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”.

ObjectivePeripheral nerve injury is a common clinical condition that can impair the regulatory function of target organs, leading to functional disability; severe injuries may even result in lifelong disability. Therefore, how to effectively promote the morphological and functional reconstruction of injured peripheral nerves has become a critical scientific issue that urgently needs to be addressed. This study biomimicked the highly oriented structure and electrophysiological properties of peripheral nerves and investigated the feasibility and effectiveness of combining electrical stimulation (ES) with oriented conductive substrates for the repair of injured peripheral nerves at the cellular level. MethodsPolydimethylsiloxane (PDMS) substrates with oriented microgroove structures of varying widths (0, 5, 10, and 20 μm) were fabricated using silicon wafers with micropatterned grooves as templates. Graphene-based conductive patterned substrates (GCPSs) with oriented microgrooves were then constructed via soft lithography and designated as GCPS 0, GCPS 5, GCPS 10, and GCPS 20 according to the groove width. The GCPSs were prepared from collagen/polycaprolactone (PCL) matrices containing 1.0 wt% graphene, and their morphology, wettability, and conductivity were characterized. Mesenchymal stem cells (MSCs) viability, proliferation, adhesion, and orientation on the substrates were evaluated before ES treatment. Then, ES at a frequency of 2 Hz and field strengths of 10, 20, or 50 mV/cm were applied to the cells for 10 min daily, over 3 or 7 consecutive days. Neural differentiation was assessed by morphological observation, immunofluorescence staining, and analysis of neural lineage-related markers. ResultsThe oriented microgroove PDMS substrates supported long-term MSCs growth, guided cells to align along the groove direction, and promoted a morphological transition toward neural-like cells, exhibiting the capacity to induce MSCs differentiation toward the neural lineage. Microgroove structures effectively guided the alignment of MSCs and promoted the development of a more elongated, neural-like morphology, with the most pronounced morphological changes observed on grooves of 10 and 20 μm in width. Meanwhile, the incorporation of graphene endowed the GCPSs with favorable electrical conductivity, though the groove width did not significantly affect the conductive performance. The GCPSs showed good cytocompatibility with MSCs, and cell proliferation on these substrates exhibited a time-dependent pattern, with no significant effect observed from groove width. The inductive effect of ES combined with GCPSs on MSCs differentiation toward specific neural lineages was co-regulated by ES intensity (0, 10, 20, and 50 mV/cm) and microgroove width. ES at 10 mV/cm and 20 mV/cm promotes MSCs differentiation toward neurons on microgrooved substrates of various widths (5, 10, and 20 μm); under the same ES intensities, microgrooves with widths of 5 μm and 10 μm are more favorable for MSCs differentiation toward glial cells/SCs. Notably, low-intensity ES (10 or 20 mV/cm) combined with GCPS 10 significantly enhanced the neural differentiation of MSCs. Neurotrophic factor secretion levels by MSCs cultured on various GCPS substrates after 3 d of ES application was further detected. There were no significant differences in nerve growth factor‑β (NGF‑β) and brain-derived neurotrophic factor (BDNF) concentrations among the GCPS 5, GCPS 10, and GCPS 20 substrates, and these levels were not affected by variations in ES intensity. Under ES at 10 mV/cm, the glial cell line-derived neurotrophic factor (GDNF) concentration in each GCPS substrate was higher than that in the corresponding non-ES group on the same substrate. However, as ES intensity increased, the GDNF concentration decreased to varying degrees. ConclusionThis study demonstrates that the combined strategy of “oriented topographical structure+ES” can effectively promote the neural differentiation of MSCs and holds promise for accelerating the repair process of peripheral nerve injuries.

Objective To explore the effects of pretreatment with rehmannia glutinosa polysaccharide(RGP)and basic fibroblast growth factor(FGF-2)alone or in combination on the differentiation of bone marrow mesenchymal stem cells(BMSCs)into cardiomyoid cells and the related mechanisms.Methods BMSCs pretreated with FGF-2 and RGP alone or in combination were cultured for 1,2,and 4 weeks.The expression levels of NKx2.5 and GATA-4 were detected by Real-time PCR.BMSCs pretreated with FGF-2 or RGP alone and PI3K/Akt signaling pathway inhibitor LY294002 were used to detect the expression levels of myocardial specific proteins and related pathway proteins by Western blotting.Results Compared with the blank control group,pretreatment with FGF-2 or RGP alone increased the expression rate of myocardial specific markers,and the effect of combined pretreatment with FGF-2 and RGP was more obvious(P＜0.05).Compared with pretreatment with FGF-2 or RGP alone,pretreatment with LY294002 combined with FGF-2 or RGP significantly down-regulated the expression of cardiac-specific proteins in BMSCs and inhibited the phosphorylation of Akt(P＜0.05).Conclusion Both FGF-2 and RGP can induce BMSCs to differentiate into cardiomyocyte-like cells by regulating PI3K/Akt signaling pathway,and the combination of FGF-2 and RGP has a better inductive effect.

Obesity, as a chronic recurrent disease, has become a major public health challenge in China. To implement the requirements of the Healthy China Initiative (2019—2030), under domestic guidelines or consensus statements on overweight and obesity, and in alignment with the latest scientific advances globally, the Quality control protocol for adult overweight and obesity screening in health management (examination) institutions (2025 edition) was developed. This protocol was drafted by the Health Management Center of Shanghai Changzheng Hospital and formulated through multiple rounds of deliberation by experts in China’s health examination quality control field. The protocol establishes unified standards for screening facilities, personnel qualifications, and measurement or testing procedures. It defines specific screening items, outlines a standardized screening pathway, and sets requirements for the final medical review, ensuring the scientific validity, effectiveness, and safety of the screening process. The implementation of this protocol will enhance the consistency of weight management practices for adults across health examination institutions and strengthen the quality control of overweight and obesity screening programs.

Objective To investigate the expression of serum interleukin(IL)-32β and long non-coding RNA-T cell leukemia/lymphoma 6(lncRNA-TCL6)in women with severe preeclampsia(sPE)and its relationship with adverse pregnancy outcomes.Methods A total of 316 women with sPE(sPE group)and 323 healthy pregnant women(control group)admitted to Sichuan Jinxin Women and Children's Hospital from November,2020 to June,2023 were enrolled.Serum IL-32β and lncRNA-TCL6 were detected,and pregnancy outcomes were followed up.Multivariate Logistic regression was used to analyze the influencing factors of adverse pregnancy outcomes in women with sPE.Receiver operating characteristic(ROC)curve was used to analyze the efficacy of IL-32β and lncRNA-TCL6 in predicting the pregnancy outcomes of sPE.Results The serum IL-32β level(4.13±1.46 pg/ml)and lncRNA-TCL6 expression(1.36±0.30)in the sPE group were higher than those in the control group(2.35±0.77 pg/ml,0.89±0.21),and the differences were statistically significant(t=19.335,22.983,all P<0.05).The level of serum IL-32β(4.65±0.39 pg/ml)and the expression of lncRNA-TCL6(1.42±0.19)in the poor outcome group were higher than those in the good outcome group(3.98±0.37pg/ml,1.34±0.11),and the differences were statistically significant(t=13.207,4.480,all P<0.05).Early onset preeclampsia,high levels of IL-32β and high expression of lncRNA-TCL6 were risk factors for adverse pregnancy outcomes in sPE patients(Wald χ2=6.138,1.921,1.702,all P<0.05).The AUC of IL-32 β,lncRNA-TCL6,and their combined prediction of sPE pregnancy outcomes were 0.815,0.825 and 0.958,respectively.The combined prediction of IL-32β and lncRNA-TCL6 was higher than the individual prediction,and the differences were statistically significant(Z=4.502,4.017,all P<0.05).Conclusion The serum levels of IL-32β and lncRNA-TCL6 are increased in patients with sPE,which are related to the adverse pregnancy outcome.Combined IL-32β and lncRNA-TCL6 have a high value in predicting the pregnancy outcome of patients with sPE.

Objective To investigate the expression of serum interleukin(IL)-32β and long non-coding RNA-T cell leukemia/lymphoma 6(lncRNA-TCL6)in women with severe preeclampsia(sPE)and its relationship with adverse pregnancy outcomes.Methods A total of 316 women with sPE(sPE group)and 323 healthy pregnant women(control group)admitted to Sichuan Jinxin Women and Children's Hospital from November,2020 to June,2023 were enrolled.Serum IL-32β and lncRNA-TCL6 were detected,and pregnancy outcomes were followed up.Multivariate Logistic regression was used to analyze the influencing factors of adverse pregnancy outcomes in women with sPE.Receiver operating characteristic(ROC)curve was used to analyze the efficacy of IL-32β and lncRNA-TCL6 in predicting the pregnancy outcomes of sPE.Results The serum IL-32β level(4.13±1.46 pg/ml)and lncRNA-TCL6 expression(1.36±0.30)in the sPE group were higher than those in the control group(2.35±0.77 pg/ml,0.89±0.21),and the differences were statistically significant(t=19.335,22.983,all P<0.05).The level of serum IL-32β(4.65±0.39 pg/ml)and the expression of lncRNA-TCL6(1.42±0.19)in the poor outcome group were higher than those in the good outcome group(3.98±0.37pg/ml,1.34±0.11),and the differences were statistically significant(t=13.207,4.480,all P<0.05).Early onset preeclampsia,high levels of IL-32β and high expression of lncRNA-TCL6 were risk factors for adverse pregnancy outcomes in sPE patients(Wald χ2=6.138,1.921,1.702,all P<0.05).The AUC of IL-32 β,lncRNA-TCL6,and their combined prediction of sPE pregnancy outcomes were 0.815,0.825 and 0.958,respectively.The combined prediction of IL-32β and lncRNA-TCL6 was higher than the individual prediction,and the differences were statistically significant(Z=4.502,4.017,all P<0.05).Conclusion The serum levels of IL-32β and lncRNA-TCL6 are increased in patients with sPE,which are related to the adverse pregnancy outcome.Combined IL-32β and lncRNA-TCL6 have a high value in predicting the pregnancy outcome of patients with sPE.

Objective To analyze the correlation of serum levels of GAT A-binding protein 3(GATA-3),T cell immunoglobulin domain and mucin domain protein-3(Tim-3)and retinoid-related orphan nuclear receptor gamma t(RORγt)with cardiac function and myocardial injury in-dicators in patients with coronary heart disease(CHD)and angina pectoris.Methods A retrospec-tive analysis was performed on 120 CHD patients with angina pectoris(observation group)in our hospital from December 2020 to December 2023.They were divided into stable angina subgroup(53 cases)and unstable angina subgroup(67 cases)according to the results of electrocardiography and coronary angiography.Another 120 healthy individuals taking physical examination in our hospital during the same period were subjected and served as the control group.ELISA was used to detect the serum levels of GATA-3,Tim-3 and RORγt.Left ventricular ejection fraction(LVEF),left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD),and left ventricular short-axis shortening rate(FS)were measured by echocardio-graphy.Creatine kinase(CK)and creatine kinase isoenzyme(CK-MB)were detected by biochemi-cal analyzer,and troponin Ⅰ(cTnⅠ)and heart fatty acid binding protein(H-FABP)were detected by electrochemiluminescence immunoassay.Pearson analysis was applied to analyze the correlation the correlation of serum levels of GATA-3,Tim-3 and RORγt with cardiac function and myocar-dial injury indicators.ROC curve was plotted to analyze the predictive value of the above three in-dicators for unstable angina.Results The serum levels of Tim-3 and ROR y t were significantly higher,and that of GATA-3 was obviously lower in the observation group than the control group(397.14±78.42 ng/L vs 246.45±56.81 ng/L,P＜0.01;93.18±19.94 ng/L vs 35.72±7.79 ng/L,P＜0.01;38.21±9.45 ng/L vs 71.62±14.98 ng/L,P＜0.01).The LVEDD and LVESD values were notably higher,while the LVEF and LVFS values were remarkably lower in the observation group than the control group(P＜0.01).The observation group has statistically higher cTnⅠ,CK,CK-MB,and H-FABP levels than the control group(P＜0.01).In the CHD patients with angina pectoris,serum GATA-3 level was positively correlated with LVEF and LVFS,and negatively with LVEDD,LVESD,cTnⅠ,CK,CK-MB,and H-FABP;Tim-3 and RORγt were negatively corre-lated with LVEF and LVFS,and positively with LVEDD,LVESD,cTnⅠ,CK,CK-MB,and H-FABP(P＜0.01).The AUC value of GATA-3,Tim-3,and RORγt alone and in combination in predicting unstable angina was 0.859,0.827,0.780,and 0.921,respectively,with that of combina-tion significantly higher than that of above indicators alone(Z=1.993,3.021,4.532,P=0.036,0.007,0.001).Conclusion As the disease progresses in CHD patients with angina pectoris,serum levels of Tim-3 and RORγt are increasing,while that of GATA-3 is decreasing,and all of them are closely associated with heart function and myocardial injury indicators.

Objective To analyze the correlation of serum levels of GAT A-binding protein 3(GATA-3),T cell immunoglobulin domain and mucin domain protein-3(Tim-3)and retinoid-related orphan nuclear receptor gamma t(RORγt)with cardiac function and myocardial injury in-dicators in patients with coronary heart disease(CHD)and angina pectoris.Methods A retrospec-tive analysis was performed on 120 CHD patients with angina pectoris(observation group)in our hospital from December 2020 to December 2023.They were divided into stable angina subgroup(53 cases)and unstable angina subgroup(67 cases)according to the results of electrocardiography and coronary angiography.Another 120 healthy individuals taking physical examination in our hospital during the same period were subjected and served as the control group.ELISA was used to detect the serum levels of GATA-3,Tim-3 and RORγt.Left ventricular ejection fraction(LVEF),left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD),and left ventricular short-axis shortening rate(FS)were measured by echocardio-graphy.Creatine kinase(CK)and creatine kinase isoenzyme(CK-MB)were detected by biochemi-cal analyzer,and troponin Ⅰ(cTnⅠ)and heart fatty acid binding protein(H-FABP)were detected by electrochemiluminescence immunoassay.Pearson analysis was applied to analyze the correlation the correlation of serum levels of GATA-3,Tim-3 and RORγt with cardiac function and myocar-dial injury indicators.ROC curve was plotted to analyze the predictive value of the above three in-dicators for unstable angina.Results The serum levels of Tim-3 and ROR y t were significantly higher,and that of GATA-3 was obviously lower in the observation group than the control group(397.14±78.42 ng/L vs 246.45±56.81 ng/L,P＜0.01;93.18±19.94 ng/L vs 35.72±7.79 ng/L,P＜0.01;38.21±9.45 ng/L vs 71.62±14.98 ng/L,P＜0.01).The LVEDD and LVESD values were notably higher,while the LVEF and LVFS values were remarkably lower in the observation group than the control group(P＜0.01).The observation group has statistically higher cTnⅠ,CK,CK-MB,and H-FABP levels than the control group(P＜0.01).In the CHD patients with angina pectoris,serum GATA-3 level was positively correlated with LVEF and LVFS,and negatively with LVEDD,LVESD,cTnⅠ,CK,CK-MB,and H-FABP;Tim-3 and RORγt were negatively corre-lated with LVEF and LVFS,and positively with LVEDD,LVESD,cTnⅠ,CK,CK-MB,and H-FABP(P＜0.01).The AUC value of GATA-3,Tim-3,and RORγt alone and in combination in predicting unstable angina was 0.859,0.827,0.780,and 0.921,respectively,with that of combina-tion significantly higher than that of above indicators alone(Z=1.993,3.021,4.532,P=0.036,0.007,0.001).Conclusion As the disease progresses in CHD patients with angina pectoris,serum levels of Tim-3 and RORγt are increasing,while that of GATA-3 is decreasing,and all of them are closely associated with heart function and myocardial injury indicators.

AIM:Bone morphogenetic protein 7(BMP7)reduces the expression of Yes-related protein 1(YAP1)by down-regulating Ajuba level and decreasing extracellular matrix(ECM)deposition.This study aimed to inves-tigate the influence of these factors on modifying the degree of renal fibrosis in rats with diabetic nephropathy.METH-ODS:Eighteen Sprague-Dawley(SD)rats were randomly divided into three groups:the normal control(NC)group,the diabetes mellitus(DM)group,and the DM group treated with BMP7 overexpressing adeno-associated virus(DM+rAAV-BMP7).Each group consisted of six rats.Diabetic kidney disease(DKD)was established in the DM and DM+rAAV-BMP7 groups by injecting 55 mg/kg streptozotocin(STZ)via the tail vein.NRK-52E cells were divided into three groups:the normal glucose(NG)group,the high glucose(HG)group,and the high glucose group treated with recombinant hu-man BMP7(HG+rhBMP7)group.Pathological changes in renal tissues were observed using hematoxylin and eosin(HE)and Sirius red staining.Immunohistochemical staining was performed to examine the expression sites of Ajuba and YAP1 in the renal cortex.Western blot analysis was conducted to determine the expression levels of BMP7,Ajuba,YAP1,colla-gen type Ⅲ(Col-Ⅲ),and fibronectin(FN)in the rat renal cortex and NRK-52E cells.RT-qPCR was used to measure the mRNA levels of Ajuba and YAP1 in the rat renal cortex.RESULTS:Biochemical indices revealed significantly ele-vated levels of blood glucose,serum creatinine,triglycerides,total cholesterol,and 24-hour urinary protein in the DM group compared to the NC group(P<0.05).In the DM+rAAV-BMP7 group,the levels of serum creatinine,24-hour uri-nary protein,triglycerides,and total cholesterol were lower than those in the DM group(P<0.05).Pathological staining demonstrated that the renal interstitium of the DM group exhibited inflammatory cell infiltration,fibrous tissue,collagen fi-ber deposition,disordered renal tubule arrangement,atrophy,and vacuolar degeneration,which were ameliorated in the DM+rAAV-BMP7 group.Immunohistochemistry revealed that Ajuba and YAP1 were mainly expressed in the cytoplasm and nucleus,with high expression in the cytoplasm of the DM group,which was significantly decreased in the DM+rAAV-BMP7 group.Western blot results indicated that the protein levels of FN,Col-Ⅲ,Ajuba,and YAP1 were up-regulated in the DM and the HG groups(P<0.05),but significantly down-regulated in the DM+rAAV-BMP7 group(P<0.05).RT-qP-CR results demonstrated that the mRNA levels of Ajuba and YAP1 were higher in the DM group and significantly lower in the DM+rAAV-BMP7 group(P<0.05).CONCLUSION:The overexpression of BMP7 can ameliorate renal fibrosis in rats with DKD.This effect is likely mediated by the down-regulation of Ajuba,reduction of YAP1 expression,and subse-quent inhibition of ECM deposition.