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 formulate Guidelines for the standardized implementation of pharmacist-managed clinics （ 2026 edition ） in response to the challenges faced by such clinics in China， including uneven development， large discrepancies in service specifications， insufficient patient awareness， and limited medical insurance coverage. METHODS Led by the Pharmaceutical Affairs Professional Committee of the Chinese Hospital Association， the Evidence-based Pharmacy Professional Committee of the Chinese Pharmaceutical Association， and the Hospital Pharmacy Professional Committee of the Cross-strait Medical and Health Exchange Association， a total of 19 domestic hospital pharmacy experts were organized. Through a systematic review of national policies and literature research， current practical experience was summarized. Consensus on the contents of the guidelines was reached after in-depth discussions. RESULTS &CONCLUSIONS The guidelines covered five sections： definition and connotation of pharmacist-managed clinics， establishment requirements， implementation and management， post competency， and practical research. Firstly， the definition and connotation included three operational forms of pharmacist-managed clinics （independent mode， physician-pharmacist joint mode， and online pharmacist-managed clinic mode） and classified service modes （specialty-specific， drug-specific， and disease-specific pharmacist-managed clinics）. The establishment requirements were further refined， covering system construction （pharmaceutical service management system， quality control and assessment mechanism）， personnel qualifications （professional credentials， continuing education and professional training， etc）， service recipients， as well as service venues and facilities. Subsequently， the implementation and management of pharmacist-managed clinics were proposed， involving service procedures， intervention measures， documentation and records， patient education and follow-up， humanistic care， as well as risk management and quality control. Finally， post competency encompassed the competency requirements for pharmacists providing services in pharmacist-managed clinics， as well as the suggestions on teaching methods； practical research encouraged the conduct of high-quality pharmaceutical practice in the setting of pharmacist-managed clinics. The guidelines provide valuable guidance for the standardized implementation of pharmacist-managed clinics in China in terms of establishment， management， teaching， and research， fill the guideline gap in this field， and can promote the high-quality development of pharmacist-managed clinics.

Objective To investigate the relationship between the appearance of ligand 1 in inflammatory fibroblasts,primary tumor-ous cells,tumor Impregna-ted immune-free cellulars and clinical pathologic signs in colorectal carcinoma.Methods The group meth-od of immunization was used,and PDGFRa and RGS5 were used to mark iCAF.The levels of PDGFRa,RGS5(mesenchyma,mesenchy-ma prior to tumor infiltration)and PD-L1 in TC and TIIC were detected in 120 CRC patients in our hospital from 2019 to 2021.To ana-lyze the relationship between the appearance of iCAF and PD-L1 in cancer tissues and clinical pathologic signs and overall survival(OS).Results The high rate of PDGFRa and RGS5 was 56.7％(68/120)and 47.5％(57/120)in the mesenchyma of 120 patients with colorectal carcinoma.The high detection rate of PDGFRa and RGS5 was 39.2％(47/120).The high surface level of PDGFRa and RGS5 is related to the emergence of neoplastic buds and the metastasis of luoba nodules in the region.The high surface reach rate of PDG-FRa was 71.7％(86/120)and that of RGS5 was 57.5％(69/120).The high detection rate of PDGFRa and RGS5 was 47.5％(57/120).The high surface level of PDGFRa and RGS5 is related to the emergence of tumor buds,the metastasis of lubaria in the region,and the phase of T.The positive rate of TC PD-L1 was 5.8％(7/120)and that of TIIC PD-L1 was 48.3％(58/120).The surface reach of TC PD-L1 is related to the regional luoba nodal transfer.Kaplan-Meier and Log-rank results were shown.The patient's OS was associated with histological grading,neoplasmic metastasis,neoplasmic budding,TIIC PD-L1,PDGFRa,and RGS5(mesenchyma,mesenchyma prior to neoplasmic infiltration).COX monofactor and polyfactor regression analysis showed:Histologic grading,luba metasta-sis,TIIC PD-L1,RGS5(mesenchyma,mesenchyma prelimbic)and PDGFRa(mesenchyma prelimbic)are independent of OS in pa-tients with CRCCreate a preemptive factor.The surface reach of PDGFRa(mesenchymal)was negatively correlated with that of TC PD-L1(r=-0.213,P=0.020).The surface reach of RGS5(mesenchymal)was negatively correlated with that of TC PD-L1(r=-0.183,P=0.046).Conclusion In CRC,iCAF has a negative correlation with TC PD-L1.Targeting the treatment of iCAF in CRC may benefit it,and the detection of TC PD-L1 and TIIC PD-L1 may provide reference for the therapeutic effect of clinical anti-epidem-ic therapy and the prediction of CRC.

Objective:To evaluate the safety and efficacy of the TiRobot assisted screw placement in patients with lumbar degenerative diseases.Methods:The clinical data of 165 patients with lumbar degenerative diseases from January 2020 to December 2022 in Beijing Friendship Hospital, Capital Medical University were retrospectively analyze, and all patients were treated with posterior lumbar decompression and instrumentation. Among them, 46 patients were used the TiRobot assisted screw placement during surgery (robotic-assisted group), and 119 patients underwent freehand screw placement by C-arm X-ray machine fluoroscopy (freehand group). The operation time, intraoperative blood loss, complication and skelalgia visual analogue scale (VAS), Oswestry disability index (ODI) before and after operation were recorded. The accuracy of screw placement and rate of proximal facet joint violation were compared between two groups.Results:There were no statistical difference in operation time, intraoperative blood loss and incidence of complication between two groups ( P>0.05). In the two groups, the 3 d VAS and ODI after operation were significantly lower than those before operation, robot-assisted group: (3.33 ± 1.40) scores vs. (6.54 ± 2.00) scores and (16.96 ± 8.03) scores vs. (43.09 ± 5.48) scores; freehand group: (3.56 ± 1.29) scores vs. (6.55 ± 1.65) scores and (18.89 ± 6.74) scores vs. (44.91 ± 4.96) scores, and there were statistical differences ( P<0.01); there were no statistical difference in VAS and ODI before operation and 3 d after operation between two groups ( P>0.05). A total of 234 screws were implanted in robot-assisted group, and 590 screws were implanted in freehand group. The accuracy of screw placement in robot-assisted group was significantly higher than that in freehand group: 80.77% (189/234) vs. 74.58% (440/590), the rate of proximal facet joint violation was significantly lower than that in freehand group: 2.56% (6/234) vs. 7.29% (43/590), and there were statistical differences ( χ2 = 3.56 and 6.68, P<0.05). Conclusions:The TiRobot assisted screw placement for lumbar degenerative diseases is safe and effective. Compared to freehand technique, the TiRobot assisted method demonstrates higher screw placement accuracy and a lower rate of proximal facet joint violation.

AKT,also known as Protein Kinase B(PKB),plays a critical role in cell proliferation and metabolism.There are three isoforms of AKT:AKT1,AKT2,and AKT3.The effects of these isoforms on the pluripotency and differentiation of mouse embryonic stem cells(mESCs)remain unclear.This study aims to explore the impact of three AKT isoform-specific knockouts on the self-renewal and differen-tiation of mouse embryonic stem cells.Using CRISPR/Cas9 gene-editing technology,AKT isoform-spe-cific knockout cell lines were established.The phenotypic and molecular changes were analyzed through Western blotting,flow cytometry,qRT-PCR,CCK-8 assays,Alkaline Phosphatase(AP)staining,and RNA-seq.The construction of AKT isoform-specific knockout cell lines was successful.The loss of AKT1 and AKT2 inhibited the proliferation of mESCs.The knockout of any single AKT isoform did not affect the expression of pluripotency genes at both mRNA or protein levels.However,during embryoid body forma-tion,the deletion of any of the three AKT isoforms affected the mRNA expression levels of genes in all three germ layers.Transcriptome analysis showed that compared to wild-type mESCs,995,547,and 429 differentially expressed genes(|log2FC|≧1,P＜0.05)were identified inAKT1,AKT2,and AKT3 isoform-specific knockout cells,respectively.There was some overlap in the differentially expressed genes regulated by these three isoforms.In conclusion,the independent knockout of AKT isoforms does not af-fect the maintenance of pluripotency in mouse embryonic stem cells,but they are crucial for differentia-tion.The three AKT isoforms can collectively regulate gene expression while retaining their own regulato-ry specificity.This study provides a foundation for understanding the unique and overlapping roles of AKT isoforms in stem cell biology,highlighting their importance in maintaining stem cell function and differen-tiation.

Objective:To analyze the learning curve of TiRobot-assisted lumbar pedicle screw fixation (LPSF) by cumulative sum (CUSUM) test method.Methods:The clinical data of 50 patients who underwent TiRobot-assisted LPSF from January 2020 to December 2022 in Beijing Friendship Hospital, Capital Medical University were retrospectively analyzed. CUSUM analysis and learning curve fitting were performed with robot usage time as the main indicator with the time for each step refined (robot registration time, path planning time and guide wire placement time), to select the best learning curve fitting model with the R2 value closest to 1. Using the turning point of the learning curve as the boundary, the learning curve was divided into two stages as learning stage and maturity stage, and then the observation indexes were compared between the two stages. Results:All 50 patients successfully completed the surgery without perioperative complications, with a total of 244 pedicle screws implanted. The total robot usage time and robot registration time showed a gradually decreasing trend with the increase of case number, and the learning curves were successfully fitted and reached their peaks at the seventeenth and thirteenth cases respectively. The entire learning process was divided into learning stage (17 cases) and maturity stage (33 cases) based on the turning point of the learning curve of total robot usage time. The path planning time and guide wire placement time did not show significant changes with the increase in the case number. The total robot usage time, robot registration time and the intraoperative blood loss in the learning stage were significantly higher than those in the maturity stage: (35.35 ± 1.58) min vs. (30.61 ± 0.43) min, (20.83 ± 1.56) min vs. (14.94 ± 0.29) min and 400 (150, 500) ml vs. 200 (110, 300) ml, the guide wire placement time of per screw was significantly lower than that in the maturity stage: 2.00 (1.83, 2.34) min/screw vs. 2.33 (2.13, 2.69) min/screw, and there were statistical differences ( P<0.05 or <0.01). There were no statistical difference in the path planning time, path planning time of per screw, guide wire placement time and the accuracy of screw placement between two stages ( P>0.05). Conclusions:TiRobot-assisted LPSF is a new technology with safety and effectiveness, and it has a relatively short learning curve. To achieve technological maturity, at least 17 surgeries are required with accumulated experience, and the robot registration is the main step of the learning process. After reaching maturity stage, the robot usage time is significantly shortened and intraoperative trauma is significantly reduced while the relatively high screw placement accuracy is ensured.

Objective To investigate the relationship between the appearance of ligand 1 in inflammatory fibroblasts,primary tumor-ous cells,tumor Impregna-ted immune-free cellulars and clinical pathologic signs in colorectal carcinoma.Methods The group meth-od of immunization was used,and PDGFRa and RGS5 were used to mark iCAF.The levels of PDGFRa,RGS5(mesenchyma,mesenchy-ma prior to tumor infiltration)and PD-L1 in TC and TIIC were detected in 120 CRC patients in our hospital from 2019 to 2021.To ana-lyze the relationship between the appearance of iCAF and PD-L1 in cancer tissues and clinical pathologic signs and overall survival(OS).Results The high rate of PDGFRa and RGS5 was 56.7％(68/120)and 47.5％(57/120)in the mesenchyma of 120 patients with colorectal carcinoma.The high detection rate of PDGFRa and RGS5 was 39.2％(47/120).The high surface level of PDGFRa and RGS5 is related to the emergence of neoplastic buds and the metastasis of luoba nodules in the region.The high surface reach rate of PDG-FRa was 71.7％(86/120)and that of RGS5 was 57.5％(69/120).The high detection rate of PDGFRa and RGS5 was 47.5％(57/120).The high surface level of PDGFRa and RGS5 is related to the emergence of tumor buds,the metastasis of lubaria in the region,and the phase of T.The positive rate of TC PD-L1 was 5.8％(7/120)and that of TIIC PD-L1 was 48.3％(58/120).The surface reach of TC PD-L1 is related to the regional luoba nodal transfer.Kaplan-Meier and Log-rank results were shown.The patient's OS was associated with histological grading,neoplasmic metastasis,neoplasmic budding,TIIC PD-L1,PDGFRa,and RGS5(mesenchyma,mesenchyma prior to neoplasmic infiltration).COX monofactor and polyfactor regression analysis showed:Histologic grading,luba metasta-sis,TIIC PD-L1,RGS5(mesenchyma,mesenchyma prelimbic)and PDGFRa(mesenchyma prelimbic)are independent of OS in pa-tients with CRCCreate a preemptive factor.The surface reach of PDGFRa(mesenchymal)was negatively correlated with that of TC PD-L1(r=-0.213,P=0.020).The surface reach of RGS5(mesenchymal)was negatively correlated with that of TC PD-L1(r=-0.183,P=0.046).Conclusion In CRC,iCAF has a negative correlation with TC PD-L1.Targeting the treatment of iCAF in CRC may benefit it,and the detection of TC PD-L1 and TIIC PD-L1 may provide reference for the therapeutic effect of clinical anti-epidem-ic therapy and the prediction of CRC.

Aim To explore the mechanism of action of Guizhi Jia Gegen decoction(GGD)in treating pneu-monia-enteritis induced by H1N1 influenza virus infec-tion in a mouse model,using network pharmacology and molecular docking techniques,followed by in vivo verification.Methods A pneumonia-enteritis mouse model was established,and the intervention effects of GGD on the model mice were evaluated using indica-tors such as body weight,rectal temperature,lung in-dex,colon length,H1N1 M gene expression,relative mRNA expression levels of inflammatory cytokines,and pathological sections of the lung and intestine.The targets of the blood-absorbed components of GGD were identified using the Swiss Target Prediction platform,and the disease targets were retrieved from the Gene-Cards platform.The intersecting targets were analyzed through PPI network analysis using the STRING data-base to identify core targets.GO analysis and KEGG pathway enrichment analysis were performed using the Metascape database.RT-qPCR was employed to vali-date the core targets and pathways.Molecular docking was conducted using AutoDock Tools software to verify the interactions between blood-absorbed components and key targets.Results GGD demonstrated signifi-cant therapeutic effects on the pneumonia-enteritis mouse model.The results of network pharmacology in-dicated that the therapeutic effects of GGD were strong-ly associated with targets such as TNF,ALB,PTGS2,MMP9,EGFR,ESR1,SRC,HSP90AA1,PPARG and MMP2.RT-qPCR results indicated that GGD could intervene in pneumonia-enteritis by regulating the targets TNF,ALB,EGFR and the related targets of the NF-κB pathway.Molecular docking results re-vealed that blood-absorbed components such as puerar-in and liquiritin could stably bind to TNF,ALB and EGFR.Conclusion Components such as puerarin and liquiritin in GGD may exert therapeutic effects on pneumonia-enteritis induced by H1N1 influenza virus infection by acting on targets such as TNF,ALB and EGFR.

Objective:To explore the role of P2Y1Receptor(P2Y1R)-activated astrocytes in delayed encephalopathy after acute carbon monoxide poisoning(DEACMP).Methods:SD rats were randomly divided into Control,DEACMP,DEACMP+DMSO,and DEACMP+MRS2179 groups.The latter three groups were exposed to static CO inhalation to establish the DEACMP model,while the Control group inhaled air.The rats in DEACMP+MRS2179 group were injec-ted with 3 μl of P2Y1R antagonist MRS2179 with concentration of 1 mmol/L via lateral ventricle,while the DEACMP+DMSO group an equal volume of DMSO.At 14 and 21 day after DEACMP model establishment,the spatial learning and memory ability of rats were observed by Morris water maze,the injury of pyramidal cells in the CA1 area of the hippo-campus was observed by HE staining.The protein levels of glial fibrillary acidic protein(GFAP)and P2Y1receptor(P2Y1R)in the hippocampal tissues of rats were detected by immunohistochemistry and Western blot.The expressions of interleukin-1β(IL-1β),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in the hippocampal tissues of rats were detected by ELISA.Results:In comparison with the Control group,the escape latency was found to be signifi-cantly prolonged in the DEACMP group and the DEACMP+DMSO group.Furthermore,the expression levels of GFAP,P2Y1R,IL-1β,IL-6,and TNF-α were significantly increased in the pyramidal cells of the CA1 region of the hippocam-pal tissue(P<0.05).In contrast,the escape latency was significantly reduced and the expression levels of GFAP,P2Y1R,IL-1β,IL-6,and TNF-α were significantly decreased in the DEACMP+MRS2179 group when compared with the DEACMP+DMSO group(P<0.05).Conclusion:P2Y1R activates astrocytes to release inflammatory factors lead-ing to the onset of cognitive impairment in DEACMP,and inhibition of P2Y1R reduces the expression of inflammatory factors and thus improves cognitive function in DEACMP.