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”.

This study employed the "disease-medicine-dose" pattern to mine the medication rules of traditional Chinese medicine(TCM) prescriptions containing Astragali Radix in ancient Chinese medical books, aiming to provide a scientific basis for the clinical application of Astragali Radix and the development of new medicines. The TCM prescriptions containing Astragali Radix were retrieved from databases such as Chinese Medical Dictionary and imported into Excel 2020 to construct the prescription library. Statical analysis were performed for the prescriptions regarding the indications, syndromes, medicine use frequency, herb effects, nature and taste, meridian tropism, dosage forms, and dose. SPSS statistics 26.0 and IBM SPSS Modeler 18.0 were used for association rules analysis and cluster analysis. A total of 2 297 prescriptions containing Astragali Radix were collected, involving 233 indications, among which sore and ulcer, consumptive disease, sweating disorder, and apoplexy had high frequency(>25), and their syndromes were mainly Qi and blood deficiency, Qi and blood deficiency, Yin and Yang deficiency, and Qi deficiency and collateral obstruction, respectively. In the prescriptions, 98 medicines were used with the frequency >25 and they mainly included Qi-tonifying medicines and blood-tonifying medicines. Glycyrrhizae Radix et Rhizoma, Angelicae Sinensis Radix, Ginseng Radix et Rhizoma, Atractylodis Macrocephalae Rhizoma, and Citri Reticulatae Pericarpium were frequently used. The medicines with high frequency mainly have warm or cold nature, and sweet, pungent, or bitter taste, with tropism to spleen, lung, heart, liver, and kidney meridians. In the treatment of sore and ulcer, Astragali Radix was mainly used with the dose of 3.73 g and combined with Glycyrrhizae Radix et Rhizoma to promote granulation and heal up sores. In the treatment of consumptive disease, Astragali Radix was mainly used with the dose of 37.30 g and combined with Ginseng Radix et Rhizoma to tonify deficiency and replenish Qi. In the treatment of sweating disorder, Astragali Radix was mainly used with the dose of 3.73 g and combined with Glycyrrhizae Radix et Rhizoma to consolidate exterior and stop sweating. In the treatment of apoplexy, Astragali Radix was mainly used with the dose of 7.46 g and combined with Glycyrrhizae Radix et Rhizoma to dispell wind and stop convulsions. Astragali Radix can be used in the treatment of multiple system diseases, with the effects of tonifying Qi and ascending Yang, consolidating exterior and stopping sweating, and expressing toxin and promoting granulation. According to the manifestations of different diseases, when combined with other medicines, Astragali Radix was endowed with the effects of promoting granulation and healing up sores, tonifying deficiency and Qi, consolidating exterior and stopping sweating, and dispelling wind and replenishing Qi. The findings provide a theoretical reference and a scientific basis for the clinical application of Astragali Radix and the development of new medicines.
Drugs, Chinese Herbal/history* ; Humans ; Medicine, Chinese Traditional/history* ; History, Ancient ; Astragalus Plant/chemistry* ; China ; Astragalus propinquus

Objective:To investigate the nutritional risk status of children in PICU and analyze its influencing factors.Methods:From July 2021 to February 2023,all children aged 1 to 18 years admitted to PICU of Beijing Children's Hospital were investigated by using the pediatric Yorkhill Malnutrition Scoring tool(PYMS)and the clinical data questionnaire.Results:A total of 492 children in PICU were enrolled.The first nutritional risk screening results showed that there were 32 cases of no/low nutritional risk(6.5%),76 cases of medium risk(15.4%),and 384 cases of high risk(78.1%).The incidence of medium/high nutritional risk was as high as 93.5%.The PYMS score of nutritional risk in PICU was(2.61±1.42).The results of multiple linear regression analysis showed that weight,fever time before admission,white blood cells,body mass index,primary diagnosis,father's education,and diet before illness were the main influencing factors of nutritional risk of children in PICU(P<0.05).Conclusion:Children in PICU are in a state of high nutritional risk.It is suggested that children in PICU should carry out nutritional screening in a standardized manner,identify children with high nutritional risk and its influencing factors early.To actively conduct nutritional assessment and nutritional intervention could improve the clinical outcome of children in PICU.

Objective To explore the effect of composite electromagnetic stimulation combined with chin tuck against resistance(CTAR)on post-stroke dysphagia.Methods From January,2021 to December,2023,156 post-stroke patients with pharyngeal dysphagia in the First Peo-ple's Hospital of Yibin were randomly divided into CTAR group(n=51),neuromuscular electrical stimulation(NMES)+repetitive transcranial magnetic stimulation(rTMS)group(n=52)and combination group(n=53).On the basis of routine swallowing function training,they received CTAR,NMES+rTMS and NMES+rTMS+CTAR,respectively,for four weeks.They were assessed with Functional Oral Intake Scale(FOIS),Standardized Swal-lowing Assessment(SSA),Swallowing-Quality of Life(SWAL-QOL),Rosenbek Penetration Aspiration Scale(PAS),video fluoroscopic swallowing study(VFSS)(including the score of VFSS,pharyngeal transit time,and upward and forward displacement of hyoid bone),average surface electromyography(AMEG)of submental and hyoid muscles,before and after treatment.Results The inter-group effect,intra-group effect and interaction effect were significant in SSA,SWAL-QOL and AMEG of the two muscles(F>6.611,P<0.001).The inter-group effect and interaction effect were significant in FOIS,VFSS,pharyngeal transit time,upward displacement of hyoid bone and forward displacement of hyoid bone(F>3.451,P<0.05).Pairwise comparison results showed that pharyngeal transit time was shorter in the combination group than in NMES+rTMS group(P=0.048),forward displacement of hyoid bone was more in the combina-tion group than in CTAR group(P=0.002),and AMEG was higher in the combination group than in CTAR group and NMES+rTMS group(P<0.001).There was significant difference in the score of PAS among three groups(Hc=8.282,P=0.016),and it was the best in the combination group.Conclusion The combination of NMES+rTMS and CTAR is superior to single electromagnetic stimulation or CTAR in the treatment of post-stroke sysphagia.

Objective To investigate the clinical efficacy and safety of facilitated percutaneous coronary intervention(PCI)with half-dose recombinant staphylokinase(r-SAK)in patients with ST-segment elevation myocardial infarction(STEMI)who are expected to undergo PCI within 120 minutes.Methods From October 2021 to August 2022,a total of 200 STEMI patients in eight centers were included and randomly assigned in a 1﹕1 ratio to either r-SAK group or control group.Patients received loading doses of aspirin and ticagrelor and intravenous heparin and were randomized to receive an intravenous bolus of either 5 mg r-SAK or normal saline prior to PCI.The outcomes were set as ST-segment resolution(STR)at 60-90 minutes after PCI,the proportion and transition of pathological Q waves on the 5th day after PCI,and the proportion of high-sensitivity cardiac troponin T(hs-cTnT)peaking within 12 hours of onset.The safety outcome was major bleeding events defined as Bleeding Academic Research Consortium(BARC)≥type 3 bleeding during hospitalization.Results Compared with the control group,the r-SAK group had a higher proportion of STR≥70%within 60-90 minutes after PCI(58.3%vs.40.3%,P=0.009);a lower proportion of pathological Q waves(59.1%vs.74.1%,P=0.040);a lower rate of Q wave progression(14.8%vs.43.2%,P＜0.001);a higher rate of Q wave disappearance(12.5%vs.3.7%,P=0.027);and a higher proportion of hs-cTnT peaking within 12 hours of symptom onset[31/40(77.5%)vs.17/33(51.5%),P=0.027].Regarding the safety outcome,no significant difference in BARC≥type 3 bleeding was found between the two groups during hospitalization(P＞0.05).Conclusions For STEMI patients who were expected to undergo primary PCI within 120 minutes of symptom onset,the facilitated PCI with half-dose r-SAK significantly increased the proportion of STR≥70%at 60-90 minutes after PCI,reduced the formation of pathological Q waves,and shortened the time to peak hs-cTnT,without increasing the risk of bleeding,which should be an alternative reperfusion strategy worthy of further study.

Objective To determine the mechanism by which 1,25-dihydroxyvitamin D3(VD3)regulates the Th17/Treg cell balance to improve sepsis-induced acute kidney injury(AKI).Methods C57BL/6 mice were assigned to a sham operation group(sham group),a sepsis group(CLP group),a VD3 group,a VD3+knockdown control group(VD3+sh-NC group),and a VD3+NLRP6 knockdown group(VD3+sh-NLRP6 group)(n=10 per group).A mouse model of sepsis-induced AKI was established by cecal ligation and puncture(CLP).Hematoxylin and eosin staining was used to observe pathological damage to renal tissue.ELISA was used to detect the serum creatinine(sCr),kidney injury molecule 1(KIM-1),interleukin(IL)-17,and IL-10 levels.Flow cytometry was used to determine the Th17 cell and Treg cell numbers in peripheral blood.Western blotting was used to detect the expression of vitamin D receptor(VDR)and NLRP6 pro-teins in renal tissue.Results Administration of VD3 alleviated the pathological kidney damage in CLP mice,reduced the sCr,KIM-1,and IL-17 levels and the proportion of Th 17 cells in peripheral blood.It also increased the proportion of Treg cells and the IL-10 level,and upregulated the expression of VDR and NLRP6 proteins.Knockdown of NLRP6 reversed the effect of VD3on the Th17/Treg cell imbal-ance and its renoprotective effects in CLP mice.Conclusion VD3 exerts protective effects in sepsis-induced AKI.Its mechanism likely involves the activation of VDR,leading to subsequent upregulation of NLRP6 expression,ultimately improving the Th17/Treg cell imba-lance.

Objective To explore the effect of composite electromagnetic stimulation combined with chin tuck against resistance(CTAR)on post-stroke dysphagia.Methods From January,2021 to December,2023,156 post-stroke patients with pharyngeal dysphagia in the First Peo-ple's Hospital of Yibin were randomly divided into CTAR group(n=51),neuromuscular electrical stimulation(NMES)+repetitive transcranial magnetic stimulation(rTMS)group(n=52)and combination group(n=53).On the basis of routine swallowing function training,they received CTAR,NMES+rTMS and NMES+rTMS+CTAR,respectively,for four weeks.They were assessed with Functional Oral Intake Scale(FOIS),Standardized Swal-lowing Assessment(SSA),Swallowing-Quality of Life(SWAL-QOL),Rosenbek Penetration Aspiration Scale(PAS),video fluoroscopic swallowing study(VFSS)(including the score of VFSS,pharyngeal transit time,and upward and forward displacement of hyoid bone),average surface electromyography(AMEG)of submental and hyoid muscles,before and after treatment.Results The inter-group effect,intra-group effect and interaction effect were significant in SSA,SWAL-QOL and AMEG of the two muscles(F>6.611,P<0.001).The inter-group effect and interaction effect were significant in FOIS,VFSS,pharyngeal transit time,upward displacement of hyoid bone and forward displacement of hyoid bone(F>3.451,P<0.05).Pairwise comparison results showed that pharyngeal transit time was shorter in the combination group than in NMES+rTMS group(P=0.048),forward displacement of hyoid bone was more in the combina-tion group than in CTAR group(P=0.002),and AMEG was higher in the combination group than in CTAR group and NMES+rTMS group(P<0.001).There was significant difference in the score of PAS among three groups(Hc=8.282,P=0.016),and it was the best in the combination group.Conclusion The combination of NMES+rTMS and CTAR is superior to single electromagnetic stimulation or CTAR in the treatment of post-stroke sysphagia.

Objective To investigate the clinical efficacy and safety of facilitated percutaneous coronary intervention(PCI)with half-dose recombinant staphylokinase(r-SAK)in patients with ST-segment elevation myocardial infarction(STEMI)who are expected to undergo PCI within 120 minutes.Methods From October 2021 to August 2022,a total of 200 STEMI patients in eight centers were included and randomly assigned in a 1﹕1 ratio to either r-SAK group or control group.Patients received loading doses of aspirin and ticagrelor and intravenous heparin and were randomized to receive an intravenous bolus of either 5 mg r-SAK or normal saline prior to PCI.The outcomes were set as ST-segment resolution(STR)at 60-90 minutes after PCI,the proportion and transition of pathological Q waves on the 5th day after PCI,and the proportion of high-sensitivity cardiac troponin T(hs-cTnT)peaking within 12 hours of onset.The safety outcome was major bleeding events defined as Bleeding Academic Research Consortium(BARC)≥type 3 bleeding during hospitalization.Results Compared with the control group,the r-SAK group had a higher proportion of STR≥70%within 60-90 minutes after PCI(58.3%vs.40.3%,P=0.009);a lower proportion of pathological Q waves(59.1%vs.74.1%,P=0.040);a lower rate of Q wave progression(14.8%vs.43.2%,P＜0.001);a higher rate of Q wave disappearance(12.5%vs.3.7%,P=0.027);and a higher proportion of hs-cTnT peaking within 12 hours of symptom onset[31/40(77.5%)vs.17/33(51.5%),P=0.027].Regarding the safety outcome,no significant difference in BARC≥type 3 bleeding was found between the two groups during hospitalization(P＞0.05).Conclusions For STEMI patients who were expected to undergo primary PCI within 120 minutes of symptom onset,the facilitated PCI with half-dose r-SAK significantly increased the proportion of STR≥70%at 60-90 minutes after PCI,reduced the formation of pathological Q waves,and shortened the time to peak hs-cTnT,without increasing the risk of bleeding,which should be an alternative reperfusion strategy worthy of further study.

Objective To determine the mechanism by which 1,25-dihydroxyvitamin D3(VD3)regulates the Th17/Treg cell balance to improve sepsis-induced acute kidney injury(AKI).Methods C57BL/6 mice were assigned to a sham operation group(sham group),a sepsis group(CLP group),a VD3 group,a VD3+knockdown control group(VD3+sh-NC group),and a VD3+NLRP6 knockdown group(VD3+sh-NLRP6 group)(n=10 per group).A mouse model of sepsis-induced AKI was established by cecal ligation and puncture(CLP).Hematoxylin and eosin staining was used to observe pathological damage to renal tissue.ELISA was used to detect the serum creatinine(sCr),kidney injury molecule 1(KIM-1),interleukin(IL)-17,and IL-10 levels.Flow cytometry was used to determine the Th17 cell and Treg cell numbers in peripheral blood.Western blotting was used to detect the expression of vitamin D receptor(VDR)and NLRP6 pro-teins in renal tissue.Results Administration of VD3 alleviated the pathological kidney damage in CLP mice,reduced the sCr,KIM-1,and IL-17 levels and the proportion of Th 17 cells in peripheral blood.It also increased the proportion of Treg cells and the IL-10 level,and upregulated the expression of VDR and NLRP6 proteins.Knockdown of NLRP6 reversed the effect of VD3on the Th17/Treg cell imbal-ance and its renoprotective effects in CLP mice.Conclusion VD3 exerts protective effects in sepsis-induced AKI.Its mechanism likely involves the activation of VDR,leading to subsequent upregulation of NLRP6 expression,ultimately improving the Th17/Treg cell imba-lance.