Clinical-mediated discovery of pyroptosis in CD8+ T cell and NK cell reveals melanoma heterogeneity by single-cell and bulk sequence.

Histologically, melanoma tissues had fewer positive cells percentage of pyroptosis-related genes (PRGs), GZMA, GSDMB, NLRP1, IL18, and CHMP4A in epidermal than in normal skin. Pyroptosis, a new frontier in cancer, affects the tumor microenvironment and tumor immunotherapy. Nevertheless, the role of pyroptosis remains controversial, which reason is partly due to the heterogeneity of the cellular composition in melanoma. In this study, we present a comprehensive analysis of the single-cell transcriptome landscape of pyroptosis in melanoma specimens. Our findings reveal dysregulation in the expression of PRGs, particularly in immune cells, such as CD8+ cells (representing CD8+ T cells) and CD57+ cells (representing NK cells). Additionally, the immunohistochemical and multiplex immunofluorescence staining experiments results further confirmed GZMA+ cells and GSDMB+ cells were predominantly expressed in immune cells, especially in CD8 + T cells and NK cells. Melanoma specimens secreted a minimal presence of GZMA+ merged CD8+ T cells (0.11%) and GSDMB+ merged CD57+ cells (0.08%), compared to the control groups exhibiting proportions of 4.02% and 0.62%, respectively. The aforementioned findings indicate that a reduced presence of immune cells within tumors may play a role in diminishing the ability of pyroptosis, consequently posing a potential risk to the anti-melanoma properties. To quantify clinical relevance, we constructed a prognostic risk model and an individualized nomogram (C-index=0.58, P = 0.002), suggesting a potential role of PRGs in malignant melanoma prevention. In conclusion, our integrated single-cell and bulk RNA-seq analysis identified immune cell clusters and immune gene modules with experiment validation, contributing to our better understanding of pyroptosis in melanoma.

Trends in the distribution of socioeconomic inequalities in smoking and cessation: evidence among adults aged 18 ~ 59 from China Family Panel Studies data.

INTRODUCTION: Cigarette smoking is usually more prevalent among those with a lower socioeconomic status (SES), which can be driven by inequalities in the initiation and cessation of smoking, giving rise to SES disparities in health. This study aimed to gauge the SES inequalities in smoking related behaviours and their evolving trends based on a nationally representative database. METHOD: Data were extracted from repeated cross-sectional China Family Panel Studies (CFPS) of adults aged ≥18 and <60 years in 2012, 2014, 2016 and 2018. SES was constructed by principal component analysis based on income, education and occupation. Regression-based odds ratios and coefficients as the relative effect index of inequality were applied to quantify the degree of socioeconomic inequality in smoking related behaviours and to adjust for possible confounding factors. Multivariable regressions were utilized to explore the temporal trends in smoking inequalities. RESULTS: The smoking prevalence among men decreased from 61.16% to 2012 to 57.88% in 2018, cigarette consumption among current smokers declined from 16.71 to 15.49 cigs/per day, and the cessation rate increased from 17.55% to 24.08%. Cigarette consumption for women decreased from 13.39 in 2012 to 11.01 cigs/per day in 2018. Smoking prevalence showed significant SES inequalities among men and women from 2012 to 2018 (men: OR2012 (95%CI)= 0.72 (0.63, 0.83), OR2014 = 0.60 (0.52, 0.69), OR2016 = 0.58 (0.50, 0.67), OR2018 = 0.56 (0.48, 0.66); women: OR2012 = 0.63 (0.41, 0.97), OR2014 = 0.50 (0.32, 0.79), OR2016 = 0.44 (0.26,  0.73), OR2018 = 0.50 (0.30,  0.85)). Cigarette consumption showed significant SES inequalities among men from 2012 to 2018 (ß2012=-1.39 (-2.22, -0.57), ß2014=-2.37 (-3.23, -1.50), ß2016=-2.35 (-3.25, -1.44), ß2018=-2.91 (-3.86, -1.97)). In 2018, inequality emerged in smoking cessation rates among men and smoking intensity among women. However, all tests for trends in changes over time were not statistically significant (P varied from 0.072 to 0.602). CONCLUSION: The smoking prevalence declined between 2012 and 2018 in China. However, SES inequalities in smoking persist, while socioeconomic inequalities in smoking were not alleviated among adults aged 18 ~ 59 in China. Tobacco control measures should be implemented by giving more attention to people with lower SES who are more vulnerable to tobacco use.

Trends in Socio-economic Inequality in Smoking Among Middle-aged and Older Adults in China: Evidence From the 2011 and 2018 China Health and Retirement Longitudinal Study.

INTRODUCTION: Socio-economic inequalities in smoking and related health problems are a public health concern worldwide. To support the development of effective tobacco control policies, this study examines trends in smoking rates according to socio-economic status (SES) in China. AIMS AND METHODS: We analyzed data from repeated cross-sectional China Health and Retirement Longitudinal Study (CHARLS) on adults aged ≥45 years for the years 2011 and 2018, which involved 16 471 participants in 2011 and 19 367 in 2018. We then estimated the SES of individuals based on four types of wealth-related variables, namely, education, occupation, household characteristics, and durable consumer goods. Principal-component analysis was conducted to measure SES, and the Erreygers normalised concentration index (ECI) was used to calculate socio-economic inequality in current smoking by gender, age, and region. RESULTS: The overall ECI (95% confidence interval) for women was -0.042 (-0.054 to -0.031) and -0.038 (-0.047 to -0.029) for 2011 and 2018, respectively. The ECI (95% confidence interval) for men was -0.077 (-0.101 to -0.050) and -0.019 (-0.042 to 0.005) for 2011 and 2018, respectively. The inequality in smoking by SES for adults aged < 60 years in the Northeast region increased during 2011-2018, from -0.069 (-0.144 to 0.006) to -0.119 (-0.199 to -0.038) for women and from 0.009 (-0.115 to 0.132) to -0.164 (-0.296 to -0.032) for men. CONCLUSIONS: smoking inequality by socio-economic among adults aged ≥45 years declined in recent years in China. However, smoking inequality by SES increased in other population groups. IMPLICATIONS: Our research indicated that socio-economic inequality of current smoking among residents aged 45 years and older declined in 2018 when compared with 2011 numbers, particularly for men aged ≥ 60 years. Women in the Northeast region displayed more significant smoking inequality by SES than women in other regions did. During the study period, there was an increase in inequality in smoking by SES for adults aged < 60 years in the Northeast region. Thus, tobacco control policies and interventions should be targeted at high-risk subpopulations with lower SES, particularly in Northeast China.

Effects of Social Participation by Middle-Aged and Elderly Residents on the Utilization of Medical Services: Evidence From China.

Objectives: Aim to evaluate the effect of social participation on utilization of medical services among middle-aged and elderly residents in China. Methods: We used data from the 2018 wave of the China Health and Retirement Longitudinal Study. Social participation is classified into three types. Furthermore, to control for confounding factors, our study computed propensity score matching (PSM) to evaluate the effect of social participation on the utilization of medical services. Result: The result of PSM indicates that social participation significantly positively affects the utilization of outpatient services, the average treatment effect on the treated (ATT = 0.038***) and the utilization of inpatient services (ATT = 0.015**) by middle-aged and elderly residents. Furthermore, the utilization of outpatient health care services was significantly positively associated with leisure activities (ATT = 0.035***), social activities to help others (ATT = 0.031***), and learning activities to gain new knowledge (ATT = 0.034***) among middle-aged and elderly residents. The utilization of inpatient health care was significantly positively associated with leisure activities (ATT = 0.015***) but had no significant association with social deeds that help others and increased new knowledge among middle-aged and elderly residents. Conclusion: Thus, social participation significantly positively affects healthcare utilization by middle-aged and elderly residents. Hence, the government and society should provide more conveniences and promote social participation among middle-aged and elderly residents.

Gene set enrichment analysis and ingenuity pathway analysis to identify biomarkers in Sheng-ji Hua-yu formula treated diabetic ulcers.

ETHNOPHARMACOLOGICAL RELEVANCE: Sheng-ji Hua-yu (SJHY) formula is a Chinese herbal prescription for diabetic ulcers (DUs) treatment, which can accelerate wound reconstruction and shorten the healing time. However, its mechanism role maintains unclear. AIM OF THE STUDY: To elucidate the molecular mechanisms of SJHY application on DUs. MATERIALS AND METHODS: To begin with, transcriptome sequencing was adopted to identified differentially expression mRNAs among normal ulcers, DUs, and DUs + SJHY treatment in vivo. Liquid chromatography-tandem mass spectrometry was applied for the quality control of SJHY formula. GO and KEGG enrichment analysis were used to identify the mechanisms underlying the therapeutic effect of SJHY formula, and then gene set enrichment analysis and ingenuity pathway analysis were conducted for functional analysis. Further, qPCR detection was performed in vivo for validation. RESULTS: SJHY administration could regulate the glucose metabolic process, AMPK and HIF-1 pathway to accelerate healing processes of DUs. Besides, CRHR1, SHH, and GAL were identified as the critical targets, and SLC6A3, GRP, FGF23, and CYP27B1 were considered as the upstream genes of SJHY treatment. Combined with animal experiments, the prediction results were validated in DUs mice model. CONCLUSIONS: This study used modular pharmacology analysis to identify the biomarkers of SJHY formula and provide the potential therapeutic targets for DUs treatment as well.

The Identification of the Biomarkers of Sheng-Ji Hua-Yu Formula Treated Diabetic Wound Healing Using Modular Pharmacology.

Sheng-Ji Hua-Yu (SJHY) formula has been proved to reduce the severity of diabetic wound healing without significant adverse events in our previous clinical trials. However, based on multi-target characteristics, the regulatory network among herbs, ingredients, and hub genes remains to be elucidated. The current study aims to identify the biomarkers of the SJHY formula for the treatment of diabetic wound healing. First, a network of components and targets for the SJHY formula was constructed using network pharmacology. Second, the ClusterONE algorithm was used to build a modular network and identify hub genes along with kernel pathways. Third, we verified the kernel targets by molecular docking to select hub genes. In addition, the biomarkers of the SJHY formula were validated by animal experiments in a diabetic wound healing mice model. The results revealed that the SJHY formula downregulated the mRNA expression of Cxcr4, Oprd1, and Htr2a, while upregulated Adrb2, Drd, Drd4, and Hrh1. Besides, the SJHY formula upregulated the kernel pathways, neuroactive ligand-receptor interaction, and cAMP signaling pathway in the skin tissue homogenate of the diabetic wound healing mice model. In summary, this study identified the potential targets and kernel pathways, providing additional evidence for the clinical application of the SJHY formula for the treatment of diabetic wound healing.

Prognostic Nomogram for Liver Metastatic Colon Cancer Based on Histological Type, Tumor Differentiation, and Tumor Deposit: A TRIPOD Compliant Large-Scale Survival Study.

OBJECTIVE: A proportional hazard model was applied to develop a large-scale prognostic model and nomogram incorporating clinicopathological characteristics, histological type, tumor differentiation grade, and tumor deposit count to provide clinicians and patients diagnosed with colon cancer liver metastases (CLM) a more comprehensive and practical outcome measure. METHODS: Using the Transparent Reporting of multivariable prediction models for individual Prognosis or Diagnosis (TRIPOD) guidelines, this study identified 14,697 patients diagnosed with CLM from 1975 to 2017 in the Surveillance, Epidemiology, and End Results (SEER) 21 registry database. Patients were divided into a modeling group (n=9800), an internal validation group (n=4897) using computerized randomization. An independent external validation cohort (n=60) was obtained. Univariable and multivariate Cox analyses were performed to identify prognostic predictors for overall survival (OS). Subsequently, the nomogram was constructed, and the verification was undertaken by receiver operating curves (AUC) and calibration curves. RESULTS: Histological type, tumor differentiation grade, and tumor deposit count were independent prognostic predictors for CLM. The nomogram consisted of age, sex, primary site, T category, N category, metastasis of bone, brain or lung, surgery, and chemotherapy. The model achieved excellent prediction power on both internal (mean AUC=0.811) and external validation (mean AUC=0.727), respectively, which were significantly higher than the American Joint Committee on Cancer (AJCC) TNM system. CONCLUSION: This study proposes a prognostic nomogram for predicting 1- and 2-year survival based on histopathological and population-based data of CLM patients developed using TRIPOD guidelines. Compared with the TNM stage, our nomogram has better consistency and calibration for predicting the OS of CLM patients.

Tartary buckwheat protein prevented dyslipidemia in high-fat diet-fed mice associated with gut microbiota changes.

As one of low-digestible proteins, tartary buckwheat protein (BWP) revealed a cholesterol-lowering activity. The relationship between the prevention of BWP on dyslipidemia and changes in the numbers of gut microbiota was investigated. The male C57BL/6 mice were separately fed on normal diet, high-fat diet (HFD) with casein, and HFD with BWP extract for 6 weeks. Quantitative PCR assay was applied to quantify the microbiota composition in feces. The levels of plasma total cholesterol (TC) and triglyceride (TG) in the mice fed on HFD with BWP were significantly lower than those on HFD with casein. BWP promoted the growth of Lactobacillus, Bifidobacterium and Enterococcus and inhibited the growth of Escherichia coli in HFD-fed mice. Moreover, Bifidobacterium population was closely related to contents of plasma lipids. Further, BWP significantly decreased the levels of plasma inflammation factors as induced by HFD, including lipopolysaccharide, tumor necrosis factor α and interleukin 6. BWP significantly increased the excretion of total bile acids and short-chain fatty acids in feces. In conlusion, BWP benefited cholesterol metabolism, which could be attributed to regulating composition of gut microbiota.