Objective To explore the relationship between circadian rhythm genes and the occurrence, development, prognosis, and tumor microenvironment (TME) of lung adenocarcinoma (LUAD). Methods The Cancer Genome Atlas data were used to evaluate the expression, copy number variation, and somatic mutation frequency of circadian gene sets in LUAD. Gene ontology, Kyoto encyclopedia of genes and genomes, and gene set enrichment analysis were used to explore the potential mechanisms by which circadian rhythm genes affected LUAD progression. Cox regression, least absolute shrinkage and selection operator regression, support vector machine recursive feature elimination, and random forest screened circadian genes and established prognostic models, and on this basis constructed nomogram to predict patients’ 1-, 3-, and 5-year survival rates. Kaplan-Meier survival curves, receiver operating characteristic (ROC) curves, and time-dependent ROC curves were drawn to evaluate the predictive ability of the model, and the external dataset of GEO further verified the prognostic value of the prediction model. In addition, we evaluated the association of the prognostic model with immune cells and immune checkpoint genes. Single cell RNA sequencing (scRNA-seq) analysis was used to explore the molecular characteristics between prognostically relevant circadian genes and different immune cell populations in TME. Results Differentially expressed circadian rhythm genes were mainly enriched in biological processes related to cGMP-PKG signaling pathway, lipid and atherosclerosis, and JAK-STAT signaling pathway. Seven circadian rhythm genes: LGR4, CDK1, KLF10, ARNTL2, RORA, NPAS2, PTGDS were screened out, and a RiskScore model was established. According to the median RiskScore, samples were divided into a high-risk group and a low-risk group. Compared with patients in the low-risk group, patients in the high-risk group showed a poorer prognosis (P<0.001). Immunological characterization analysis showed that there were differences in the infiltration of multiple immune cells between the low-risk group and high-risk group. Most immune checkpoint genes had higher expression levels in the high-risk group than those in the low-risk group, and RiskScore was positively correlated with the expression of CD276, TNFSF4, PDCD1LG2, CD274, and TNFRSF9, and negatively correlated with the expression of CD40LG and TNFSF15. The scRNA-seq analysis showed that RORA and KLF10 were mainly expressed in natural killer cells. Conclusion The prognostic model based on seven feature circadian rhythm genes has certain predictive value for predicting survival of LUAD patients. Dysregulated expression of circadian genes may regulate the occurrence, progression as well as prognosis of LUAD through affecting TME, which provides a possible direction for finding potential strategies for treating LUAD from the perspective of mechanism by which circadian disorder affects immune cells.

ObjectiveSpinal cord injury (SCI) directly impairs the regulatory function of the autonomic nervous system, induces intestinal dysfunction, and significantly reduces patients’ quality of life. Preclinical studies have shown that electroacupuncture (EA) therapy can regulate the brain-gut axis and is used to treat central nervous system diseases such as major depressive disorder, Alzheimer’s disease and Parkinson’s disease. Recent research has established that fecal microbiota transplantation (FMT) from EA-treated SCI rats restored intestinal motility and colonic morphology. However, it remains unclear whether the regulation of gut microbiota by EA therapy directly contributes to neural repair after SCI. This study aims to explore whether gut microbiota mediates the neuroprotective effect of EA in the treatment of SCI and its possible mechanism. MethodsThe study employed RNA transcriptome analysis of spinal cord tissue to characterize gene expression profiles and to identify key signaling pathways following EA treatment for SCI. Hematoxylin-Eosin (HE) staining and Nissl staining were used to observe the morphological changes in spinal cord tissue. Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) were applied to detect the effects of EA on the expression of proteins related to nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) -dependent pyroptosis. Using 16S rDNA sequencing, the study observed alterations in gut microbiota diversity and community composition in SCI rats. Prior to establishing SCI models, rats were pretreated with an antibiotic cocktail to induce gut dysbiosis, and the effects on intestinal function and spinal cord neural repair were evaluated. FMT was performed to investigate the regulatory effects of post-EA FMT on motor function, general status, liver and spleen indices, and NLRP3-mediated pyroptosis in SCI rats. ResultsEA improved motor function and reduced regulated neuronal cell death in SCI rats. Transcriptomic analysis demonstrated the activation of immune- and inflammation-related pathways post-SCI, including NOD-like receptors, nuclear factor-kappa B(NF-κB), and Toll-like receptor (TLR) pathways. EA primarily influenced intestinal inflammation and autoimmune functions. 16S rDNA sequencing illustrated that EA did not alter the diversity of gut microbiota. However, EA altered the gut microbiota composition in SCI rats, increasing Lactobacillus and Akkermansia genera while rebalancing the Firmicutes/Bacteroidetes ratio. Furthermore, depletion of gut microbiota by antibiotics disrupted the intestinal barrier, reduced the expression of intestinal barrier proteins Zonula Occludens-1 (ZO-1) and Occludin, elevated serum lipopolysaccharide-binding protein (LBP) levels, exacerbated spinal cord tissue damage, and hindered motor function recovery in SCI rats. FMT from donors treated with EA reduced LBP levels in the intestine, blood, and spinal cord of rats, inhibited the TLR4 myeloid differentiation primary response protein 88 (MyD88)-NF‑κB pathway and NLRP3-dependent pyroptosis, and improved motor function. On the other hand, FMT treatment resulted in decreased body weight and food intake, whereas FMT using EA-treated donors effectively alleviated these alterations. ConclusionEA effectively alleviated neuroinflammatory responses in rats with SCI, primarily through regulating the gut microbiota and suppressing the NLRP3-dependent pyroptosis signaling pathway.

ObjectiveSpinal cord injury (SCI) directly impairs the regulatory function of the autonomic nervous system, induces intestinal dysfunction, and significantly reduces patients’ quality of life. Preclinical studies have shown that electroacupuncture (EA) therapy can regulate the brain-gut axis and is used to treat central nervous system diseases such as major depressive disorder, Alzheimer’s disease and Parkinson’s disease. Recent research has established that fecal microbiota transplantation (FMT) from EA-treated SCI rats restored intestinal motility and colonic morphology. However, it remains unclear whether the regulation of gut microbiota by EA therapy directly contributes to neural repair after SCI. This study aims to explore whether gut microbiota mediates the neuroprotective effect of EA in the treatment of SCI and its possible mechanism. MethodsThe study employed RNA transcriptome analysis of spinal cord tissue to characterize gene expression profiles and to identify key signaling pathways following EA treatment for SCI. Hematoxylin-Eosin (HE) staining and Nissl staining were used to observe the morphological changes in spinal cord tissue. Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) were applied to detect the effects of EA on the expression of proteins related to nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) -dependent pyroptosis. Using 16S rDNA sequencing, the study observed alterations in gut microbiota diversity and community composition in SCI rats. Prior to establishing SCI models, rats were pretreated with an antibiotic cocktail to induce gut dysbiosis, and the effects on intestinal function and spinal cord neural repair were evaluated. FMT was performed to investigate the regulatory effects of post-EA FMT on motor function, general status, liver and spleen indices, and NLRP3-mediated pyroptosis in SCI rats. ResultsEA improved motor function and reduced regulated neuronal cell death in SCI rats. Transcriptomic analysis demonstrated the activation of immune- and inflammation-related pathways post-SCI, including NOD-like receptors, nuclear factor-kappa B(NF-κB), and Toll-like receptor (TLR) pathways. EA primarily influenced intestinal inflammation and autoimmune functions. 16S rDNA sequencing illustrated that EA did not alter the diversity of gut microbiota. However, EA altered the gut microbiota composition in SCI rats, increasing Lactobacillus and Akkermansia genera while rebalancing the Firmicutes/Bacteroidetes ratio. Furthermore, depletion of gut microbiota by antibiotics disrupted the intestinal barrier, reduced the expression of intestinal barrier proteins Zonula Occludens-1 (ZO-1) and Occludin, elevated serum lipopolysaccharide-binding protein (LBP) levels, exacerbated spinal cord tissue damage, and hindered motor function recovery in SCI rats. FMT from donors treated with EA reduced LBP levels in the intestine, blood, and spinal cord of rats, inhibited the TLR4 myeloid differentiation primary response protein 88 (MyD88)-NF‑κB pathway and NLRP3-dependent pyroptosis, and improved motor function. On the other hand, FMT treatment resulted in decreased body weight and food intake, whereas FMT using EA-treated donors effectively alleviated these alterations. ConclusionEA effectively alleviated neuroinflammatory responses in rats with SCI, primarily through regulating the gut microbiota and suppressing the NLRP3-dependent pyroptosis signaling pathway.

ObjectiveTo investigate the association of the single nucleotide polymorphism （SNP） of the insulin-like growth factor binding protein-3 （IGFBP3） gene at rs10225396 （A>G） locus with the susceptibility to hepatocellular carcinoma， as well as its potential molecular regulatory mechanisms， and to provide novel genetic biomarkers and a theoretical basis for early screening and precise targeted therapy for hepatocellular carcinoma. MethodsA total of 192 patients with hepatocellular carcinoma who were admitted to The Affiliated Hospital of Yanbian University and Yanbian Cancer Hospital from January 2009 to August 2016 were enrolled as experimental group， and 190 healthy individuals who underwent physical examination in Physical Examination Center of Yanbian Hospital during the same period of time were enrolled as control group. Peripheral blood samples were collected from all subjects， and after DNA was extracted from whole blood， the DNA samples meeting quality standards were sent to Beijing Genomics Institute Research Center Co.， Ltd.， for Mass ARRAY mass spectrometry， while genotyping was completed. The independent-samples t test was used for comparison of continuous data between two groups， and the chi-square test was used for comparison of categorical data between two groups. A binary Logistic regression model was used to analyze the association of the SNP of the IGFBP3 gene at rs10225396 locus with the susceptibility to hepatocellular carcinoma， and odds ratio （OR） and 95% confidence interval （CI） were calculated to assess the risk of developing hepatocellular carcinoma in individuals carrying different genotypes. ResultsThere were two alleles （G and A） at the rs10225396 locus of the IGFBP3 gene， yielding the three genotypes of AA， AG， and GG， and its genotype distribution was consistent with the Hardy-Weinberg equilibrium （χ²=0.072， P=0.789）. The stratified genetic analysis showed that carriers of the IGFBP3 rs10225396 AA genotype had a significant increased risk of hepatocellular carcinoma among individuals of age <63 years， male sex， smoking history， drinking history， and the Chinese Korean population （all P<0.001）. The binary Logistic regression analysis showed that after adjustment for related risk factors in the codominant model， the population with genotype AG and GG had a significant reduction in the risk of hepatocellular carcinoma compared with the population with genotype AA （P<0.001）， and in the dominant model， the population with genotype AG+GG had a significant reduction in the risk of hepatocellular carcinoma compared with the population with genotype AA （P<0.001）. ConclusionThrough a genotyping analysis of hepatocellular carcinoma patients and healthy individuals in Yanbian Korean Autonomous Prefecture of Jilin Province， China， this study shows that genotype AA at rs10225396 locus of the IGFBP3 gene is significantly associated with the susceptibility to hepatocellular carcinoma， and this genotype can significantly increase the risk of developing hepatocellular carcinoma with the presence of specific risk factors， which provides a potential genetic marker for early screening and precise treatment of hepatocellular carcinoma.

Objective: To explore the potential category patterns of risk prevention and control behaviors of HIV infection among young students who have sex with men (MSM) and their impact on HIV infection and late detection, aiming to optimize intervention strategies. Methods: From September 2017 to December 2024, a total of 1 637 MSM young students in Tianjin were recruited through both online and offline channels. Latent class analysis was applied to classify 11 HIV risk prevention and control behaviors [condom use during the most recent anal sex in the past 6 months, consistent condom use, use of water based lubricants, abstinence from recreational drugs, regular on site professional testing, fixed sexual partners, partner testing, awareness of partner s HIV testing results, testing before sexual activity, nucleic acid testing, and use of pre exposure prophylaxis (PrEP) or post exposure prophylaxis (PEP)]. Multivariate Logistic regression analyzed associations between demographic characteristics/intervention services factors and latent classes. Differences in HIV infection and late detection across behavior patterns were compared. Results: HIV risk prevention and control behaviors among MSM students were classified into three latent classes:condom dependent group (38.42%), low prevention group (27.73%), and comprehensive prevention group (33.85%). Students who received condom promotion/testing services were more likely to belong to the comprehensive prevention group ( OR =5.58), while those who received peer education were less likely to the comprehensive prevention group ( OR =0.43) (both P <0.01). Among the MSM student population, the HIV infection rate was 4.83%, with 2.26% of cases detected late. The HIV infection rate (1.45%) and late detection proportion (0.82%) in the comprehensive prevention group were lower than those in the low prevention group (7.89% and 3.83%, respectively) ( χ 2=16.20, 7.31, both P <0.01). Conclusions HIV risk prevention and control behaviors among MSM young students exhibit significant heterogeneity. Comprehensive prevention strategies can effectively reduce HIV infection and late detection risks. It is necessary to optimize peer education content and improve the accessibility of diversified prevention measures such as PrEP/PEP to enhance HIV prevention and control.

OBJECTIVE To investigate the inhibitory effect and mechanism of Modified qifang weitong granules on gastric cancer based on in vitro and in vivo experiments. METHODS Human gastric cancer HGC-27 cells were divided into the following groups: control group （treated with fetal bovine serum）， 10% drug-containing serum group， 15% drug-containing serum group， 20% drug-containing serum group， and 5-fluorouracil （5-Fu） group （positive control， 3.90 μg/mL）. After culturing the cells in each group with the corresponding serum/drug solution， their proliferation， migratory and invasive abilities， as well as the cell cycle， were assessed. Additionally， the expression levels of epithelial-mesenchymal transition （EMT）-related proteins [E-cadherin， N-cadherin， and vimentin] in the cells were measured. Logarithmic-phase HGC-27 cells were harvested and subcutaneously injected into the right axillary region of nude mice to establish a subcutaneous xenograft tumor model in nude mice. The successfully modeled tumor-bearing nude mice were randomly divided into model group， low-， medium- and high-dose groups of Modified qifang weitong granules （17.65， 35.29 and 70.58 g/kg， respectively）， and 5-Fu group （25 mg/kg）， with 5 mice in each group. After 14 days of treatment with the corresponding drugs in each group， the histopathological morphology of the tumor tissues in the nude mice was observed. Immunohistochemistry and Western blot assay were employed to detect the expression levels of EMT- related proteins in the tumor tissues of the nude mice. RESULTS In the cell experiment， compared with the control group， the cell proliferation rate， migration rate， number of invasive cells， as well as the expression levels of N-cadherin and vimentin proteins， and the percentage of cells in the G2/M phase were all significantly decreased/reduced in the 15% drug-containing serum group， 20% drug-containing serum group （P＜0.05）. Conversely， the percentage of cells in the G0/G1 phase and the expression level of E- cadherin protein were significantly increased （P＜0.05）. In animal experiment， compared with the model group， the high-dose group of Modified qifang weitong granules exhibited significantly reduced tumor mass and expression levels of N-cadherin and vimentin proteins in the tumor tissues of nude mice （P＜0.05）， while the expression level of E-cadherinprotein in the tumor tissues was significantly increased （P＜0.05）. Additionally， the tumor cells varied in size and showed extensive necrosis. CONCLUSIONS Modified qifang weitong granules effectively inhibit gastric cancer in both in vitro and in vivo models， and the mechanism of action is related to the suppression of EMT.

OBJECTIVE: To observe the clinical effect of acupuncture at yinsanzhen combined with auricular point sticking on primary dysmenorrhea (PDM). METHODS: Sixty patients with PDM were randomly divided into an observation group and a control group, with 30 cases in each group. Patients in the observation group were treated with acupuncture at yinsanzhen combined with auricular point sticking. The acupuncture was given at yinsanzhen (Guanyuan [CV4] and bilateral Guilai [ST29], Sanyinjiao [SP6]) once daily for 5 consecutive days. Auricular point sticking was applied to gan (CO₁₂), shen (CO₁₀), neifenmi (CO₁₈), etc. every other day, alternated between ears, totaling 3 sessions. All treatments were started 5 days before menstruation. Patients in the control group were treated with ibuprofen sustained-release capsules on the first day of menstruation for 3 consecutive days. Both groups were treated for 3 menstrual cycles. The scores of Cox menstrual symptom scale (CMSS) and visual analogue scale (VAS) were compared between the two groups before and after treatment and at the second menstrual cycle after treatment completion (follow-up). The serum contents of prostaglandin (PG) F2α and PGE2 were detected before and after treatment, and the clinical effect and safety of the two groups were evaluated. RESULTS: After treatment and during follow-up, the CMSS severity and duration scores and VAS scores of the two groups were lower than those before treatment (P<0.05 ), and the scores in the observation group were lower than those in the control group (P<0.05). After treatment, the serum contents of PGF2α were decreased, and the contents of PGE2 were increased (P<0.05) in the two groups. The total effective rate of the observation group was 93.3% (28/30), which was higher than 80.0% (24/30) of the control group (P<0.05). There were no adverse reactions in both groups. CONCLUSION Yinsanzhen combined with auricular point sticking can effectively improve the pain symptoms, relieve the degree of pain and shorten the duration of pain in patients with PDM, which may play a therapeutic role by reducing the content of serum PGF2α and increasing the content of serum PGE2.
Humans ; Female ; Dysmenorrhea/therapy* ; Acupuncture Points ; Adult ; Young Adult ; Acupuncture, Ear ; Treatment Outcome ; Adolescent ; Acupuncture Therapy ; Combined Modality Therapy

Objective To observe the value of the integrated model of MR high-resolution vascular wall imaging(HR-VWI)and attention mechanism for predicting stroke recurrence in symptomatic intracranial atherosclerotic stenosis(sICAS)patients.Methods A total of 363 patients with sICAS who underwent HR-VWI were enrolled and stratified into training set(n=254)and validation set(n=109)according to their origins.Employing a radiomics model that utilized HR-VWI T1 and contrast-enhanced sequences for feature extraction,image data were captured from relevant plaques.Subsequently,a Trans model was developed by integrating the Transformer attention mechanism.The predictive performance and clinical utility of conventional radiomics models and Trans models for forecasting stroke recurrence among patients with sICAS were evaluated.Results In training set and validation set,the area under the curve of Trans model for predicting stroke recurrence in sICAS patients was 0.992 and 0.988,respectively,both superior to that of T1 model,T1 enhanced model and dual sequence model(all P＜0.05).The calibration curve and decision curve analysis showed that Trans model had good predictive probability and clinical practicality.Conclusion The obtained integrated model of HR-VWI radiomics combined with attention mechanism had certain value for predicting stroke recurrence in patients with sICAS.

microRNA(miRNA),as a short non-coding RNA molecule,regulates bone-immune signal and stress response through transcription,translation as well as the combination of shear process,exerting the effect of promoting angiogenesis and repairing damage bone;type H vessels play a role in the cou-pling of angiogenesis and osteogenesis in the formation of new bone and promoting bone regeneration at the defect site.In re-cent years,miRNA regulates hypoxia inducible factor 1α(HIF-1 α)/vascular endothelial growth factor(VEGF),Notch,plate-let-derived growth factor-BB(PDGF-BB),Wnt/glycogen syn-thase kinase-3 β(GSK3 β)to affect the migration,recruitment,proliferation and differentiation of endothelial cells(ECs)and osteoblast(OB),promote the coupling of H-type vessels and os-teoblasts to improve bone homeostasis,and then prevent and treat bone metabolic diseases.This article explains the role of miRNA in regulating H-type vascular osteogenic coupling to provide tar-gets and pathways for improving bone homeostasis.In addition,the active ingredients of traditional Chinese medicine affect the differential expression of miRNA to promote H-type angiogenesis and provide strategies for clinical prevention and treatment of bone metabolic diseases.

Objective To evaluate the efficacy and safety of high-power,short-duration radiofrequency catheter ablation for the treatment of persistent atrial fibrillation.Methods This retrospective study included 392 patients diagnosed with persistent atrial fibrillation who underwent catheter radiofrequency ablation at Suzhou Kowloon Hospital,Shanghai Jiao Tong University School of Medicine,from January 2019 to December 2023.Of these,256 patients were treated with high-power,short-duration ablation,and 136 patients with low-power,long-duration ablation.The following parameters were compared:radiofrequency ablation time,total procedure time,single-circle pulmonary vein isolation rate,immediate procedural success rate,number of ablation points,and perioperative complications(including pericardial tamponade,pseudoaneurysm,arteriovenous fistula,stroke,etc.).Follow-up assessments were conducted at 3,6,and 12 months post-surgery to evaluate the 12-month sinus rhythm maintenance rate.Results The ablation time in the high-power group was significantly shorter than that in the low-power group[(14.6±2.3)min vs.(30.3±4.2)min,P＜0.001],as was the total procedure time[(113.8±24.8)min vs.(128.5±26.7)min,P=0.001].There were no significant differences between the two groups in terms of pulmonary vein isolation rate(97.7％vs.94.9％,P=0.823),number of ablation points[(71.2±8.0)vs.(74.3±14.3),P=0.168],or perioperative complications(3.1％vs.4.4％,P=0.571).Regarding the maintenance rate of sinus rhythm at 12 months post-operation,the high-power group showed a higher rate than the low-power group,but no statistically significant difference was observed(82.8％vs.79.4％,P=0.399).Conclusions High-power,short-duration radiofrequency catheter ablation can improve procedural efficiency in the treatment of persistent atrial fibrillation.Its efficacy and safety are similar to those of the low-power,long-duration technique.