ObjectiveTo clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology. MethodsThe ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m⁶A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture. ResultsBehaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m⁶A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01). ConclusionMETTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m⁶A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of “opening Xuanfu and dredging myelin”.

ObjectiveTo clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology. MethodsThe ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m⁶A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture. ResultsBehaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m⁶A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01). ConclusionMETTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m⁶A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of “opening Xuanfu and dredging myelin”.

Low anterior resection syndrome （LARS） is a common complication of sphincter-preserving rectal cancer surgery， and its manifestations change dynamically at different stages. By analyzing and summarizing the clinical symptoms and pathomechanism evolution of LARS across different stages， this paper proposes a staged prevention and treatment strategy using acupuncture. During perioperative stage， the main principle is activating the transport function of the body， supplemented by regulating the mind， with the use of dredging the bowels as needed. During the stoma reversal stage， treatment focuses primarily on fortifying the spleen， with draining dampness as a supplementary method， to help consolidate the intestines. During the radiotherapy or chemotherapy stage， the main focus is reinfor-cing healthy qi， with reducing toxin as an adjunct， to achieve the effect of activating the transport of the pivot. During the survival management stage， treatment primarily focuses on tonifying the kidneys and secondarily on fortifying the spleen， with regulating the corporeal soul as the therapeutic emphasis. Acupoints are selected and combined in accordance with the treatment principles at each stage， and different stimulation methods such as electroacupuncture and moxibustion are applied. An analysis of the mechanisms underlying acupuncture for LARS is also provided， offering a theoretical basis and practical approach for the prevention and treatment of LARS with acupuncture.

ObjectiveThis paper aims to observe the syndrome improvement and negative antigen conversion rate of Qingxuan Daozhi formula in the treatment of influenza in children （heat accumulation in the lung and stomach syndrome）. MethodsThrough a multi-center randomized controlled methodology design，confirmed influenza cases were collected from October 2022 to April 2023 in the pediatrics department of eight hospitals，such as Dongfang Hospital of Beijing University of Chinese Medicine. A total of 180 children with influenza and heat accumulation in the lung and stomach syndrome conforming to the standard were recruited through the clinic. The sick children meeting the inclusion criteria were randomly divided into groups by a block-randomized method. The children in the experimental group were treated with Qingxuan Daozhi formula for five days，and those in the control group were treated with Oseltamivir Phosphate Granules for five days. The primary efficacy indicator was the negative conversion rate of influenza antigen detection. Secondary efficacy indicators were the Canadian acute respiratory illness and flu scale （CARIFS） and the incidence of complications，severe cases， and critical cases. Follow-up observation was conducted on the day of enrollment，48 hours after medication，72 hours after medication， and （6+1） d after medication. ResultsOne hundred and eighty participants were randomly assigned to the experimental group （90 cases） or the control group （90 cases）. All participants were followed up during the study. Comparison of influenza antigen detection results in the primary efficacy indicators showed that the average time of negative influenza antigen conversion in the experimental group was （5.29±1.25） d，and that in the control group was （5.40±1.68） d，without a statistically significant difference. After five days of intervention，52 cases in the experimental group and 51 cases in the control group converted to negative，without a statistically significant difference. CARIFS score results in the secondary efficacy indicators showed that during 72 hours after intervention，there were statistically significant differences between the experimental group and the control group in three dimensions， including headache，muscle soreness， and the need for extra care （P<0.05）. On the （6+1） days after the intervention，the differences in both the experimental group and the control group were statistically significant in 10 dimensions， including sore throat，bad sleep，uncomfortable feeling，poor spirit and fatigue，crying more than usual，the need for extra care，symptom，function，influence on parents，and total score （P<0.05）. The comparison results within the group in the dimensional scores of symptom， function， and influence on parents，as well as the CARIFS total score showed that with the delay of follow-up time，scores of both groups decreased significantly，with a statistically significant difference （P<0.01）. Inter-group comparison results showed that the mean score of the experimental group was higher than that of the control group at the time of enrollment. With the progress of intervention，the score of the experimental group was significantly decreased compared with that of the control group. At the end of follow-up，the mean score of the experimental group was lower than that of the control group，with no statistically significant difference. In terms of the incidence of complications，severe cases， and critical cases， there were no complications，severe cases， and critical cases in the two groups，without a statistically significant difference. ConclusionThe symptom improvement effect and negative antigen conversion rate of Qingxuan Daozhi formula in the treatment of influenza in children （heat accumulation in the lung and stomach syndrome） are not inferior to Oseltamivir Phosphate granules， and children's acceptance is better. It can be more widely used in clinical treatment of influenza in children （heat accumulation in the lung and stomach syndrome）.

OBJECTIVE To analyze the chemical constituents and absorbed prototype components in blood of Zhenyang jiuxin decoction， and explore its potential active substances in the treatment of chronic heart failure （CHF）. METHODS SD rats were divided into a blank group （distilled water） and administration groups with blood collection at 0， 15， 30 and 60 min after dosing （14.96 g/kg calculated by crude drug）， with 2 rats in each group. Intragastric administration was conducted once every 12 hours for 3 consecutive days. After the last administration， rat blood was collected to prepare drug-containing serum. UPLC-Q-Exactive Orbitrap HRMS combined with the Global Natural Products Social Molecular Networking technology was adopted to identify the chemical constituents of Zhenyang jiuxin decoction and the absorbed prototype components in blood in drug-containing serum. Moreover， network pharmacology， molecular docking and molecular dynamics simulation were used to predict the potential active components and action targets for CHF treatment. RESULTS A total of 116 chemical constituents were identified from Zhenyang jiuxin decoction， including flavonoids （29 species）， terpenoids （26 species）， saponins （22 species） and other compounds. Twenty absorbed prototype components in blood were detected in drug-containing serum， including icariin， ginsenoside Rg 1 and atractylenolide Ⅰ. Six key active components （icariin， ginsenoside Rg 1 ， atractylenolide Ⅰ， hexadecanamide， etc.） and six core k ey targets such as glycogen synthase kinase-3β （GSK-3β）， amyloid precursor protein （APP） and epidermal growth factor receptor （EGFR） were screened out via network pharmacology. Molecular docking results showed that all key active components had favorable binding activity with core targets； among them， icariin had the strongest binding capacity to APP. Molecular dynamics simulation verified that the complex formed by icariin and APP possessed excellent stability. CONCLUSIONS The potential active substances of Zhenyang jiuxin decoction against CHF are icariin， ginsenoside Rg₁， atractylenolide Ⅰ and other ingredients， which act on APP， EGFR， GSK-3β and other targets. APP may serve as one of the important action targets of icariin.

OBJECTIVE To analyze the chemical constituents and absorbed prototype components in blood of Zhenyang jiuxin decoction， and explore its potential active substances in the treatment of chronic heart failure （CHF）. METHODS SD rats were divided into a blank group （distilled water） and administration groups with blood collection at 0， 15， 30 and 60 min after dosing （14.96 g/kg calculated by crude drug）， with 2 rats in each group. Intragastric administration was conducted once every 12 hours for 3 consecutive days. After the last administration， rat blood was collected to prepare drug-containing serum. UPLC-Q-Exactive Orbitrap HRMS combined with the Global Natural Products Social Molecular Networking technology was adopted to identify the chemical constituents of Zhenyang jiuxin decoction and the absorbed prototype components in blood in drug-containing serum. Moreover， network pharmacology， molecular docking and molecular dynamics simulation were used to predict the potential active components and action targets for CHF treatment. RESULTS A total of 116 chemical constituents were identified from Zhenyang jiuxin decoction， including flavonoids （29 species）， terpenoids （26 species）， saponins （22 species） and other compounds. Twenty absorbed prototype components in blood were detected in drug-containing serum， including icariin， ginsenoside Rg 1 and atractylenolide Ⅰ. Six key active components （icariin， ginsenoside Rg 1 ， atractylenolide Ⅰ， hexadecanamide， etc.） and six core k ey targets such as glycogen synthase kinase-3β （GSK-3β）， amyloid precursor protein （APP） and epidermal growth factor receptor （EGFR） were screened out via network pharmacology. Molecular docking results showed that all key active components had favorable binding activity with core targets； among them， icariin had the strongest binding capacity to APP. Molecular dynamics simulation verified that the complex formed by icariin and APP possessed excellent stability. CONCLUSIONS The potential active substances of Zhenyang jiuxin decoction against CHF are icariin， ginsenoside Rg₁， atractylenolide Ⅰ and other ingredients， which act on APP， EGFR， GSK-3β and other targets. APP may serve as one of the important action targets of icariin.

Objective:To investigate the current situation and correlation between mental health and stressors of new professional master's degree graduates in clinical medicine, and to provide empirical support for improving student mental health.Methods:An online questionnaire survey was conducted among 3 587 fresh graduate students with master's degree in clinical medicine from 65 training institutions in China. SPSS 26.0 was used to compare and analyze the measurement data. Descriptive statistics were used to analyze the mental health using Symptom Checklist-90 (SCL-90) and the current status of stressors. The influence factors and correlation between mental health and stressors were analyzed by means of differentiation analysis and correlation analysis.Results:The SCL-90 overall mean score as well as the scores of obsessive-compulsive symptoms, interpersonal sensitivity, depression, terror, anxious, psychoticism, and hostility of new professional master's degree graduates in clinical medicine were all higher than the national norm ( P<0.05). Among the participants, 58.21% (2 088/3 587) exhibited varying degrees of symptoms listed in the SCL-90. Students from families with low per capita income and low parental education level have more prominent psychological problems. The top three stressors perceived were job seeking, research tasks, and self-ability. Students with positive SCL-90 symptoms reported significantly higher perceived stress in all three stressor dimensions compared to their symptom-negative counterparts ( P<0.001), indicating a significant correlation between mental health status and stressors. Conclusions:The mental health status of new professional master's degree graduates in clinical medicine is concerning and warrants focused attention from educational authorities, academic institutions, and families.

This study explored the effects and underlying mechanism of Raf kinase inhibitory protein(RKIP)on apoptosis in mast cells sensitized by Echinococcus granulosus cyst fluid.Bone marrow-derived mast cells(BMMCs)were isolated and cultured from RKIP knockout(KO)and wild-type(WT)C57BL/6 mice.Cells were divided into control and sensitized groups.The sensitized group was incubated for 24 h in RPMI1640 medium containing 10%serum from mice infected with E.granulosus,then activated for 3 h or 6 h with E.granulosus cyst fluid.The control group was incubated for 24 h in RPMI1640 medium,and then received an equal vol-ume of PBS.Cells and supernatants were collected for analysis.Flow cytometry was used to detect the expression of CD117 and FcεRⅠα on BMMCs.The levels of β-hexosaminidase,IL-4,and TNF-α in the supernatant were quantified with ELISA.Western blot analy-sis was used to assess expression changes in RKIP,apoptosis-related proteins,and pathway proteins in BMMC before and after sensi-tization.Flow cytometry analysis revealed that after 4 weeks of induction,the CD117 and FcεRⅠα double-positivity rates on both WT and KO BMMC exceeded 90%.ELISA indicated that the E.granulosus cyst fluid resulted in significantly greater β-hexosaminidase re-lease(F=16.88,P<0.05),and levels of IL-4(F=16.51,P<0.05)and TNF-α(F=9.78,P<0.05)in the KO sensitized group than the WT sensitized group.With respect to the WT control group,the WT sensitized group showed significantly down-regulated pro-tein expression levels of RKIP(F=8.20,P<0.05)and Bcl-2(F=101.40,P<0.01)after 3 h,but significantly up-regulated levels of p-PI3K(F=8.04,P<0.05),p-Akt(F=32.52,P<0.01),p-P65(F=13.29,P<0.05),and cleaved-caspase-3(F=46.34,P<0.01).With respect to the WT sensitized group,the KO sensitized group showed significantly up-regulated protein expression of p-PI3K(F=8.45,P<0.05),p-Akt(F=8.58,P<0.05),p-P65(F=11.02,P<0.05),and Bcl-2(F=84.50,P<0.001)after 3 h,but significantly down-regulated expression of cleaved-caspase-3(F=15.66,P<0.05).In conclusion,RKIP may inhibit the PI3K/Akt/NF-κB pathway,thereby inducing apoptosis in mast cells sensitized by E.granulosus cyst fluid.This process may help ease aller-gic reactions caused by mast cells in echinococcosis,thus offering a promising new approach for preventing and treating such reactions.

Objective:To investigate the clinical characteristics and prognosis of acute pancreatitis (AP) with different etiologies.Methods:The retrospective cohort study was conducted. The clinical data of 702 patients with AP who were admitted to Xuanwu Hospital of Capital Medical University from January 2018 to December 2022 were collected. There were 451 males and 251 females,aged (52±17)years. Observation indicators: (1) clinical characteristics of AP patients with different etiologies; (2) complications and prognosis of AP patients with different etiologies; (3) complications and prognosis of moderately severe and severe AP patients with different etiologies. Comparison of measurement data with normal distribution among multiple groups was conducted using the one-way analysis of variance. Comparison of measurement data with skewed distribution among multiple groups was conducted using the Kruskal-Wallis H test. Comparison of count data among multiple groups was conducted using the chi-square test. Results:(1) Clinical characteris-tics of AP patients with different etiologies. Of the 702 AP patients, 434 cases were biliary AP, 199 cases were hypertriglyceridemic-induced AP,29 cases were alcoholic AP, and 40 cases were idiopathic AP. There were significant differences in gender,age,body mass index,combined cardiovascular disease,combined diabetes,combined nephrosis,disease severity and modified computed tomography severity index among AP patients with different etiologies ( χ2=24.61, F=48.65, 12.24, χ2=13.67, 12.90, 14.12, 15.56, H=17.62, P<0.05). (2) Complications and prognosis of AP patients with different etiologies. There was no significant difference in infectious pancreatic necrosis,duration of intensive care unit stay,total duration of hospital stay, and death of patients during hospitalization among AP patients with different etiologies ( P>0.05). (3) Complications and prognosis of moderately severe and severe AP patients with different etiologies. Of the 395 patients with moderately severe and severe AP, 217 cases were biliary AP, 128 cases were hypertriglyceridemic-induced AP, 19 cases were alcoholic AP, and 31 cases were idiopathic AP. There were significant differences in renal injury, multiple organ dysfunction syndrome and walled-off necrosis among moderately severe and severe AP patients with different etiologies ( χ2=12.62, 8.25, 14.33, P<0.05), and there was no significant difference in infectious pancreatic necrosis, lung injury, circulation system injury, duration of intensive care unit stay, total duration of hospital stay, or death of patients during hospitalization among moderately severe and severe AP patients with different etiologies ( P>0.05). Conclusions:The clinical charac-teristics vary among AP patients with different etiologies. The risk of complications varies among moderately severe and severe AP patients with different etiologies.

Objective To investigate the molecular mechanism by which super enhancers(SEs)regulate the expression of the genetic suppressor element 1(GSE1)and influence the proliferation of breast cancer cell line MCF-7.Methods The oncogene GSE1,driven by SEs,was identified through analysis of the ChIP-seq dataset of MCF-7 cells obtained from the GEO database.The protein expression level of GSE1 was assessed via Western blot following treatment with the bromodomain-containing protein 4(BRD4)inhibitor JQ1.The expression of GSE1 across various cancers and different breast cancer subtypes was analyzed using the Human Protein Atlas(HPA)and The Cancer Genome Atlas(TCGA)databases,respectively.Overall survival(OS)of breast cancer patients was compared between the GSE1 high-expression and low-expression groups using the GEPIA database.Immunohis-tochemistry(IHC)was performed to evaluate GSE1 expression in breast cancer tissue samples.The mRNA expres-sion levels of GSE1 in different breast cancer cell lines were validated by RT-qPCR.A GSE1 interference vector was constructed and transfected into MCF-7 cells,and the knockdown efficiency was assessed using Western blot.Cell proliferation and migration were evaluated using CCK-8 assay,colony formation assay,wound healing assay,and Transwell migration assay.Potential GSE1-interacting proteins were predicted using the STRING database.Finally,Western blot analysis was conducted to assess changes in epithelial-mesenchymal transition(EMT)-related proteins and key components of the Wnt/β-catenin signaling pathway.Results The integrative genomics viewer(IGV)was used to visualize ChIP-seq data from MCF-7 cells,and the rank ordering of super enhancers(ROSE)algorithm was applied to predict the SE region of GSE1 in the MCF-7 genome.GSE1 and BRD4 expression levels were significantly reduced following JQ1 treatment(P<0.05).High expression levels of GSE1 in breast cancer were confirmed through analyses using the HPA,TCGA,and GEPIA databases,as well as IHC,and these find-ings were associated with poor prognostic outcomes in breast cancer patients(P<0.05).RT-qPCR results further demonstrated that GSE1 is significantly upregulated in breast cancer cells(P<0.05).Analysis of the STRING database revealed a strong correlation between GSE1 and Snail(Snai1).Transfection of a GSE1-specific interference vector significantly inhibited the proliferation and migration of MCF-7 cells compared to the control group,upregu-lated E-cadherin and Occludin expression,and downregulated N-cadherin and Snail expression(P<0.05).Addi-tionally,knockdown of GSE1 resulted in decreased expression of Wnt/β-catenin signaling pathway-related proteins,including β-catenin,Wnt-5a,and Cyclin-D1,along with increased Axin1 expression(P<0.05).Conclusion SE driven GSE1 promotes the proliferation,migration and EMT of MCF-7 cells via the Wnt/β-catenin signaling pathway.