Over the past two decades, genome-wide association study(GWAS) has identified numerous genetic variants and loci associated with heritable diseases. With the gradual maturation and saturation of GWAS methodologies, transcriptome-wide association study(TWAS) offers a novel perspective by linkinggenetic phenotypes to gene expression levels. By integrating TWAS with other multi-omics analyses, researchers can gain a deeper understanding of heritable diseases. This article provides an overview of recent groundbreaking and representative TWAS methods and tools, analyzes their strengths and limitations, and discusses future trends in TWAS development.

Based on the theory that Shaoyang （少阳） and Shaoyin （少阴） function as the principal pivots， and integrating the laws of qi-fire transformation， yin-yang alternation， and the waxing and waning of healthy qi and pathogenic factors， as well as the action characteristics of immune checkpoint inhibitors （ICIs）， this study systematically expounds the dynamic evolution laws of the Shaoyang and Shaoyin pivots and constructs a stage- and phase-based differentiation and treatment framework for gastric cancer immunotherapy. In the neoadjuvant treatment stage， the core pathogenesis is the dysfunction of the Shaoyang pivot and the disorder of qi and fire transformation， often accompanied by stagnation of cold-deficiency in the middle jiao （焦）. In the postoperative adjuvant treatment stage， the dominant factors are the depletion of Shaoyin fire and the damage of yin fluid， accompanied by the lingering of residual pathogen in the Shaoyang level and the obstruction of collaterals by phlegm and stasis. In the advanced stage， the critical conditions are the exhaustion of the Shaoyin pivot， the blockage of orifices by turbid toxins， and the separation of yin and yang， leading to the rampant of pathogenic toxins and the emergence of various complicated syndromes. The treatment should be guided by the principle of regulating the double pivot. In the neoadjuvant treatment stage， it is recommended to activate the Shaoyang pivot and warm and promote the qi movement in the middle jiao. In the postoperative adjuvant treatment stage， clearing the residual pathogenic factors from the pivot and nourishing the Shaoyin. In the advanced stage， rescuing the Shaoyin pivot and opening the orifices to transform turbidity. Furthermore， stage-based treatment should serve as the foundation for dynamically assessing the patient's immune status and phase-specific changes in immunity， thereby promoting immune activation while preventing and managing immune overactivation and drug resistance.

ObjectiveThis study aims to elucidate the potential mechanism of Zuojinwan in improving liver fibrosis through hepatic tissue metabolomics analysis. MethodsTwenty-four mice were randomly allocated into normal group， model group ， positive drug group （silymarin， 100 mg·kg^-1）， and Zuojinwan group （Zuojinwan solution， 2.5 g·kg^-1）， with per group six mice. Liver fibrosis model was induced via intraperitoneal injection of olive oil solution with 10% carbon tetrachloride （CCl₄） （0.5 μL·g^-1， three times weekly for 8 weeks） in all groups except the normal group. During the final 4 weeks， the silymarin group received silymarin （100 mg·kg^-1） by gavage thrice weekly， while the Zuojinwan group was administered Zuojinwan solution （2.5 g·kg^-1） under the same regimen. After the last administration， the levels of liver fibrosis indicators and liver injury markers in serum were detected. The pathological morphological changes of the liver tissues were observed. The levels of liver fibrosis markers α-smooth muscle actin （α-SMA） and Collagen Ⅰ（ColⅠ） were detected. Metabolomics was analyzed on mice's liver tissues. The mice's serum was collected for metabolomics analysis. ResultsCompared with the model group， Zuojinwan significantly improved indicators related to liver fibrosis and liver injury. Compared with the normal group， the model group showed significantly elevated levels of fibrosis markers such as laminin （LN）， hyaluronic acid （HA）， procollagen typeⅢ （PC-Ⅲ）， and type Ⅳ Collagen （Ⅳ-C）， while liver injury indicators such as alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， alkaline phosphatase （ALP）， lactate dehydrogenase （LDH）， and total bilirubin （TBIL）， exhibited a marked upward trend （P<0.05）. Compared with the model group， the silymarin group showed a significant decrease in the aforementioned indicators （P<0.05）. Notably， compared with the model group， the Zuojinwan group exhibited a significant reduction in all these indicators （P<0.05）， with efficacy comparable to that of the silymarin group. Zuojinwan reduced mRNA and protein levels of α-SMA and ColⅠ in the liver tissue. Metabolomics results revealed that compared with the model group， Zuojiinwan significantly reduced levels of glucose metabolism-related metabolites such as D-fructose 1，6-bisphosphate （FBP）， nicotinamide adenine dinucleotide phosphate （NADPH）， sodium beta-D-fructose 6-phosphate （F6P）， dihydroxyacetone phosphate （DHAP）， fumaric acid， and D-glucose 6-phosphate （G6P） （P<0.05）. Serum enzyme-linked immunosorbent assay （ELISA） was used to detect glucose metabolism indicators and further validate the regulatory effect of Zuojinwan on glucose metabolism. ConclusionThese results suggest that Zuojinwan may improve liver fibrosis by regulating the dysregulated levels of glucose metabolism during the progression of liver fibrosis.

BACKGROUND:The stress effect of autophagy on epithelial cells,fibroblasts and myofibroblasts is closely related to the formation process of pulmonary fibrosis.OBJECTIVE:To screen the genes related to autophagy in patients with pulmonary fibrosis,and explore their correlation with the prognosis of patients with pulmonary fibrosis,in order to provide a new target for clinical intervention in pulmonary fibrosis.METHODS:The gene expression profiling dataset downloaded from GSE70866 was used as a training set,differentially expressed genes between pulmonary fibrosis patients and normal healthy individuals was analyzed using the R language and intersected with autophagy-related genes to identify the differentially expressed genes with the most significant changes.Multiple analysis methods were used to identify key prognostic genes and construct genetic prognostic models.Patients with pulmonary fibrosis were divided into high-risk and low-risk groups according to their risk scores,and the validity of the prognostic model was verified using the Siena cohort and Leuven cohort validation sets.A cell model of pulmonary fibrosis was established by inducing HFL-1 cells(human embryonic lung fibroblasts)with transforming growth factor-β1,and an animal model of pulmonary fibrosis was established in mice by tracheal instillation of bleomycin to validate the expressions of prognostic genes.RESULTS AND CONCLUSION:(1)There were 2 650 differentially expressed genes between fibrotic tissue and normal tissue.Among them,34 genes related to autophagy showed significant expression changes.(2)Kaplan-Meier survival analysis curves for the Siena cohort and Leuven cohort validation sets showed significantly lower survival in the high-risk group than in the low-risk group.(3)Three autophagy genes related to prognosis were screened out:myelocytomatosis viral oncogene(MYC),C-C motif chemokine ligand 2(CCL2),and GABA type a receptor associated protein like 1(GABARAPL1).(4)Both in vivo and in vitro studies showed that compared with the control group,the expression levels of myelocytomatosis viral oncogene and C-C motif chemokine ligand 2 mRNA and protein were significantly higher in the lung fibrosis model group(P＜0.01,P＜0.05),while the expression levels of GABA type a receptor associated protein like 1 mRNA and protein were lower(P＜0.001).To conclude,bioinformatics methods are used to analyze the expression of three autophagy-related genes in pulmonary fibrosis and their correlation with the prognosis of patients with pulmonary fibrosis.The constructed prognostic model has good predictive ability for the 1-,2-,and 3-year survival rates of patients with pulmonary fibrosis.Moreover,in vivo and in vitro models have been used to verify that myelocytomatosis viral oncogene and C-C motif chemokine ligand 2 are highly expressed in lung fibroblasts and tissues,and that GABA type a receptor associated protein like 1 is lowly expressed.

BACKGROUND:The stress effect of autophagy on epithelial cells,fibroblasts and myofibroblasts is closely related to the formation process of pulmonary fibrosis.OBJECTIVE:To screen the genes related to autophagy in patients with pulmonary fibrosis,and explore their correlation with the prognosis of patients with pulmonary fibrosis,in order to provide a new target for clinical intervention in pulmonary fibrosis.METHODS:The gene expression profiling dataset downloaded from GSE70866 was used as a training set,differentially expressed genes between pulmonary fibrosis patients and normal healthy individuals was analyzed using the R language and intersected with autophagy-related genes to identify the differentially expressed genes with the most significant changes.Multiple analysis methods were used to identify key prognostic genes and construct genetic prognostic models.Patients with pulmonary fibrosis were divided into high-risk and low-risk groups according to their risk scores,and the validity of the prognostic model was verified using the Siena cohort and Leuven cohort validation sets.A cell model of pulmonary fibrosis was established by inducing HFL-1 cells(human embryonic lung fibroblasts)with transforming growth factor-β1,and an animal model of pulmonary fibrosis was established in mice by tracheal instillation of bleomycin to validate the expressions of prognostic genes.RESULTS AND CONCLUSION:(1)There were 2 650 differentially expressed genes between fibrotic tissue and normal tissue.Among them,34 genes related to autophagy showed significant expression changes.(2)Kaplan-Meier survival analysis curves for the Siena cohort and Leuven cohort validation sets showed significantly lower survival in the high-risk group than in the low-risk group.(3)Three autophagy genes related to prognosis were screened out:myelocytomatosis viral oncogene(MYC),C-C motif chemokine ligand 2(CCL2),and GABA type a receptor associated protein like 1(GABARAPL1).(4)Both in vivo and in vitro studies showed that compared with the control group,the expression levels of myelocytomatosis viral oncogene and C-C motif chemokine ligand 2 mRNA and protein were significantly higher in the lung fibrosis model group(P＜0.01,P＜0.05),while the expression levels of GABA type a receptor associated protein like 1 mRNA and protein were lower(P＜0.001).To conclude,bioinformatics methods are used to analyze the expression of three autophagy-related genes in pulmonary fibrosis and their correlation with the prognosis of patients with pulmonary fibrosis.The constructed prognostic model has good predictive ability for the 1-,2-,and 3-year survival rates of patients with pulmonary fibrosis.Moreover,in vivo and in vitro models have been used to verify that myelocytomatosis viral oncogene and C-C motif chemokine ligand 2 are highly expressed in lung fibroblasts and tissues,and that GABA type a receptor associated protein like 1 is lowly expressed.

A 20-year-old male patient presented to the Department of Dermatology of Peking Union Medical College Hospital with complaints of an 8-year history of facial scarring, swelling of the lower limbs, and a 4-year history of scalp thickening. Physical examination showed thickening furrowing wrinkling of the skin on the face and behind the ears, ciliary body hirsutism, blepharoptosis, and cutis verticis gyrate. Both lower limbs were swollen, especially the knees and ankles. The skin of the palms and soles of the feet was keratinized and thickened. Laboratory examination using bone and joint X-ray showed periostosis of the proximal middle phalanges and metacarpals of both hands, distal ulna and radius, tibia and fibula, distal femurs, and metatarsals.Genetic testing revealed two variants in SLCO2A1, c.1658T > A and c.96+4A > C.Through multidisciplinary consultation, we confirmed the diagnosis of pachydermoperiostosis.

BACKGROUND: Double-stranded RNA-specific adenosine deaminase 1 (ADAR1) binds to double-stranded RNA and catalyzes the deamination of adenosine (A) to inosine (I). The functional mechanism of ADAR1 in non-small cell lung cancer (NSCLC) remains incompletely understood. This study aimed to investigate the prognostic significance of ADAR1 in NSCLC and to elucidate its potential role in regulating tumor cell proliferation and migration. METHODS: Data from The Cancer Genome Atlas (TCGA) and cBioPortal were analyzed to assess the correlation between high ADAR1 expression and clinicopathological features as well as prognosis in lung cancer. We performed Western blot (WB), cell proliferation assays, Transwell invasion/migration assays, and nude mouse xenograft modeling to examine the phenotypic changes and molecular mechanisms induced by ADAR1 knockdown. Furthermore, the ADAR1 p150 overexpression model was utilized to validate the proposed mechanism. RESULTS: ADAR1 expression was significantly elevated in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) tissues compared with adjacent non-tumor tissues (LUAD: P=3.70×10-15, LUSC: P=0.016). High ADAR1 expression was associated with poor prognosis (LUAD: P=2.03×10-2, LUSC: P=2.81×10-2) and distant metastasis (P=0.003). Gene Set Enrichment Analysis (GSEA) indicated that elevated ADAR1 was associated with mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway activation, matrix metalloproteinase-9 (MMP-9) expression, and cell adhesion. ADAR1 and MMP-9 levels showed a strongly positive correlation (P=6.45×10-34) in 10 lung cancer cell lines, highest in H1581. Knockdown of ADAR1 in H1581 cells induced a rounded cellular morphology with reduced pseudopodia. Concomitantly, it suppressed cell proliferation, invasion, migration, and in vivo tumorigenesis. It also suppressed ERK phosphorylation and downregulated cellular Finkel-Biskis-Jinkins murine osteosarcoma viral oncogene homolog (c-FOS), MMP-9, N-cadherin, and Vimentin. Conversely, ADAR1 p150 overexpression in PC9 cells enhanced ERK phosphorylation and increased c-FOS and MMP-9 expression. CONCLUSIONS High ADAR1 expression is closely associated with poor prognosis and distant metastasis in NSCLC patients. Mechanistically, ADAR1 may promote proliferation, invasion, migration, and tumorigenesis in lung cancer cells via the ERK/c-FOS/MMP-9 axis.
Humans ; Lung Neoplasms/physiopathology* ; Adenosine Deaminase/genetics* ; Matrix Metalloproteinase 9/genetics* ; Cell Proliferation ; Carcinoma, Non-Small-Cell Lung/physiopathology* ; Cell Movement ; Animals ; Mice ; RNA-Binding Proteins/genetics* ; Female ; Male ; Cell Line, Tumor ; Proto-Oncogene Proteins c-fos/genetics* ; Middle Aged ; MAP Kinase Signaling System ; Gene Expression Regulation, Neoplastic ; Mice, Nude ; Extracellular Signal-Regulated MAP Kinases/genetics*

OBJECTIVES: The prevalence of overweight and obesity among children and adolescents continues to rise, becoming one of the most serious global public health issues of the 21st century. Given the differing growth and development environments between urban and rural children, associated risk factors also vary. This study aims to explore the distribution characteristics and influencing factors of overweight and obesity among urban and rural primary and secondary school students in Hunan Province, providing scientific evidence for targeted interventions. METHODS: A stratified, randomized cluster sampling method was used to select participants. A total of 197 084 students from primary and secondary schools across 14 prefectures in Hunan Province underwent physical examinations and questionnaire surveys. Population and spatial distribution characteristics of overweight and obesity were analyzed. Spatial distribution maps and spatial autocorrelation analyses were conducted using ArcGIS. Multivariate Logistic regression was used to identify influencing factors for overweight and obesity. RESULTS: The overall overweight and obesity rates among students in Hunan Province were 14.7% and 10.9%, respectively. Both rates were higher in urban areas than in rural counties (16.0% vs 13.9% for overweight; 12.1% vs 10.2% for obesity). Among both urban and rural students, boys had higher rates of overweight and obesity than girls. Higher-grade students had a higher overweight rate but a lower obesity rate than lower-grade students. In urban areas, the overweight and obesity rates of Han Chinese primary and secondary school students are lower than those of ethnic minority students (both P<0.05). In rural areas, the obesity rate of Han primary and secondary school students is lower than that of ethnic students (P<0.05). Across cities and prefectures, urban overweight and obesity rates ranged from 14.7% to 18.7% and 8.4% to 20.6% respectively, while rural rates ranged from 10.9% to 17.2% and 6.6% to 13.7% respectively. Spatial autocorrelation analysis revealed high-value clusters of overweight/obesity in urban areas of Changde and Zhangjiajie, and in rural areas of Loudi, Huaihua, and Shaoyang. Multivariate Logistic regression showed that gender, school stage, ethnicity, frequency of fresh vegetable intake, and sleep duration were associated with overweight and/or obesity in both urban and rural students. In urban students, frequency of fried food and fresh fruit intake, breakfast habits, physical activity on weekdays and holidays, and screen time on computers were also significant. In rural students, TV viewing time and sedentary duration were additional relevant factors. CONCLUSIONS The situation of overweight and obesity among primary and secondary school students in Hunan Province remains concerning. Greater attention should be paid to regions with high-value clusters of overweight/obesity, and targeted interventions should be developed based on urban-rural differences in influencing factors.
Humans ; China/epidemiology* ; Adolescent ; Male ; Female ; Rural Population/statistics & numerical data* ; Child ; Overweight/epidemiology* ; Students/statistics & numerical data* ; Urban Population/statistics & numerical data* ; Risk Factors ; Prevalence ; Obesity/epidemiology* ; Surveys and Questionnaires ; Pediatric Obesity/epidemiology* ; Schools

Hepatic ischemia-reperfusion injury (HIRI) has been considered as an inevitable process of liver transplantation. Hepatocyte ferroptosis is a key factor in HIRI development, yet precise mechanism and potential therapies are still unclear. Here, we demonstrated a strong correlation between hepatocyte ferroptosis and the downregulation of poly(rC)-binding protein (PCBP2), which compromised the stability of antiporter system Xc^- (consisted of SL3A2/SLC7A11). Besides, inhibiting PCBP2 contributed to facilitating cofactor p300 to enhance the transcriptional activity of HIF1α, leading to the expression and secretion of HMGB1. Then, released HMGB1 from ferroptotic hepatocytes worsened M1 macrophage recruitment and immune response during HIRI. Additionally, acteoside (ACT) was shown to assist PCBP2 in stabilizing the mRNA stability of Slc3a2 and Slc7a11, as well as enhance the binding affinity of PCBP2-system Xc^-. Beyond that, ACT also supported PCBP2 to limit HMGB1-induced M1 macrophage recruitment through imposing restrictions on p300 and HIF1α. Furthermore, specific knockdown of PCBP2 in hepatocytes directly interposed the therapeutic efficacy of ACT on HIRI mice. In conclusion, ACT alleviated hepatocyte ferroptosis and HIRI via promoting PCBP2 to maintain the stability of system Xc^- and limit HIF1α/p300-HMGB1 signaling. These findings highlight the therapeutic benefits of ACT in treating HIRI and offer insights into innovative therapeutic strategies.

Malignant tumors (commonly referred to as cancer) represent a major global public health challenge and contribute significantly to the worldwide disease burden. Early screening plays a critical role in improving detection rates, enabling timely intervention, and enhancing patient survival rates. However, current cancer screening guidelines primarily focus on site-specific screening, which may not fully address the need for comprehensive early detection. A scientifically rational, multi-cancer screening approach offers several advantages: it optimizes the use of biological samples, reduces time costs for participants, enhances the efficiency and comprehensiveness of screening, and minimizes overall expenses. Such an approach also facilitates the rational allocation of healthcare resources, ultimately helping to reduce the societal burden of cancer. To address this need, the Cancer Epidemiology Committee of the Chinese Anti-Cancer Association has developed the Expert Consensus on Combined Screening for Common Cancers in China. This consensus integrates multidisciplinary expertise and synthesizes the latest domestic and international researches on cancer screening, early detection, and treatment for prevalent malignancies. Drawing upon China's unique demographic and healthcare context, as well as practical screening experiences, the consensus provides evidence-based recommendations on target populations, screening technologies, and procedural workflows for multi-cancer screening. These guidelines align with the principles and methodologies established by the World Health Organization (WHO), aiming to enhance the effectiveness of combined cancer screening in China, improve early detection rates, and provide a scientific foundation for national cancer prevention and control strategies.