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 evaluate the diagnostic performance of the minimum-cost-path-based CT angiography-derived fractional flow reserve(MCP-FFR)and the deep learning-driven CT angiography-derived fractional flow reserve(DeepCL-FFR),and to particularly explore the potential value of the DeepCL algorithm in improving diagnostic accuracy within the"gray zone."Methods A retrospective analysis was conducted on 151 coronary vessels from 109 patients with coronary artery disease,who were hospitalized at the General Hospital of the People's Liberation Army between January 2020 and June 2021.Pearson correlation and Bland-Altman plots were employed to assess the correlation and agreement of the two CT-FFR methods with invasive FFR.A CT-FFR range of 0.70-0.80 was defined as the diagnostic"gray zone."The accuracy,sensitivity,specificity,positive predictive value,and negative predictive value for detecting hemodynamic abnormalities were calculated and analyzed.The DeLong test was used to compare the areas under the receiver operating characteristic curves(AUC)between the two CT-FFR calculation methods.Results Both CT-FFR methods exhibited a positive correlation with invasive FFR(MCP-FFR:r=0.75,P＜0.001;DeepCL-FFR:r=0.86,P＜0.001)and showed good agreement(MCP-FFR:mean difference=0.010,P=0.351;DeepCL-FFR:mean difference=-0.003,P=0.772).Both DeepCL-FFR(AUC 0.97,95％CI 0.94-0.99)and MCP-FFR(AUC 0.92,95％CI 0.88-0.97)demonstrated favorable diagnostic performance for detecting hemodynamic abnormalities(P=0.122).In the"gray zone"for hemodynamic abnormality,the diagnostic accuracy of MCP-FFR was 68.8％,whereas DeepCL-FFR increased it to 89.7％.DeepCL-FFR also exhibited superior diagnostic performance(AUC 0.89,95％CI 0.73-0.99)within the"gray zone,"which was significantly higher than that of MCP-FFR(AUC 0.71,95％CI 0.54-0.87)(P＜0.001).Conclusions The deep learning-driven coronary centerline extraction algorithm,DeepCL,demonstrates superior diagnostic performance in CT-FFR for detecting hemodynamic abnormalities,particularly by significantly improving diagnostic accuracy in the"gray zone."

Objective To investigate the therapeutic efficacy of the combination of upper limb rehabilitation robots(ULRR)and ultrasound-guided extracorporeal shock wave(ESW)in the treatment of mild to moderate rotator cuff injuries.Methods A total of 90 patients with mild to moderate rotator cuff injuries,who were admitted to the Second Affiliated Hospital of Anhui Medical University between January 2023 and March 2024,were selected and randomly assigned to three groups:Group A(basic rehabilitation),Group B(basic rehabilitation+ESW),and Group C(basic rehabilitation+ESW+ULRR),with 30 patients in each group.Before and after treatment,shoulder pain was assessed using the visual analogue scale(VAS).Proprioception was evaluated based on shoulder joint position reproduction error.The integral electromyography(iEMG)and root mean square(RMS)values of the median tract,supraspinatus,infraspinatus,and deltoid muscles on the affected side were measured using surface electromyography(sEMG).The active range of motion(AROM)for external rotation,abduction,and forward flexion of the affected shoulder was measured using a standard goniometer.Overall shoulder function was evaluated using the Constant-Murley Shoulder Score(CMS).The clinical efficacy rate and patient satisfaction levels were compared among the three groups.Results After treatment,the VAS score and shoulder position reproduction deviation angle in all three groups significantly decreased.Additionally,the iEMG and RMS values of the middle deltoid,supraspinatus,and infraspinatus muscles;the AROM of shoulder external rotation,abduction,and forward flexion;and the CMS scores(including pain,activities of daily living,active range of motion,and muscle strength)all showed significant improvement.Furthermore,the degree of improvement in all evaluated parameters in Group C was significantly greater than that observed in Groups A and B(P<0.05).Moreover,Group C demon-strated a significantly higher clinical response rate and greater patient satisfaction compared to Groups A and B(P<0.05).Conclusion For patients with mild to moderate rotator cuff injuries,the integration of an upper limb rehabilitation robot with extracorporeal shock wave therapy demonstrates superior therapeutic outcomes,effectively alleviating pain,enhancing motor function and proprioception,and increasing overall patient satisfaction.

Objective To investigate the correlation between peripheral blood CD14+CD16+monocytes and IgG N-glycosyl levels and disease activity in patients with systemic lupus erythematosus(SLE).Methods A total of 109 SLE patients admitted to Xingtai Cental Hospital from August 2021 to November 2024 were retrospectively selected as the study objects.According to SLE disease activity index(SLE-DAI),the patients were divided into active group(n=52)and stable group(n=57).In addition,56 patients who underwent physical examination during the same period were selected as the control group.Clinical data of patients were collected,CD14+CD16+mononuclear cells were detected by flow cytometry(FCM),and IgG N-glycosyl levels were detected by hydrophilic interaction chromatography-mass spectrometry.Logistic regression analysis was performed to analyze the influencing factors of SLE-DAI,and multicollinearity test[variance inflation factor(VIF)]was performed for independent variables.The prediction model of disease activity was constructed.The effectiveness of the predictive model was evaluated by describing the recviver operator characteristic(ROC)curve and calculating the area under the curve(AUC)value.Hosmer-Lemeshow goodness-of-fit test predicted the calibration degree of the model.Results The levels of WBC,Hb,PLT,ALB,complement C3 and complement C4 in control group were higher than those in SLE group(t=8.917～22.171),and the levels of CRP were lower than those in SLE group(t=-17.359),with differences were statistical significance(all P<0.05).The CRP level and the proportion of CD14+CD16+mononuclear cells in the active group were higher than those in the stable group,and the differences were statistically significant(t=5.449,11.112,all P<0.05).The IgG glycosylation characteristics of galactosylation,sialylation and N-acetylglucosamine modification were lower than those in the stable group,and the differences were statistical significance(Z=-2.432～-0.158,all P<0.05).Spearman correlation analysis showed that the proportion of CD14+CD16+monocytes was significantly negatively correlated with IgG galactosylation,sialylation level and bisection N-acetylglucosamine modification(r=-0.656,-0.531,-0.608,all P<0.01).CD14+CD16+monocyte ratio was positively correlated with SLE-DIA(r=0.581,all P<0.01).IgG galactosylation,sialylation levels and bisection N-acetylglucosamine modification were negatively correlated with SLE-DIA(r=-0.645,-0.609,-0.503,all P<0.01).Logistic regression analysis showed that CRP>8.21mg/L,CD14+CD16+≥16.17%,sialylation<22.05%and isotropic N-acetylglucosamine modification<16.53%were independent risk factors for disease activity in SLE patients(Wald χ2=4.471～12.811,all P<0.05).The VIF values of the above independent variables were all less than 10.By establishing the Logistic regression prediction model and drawing the ROC curve,the AUC value for diagnosing SLE disease activity was 0.821(95%CI:0.733～0.905),the sensitivity,specificity and the Yodon index were 85.37%,75.67%,0.677,respectively.and the P values of Hosmer-Lemeshow goodness-of-fit test models were 0.568,respectively.Conclusion The proportion of CD14+CD16+monocytes in peripheral blood of SLE patients increase significantly,and the level of IgG glycosylation characteristics decrease,both of which are correlated with SLE-DIA.The predictive model constructed based on the two had a good ability to distinguish SLE-DIA from inactive state,with high sensitivity and moderate specificity conducive to early clinical recognition,and the model fitting effect is good.SLE-DIA can be evaluated more accurately.

Objective To evaluate the diagnostic performance of the minimum-cost-path-based CT angiography-derived fractional flow reserve(MCP-FFR)and the deep learning-driven CT angiography-derived fractional flow reserve(DeepCL-FFR),and to particularly explore the potential value of the DeepCL algorithm in improving diagnostic accuracy within the"gray zone."Methods A retrospective analysis was conducted on 151 coronary vessels from 109 patients with coronary artery disease,who were hospitalized at the General Hospital of the People's Liberation Army between January 2020 and June 2021.Pearson correlation and Bland-Altman plots were employed to assess the correlation and agreement of the two CT-FFR methods with invasive FFR.A CT-FFR range of 0.70-0.80 was defined as the diagnostic"gray zone."The accuracy,sensitivity,specificity,positive predictive value,and negative predictive value for detecting hemodynamic abnormalities were calculated and analyzed.The DeLong test was used to compare the areas under the receiver operating characteristic curves(AUC)between the two CT-FFR calculation methods.Results Both CT-FFR methods exhibited a positive correlation with invasive FFR(MCP-FFR:r=0.75,P＜0.001;DeepCL-FFR:r=0.86,P＜0.001)and showed good agreement(MCP-FFR:mean difference=0.010,P=0.351;DeepCL-FFR:mean difference=-0.003,P=0.772).Both DeepCL-FFR(AUC 0.97,95％CI 0.94-0.99)and MCP-FFR(AUC 0.92,95％CI 0.88-0.97)demonstrated favorable diagnostic performance for detecting hemodynamic abnormalities(P=0.122).In the"gray zone"for hemodynamic abnormality,the diagnostic accuracy of MCP-FFR was 68.8％,whereas DeepCL-FFR increased it to 89.7％.DeepCL-FFR also exhibited superior diagnostic performance(AUC 0.89,95％CI 0.73-0.99)within the"gray zone,"which was significantly higher than that of MCP-FFR(AUC 0.71,95％CI 0.54-0.87)(P＜0.001).Conclusions The deep learning-driven coronary centerline extraction algorithm,DeepCL,demonstrates superior diagnostic performance in CT-FFR for detecting hemodynamic abnormalities,particularly by significantly improving diagnostic accuracy in the"gray zone."

Aim To investigate the protective effects and potential mechanisms of Radix Angelica sinensis and Radix Hedysari ultrafiltrate(RAS-RH)on X-ray-induced cellular damage in Raw264.7 macrophages.Methods An integrated approach combining network pharmacology,molecular docking,and bioinformatics a-nalysis was employed to predict therapeutic targets and signaling pathways of RAS-RH in coronary heart dis-ease(CHD).Subsequent in vitro validation was per-formed using an X-ray(6 Gy)-induced macrophage in-jury model with four experimental groups:control,radi-ation-only model,and three RAS-RH-treated groups at varying concentrations.Cell viability was assessed by CCK-8 assay,apoptosis by flow cytometry with Annexin V-FITC/PI staining,mitochondrial membrane potential by JC-1 fluorescence,and inflammatory cytokine levels(IL-1 β,IL-6,IL-18,TNF-α)by ELISA.Molecular mechanisms were investigated through Western blot and qRT-PCR analyses of TLR4/NLRP3/Caspase-1 sig-naling pathway components and Bcl-2 family proteins.Results Network pharmacology revealed RAS-RH's multi-target action on apoptosis and inflammation-relat-ed pathways,particularly NF-κB and Bcl-2 signaling.Molecular docking identified strong binding affinities between RAS-RH components and TLR4/NLRP3 pro-teins.In vitro studies demonstrated that RAS-RH treat-ment significantly improved cell viability(P＜0.01),reduced apoptosis(P＜0.01),restored mitochondrial membrane potential(P＜0.05),and attenuated radia-tion-induced ultrastructural damage including mem-brane disruption and cytoplasmic vacuolization.ELISA showed marked suppression of pro-inflammatory cyto-kines(P＜0.01).Transmission electron microscopy(TEM)analysis revealed that RSA-RH ameliorated pyroptosis-associated ultrastructural alterations,inclu-ding plasma membrane disruption and cytoplasmic vac-uolization.Protein and gene expression analyses con-firmed downregulation of TLR4/NLRP3/Caspase-1 pathway and modulation of Bcl-2/Bax ratio.Conclu-sion RAS-RH exerts radioprotective effects through dual regulation of pyroptosis and apoptosis pathways,suggesting its potential as an adjuvant therapy for radia-tion-induced cardiovascular complications in CHD pa-tients.

Aim To investigate the protective effects and potential mechanisms of Radix Angelica sinensis and Radix Hedysari ultrafiltrate(RAS-RH)on X-ray-induced cellular damage in Raw264.7 macrophages.Methods An integrated approach combining network pharmacology,molecular docking,and bioinformatics a-nalysis was employed to predict therapeutic targets and signaling pathways of RAS-RH in coronary heart dis-ease(CHD).Subsequent in vitro validation was per-formed using an X-ray(6 Gy)-induced macrophage in-jury model with four experimental groups:control,radi-ation-only model,and three RAS-RH-treated groups at varying concentrations.Cell viability was assessed by CCK-8 assay,apoptosis by flow cytometry with Annexin V-FITC/PI staining,mitochondrial membrane potential by JC-1 fluorescence,and inflammatory cytokine levels(IL-1 β,IL-6,IL-18,TNF-α)by ELISA.Molecular mechanisms were investigated through Western blot and qRT-PCR analyses of TLR4/NLRP3/Caspase-1 sig-naling pathway components and Bcl-2 family proteins.Results Network pharmacology revealed RAS-RH's multi-target action on apoptosis and inflammation-relat-ed pathways,particularly NF-κB and Bcl-2 signaling.Molecular docking identified strong binding affinities between RAS-RH components and TLR4/NLRP3 pro-teins.In vitro studies demonstrated that RAS-RH treat-ment significantly improved cell viability(P＜0.01),reduced apoptosis(P＜0.01),restored mitochondrial membrane potential(P＜0.05),and attenuated radia-tion-induced ultrastructural damage including mem-brane disruption and cytoplasmic vacuolization.ELISA showed marked suppression of pro-inflammatory cyto-kines(P＜0.01).Transmission electron microscopy(TEM)analysis revealed that RSA-RH ameliorated pyroptosis-associated ultrastructural alterations,inclu-ding plasma membrane disruption and cytoplasmic vac-uolization.Protein and gene expression analyses con-firmed downregulation of TLR4/NLRP3/Caspase-1 pathway and modulation of Bcl-2/Bax ratio.Conclu-sion RAS-RH exerts radioprotective effects through dual regulation of pyroptosis and apoptosis pathways,suggesting its potential as an adjuvant therapy for radia-tion-induced cardiovascular complications in CHD pa-tients.

Objective To investigate the therapeutic efficacy of the combination of upper limb rehabilitation robots(ULRR)and ultrasound-guided extracorporeal shock wave(ESW)in the treatment of mild to moderate rotator cuff injuries.Methods A total of 90 patients with mild to moderate rotator cuff injuries,who were admitted to the Second Affiliated Hospital of Anhui Medical University between January 2023 and March 2024,were selected and randomly assigned to three groups:Group A(basic rehabilitation),Group B(basic rehabilitation+ESW),and Group C(basic rehabilitation+ESW+ULRR),with 30 patients in each group.Before and after treatment,shoulder pain was assessed using the visual analogue scale(VAS).Proprioception was evaluated based on shoulder joint position reproduction error.The integral electromyography(iEMG)and root mean square(RMS)values of the median tract,supraspinatus,infraspinatus,and deltoid muscles on the affected side were measured using surface electromyography(sEMG).The active range of motion(AROM)for external rotation,abduction,and forward flexion of the affected shoulder was measured using a standard goniometer.Overall shoulder function was evaluated using the Constant-Murley Shoulder Score(CMS).The clinical efficacy rate and patient satisfaction levels were compared among the three groups.Results After treatment,the VAS score and shoulder position reproduction deviation angle in all three groups significantly decreased.Additionally,the iEMG and RMS values of the middle deltoid,supraspinatus,and infraspinatus muscles;the AROM of shoulder external rotation,abduction,and forward flexion;and the CMS scores(including pain,activities of daily living,active range of motion,and muscle strength)all showed significant improvement.Furthermore,the degree of improvement in all evaluated parameters in Group C was significantly greater than that observed in Groups A and B(P<0.05).Moreover,Group C demon-strated a significantly higher clinical response rate and greater patient satisfaction compared to Groups A and B(P<0.05).Conclusion For patients with mild to moderate rotator cuff injuries,the integration of an upper limb rehabilitation robot with extracorporeal shock wave therapy demonstrates superior therapeutic outcomes,effectively alleviating pain,enhancing motor function and proprioception,and increasing overall patient satisfaction.

Objective To investigate the correlation between peripheral blood CD14+CD16+monocytes and IgG N-glycosyl levels and disease activity in patients with systemic lupus erythematosus(SLE).Methods A total of 109 SLE patients admitted to Xingtai Cental Hospital from August 2021 to November 2024 were retrospectively selected as the study objects.According to SLE disease activity index(SLE-DAI),the patients were divided into active group(n=52)and stable group(n=57).In addition,56 patients who underwent physical examination during the same period were selected as the control group.Clinical data of patients were collected,CD14+CD16+mononuclear cells were detected by flow cytometry(FCM),and IgG N-glycosyl levels were detected by hydrophilic interaction chromatography-mass spectrometry.Logistic regression analysis was performed to analyze the influencing factors of SLE-DAI,and multicollinearity test[variance inflation factor(VIF)]was performed for independent variables.The prediction model of disease activity was constructed.The effectiveness of the predictive model was evaluated by describing the recviver operator characteristic(ROC)curve and calculating the area under the curve(AUC)value.Hosmer-Lemeshow goodness-of-fit test predicted the calibration degree of the model.Results The levels of WBC,Hb,PLT,ALB,complement C3 and complement C4 in control group were higher than those in SLE group(t=8.917～22.171),and the levels of CRP were lower than those in SLE group(t=-17.359),with differences were statistical significance(all P<0.05).The CRP level and the proportion of CD14+CD16+mononuclear cells in the active group were higher than those in the stable group,and the differences were statistically significant(t=5.449,11.112,all P<0.05).The IgG glycosylation characteristics of galactosylation,sialylation and N-acetylglucosamine modification were lower than those in the stable group,and the differences were statistical significance(Z=-2.432～-0.158,all P<0.05).Spearman correlation analysis showed that the proportion of CD14+CD16+monocytes was significantly negatively correlated with IgG galactosylation,sialylation level and bisection N-acetylglucosamine modification(r=-0.656,-0.531,-0.608,all P<0.01).CD14+CD16+monocyte ratio was positively correlated with SLE-DIA(r=0.581,all P<0.01).IgG galactosylation,sialylation levels and bisection N-acetylglucosamine modification were negatively correlated with SLE-DIA(r=-0.645,-0.609,-0.503,all P<0.01).Logistic regression analysis showed that CRP>8.21mg/L,CD14+CD16+≥16.17%,sialylation<22.05%and isotropic N-acetylglucosamine modification<16.53%were independent risk factors for disease activity in SLE patients(Wald χ2=4.471～12.811,all P<0.05).The VIF values of the above independent variables were all less than 10.By establishing the Logistic regression prediction model and drawing the ROC curve,the AUC value for diagnosing SLE disease activity was 0.821(95%CI:0.733～0.905),the sensitivity,specificity and the Yodon index were 85.37%,75.67%,0.677,respectively.and the P values of Hosmer-Lemeshow goodness-of-fit test models were 0.568,respectively.Conclusion The proportion of CD14+CD16+monocytes in peripheral blood of SLE patients increase significantly,and the level of IgG glycosylation characteristics decrease,both of which are correlated with SLE-DIA.The predictive model constructed based on the two had a good ability to distinguish SLE-DIA from inactive state,with high sensitivity and moderate specificity conducive to early clinical recognition,and the model fitting effect is good.SLE-DIA can be evaluated more accurately.