ObjectiveTo investigate the epigenetic mechanism of Diwu Yanggan Capsule in improving liver regeneration microenvironment in a rat model of liver cancer by regulating DNA methylation， and to provide a basis for scientific clinical medication. MethodsA total of 48 specific pathogen-free Sprague-Dawley rats were divided into normal group， model group， and Diwu Yanggan Capsule group using a random number table， with 16 rats in each group. The Solt-Farber two-step method was used to establish a rat model of liver cancer. The rats in the Diwu Yanggan Capsule group were given Diwu Yanggan Capsule at a dose of 750 mg/kg/d by gavage， and those in the normal group and the model group were given an equal volume of normal saline by gavage. Liver tissue samples were collected from each group of rats after 16 weeks of continuous intervention； DNA methylation chips were used to analyze the change in DNA methylation in liver tissue， and gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） analyses were used for data analysis. In addition， the MeDIP-PCR technique was used to detect the changes in candidate differentially methylated genes such as YWHAB， ADCK2， ERLIN2， SEMA3B， and TPH2 in the liver tissue of rats， and Western blot and RT-qPCR were used to verify the expression of key methylated genes. The independent-samples t test was used for comparison of continuous data between two groups， and a one-way analysis of variance was used for comparison between multiple groups， while the least significant difference t-test was used for further comparison between two groups. ResultsThe DNA methylation chip analysis showed that compared with the normal group， the model group had significant methylation changes in the promoter region of 2 422 genes in liver tissue of rats. The GO functional enrichment analysis and the KEGG pathway enrichment analysis showed that these differentially methylated genes were significantly enriched in metabolic pathways such as steroid hormone biosynthesis and drug metabolism-cytochrome P450. Compared with the model group， the Diwu Yanggan Capsule group had significant reversal of promoter methylation in 1 650 genes， and the KEGG enrichment analysis showed that these genes were mainly involved in the pathways closely associated with cell proliferation， apoptosis， and microenvironment regulation， such as the calcium ion signaling pathway， the cAMP signaling pathway， and the extracellular factor signaling pathway. Compared with the model group， the Diwu Yanggan Capsule group had a significant increase in the promoter methylation level of the ADCK2 gene （P<0.05） and significant reductions in the promoter methylation levels of the ERLIN2 and TPH2 genes （all P<0.05）. Compared with the model group， the Diwu Yanggan Capsule group had significant reductions in the mRNA expression levels and the protein expression levels of the ADCK2 （all P<0.05）. ConclusionAbnormal DNA methylation in liver tissue participates in the development and progression of liver cancer. The effect of Diwu Yanggan Capsule on DNA methylation level is an important epigenetic mechanism for its effect in the prevention and treatment of liver cancer.

Autism spectrum disorder （ASD）， also known as autism， is a series of neurodevelopmental disorders characterized by social disorders and repetitive stereotyped behaviors/narrow interests. Its pathogenesis is complex， and there is a lack of effective treatment drugs， with some cases having adverse outcomes. Recent studies have consistently revealed that individuals with autism spectrum disorder （ASD） commonly exhibit characteristics such as gut microbiota dysbiosis （abnormal Bacteroidetes/Firmicutes ratio）， impaired intestinal barrier function （elevated serum levels of zonulin and LPS）， and intestinal immune dysregulation （increased pro-inflammatory cytokines including IL-6 and TNF-α）， suggesting that gastrointestinal abnormalities may influence central nervous system development through neuroendocrine， immunoregulatory， and metabolic pathways. Consequently， growing scholarly attention has focused on dietary interventions as potential approaches to alleviate clinical symptoms in children with ASD. This review systematically summarizes the role of gut microbiota and their metabolite alterations in ASD pathogenesis， along with recent advancements in understanding the microbiota-gut-brain axis mechanisms. Additionally， it elaborates on the therapeutic effects and underlying biological basis of restrictive diet therapy， modified diet therapy， and nutritional supplementation therapy in promoting the health of children with ASD. This systematic review reveals that children with ASD exhibit significant gut microbiota dysbiosis （e.g.， increased Clostridium， decreased Faecalibacterium） and abnormal metabolite profiles （e.g.， altered short-chain fatty acid spectra， elevated 4EPS levels）. These alterations exacerbate neuroinflammation and immune dysregulation through the microbiota-gut-brain axis， thereby impacting nervous system development and function. Furthermore， interventions such as ketogenic diets， camel milk， and specific nutritional supplements can alleviate certain ASD symptoms by modulating gut microbiota， restoring intestinal barrier function， and improving metabolic pathways. Future investigations should aim to create multi-omics evaluation systems for pinpointing potential beneficiaries， devise individualized intervention strategies rooted in microbiome characteristics， and verify their therapeutic value and safety in large-scale randomized controlled trials. These efforts are crucial to transitioning ASD treatment from symptomatic control to address disease etiology， thereby paving the way for improving prognoses.

This study aims to describe the population and regional distribution characteristics of smoking behavior among adult twins in the China Twin Registry (CNTR), as well as the concordance rates for smoking behavior in monozygotic and dizygotic twins, and estimate the heritability. The study population included adult twins in CNTR who had smoking questionnaire data. A random-effects regression model was used to describe the distribution of smoking behavior among different subgroups based on various characteristics. The concordance of smoking behavior between different zygosity groups was calculated, and heritability was estimated. A total of 28 444 twin pairs were included in this study, with an average age of (36.6±12.0) years. Among male twins, 41.2% were current smokers, while only 1.2% of females smoked. Higher smoking rates were observed among male smokers in the 50-59 age group ( z=23.0, P<0.001), northern regions ( z=2.9, P<0.01), rural areas ( z=-5.2, P<0.001), those who were divorced/widowed ( z=3.8, P<0.001), and first-born twins ( z=-4.3, P<0.001), while lower smoking rates were found in those with higher education ( z=-16.1, P<0.001) and unmarried individuals ( z=-16.0, P<0.001). The smoking concordance rate for male monozygotic twins was 69.6%, significantly higher than the 57.3% concordance rate for dizygotic twins ( χ 2=105.0, P<0.05). The heritability of smoking behavior in male twins was estimated at 28.9% (95% CI: 24.3%-33.4%). Stratified analyses showed differences in heritability across regions and age groups: the heritability in northern regions was 32.6% (95% CI: 27.3%-38.0%), higher than the 21.0% (95% CI: 12.4%-29.5%) observed in southern regions; the highest heritability of 35.1% (95% CI: 26.3%-43.9%) was found in the 18-29 age group, with heritability decreasing with age. In conclusion, the smoking rate and influencing factors in the twin population are similar to those in the general population, with unique characteristics, such as higher smoking rates in first-born twins. Genetic factors have a significant impact on smoking behavior.

Alzheimer's disease(AD)is a preva-lent neurodegenerative disorder,characterized by the accumulation of beta-amyloid plaques,the phosphorylation of Tau proteins,and neuronal loss.As the global population ages,the incidence of AD is rising,and there is currently no effective cure.Herbal monomers have garnered interest due to their multifaceted pharmacological effects and low toxicity.This paper aims to provide a comprehen-sive overview of the mechanisms of Nrf2,NF-κB,PI3K/Akt,MAPK and other signalling pathways in the pathogenesis of AD.It also explores the modu-lation of these pathways by various TCM mono-mers,such as leptomeningine and tanshinone ⅡA,and details the research progress to date.For in-stance,Leptosine has been shown to activate Nrf2,thereby reducing oxidative stress,while Tanshinone ⅡA has been observed to inhibit the NF-κB path-way,leading to a reduction in inflammation.Not-withstanding the encouraging indications for the treatment of AD with TCM monomers,there are several challenges that must be addressed.Firstly,the precise mechanism of action remains to be ful-ly elucidated.Secondly,there are significant chal-lenges related to pharmacokinetics and bioavailabil-ity.Thirdly,the sample size of clinical studies is lim-ited and of variable quality.Fourthly,the quality control process is complex.Finally,interactions with other drugs must be taken into account.

This study explored a novel digital design and fabrication method for a double constrained split orthodontic miniscrew guide to improve the accuracy and safety of clinical miniscrew implantation and reduce related complications. A patient requiring miniscrew implantation was selected, and data were acquired using cone beam computed tomography (CBCT) and intraoral optical scanning. For the construction of a double constrained split guide including a screw-hole guide and an insertion rod guide, different types of software such as Mimics 24.0, Geomagic wrap 2021, and Materialise magics 21.0 were utilized for 3D reconstruction, model integration, and guide design. The guide was then fabricated via laser metal 3D printing. Model and intraoral try-in results demonstrated that the guide fitted well and was stable. Postoperative CBCT verified that the final miniscrew implantation site was consistent with the preoperative design, and no related complications occurred. This double constrained split orthodontic miniscrew guide provides a precise and safe digital solution for clinical miniscrew implantation.
Humans ; Bone Screws ; Cone-Beam Computed Tomography ; Printing, Three-Dimensional ; Orthodontic Anchorage Procedures/instrumentation* ; Imaging, Three-Dimensional ; Computer-Aided Design

In the earliest existing pharmacological monograph in China,Shennong Bencao Jing,there is a record of a parasitic plant known as"desert ginseng"——Cistanche deserticola.It exerts beneficial effects on the function of the heart,kidney,spleen,and lung.As research into its pharmacological properties has progressed,the active components of Cistanche deserticola also have shown promise for treating intestinal disorders.Due to the complex pathological mechanisms of sepsis,which often accompany multiple organ dysfunction,aligns closely with the multi-target pharmacological characteristics of Cistanche deserticola's.Furthermore,because Cistanche deserticola is both edible and medicinal,and exhibits a wide therapeutic window,it has a natural advantage in the clinical transformation and application after drug development.Currently,to expand its medicinal range,a review of the main active components of Cistanche deserticola and their related pharmacological effects is conducted,and combined with the specific characteristics of intestinal injury in sepsis,the potential value of Cistanche deserticola in intestinal barrier protection,microbial coordination,and intestinal motility regulation is further elaborated,providing new ideas and a theoretical basis for the application of Cistanche deserticola in sepsis-induced intestinal injury.

Objective:To describe the distribution characteristics of alcohol consumption in adult twins in the Chinese National Twin Registry (CNTR), and further explore the influence of genetic factors on alcohol consumption in adult twins.Methods:The subjects of the study were twins registered by CNTR in 11 project areas across China from 2010 to 2018. A total of 56 966 twins (28 483 pairs) aged 18 years and above who answered questions about drinking behavior were included, and the random effect model was used to describe the population and regional distribution characteristics of alcohol consumption. Intra-pair analysis was performed to calculate the concordance rate and heritability of their alcohol consumption.Results:The age of all subjects was (36.6±12.0) years, and current drinkers accounted for 16.6% (9 461/56 966) of all subjects. In men, those aged 50-59 years, those in northern China, those living in rural area, those with low education level and those with high BMI, the proportions of current drinkers were higher. After excluding 468 pairs of twins who had stopped alcohol use and 21 764 pairs of twins who had no drink or had small amount drink, an intra-pair analysis was conducted in 4 929 pairs of same-sex twins, and found that the concordance rate of alcohol consumption was 64.0% (2 059/3 215) in monozygotic twins, and 52.6% (902/1 714) in dizygotic twins, the difference was significant ( P<0.001), and the heritability of alcohol consumption was 24.1% (95% CI: 18.9%- 29.3%). The further stratified analysis found that in southern men, the heritability was highest in those aged 40-49 years (36.1%, 95% CI: 21.6%-50.7%), while in northern men, the heritability was highest in those aged 50-59 years (34.2%, 95% CI: 18.1%-50.3%). Conclusions:In adult twins in China, there were population and regional differences in the distribution of alcohol consumption behavior, and alcohol consumption was influenced by genetic factors, and gender, age and region had potential modifying effects.

Objective To investigate the impact of the dominant deafness point mutation p.D179N on the oli-gomeric equilibrium state of Connexin 26(Cx26).Methods The wild-type Cx26 fusion protein(Cx26-WT-GFP)and mutant fusion proteins(Cx26-D179N-GFP,Cx26-D179C-GFP)were expressed in HEK293F cells.By using Fluorescence-detection size-exclusion chromatography(FSEC)and size-exclusion chromatography(SEC)to analysis the oligomeric state of the target protein based on malecular weight under the condition of solubilization and purifica-tion respectively.Cryo-electron microscopy(Cryo-EM)single particle analysis(SPA)was conducted to analysis the target protein's oligomeric states based on the 2D classification morphology of the protein particles.Results In vitro,the wild-type Cx26 protein(Cx26-WT)is almost exclusively dodecameric.The deafness mutation p.D179N protein(Cx26-D179N)exists as both dodecamers and hexamers,whereas the artificial mutation p.D179C protein(Cx26-D179C)does not form dodecamers.Conclusion The dominant deafness mutation GJB2 p.D179N could weaken the ability of docking between hexameric proteins,which could affect the balance between hexamers and do-decamers.

Objective To investigate the damage mechanism of extracellular vesicles(EVs)derived from ves-tibular schwannoma(VS)on HEI-OC1 cells and the protective effect of the ferroptosis inhibitor ferrostatin-1(Fer-1).Methods Tumor tissues and clinical data were collected from four patients with stage Ⅱ or Ⅲ VS,classified as grade D according to the AAO-HNS hearing classification.Primary VS cells were extracted,and their conditioned supernatant was collected.EVs were isolated using ultracentrifugation and identified.HEI-OC1 cells were cultured in vitro and divided into three groups:the control group(no treatment),the EVs group(treated with 3000 parti-cles/cell VS-EVs for 24 hours),and the EVs+Fer-1 group(pretreated with 20 μmol/L Fer-1 for 2 hours followed by co-culture with 3000 particles/cell VS-EVs for 24 hours).Cell viability was assessed using the CCK-8 assay,re-active oxygen species(ROS)levels were quantified using the DCFH-DA fluorescent probe,and lipid peroxidation was evaluated using the BODIPY 581/591 C11 probe.Results Compared with the control group,the EVs group showed significantly reduced cell viability(P＜0.001)and increased levels of ROS(P＜0.001)and lipid peroxides(P＜0.001).However,the EVs+Fer-1 group exhibited significantly restored cell viability(P＜0.001)and re-duced levels of ROS and lipid peroxidation(P＜0.001).Conclusion VS-derived EVs disrupts redox homeostasis,promotes intracellular accumulation of lipid peroxides and ROS,and induces ferroptosis in HEI-OC1 cells.Fer-1 significantly alleviates VS-EVs-induced ferroptosis,thereby protecting HEI-OC1 cells from damage.

Alzheimer's disease(AD)is a preva-lent neurodegenerative disorder,characterized by the accumulation of beta-amyloid plaques,the phosphorylation of Tau proteins,and neuronal loss.As the global population ages,the incidence of AD is rising,and there is currently no effective cure.Herbal monomers have garnered interest due to their multifaceted pharmacological effects and low toxicity.This paper aims to provide a comprehen-sive overview of the mechanisms of Nrf2,NF-κB,PI3K/Akt,MAPK and other signalling pathways in the pathogenesis of AD.It also explores the modu-lation of these pathways by various TCM mono-mers,such as leptomeningine and tanshinone ⅡA,and details the research progress to date.For in-stance,Leptosine has been shown to activate Nrf2,thereby reducing oxidative stress,while Tanshinone ⅡA has been observed to inhibit the NF-κB path-way,leading to a reduction in inflammation.Not-withstanding the encouraging indications for the treatment of AD with TCM monomers,there are several challenges that must be addressed.Firstly,the precise mechanism of action remains to be ful-ly elucidated.Secondly,there are significant chal-lenges related to pharmacokinetics and bioavailabil-ity.Thirdly,the sample size of clinical studies is lim-ited and of variable quality.Fourthly,the quality control process is complex.Finally,interactions with other drugs must be taken into account.