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”.

Objective To construct a predictive model for patients with verte-brobasilar dolichoectasia(VBD)complicated by posterior circulation acute cerebral infarction(ACI)based on the characteristics of verte-brobasilar artery magnetic resonance vascular wall imaging(MR-VWI),thereby providing a reference for clinical prevention.Methods A total of 102 patients with VBD complicated by posterior circulation ACI who were admitted to our hospital between March 2016 and January 2023 were selected as the concurrent group.An addi-tional 102 patients with VBD without concurrent posterior circulation ACI were selected as the non-concurrent group at a 1∶1 ratio.The MR-VWI characteristics(basilar artery[BA]diameter,BA length,vertebral artery[VA]intracranial segment length,BA deviation grade,and BA bifurcation height grade)and clinical data of the two groups were compared,and the factors influencing posterior circulation ACI in patients with VBD were analyzed.Based on the MR-VWI features and related influencing factors,a nomogram prediction model of pos-terior circulation ACI in patients with VBD was constructed,and the clinical significance of the nomogram prediction model was evaluated using receiver operating characteristic(ROC)and calibration curves.Results A significant difference were observed in the D-dimer levels between the two groups(P＜0.05).BA diameter,BA length,and VA intracranial length were greater in the concurrent group than in the non-concurrent group.The proportion of BA bifurcation height grade ≥ 2 and BA deviation grade≥2 in the concurrent group was higher than that in the non-concurrent group(P＜0.05).BA diameter,BA length,BA bifurcation height classification,BA deviation classi-fication,and D-dimer level were all independent risk factors for posterior circulation ACI in patients with VBD(P＜0.05).The area under the curve of the posterior circulation ACI predicted using the nomogram model was 0.900(95％CI:0.858-0.943),and good calibration was noted.Conclusion BA diameter,BA length,BA bifurcation height classification,BA offset classification,and D-dimer level were inde-pendent influencing factors of posterior circulation ACI in patients with VBD.Based on these factors,a nomogram prediction model was constructed with high prediction efficiency and good calibration.

AIM To investigate the effects of Rutong Ruanjian Tablets on angiogenesis in cancer tissues of rats with preneoplastic breast cancer(PBC).METHODS 60 female SD rats were randomly divided into a blank group of 10 rats and a model group of 50 rats for the establishment of the PBC models of Liver-Qi Stagnation and Blood Stasis Pattern with 9 weeks of oral administration of 7,12-dimethylbenz[a]anthracene(DMBA)and cervical ligation.After successful modeling,the rats were randomly divided into the model group,the tamoxifen group(3.2 mg/kg),the Rutong Ruanjian Tablets group(128 mg/kg),the 3,5-difluorobenzoyl group(DAPT,5 mg/kg),and the Rutong Ruanjian Tablets(128 mg/kg via gavage)+DAPT(5 mg/kg intraperitoneal injection)group,for 1 month corresponding drug administration,with 10 rats in each group.Then the rats had their cancer progression and syndrome scores observed;their angiogenesis evaluated by assessment of microvascular density(MVD);their vascular endothelial growth factor(VEGF)expression assessed by immunohistochemistry;and their mRNA and protein expressions of proteins related to the DLL4/Notch1/Hes1 pathway measured using RT-qPCR,immunohistochemistry and Western blot.RESULTS During carcinogenesis of rats induced by DMBA,there was gradual disappearance of E-cadherin expression and consistency of HE staining result with the PBC progression confirming the success of the modeling.Compared with the blank group,the model group showed increased MVD values,mRNA expression of Notch1 and Hes1,and protein expressions of VEGF,DLL4,Notch1 and Hes1(P＜0.05,P＜0.01).Compared with the model group,the Rutong Ruanjian Tablets group exhibited reduced MVD values,mRNA expression of Notch1 and Hes1,and protein expressions of VEGF,DLL4,Notch1 and Hes1(P＜0.05,P＜0.01).The Rutong Ruanjian Tablets+DAPT group showed reduced mRNA expression of Notch1 and Hes1,and protein expressions of DLL4,Notch1 and Hes1 compared to the Rutong Ruanjian Tablets group(P＜0.05,P＜0.01).CONCLUSION Rutong Ruanjian Tablets can inhibit angiogenesis and attenuate cancer progression in PBC rats of Liver-Qi Stagnation and Blood Stasis Pattern,and the mechanism may lie in the downregulation of DLL4/Notch1/Hes1 signaling pathway related proteins.

Objective:To analyze the perioperative risks and prognosis in patients with differentiated thyroid cancer (DTC) complicated by Graves' disease (GD) .Methods:Clinical data of 880 DTC patients who underwent radical thyroidectomy at the Department of General Surgery, the First Medical Center of Chinese PLA General Hospital between Jan. 2016 and Dec. 2022 was retrospectively collected. The cohort included 119 DTC patients with GD (hyperthyroid group) and 761 DTC patients with normal thyroid function without GD (normal group) . Use propensity score matching (PSM) was used to obtain cohorts with balanced baseline characteristics between the hyperthyroid and normal groups. Perioperative risks and prognos were compared between the two groups.Results:For perioperative risk comparison, after 1∶2 matching, 116 patients were included in the hyperthyroid group and 232 patients in the normal group. The operation time was longer in the hyperthyroid group compared to normal group ([107.26±30.27]min vs. [96.45±32.51]min, P=0.003) . There were no statistical differences between the two groups in intraoperative blood loss, postoperative drainage volume or postoperative hospital stay ( P>0.05) . There were no statistical difference in the incidence of complications such as temporary or permanent hypoparathyroidism, temporary or permanent recurrent laryngeal nerve injury, and lymphatic leakage between the two groups. For prognostic comparison, after 1∶2 matching, 117 cases were included in the hyperthyroid group and 234 cases in the normal group. During follow-up period, 16 patients experienced DTC recurrence, including 5 cases in the hyperthyroid group and 11 cases in the normal group. There was no statistical difference in reccurence-free survival rates between the two groups (5-year RFS: 95.7% vs. 95.3%, P=0.907) . Multivariate Cox regression analysis revealed GD was not the risk factor for DTC recurrence. Conclusions:With standardized and adequate preoperative preparation, the perioperative risks of DTC patients complicated by GD were not increased compared to patients without GD. Additionally, GD does not increase the risk of postoperative tumor recurrence in DTC patients.

Objective To construct a predictive model for patients with verte-brobasilar dolichoectasia(VBD)complicated by posterior circulation acute cerebral infarction(ACI)based on the characteristics of verte-brobasilar artery magnetic resonance vascular wall imaging(MR-VWI),thereby providing a reference for clinical prevention.Methods A total of 102 patients with VBD complicated by posterior circulation ACI who were admitted to our hospital between March 2016 and January 2023 were selected as the concurrent group.An addi-tional 102 patients with VBD without concurrent posterior circulation ACI were selected as the non-concurrent group at a 1∶1 ratio.The MR-VWI characteristics(basilar artery[BA]diameter,BA length,vertebral artery[VA]intracranial segment length,BA deviation grade,and BA bifurcation height grade)and clinical data of the two groups were compared,and the factors influencing posterior circulation ACI in patients with VBD were analyzed.Based on the MR-VWI features and related influencing factors,a nomogram prediction model of pos-terior circulation ACI in patients with VBD was constructed,and the clinical significance of the nomogram prediction model was evaluated using receiver operating characteristic(ROC)and calibration curves.Results A significant difference were observed in the D-dimer levels between the two groups(P＜0.05).BA diameter,BA length,and VA intracranial length were greater in the concurrent group than in the non-concurrent group.The proportion of BA bifurcation height grade ≥ 2 and BA deviation grade≥2 in the concurrent group was higher than that in the non-concurrent group(P＜0.05).BA diameter,BA length,BA bifurcation height classification,BA deviation classi-fication,and D-dimer level were all independent risk factors for posterior circulation ACI in patients with VBD(P＜0.05).The area under the curve of the posterior circulation ACI predicted using the nomogram model was 0.900(95％CI:0.858-0.943),and good calibration was noted.Conclusion BA diameter,BA length,BA bifurcation height classification,BA offset classification,and D-dimer level were inde-pendent influencing factors of posterior circulation ACI in patients with VBD.Based on these factors,a nomogram prediction model was constructed with high prediction efficiency and good calibration.

Objective:To analyze the perioperative risks and prognosis in patients with differentiated thyroid cancer (DTC) complicated by Graves' disease (GD) .Methods:Clinical data of 880 DTC patients who underwent radical thyroidectomy at the Department of General Surgery, the First Medical Center of Chinese PLA General Hospital between Jan. 2016 and Dec. 2022 was retrospectively collected. The cohort included 119 DTC patients with GD (hyperthyroid group) and 761 DTC patients with normal thyroid function without GD (normal group) . Use propensity score matching (PSM) was used to obtain cohorts with balanced baseline characteristics between the hyperthyroid and normal groups. Perioperative risks and prognos were compared between the two groups.Results:For perioperative risk comparison, after 1∶2 matching, 116 patients were included in the hyperthyroid group and 232 patients in the normal group. The operation time was longer in the hyperthyroid group compared to normal group ([107.26±30.27]min vs. [96.45±32.51]min, P=0.003) . There were no statistical differences between the two groups in intraoperative blood loss, postoperative drainage volume or postoperative hospital stay ( P>0.05) . There were no statistical difference in the incidence of complications such as temporary or permanent hypoparathyroidism, temporary or permanent recurrent laryngeal nerve injury, and lymphatic leakage between the two groups. For prognostic comparison, after 1∶2 matching, 117 cases were included in the hyperthyroid group and 234 cases in the normal group. During follow-up period, 16 patients experienced DTC recurrence, including 5 cases in the hyperthyroid group and 11 cases in the normal group. There was no statistical difference in reccurence-free survival rates between the two groups (5-year RFS: 95.7% vs. 95.3%, P=0.907) . Multivariate Cox regression analysis revealed GD was not the risk factor for DTC recurrence. Conclusions:With standardized and adequate preoperative preparation, the perioperative risks of DTC patients complicated by GD were not increased compared to patients without GD. Additionally, GD does not increase the risk of postoperative tumor recurrence in DTC patients.

Objective Investigate the clinical efficacy and application value of gasless trans-subclavian approach endoscopic surgery(GTAES)for the treatment of primary hyperparathyroidism(PHPT).Methods 55 PHPT patients who underwent unilateral parathyroid adenoma resection from January 2023 to October 2024 were absorbed.Based on surgical approaches,31 patients were divided into the traditional low-collar arc incision open surgery group(open group)and 24 patients were divided into gasless trans-subclavian approach endoscopic surgery group(endoscopic group).The perioperative data,anterior neck function recovery,and incision healing were compared between two groups.Results There were no significant differences in operation time,intraoperative blood loss,postoperative drainage volume and postoperative hospital stay between the two groups(P＞0.05).The preoperative blood calcium levels of the endoscopic group and the open group were(2.68±0.10)mmol/L and(2.63±0.16)mmol/L,respectively.The blood calcium levels on the first day after surgery were(2.39±0.12)mmol/L and(2.36±0.16)mmol/L,respectively.Preoperative parathyroid hormone(PTH)was 27.55(110.05,155.75)pg/ml in endoscopic group and 137.00(107.00,210.30)pg/ml in open group,respectively.PTH on the first day after surgery was 18.98(12.64,23.53)pg/ml and 13.85(7.36,23.23)pg/ml,respectively.Blood calcium and PTH in each group 1 day after surgery decreased compared with that before surgery,and the difference was statistically significant(P＜0.05).There were no postoperative complications such as hoarseness,bleeding and incision infection in both groups.Postoperative hand and foot numbness:5 cases(20.8％)in endoscopic group and 8 patients(25.8％)in the open group,there was no significant difference between the two groups(P=0.667).There was no significant difference in the score of neck pain 3 days after surgery between the two groups(P＞0.05).The score of swallowing tension[(1.92±1.06)points compared with(2.77±1.38)points,P=0.015]and the score of anterior neck discomfort[(1.63±0.88)points compared with(2.35±1.33)points,P=0.018]in the endoscopic group one month after surgery were better than those in the open group.There was no significant difference in the score of neck scar scale between the two groups 3 months after surgery(P＞0.05).VAS score of incision satisfaction was(9.08±0.88)in endoscopic group and(8.19±1.30)in open group,and the endoscopic group was better than the open group,the difference was statistically significant(P=0.004).Conclusion For PHPT patients caused by unilateral parathyroid adenoma confirmed through preoperative localization examination,GTAES is a safe and effective surgical approach,which offers unique advantages in intraoperative lesion identification,cosmetic outcomes of the incision,and preservation of anterior neck function,providing a new surgical option for PHPT patients.

Objective:To translate and adapt the postoperative recovery in children (PRiC) scale, developing a Chinese version for children undergoing dental treatment under general anesthesia (PRiC-DTGA) with validated psychometric properties.Methods:The PRiC scale underwent forward-backward translation using Brislin′s model. A convenience sample of DTGA patients from the Department of Anesthesiology, School of Stomatology, The Fourth Mility Force Medical University was enrolled for a cross-sectional survey on postoperative complications. Delphi expert consultation informed cultural adaptation based on survey findings to develop the PRiC-DTGA Chinese version. Psychometric validation included reliability and validity testing in a separate DTGA cohort at the same center (April-October 2024).Results:Results from the cross-sectionalsurvey of 231 children showed that 82.7% (191/231) of them hadat least one postoperative complication within 72 hours, and these complications were mainly mild local symptoms. Additionally, 358 copies of the Chinese version of the PRiC-DTGA scale were distributed; 21 invalid questionnaires with incomplete information were excluded, and a total of 337 cases were included inthe study. The final PRiC-DTGA comprised 22 items across three dimensions including physical comfort, social ability, and negative emotional. Exploratory factor analysis (EFA) confirmed all factor loadings>0.4. Confirmatory factor analysis (CFA) demonstrated adequate fit: χ 2/df=1.665, tucker-Lewis index (TLI)=0.924, comparative fit index (CFI)=0.896, standardized root mean square residual (SRMR)=0.041, and root mean square error of approximation (RMSEA)=0.044 (90% CI: 0.035-0.053). Reliability was strong with Cronbach′s α (total scale)=0.853, subscale α=0.632-0.723, split-half reliability=0.824. Validity indices met standards: scale-content validity index (S-CVI)=0.909, Item-CVI range=0.944-1.000, average variance extracted (AVE)=0.473-0.501, composite reliability (CR)=0.830-0.913. Conclusions:The systematically adapted PRiC-DTGA demonstrates robust reliability and validity, serving as an effective tool for assessing postoperative recovery quality in Chinese children following DTGA.

Aim To investigate the antidepressant mechanism of hyperforin via the utilization of single-cell sequencing technology.Methods C57BL/6 mice were randomly divided into the control group,depres-sion model group,and hyperforin intervention group.The chronic unpredictable mild stress(CUMS)model was induced and drug interventions were administered for 28 d.Behavioral experiments were conducted to as-sess depressive symptoms,and hippocampal tissue was collected for single-cell RNA sequencing.Key cell populations and differentially expressed genes across groups were identified,followed by PPI network,GO,and KEGG enrichment analysis.Results Behavioral experiments indicated that CUMS successfully induced depressive symptoms in mice,while hyperforin im-proved depressive behavior.In the depression model group,the proportion of brain perivascular macrophages(PVM)increased,and this proportion decreased after hyperforin intervention,approaching the level seen in the control group.The top 20 common differentially ex-pressed genes in the PVM subpopulation were Saa3,Hbb-bs and Ccl24.PPI network analysis identified core targets,including Ccl2,Dhx9,C3,Msr1,Cxcl2 and Cx3cr1.KEGG enrichment analysis revealed pathways related to chemokines,phagosome formation,and inosi-tol phosphate metabolism.Conclusion The antide-pressant mechanism of hyperforin may be related to the regulation of Ccl24 and its related chemokine signaling pathway by PVM.