in diabetic db/db mice and downregulated Sp1 transcription and TGF-β1/SMAD3 pathway activation. In human proximal tubular epithelial cells, high glucose-induced high expression of SP1 and TGF-β1/SMAD3 pathway activation as well as overaccumulation of extracellular matrix (ECM) were abrogated by MYCT1 overexpression. Mechanistically, the binding of VDR to the MYCT1 promoter was predicted and confirmed using dual-luciferase reporter and ChIP analysis. VDR transcriptionally upregulates MYCT1. Our data reveal MYCT1 as a new and potential therapeutic target in treating DKD.

Objective To explore the characteristics of steroid hormone spectrum in patients with primary aldoste- ronism (PA) and to evaluate the diagnostic efficacy of 18-oxocortisol and 18-hydroxycortisol. Methods Fifty-nine PA patients including 41 cases of aldosterone-producing adenoma (APA) and 18 cases of idiopathic hyperaldosteronism (IHA)and 42 essential hypertension (EH) patients as control group diagnosed in Department of Endocrinology of Peking Union Medical College Hospital from November 2020 to November 2021 were enrolled in this study. The basic information and clinical characteristics of patients were collected and 8 kinds of steroid hormones,including mineral corticoids (pregnenolone,progesterone, 11-deoxycorticosterone, corticosterone, 18-OH-corticoster-one) and aldosterone, and metabolites 18-oxocortisol as well as 18-hydroxycortisol were detected by liquid chromatography-tandem mass spectrometry(LC-MS/MS).The differences of steroid hormone level from different groups were compared and their diagnostic efficacy was evaluated. Results The average level of progesterone in APA group was higher than that in EH group (P＜0.05). The average level of 11-deoxycorticosterone in APA and IHA group was higher than that in EH group (both P＜0.001). The average level of 18-OH-corticosterone in APA group was higher than that in EH group (P＜0.001). The average level of aldosterone in APA and IHA group were higher than that in EH group(P＜0.01). The average level of 18-oxocortisol in APA group was higher than that in IHA group and higher than that in EH group (all P＜0.05). The average level of 18-hydroxycortisol in APA group was higher than that in EH group (P＜0.001). As for the diagnostic efficiency of 18-oxocortisol and 18-hydroxycortisol differentiating PA from EH, when the cut-off value of 18-oxocortisol was 0.06 ng/mL, the sensitivity reached 74.6% with specificity as 88.1%; when the cut-off value of 18-hydroxycortisol was 1.44 ng/mL, the sensitivity was 71.2% and the specificity was 69.0%. As for the diagnostic efficiency of 18-oxocortisol and 18-hydroxycortisol recognizing APA, when the cut-off value of 18-oxocortisol was 0.11 ng/mL, the sensitivity was 73.2% and the specificity was 95.2%; when the cut-off value of 18-hydroxycortisol was 1.77 ng/mL, the sensitivity was 68.3% and the specificity was 78.6%. Conclusions The level of various steroid hormone precursors and metabolites in PA patients is significantly higher than those in EH patients. The detection of steroid hormone profiling by LC-MS/MS plays an important role in the diagnosis and typing of PA. In particular, 18-oxocortisol and 18-hydroxycortisol show unique advantages in the recognition of APA.