ObjectiveOur recent study has demonstrated that extracellular acidification promotes lipid accumulation in macrophages via the activation of acid sensing ion channel 1 (ASIC1), but the underlying mechanism remains unclear. This study aims to explore the effect of extracellular acidification on macrophage lipophagy and the underlying mechanism. MethodsRAW264.7 macrophages were incubated with 25 mg/Lox-LDL in a pH 6.5 culture medium for 24 h to build macrophage-derived foam cell models induced by extracellular acidification. Then, RAW264.7 macrophages were cultured in the acidic medium of pH 6.5 with or without PcTx-1 (ASIC1 specific blocker, 10 μg/L) or Nec-1 (RIP1 specific inhibitor, 20 μmol/L) for 24 h, intracellular lipid accumulation was observed by oil red O staining. The expressions of total ASIC1, plasma membrane ASIC1, RIP1, p-RIP1 Ser166, TFEB, p-TFEB Ser142, LC3 and p62 were measured by Western blot. The co-localization of lipids (indicated by Bodipy) with LC3II (autophagosomes) and LAMP1 (lysosomes) was analyzed by a confocal laser scanning microscopy, respectively. Morphological changes of lipophagy in the cells were observed by using transmission electron microscopy. ABCA1-mediated cholesterol efflux was determined by cholesterol fluorescence kits. ResultsCompared with pH 7.4+ox-LDL group, the intracellular lipid accumulation in the pH 6.5+ox-LDL group was significantly increased. Meanwhile, the expressions of plasma membrane ASIC1, p-RIP1 Ser166, p-TFEB Ser142, and p62 proteins were elevated significantly, while LC3II protein level and LC3II/LC3I ratio were decreased. Accordingly, compared with pH 7.4+ox-LDL group, the macrophage lipophagy of the pH 6.5+ox-LDL group was inhibited as indicated by the decreased localization of lipid droplets with LC3 and LAMP1, a decrease in the number of lipophagosomes as well as an increase in lipid droplets. Furthermore, ATP binding cassette transporter A1 (ABCA1)-dependent cholesterol efflux from the macrophages of pH 6.5+ox-LDL group reduced dramatically. However, these above effects of extracellular acidification on RAW264.7 macrophages were abolished by PcTx-1 and Nec-1, respectively. ConclusionThese findings suggest extracellular acidification promotes the phosphorylation of TFEB at Ser142 via activating ASIC1/RIP1 pathway, thereby impeding lipophagy in RAW 264.7 macrophages, and that ASIC1 may be a new potential target for preventing aberrant lipid accumulation diseases including atherosclerosis.

Objective:To explore diagnostic value of echocardiography indexes for moderate-severe stenosis in CHD patients and their correlation with cardiac function class.Methods:A total of 90 CHD patients admitted to our hos-pital from Dec 2020 to Dec 2022 were selected as CHD group.According to NYHA classification,CHD group was divided into class Ⅰ～Ⅱ group(n=38)and class Ⅲ～Ⅳ group(n=52).The patients were divided into mild group(n=35)and moderate-severe group(n=55)according to Gensini score.In addition,90 healthy people who sim-ultaneously received physical examination in our hospital were enrolled as control group.LVEF,LVEDd and LVESd were measured in two groups by color ultrasound diagnostic instrument.Cardiac function indexes were com-pared between control group and CHD group,mild group and moderate-severe group,class Ⅰ～Ⅱ group and classⅢ～Ⅳ group.ROC curve was used to analyze the diagnostic value of above cardiac function indexes for moderate-severe stenosis in CHD patients.The correlation among LVEF,LVEDd,LVESd,different stenosis degree and NYHA class in CHD patients was analyzed by Spearman correlation analysis.Results:Compared with the control group,there was significant reduction in LVEF,and significant rise in LVEDd and LVESd in CHD group,P=0.001 all;compared with the mild group,there was significant reduction in LVEF[(45.31±5.08)％vs.(40.34±3.01)％],and significant rise in LVEDd[(53.92±5.09)mm vs.(61.68±4.79)mm]and LVESd[(52.72±4.72)mm vs.(58.06±3.50)mm]in moderate-severe group,P=0.001 all;compared with the class Ⅰ～Ⅱgroup,there was significant reduction in LVEF[(45.07±4.95)％vs.(40.23±3.06)％],and significant rise in LVEDd[(54.50±5.30)mm vs.(61.71±4.91)mm]and LVESd[(52.92±4.63)mm vs.(58.22±3.43)mm]in class Ⅲ～Ⅳ group,P=0.001 all.AUC of combined detection of LVEF,LVEDd and LVESd diagnosing moder-ate-severe stenosis in CHD was 0.909,which was significantly higher than those of single detections(0.733,0.787,0.789)(Z=2.925,2.125,2.043,P＜0.05 or＜0.01).Spearman correlation analysis showed a significant negative correlation between LVEF and NYHA class(r=-0.514),LVEDd and LVESd showed a significant posi-tive correlation with NYHA class(r=0.538,0.546,P=0.001 both),and it showed a significant positive correla-tion between different degrees of coronary stenosis and NYHA class in CHD patients(r=0.875,P=0.001).Con-clusion:The combined detection of LVEF,LVEDd and LVESd possesses high diagnostic value for moderate-se-vere stenosis in CHD patients,and it is significantly correlated with NYHA cardiac function class,which can be used as one of assessing methods for coronary stenosis severity in CHD patients.

Humans ; DNA, Mitochondrial ; Cross-Sectional Studies ; East Asian People ; DNA Methylation ; Hypertension/epidemiology* ; China/epidemiology*

Objective:To investigate the feasibility of anthocyanins(C3G)antioxidant inhibition of autophagy to al-leviate epilepsy.Methods:Seventy-five SD rats were randomly divided into 5 groups:Control group,pentetrazole(PTZ)group,hydrogen peroxide(H2O2)intervention group,3-methyladenine(3-MA)intervention group,and C3G intervention group.The seizure grade,latency,and frequency were documented.Electroencephalography was employed to detect abnormal electrical discharges in the brain across.Patch clamp technique was utilized to measure action poten-tials in hippocampal neurons for each group.The concentration of 4-hydroxynonenoic(4-HNE)hippocampus was deter-mined using a specific kit.Ultrastructural alterations in hippocampal neurons were examined through electron microsco-pyissl staining was performed to assess neuronal damage within the hippocampus.Immunohistochemical staining and Western Blot were conducted to evaluate expression levels of 15-LOX,GPX4,and LC3 proteins within the hippocampus of rats.Results:Compared with the control group,the PTZ group was completely ignited and the modeling was success-ful.Compared with other epilepsy groups,the seizure grade of C3G group decreased,abnormal discharge decreased,latency increased,hippocampal neuron excitability decreased,nishi content increased,4-HNE content,15-LOX expression and LC3Ⅱ/LC3Ⅰ ratio decreased,but GPX4 expression increased(P＜0.05).Conclusion:The oxidative stress induced by epilepsy can induce excessive autophagy of neurons,and C3G can alleviate the occurrence and devel-opment of epilepsy by anti-oxidation and inhibition of autophagy.

Objective: To investigate the differences of immune microenvironment between stage T1N3 and stage T3N0 breast cancer patients and explore the relationship between M1 macrophage infiltration and lymph node metastasis in breast cancer. Methods: Clinical information and RNA-sequencing (RNA-Seq) expression data of stage T1N3 (n=9) and stage T3N0 (n=11) breast cancer patients were extracted from Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) databases. Using CIBERSORT, the proportions of 22 types of immune cells were calculated, and then the differences of immune cell infiltration between stage T1N3 and T3N0 patients were compared. From 2011 to 2022, pathologic specimens were collected from breast cancer patients who underwent curative resection at the Cancer Hospital, Chinese Academy of Medical Sciences, including 77 at stage T1N3 and 58 at stage T3N0.The METABRIC database analysis results were verified by examining the density of M1 macrophages in tissues using dual-staining immunohistochemistry. Results: METABRIC data analysis showed M1 macrophage was the highest proportion, 15.85% in stage T1N3 breast cancer; M2 macrophage was the highest proportion, 13.07% in stage T3N0 breast cancer.M1 macrophage proportions were statistically different between patients with stage T1N3 and stage T3N0 (P=0.010). The dual-staining immunohistochemistry analysis of breast cancer tissues showed M1 macrophage density (median) of 62.0 and 38.0 cells/mm(2) for stage T1N3 and T3N0, respectively. The difference was statistically significant (P=0.002). Conclusion: The density of M1 macrophages is notably higher in stage T1N3 patients and is associated with lymph node metastasis.
Humans ; Female ; Breast Neoplasms/pathology* ; Lymphatic Metastasis/pathology* ; Macrophages/metabolism* ; Tumor Microenvironment

Objective: To describe and compare blood glucose levels in adults aged 18 years old and above in China and explore the relationship between BMI and waist circumference with blood glucose. Methods: China Chronic Disease and Risk Factor Surveillance were conducted in 298 counties/districts in China in 2018, covering 31 provinces (autonomous regions, municipalities). A multi-stage stratified cluster random sampling method selected permanent residents aged 18 years and above. Information on demographics, behavior-related risk factors, BMI, waist circumference, and blood glucose were collected through a face-to-face questionnaire, physical measurement, and laboratory examination. After complex weighting of data, they described the blood glucose levels of people with different characteristics and explored the relationship of BMI and waist circumference with blood glucose by multiple linear regression model analysis. Results: A total of 177 816 adults were included in the study. The average fasting blood glucose and average glycosylated hemoglobin were (5.73±1.46) mmol/L and (5.37±0.83) %, with people aged 60 years old and above group highest than that of other, with males higher than females (P<0.001); and urban was higher slightly than rural for the average of average glycosylated hemoglobin (P<0.001). The average fasting blood glucose and average glycosylated hemoglobin increased with increased BMI and waist circumference (P<0.001). Results from multiple linear regression model analysis showed that: 1) for each increase in BMI unit and waist circumference, the fasting glucose levels increased by 0.019 mmol/L and 0.008 mmol/L (all P<0.001) in those not diagnosed with diabetes, 2) by 0.021 mmol/L (P=0.163) and 0.014 mmol/L (P=0.004) in those newly detected as diabetes, and 3) by 0.028 mmol/L (P=0.088) and 0.023 mmol/L (P<0.001) in those self-reported as having been diagnosed as diabetes, respectively. However, glycosylated hemoglobin levels increased: 1) by 0.015% and 0.006% in those not diagnosed as diabetes (all P<0.001), 2) by 0.050% and 0.019% in those newly detected as diabetes (all P<0.001), and 3) by 0.033% and 0.019% in those self-reported as having been diagnosed as diabetes (all P<0.001), respectively. These associations with waist circumference were more robust than with BMI. Conclusions: Adults not diagnosed with diabetes with abnormal BMI or waist circumference are the key population for prevention and control. Measures improving the awareness rate of waist circumference should be taken to maintain average blood glucose in various groups.
Adolescent ; Adult ; Blood Glucose/analysis* ; Body Mass Index ; China/epidemiology* ; Diabetes Mellitus/epidemiology* ; Female ; Glycated Hemoglobin/analysis* ; Humans ; Male ; Middle Aged ; Risk Factors ; Waist Circumference

The classical famous prescription Dajianzhong Decoction is recorded in Synopsis of the Golden Chamber written by Zhang Zhongjing in the Eastern Han Dynasty. It has a long history and definite clinical effects, while this prescription has not been manufactured into Chinese patent medicine preparation. We collected many ancient books of traditional Chinese medicine(TCM) by using the method of bibliometrics and got 211 valid data terms which involved 67 ancient books. The history, main treated syndromes, formulation principle, origins and processiong of medicinal materials, and decoction method of Dajianzhong Decoction were analyzed. Despite the different views of various generations of medical experts toward the composition of this prescription, the compatibility ratio of Ginseng Radix et Rhizoma to Zingiberis Rhizoma Recens is constant. Furthermore, we explored the origins of synonyms of Dajianzhong Decoction. On the basis of this study, we hope to gain an insight into the research and development of the compound preparations of Dajianzhong Decoction and provide reference for the heritage and innovation of other classical prescriptions.
Drugs, Chinese Herbal ; Medicine, Chinese Traditional ; Prescriptions ; Rhizome

OBJECTIVES: To study the association between paternal age at childbirth and the risk of autism spectrum disorder (ASD) in offspring. METHODS: In this cross-sectional study, 71 children with ASD who were diagnosed in the Department of Child Healthcare in six hospitals in Guangzhou, Foshan, Beijing, Wuhan, Hangzhou, and Chongqing of China from August 2016 to March 2017 were enrolled as subjects, and 284 typically developing children matched for age, sex, and maternal age at childbirth with the ASD children served as controls. A self-design questionnaire was used to collect the data on social demography, maternal pregnancy, and delivery. The association between paternal age at childbirth and the development of ASD in offspring was evaluated by the logistic regression analysis. RESULTS: After control for demographic factors and pregnancy- and delivery-related factors, the logistic regression analysis showed that a relatively high paternal age at childbirth was significantly associated with the increased risk of ASD in offspring (OR=1.12, 95%CI: 1.02-1.23, P<0.05). After grouping based on the paternal age, the logistic regression analysis showed that paternal age at childbirth of ≥40 years was significantly associated with the risk of ASD in offspring (before adjustment: OR=7.08, 95%CI: 1.77-28.32, P<0.05; after adjustment: OR=8.50, 95%CI: 1.71-42.25, P<0.05). CONCLUSIONS High paternal age at childbirth is significantly associated with the increased risk of ASD in offspring, and paternal age at childbirth ≥40 years may be the high-risk age group for ASD in offspring.
Adult ; Autism Spectrum Disorder ; Child ; China ; Cross-Sectional Studies ; Female ; Humans ; Maternal Age ; Paternal Age ; Pregnancy ; Risk Factors

AR (androgen receptor) and CCAT2 are two prostate cancer (PCa)-related genes whereas their relationship is not yet reported. AR is the classical major functional gene in PCa progression. CCAT2, a non-coding gene, was identified based on big-data GWAS (Genome-Wide Association Studies) in the year of 2013. Androgen deprivation therapy (ADT) is usually used to treat PCa in the early stage. After persistent androgen deprivation, PCa would generally lead to castration resistant prostate cancer (CRPC), whereas the mechanism is yet unclear. Here we explore the function of AR and CCAT2 in PCa progression, especially their relation in androgen sensitive and insensitive cell model LNCap and DU145. We found a loop between AR and CCAT2 transcription by over-expression and knock-down strategies. In DU145 cells, G-CCAT2 activated AR mRNA level 2. 6 times, while T-CCAT2 inhibited it to 0. 2 times (P<0. 05). In LNCaP cells, G-CCAT2 could activate AR mRNA levels 1. 5 times, and TCCAT2 had no significant effect (P<0. 05). Under overexpression of AR in DU145 cells, the expression of CCAT2 increased 2. 9 times (P < 0. 05). The abundance of CCAT2 decreased to 0. 48 (P < 0. 05) in LNCaP cells by AR knock-down. Reporter gene analysis showed that CCAT2 could function on the AR promoter. We then performed CCK8 assays and AR protein level detection as supplement for the new gene CCAT2 studies. Finally we primarily studied some target genes that are related to AR and CCAT2 . The results showed that the G-CCAT2 transcript could activate AR expression in LNCap cells while UCCAT2 had no significant effect. In DU145 cells, G-CCAT2 exhibited a more relative stronger activation effect on AR, and U-CCAT2 could inhibit AR transcription. AR activates the transcriptional activity of CCAT2 in both cell lines, suggesting a feedback regulation between them. Our data showed that there would be a feedback loop between CCAT2 and AR, which may indicate a new method for PCa treatment.

OBJECTIVE: To study the molecular connection among cardiovascular diseases (CVD) subtypes defined by the International Classification of Diseases (ICD) version 10 (ICD-10). METHODS: Both phenotypic data and genotypic data used in this study were obtained from the UK Biobank. A total of 380 083 participants aged between 40 and 69 years were included. Those without any cardiovascular disease (either no ICD-10 code at all or no ICD-10 code containing letter I) were assigned to the control group. The five CVD subtypes were: ischaemic heart diseases (IHD), pulmonary heart disease and diseases of pulmonary circulation (PHD), cerebrovascular diseases (CRB), diseases of arteries, arterioles and capillaries (AAC), diseases of veins, lymphatic vessels and lymph nodes, and diseases not elsewhere classified (VLL). We first performed a genome-wide association study (GWAS) for each of the five subtypes. We summarized novel loci using genome-wide significance threshold P=5×10^-8. Next, we used linkage disequilibrium score regression (LDSC) method to assess genetic correlation among the five subtypes. Lastly, we applied mendelian randomization (MR) approach to assess the causal relationship among the subtypes. The particular software that we used was generalised summary-data-based mendelian randomisation (GSMR). RESULTS: Through GWAS, we identified hundreds of genome-wide significant SNPs: 672 for IHD, 241 for PHD, 31 for CRB, 48 for AAC, and 193 for VLL. By comparing with published literature, we found 28 novel loci, for PHD (n=14), CRB (n =7) and AAC (n =7). Eight of these 28 loci were rare, where the lead SNP had minor allele frequency (MAF) less than 1%. LDSC analyses indicated IHD had significant genetic correlation with VLL (P=2.52×10^-7), PHD (P=3.77×10^-3) and AAC (P=4.90×10^-3), respectively. Bidrectional GSMR analyses showed that IHD had a positive causal relationship with VLL (P=7.40×10^-5) and AAC (P=1.50×10^-3), while reverse causality was not supported. CONCLUSION This study adopted an innovative approach to study the molecular connection among CVD subtypes that are defined by ICD. We identified potentially positive genetic correlation and causal effects among some of these subtypes. Research along this line will provide scientific insights and serve as a guidance for future ICD standards.
Adult ; Aged ; Cardiovascular Diseases/genetics* ; Genome-Wide Association Study ; Humans ; International Classification of Diseases ; Mendelian Randomization Analysis ; Middle Aged ; Polymorphism, Single Nucleotide