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”.

ObjectiveTo investigate the attitudes and competence of nursing interns toward hospice care， analyze the key factors influencing their performance， and provide targeted improvement suggestions to enhance their comprehensive quality. MethodsA total of 273 undergraduate nursing interns were investigated. General demographic data were collected， and the Chinese version of the Frommelt Attitude Toward Care of the Dying Scale， Form B （FATCOD-B） and the Hospice Care Competency Assessment Questionnaire were administered to evaluate their attitudes and competence in hospice care. A total of 270 valid questionnaires were recovered. Multiple linear regression was performed to analyze the influencing factors of hospice care attitudes and competence among nursing interns. ResultsThe total score of hospice care attitudes was （76.04±12.18） among these 270 nursing interns. Having cared for critically ill patients， receiving hospice care-related education， and experiencing the loss of relatives or friends in the past year were positive influencing factors for their hospice care attitudes， whereas avoidance of talking about death was a negative influencing factor. The total score of hospice care competence was （42.75±4.68）. Having cared for critically ill patients and receiving hospice care-related education represented positive influencing factors for their hospice care competence. ConclusionThe hospice care attitudes and competence of nursing interns are all at moderate levels. It is necessary to strengthen education and training on hospice care for nursing interns， improve the ability of nurses， and provide assistance for the development of hospice care.

Objective:To study the expression of T follicular helper (Tfh) cells, B cells and their subsets in the peripheral blood of patients with systemic lupus erythematosus (SLE), and to preliminarily explore the relationship between Tfh cells and their subsets and the occurrence and development of SLE.Methods:A total of 38 SLE patients who were hospitalized in the Nanjing Drum Tower Hospital from January 2020 to January 2021 and 23 healthy volunteers matched by age and gender were included. The expression of Tfh cells, B cells, their subsets and cytokines in the peripheral blood of healthy volunteers and SLE patients was analyzed by flow cytometry. The clinical data of SLE patients were collected to evaluate the correlation between Tfh cells and their subsets and disease activity.Results:The proportions of Tfh cells, primary B cells, memory B cells and plasma blasts in the peripheral blood of SLE patients and the level of interleukin-21 (IL-21) were significantly higher than those in healthy volunteers (all P<0.05). Among them, Tfh cells were mainly characterized by an increase in the proportion of Tfh17 cell subsets, and the proportion of Tfh17 cells was positively correlated with the systemic lupus erythematosus disease activity index 2000 (SLEDAI-2K), primary B cells, and plasmatocytes ( r=0.592, 0.300, 0.379, all P<0.05). The proportion of Tfh cells was also positively correlated with SLEDAI-2K ( r=0.469, P<0.05). The proportions of Tfh cells and Tfh17 cells in the peripheral blood of SLE patients with medium to high disease activity were significantly higher than those in the low disease activity group (all P<0.05). After 3 months of treatment for SLE patients with medium to high disease activity, the proportions of Tfh ( P=0.04) and Tfh17 cells ( P=0.003) in the peripheral blood decreased compared with before. Conclusions:The disorder of Tfh cells and B cell subsets is involved in the occurrence and development of SLE. Peripheral blood Tfh17 cells can be used as biomarkers for evaluating the disease activity of SLE patients.

Objective To investigate the protective effect of benzoylaconine(BAC)on H9c2 cardio-myocytes after oxygen glucose deprivation/reoxyenation(OGD/R)injury.Methods After an in vitro model of OGD/R injury was established in H9c2 cells,the cells were treated with BAC at different concentrations(0,25,50,75,100 μmol/L)to determine its optimal dose.Then,H9c2 cells were randomly divided into control group,OGD/R group,OGD/R+BAC group(75 μmol),OGD/R+LY294002 group(PI3K/Akt inhibitor),and OGD/R+LY294002+BAC group.Corre-sponding reagent kits were used to determine cell viability and LDH level,as well as the expres-sion levels of TNF-α,IL-6,IL-1β,MDA and GSH-Px in the cells.Western blotting was applied to detect the expression of the PI3K/Akt pathway proteins,as well as autophagic proteins such as LC3,Beclin1,and P62.Results Compared to the control group,the cell viability was significantly decreased,and LDH level was obviously increased in the OGD/R group(P＜0.01).Treatment of 75 μmol/L BAC significantly increased the cell viability(0.87±0.06 vs 40.49±0.06,P＜0.01)and decreased the LDH level(86.75±7.79 U/L vs 234.42±6.20 U/L,P＜0.01)when compared to the levels of the OGD/R group.OGD/R injury induced notable increases in TNF-α,IL-6,IL-1β,and MDA expression levels,while decrease of GSH-Px expression level(P＜0.01),and down-regulation of p-PI3K,p-Akt and P62 and up-regulation of LC3 Ⅱ/LC3 Ⅰ and Beclin-1(P＜0.01)when compared with the control group.Treatment of 75 μmol/L BAC increased the levels of p-PI3K,p-Akt,and P62 proteins(0.90±0.07 vs 0.58±0.04,1.02±0.02 vs 0.49±0.01,1.48±0.05 vs 0.87±0.04)and decreased those of LC3 Ⅱ/LC3 Ⅰ and Beclin-1(0.52±0.01 vs 1.24±0.04,0.12±0.01 vs 0.32±0.02)when compared with the OGD/R group(P＜0.01).Conclusions BAC attenu-ates the inflammatory response and oxidative stress of myocardial cells after OGD/R injury,regu-lates autophagy homeostasis,and reduces myocardial cell damage.Its regulatory effect on myocar-dial autophagy homeostasis may be related to the activation of the PI3K/Akt pathway.

AIM To investigate the improvement effects of Poria cocos polysaccharides(PCPs)on mouse nonalcoholic fatty liver disease(NAFLD).METHODS Forty-eight C57BL/6 mice were randomly divided into the blank group,the model group,the simvastatin group(4 mg/kg)and the high,medium and low dose PCPs groups(200,100 and 50 mg/kg),with 8 mice in each group.The NAFLD model was reproduced by 16 weeks feeding of high-fat and high-cholesterol diet,followed by 8 weeks administration of corresponding drug by gavage.The mice had their body mass and liver coefficient assessed;their levels of hepatic free fatty acid(FFA),and serum total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),aspartate aminotransferase(AST),alanine aminotransferase(ALT),γ-glutamyltransferase(γ-GT)and malondialdehyde(MDA)detected;their hepatic pathological changes and lipid deposition observed using HE staining,NAFLD activity score(NAS)and oil red O staining;and their hepatic protein expressions of Akt,mTOR,p-Akt,p-mTOR and SREBP-1c detected by Western blot.RESULTS Compared with the blank group,the model group demonstrated all increased body weight,liver coefficient,hepatic FFA level,and serum TC,TG,LDL-C,AST,ALT,γ-GT,MDA,IL-1β and TNF-α.levels(P＜0.05,P＜0.01);decreased HDL-C level and activities of SOD and GSH-Px(P＜0.05,P＜0.01);more obvious hepatic pathological damage as revealed by increased NAS score(P＜0.01)and increased lipid deposition area(P＜0.01).Compared with the model group,the groups intervened with high or medium dose PCPs,or simvastatin displayed decreased body weight,liver coefficient,hepatic FFA level,and serum TC,TG,LDL-C,AST,ALT,γ-GT,MDA,IL-1β and TNF-α levels(P＜0.05,P＜0.01);increased HDL-C level and SOD,GSH-Px activities(P＜0.05,P＜0.01);decreased hepatic pathological damage as revealed by the decreased NAS score and lipid deposition area(P＜0.05,P＜0.01);and decreased hepatic protein expressions of p-Akt,p-mTOR and SREBP-1c protein(P＜0.05)as well.CONCLUSION PCPs can improve mouse NAFLD,and its mechanism may lie in their function in reversing abnormal lipid metabolism via Akt/mTOR/SREBP-1c signaling pathway.

Objective To investigate the protective effect of benzoylaconine(BAC)on H9c2 cardio-myocytes after oxygen glucose deprivation/reoxyenation(OGD/R)injury.Methods After an in vitro model of OGD/R injury was established in H9c2 cells,the cells were treated with BAC at different concentrations(0,25,50,75,100 μmol/L)to determine its optimal dose.Then,H9c2 cells were randomly divided into control group,OGD/R group,OGD/R+BAC group(75 μmol),OGD/R+LY294002 group(PI3K/Akt inhibitor),and OGD/R+LY294002+BAC group.Corre-sponding reagent kits were used to determine cell viability and LDH level,as well as the expres-sion levels of TNF-α,IL-6,IL-1β,MDA and GSH-Px in the cells.Western blotting was applied to detect the expression of the PI3K/Akt pathway proteins,as well as autophagic proteins such as LC3,Beclin1,and P62.Results Compared to the control group,the cell viability was significantly decreased,and LDH level was obviously increased in the OGD/R group(P＜0.01).Treatment of 75 μmol/L BAC significantly increased the cell viability(0.87±0.06 vs 40.49±0.06,P＜0.01)and decreased the LDH level(86.75±7.79 U/L vs 234.42±6.20 U/L,P＜0.01)when compared to the levels of the OGD/R group.OGD/R injury induced notable increases in TNF-α,IL-6,IL-1β,and MDA expression levels,while decrease of GSH-Px expression level(P＜0.01),and down-regulation of p-PI3K,p-Akt and P62 and up-regulation of LC3 Ⅱ/LC3 Ⅰ and Beclin-1(P＜0.01)when compared with the control group.Treatment of 75 μmol/L BAC increased the levels of p-PI3K,p-Akt,and P62 proteins(0.90±0.07 vs 0.58±0.04,1.02±0.02 vs 0.49±0.01,1.48±0.05 vs 0.87±0.04)and decreased those of LC3 Ⅱ/LC3 Ⅰ and Beclin-1(0.52±0.01 vs 1.24±0.04,0.12±0.01 vs 0.32±0.02)when compared with the OGD/R group(P＜0.01).Conclusions BAC attenu-ates the inflammatory response and oxidative stress of myocardial cells after OGD/R injury,regu-lates autophagy homeostasis,and reduces myocardial cell damage.Its regulatory effect on myocar-dial autophagy homeostasis may be related to the activation of the PI3K/Akt pathway.

AIM To investigate the improvement effects of Poria cocos polysaccharides(PCPs)on mouse nonalcoholic fatty liver disease(NAFLD).METHODS Forty-eight C57BL/6 mice were randomly divided into the blank group,the model group,the simvastatin group(4 mg/kg)and the high,medium and low dose PCPs groups(200,100 and 50 mg/kg),with 8 mice in each group.The NAFLD model was reproduced by 16 weeks feeding of high-fat and high-cholesterol diet,followed by 8 weeks administration of corresponding drug by gavage.The mice had their body mass and liver coefficient assessed;their levels of hepatic free fatty acid(FFA),and serum total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),aspartate aminotransferase(AST),alanine aminotransferase(ALT),γ-glutamyltransferase(γ-GT)and malondialdehyde(MDA)detected;their hepatic pathological changes and lipid deposition observed using HE staining,NAFLD activity score(NAS)and oil red O staining;and their hepatic protein expressions of Akt,mTOR,p-Akt,p-mTOR and SREBP-1c detected by Western blot.RESULTS Compared with the blank group,the model group demonstrated all increased body weight,liver coefficient,hepatic FFA level,and serum TC,TG,LDL-C,AST,ALT,γ-GT,MDA,IL-1β and TNF-α.levels(P＜0.05,P＜0.01);decreased HDL-C level and activities of SOD and GSH-Px(P＜0.05,P＜0.01);more obvious hepatic pathological damage as revealed by increased NAS score(P＜0.01)and increased lipid deposition area(P＜0.01).Compared with the model group,the groups intervened with high or medium dose PCPs,or simvastatin displayed decreased body weight,liver coefficient,hepatic FFA level,and serum TC,TG,LDL-C,AST,ALT,γ-GT,MDA,IL-1β and TNF-α levels(P＜0.05,P＜0.01);increased HDL-C level and SOD,GSH-Px activities(P＜0.05,P＜0.01);decreased hepatic pathological damage as revealed by the decreased NAS score and lipid deposition area(P＜0.05,P＜0.01);and decreased hepatic protein expressions of p-Akt,p-mTOR and SREBP-1c protein(P＜0.05)as well.CONCLUSION PCPs can improve mouse NAFLD,and its mechanism may lie in their function in reversing abnormal lipid metabolism via Akt/mTOR/SREBP-1c signaling pathway.

AIM To investigate the mechanism of Jiedu Yizhi Formula on cognitive function and neuroinflammation in a mouse model of Alzheimer's disease(AD).METHODS 50 APP/PS1 double transgenic mice were randomly divided into the model group,the donepezil group,and the low-dose,moderate-dose,and high-dose Jiedu Yizhi Formula group(1.78,3.56 and 7.12 g/kg),with 10 mice in each group,in contrast to the 10 WT mice of the blank group.Following anesthesia administration and 8-week oral gavage regimen with respective drugs,all mice underwent final tissue sample collection.The mice had their learning and memory ability assessed by Morris water maze and nesting behavior scores;their pathology of brain tissue and Aβ expression observed using HE,Nissl and IHC staining;their polarization of microglia and the expression of inflammatory factors in hippocampal tissue detected by IF and ELISA;their hippocampal expression of PI3K/Akt/mTOR signaling pathway detected by RT-qPCR and Western blot.RESULTS Compared with the blank group,the model group had lower scores in total swimming distance,frequency in crossing the platform,residence time in the target quadrant,and nesting behavior scores(P＜0.05,P＜0.01);prolonged evasion latency(P＜0.01);more disorganized arrangement of pyramidal cells,solidification and deep staining,unclear demarcation,irregular cell shapes,reduction of Nyctinidia,and increased Aβ deposition in the brain tissue(P＜0.01);elevated expression of hippocampal microglia M1-type markers CD16/32 and lba-1(P＜0.01);decreased levels of M2-type marker CD206(P＜0.05);elevated levels of TNF-α and IL-1β(P＜0.01);decreased expressions of IL-13 and IL-4(P＜0.01);and decreased levels of PI3K,Akt and mTOR mRNA,and reduced p-PI3K,p-Akt and p-mTOR protein expressions(P＜0.01).Compared to the model group,the donepezil group and the Jiedu Yizhi Formula groups showed statistically significant improvements in the aforementioned indexes(P＜0.05,P＜0.01),with the magnitude of improvement being higher in the high-dose Jiedu Yizhi Formula group.CONCLUSION Jiedu Yizhi Formula suppresses microglia Ml-type polarization while enhancing M2-type polarization via activation the PI3K/Akt/mTOR signaling pathway,which subsequently reduces inflammatory cytokine secretion.This mechanism attenuates Aβ deposition in brain tissues and ameliorates cognitive dysfunction in AD mouse models.

To formulate an expert consensus on the principles of preoperative hair removal and provide scientific guidance for standardized removal of hair before surgical procedures so as to reduce the incidence of surgical site infections.METHODS Led by the Hospital Management Institute of National Health Commission of the People's Republic of China,this consensus was reached with the joint efforts from the expects of relevant fields such as surgeries,interventional therapies,nursing,and infection prevention and control.The consensus facilitates the classification and evaluation of literatures by following the evidence grade formulated by Oxford Evidence-based Medicine Center and focuses on the association of preoperative hair removal with surgical site infection,it reaches the evidence grade of expert consensus and recommendation intensity by integrating with discussions on meetings and clinical experience of the expects from relevant fields.RESULTS A total of 6 items of consensus were reached by summarizing the latest evidence on the aspects including the indications for preoperative hair removal,tools,range,timing and places.CONCLUSION The consensus,to some extent,make supplements to and complete the exiting regulations and standards.It provides guidance for the medical institutions to carry out the preoperative hair removal.