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 analyze differences of purified water,softened water,and concentrated softened water on the decontamination effects for surgical instruments,so as to provide a reliable reference for the selection of cleaning water for surgical instruments.Methods The physical and chemical indexes and components of concentrated softened water,softened water,and purified water were detected,and their cleaning effects for instrument were compared.The decontamination cleaning experiment was conducted in three hospitals with different types of cleaning water using mini cleaning machines,and the differences in the decontamination time were analyzed.The cleaning and decontamination experiments on contaminated instruments was performed using a spray cleaning and disinfection device,and the cleaning effects of instruments with different types of cleaning water were analyzed.Results Calcium content and hardness of the three types of cleaning water were all at a low level,which can avoid scale adhesion for cleaning surgical instruments.There were statistically significant differences in the pH value,conductivity,and contents of calcium,sodium and chloride among the three cleaning waters(P<0.05).The purified water was weakly acidic,and its conductivity,hardness,and contents of calcium,sodium and chloride were all at low levels.The softened water and concentrated softened water were weakly alkaline,with high levels of conductivity and sodium content and low level of chloride.There were statistically significant differences in the decontamination time of the purified water,softened water and concentrated softened water among hospitals(P<0.05).Under the same contamination condition of surgical instruments,there was a statistically significant difference in the qualified rate of instruments cleaning in the upper and lower cleaning baskets with different cleaning waters(P<0.05).Conclusion Concentrated softened water and softened water have high sodium content,and their decontamination and cleaning abilities are significantly stronger than those of the purified water.Purified water has poorer effects in instruments cleaning for its deionized property.Using concentrated softened water or softened water in the surgical instrument cleaning process can achieve effects of reducing consumption and increasing efficiency,which is conducive to improving cleaning efficiency and quality.

Aim To investigate the possible mechanism of the myocardial protective effect of Danzhi Jiangtang capsules(DJC)on db/db mice based on NLRP3 in-flammasome-mediated pyroptosis.Methods The db/db mice were randomly divided into the model group,DJC low,medium,and high dose groups,and the met-formin group,and the db/m mice were taken as the blank group.The administration lasted for eightweeks.At the end of drug administration,blood glucose,blood lipids,cardiac enzymes and inflammatory factors were detected in each group of mice.HE and Masson stai-ning was performed to observe the morphology and fi-brosis of myocardial tissue.TUNEL staining was per-formed to detect apoptosis.RT-qPCR was performed to detect the mRNA expression of ANP,BNP and β-MHC,and Western blot was performed to detect the protein expression of NLRP3,ASC,caspase-1,cleaved-caspase-1,GSDMD and GSDMD-NT in myocardial tis-sue.Results DJC could alleviate myocardial patho-logical damage,reduce collagen deposition and apopto-sis,reduce the levels of blood glucose,blood lipid,myo-cardial enzyme and inflammatory factors in db/db mice.DJC could reduce the mRNA expressions of ANP,BNP and β-MHC,and the protein expressions of NLRP3,ASC,caspase-1,cleavedcaspase-1,GSDMD and GSDMD-NT in myocardial tissues.Conclusion DJC attenuates myocardial injury in db/db mice,prob-ably by inhibiting the activation of NLRP3 inflamma-somes,attenuating cardiomyocyte pyroptosis,and amel-iorating the inflammatory state.

AIM To investigate the effects of Rutong Ruanjian Tablets on angiogenesis in cancer tissues of rats with preneoplastic breast cancer(PBC).METHODS 60 female SD rats were randomly divided into a blank group of 10 rats and a model group of 50 rats for the establishment of the PBC models of Liver-Qi Stagnation and Blood Stasis Pattern with 9 weeks of oral administration of 7,12-dimethylbenz[a]anthracene(DMBA)and cervical ligation.After successful modeling,the rats were randomly divided into the model group,the tamoxifen group(3.2 mg/kg),the Rutong Ruanjian Tablets group(128 mg/kg),the 3,5-difluorobenzoyl group(DAPT,5 mg/kg),and the Rutong Ruanjian Tablets(128 mg/kg via gavage)+DAPT(5 mg/kg intraperitoneal injection)group,for 1 month corresponding drug administration,with 10 rats in each group.Then the rats had their cancer progression and syndrome scores observed;their angiogenesis evaluated by assessment of microvascular density(MVD);their vascular endothelial growth factor(VEGF)expression assessed by immunohistochemistry;and their mRNA and protein expressions of proteins related to the DLL4/Notch1/Hes1 pathway measured using RT-qPCR,immunohistochemistry and Western blot.RESULTS During carcinogenesis of rats induced by DMBA,there was gradual disappearance of E-cadherin expression and consistency of HE staining result with the PBC progression confirming the success of the modeling.Compared with the blank group,the model group showed increased MVD values,mRNA expression of Notch1 and Hes1,and protein expressions of VEGF,DLL4,Notch1 and Hes1(P＜0.05,P＜0.01).Compared with the model group,the Rutong Ruanjian Tablets group exhibited reduced MVD values,mRNA expression of Notch1 and Hes1,and protein expressions of VEGF,DLL4,Notch1 and Hes1(P＜0.05,P＜0.01).The Rutong Ruanjian Tablets+DAPT group showed reduced mRNA expression of Notch1 and Hes1,and protein expressions of DLL4,Notch1 and Hes1 compared to the Rutong Ruanjian Tablets group(P＜0.05,P＜0.01).CONCLUSION Rutong Ruanjian Tablets can inhibit angiogenesis and attenuate cancer progression in PBC rats of Liver-Qi Stagnation and Blood Stasis Pattern,and the mechanism may lie in the downregulation of DLL4/Notch1/Hes1 signaling pathway related proteins.

OBJECTIVE: Observational studies have found associations between inflammatory bowel disease (IBD) and the risk of dementia, including Alzheimer's dementia (AD) and vascular dementia (VD); however, these findings are inconsistent. It remains unclear whether these associations are causal. METHODS: We conducted a meta-analysis by systematically searching for observational studies on the association between IBD and dementia. Mendelian randomization (MR) analysis based on summary genome-wide association studies (GWASs) was performed. Genetic correlation and Bayesian co-localization analyses were used to provide robust genetic evidence. RESULTS: Ten observational studies involving 80,565,688 participants were included in this meta-analysis. IBD was significantly associated with dementia (risk ratio [ RR] =1.36, 95% CI = 1.04-1.78; I ² = 84.8%) and VD ( RR = 2.60, 95% CI = 1.18-5.70; only one study), but not with AD ( RR = 2.00, 95% CI = 0.96-4.13; I ² = 99.8%). MR analyses did not supported significant causal associations of IBD with dementia (dementia: odds ratio [ OR] = 1.01, 95% CI = 0.98-1.03; AD: OR = 0.98, 95% CI = 0.95-1.01; VD: OR = 1.02, 95% CI = 0.97-1.07). In addition, genetic correlation and co-localization analyses did not reveal any genetic associations between IBD and dementia. CONCLUSION Our study did not provide genetic evidence for a causal association between IBD and dementia risk. The increased risk of dementia observed in observational studies may be attributed to unobserved confounding factors or detection bias.
Humans ; Mendelian Randomization Analysis ; Inflammatory Bowel Diseases/complications* ; Dementia/etiology* ; Observational Studies as Topic ; Genome-Wide Association Study

Objective To explore the activity and mechanism of polymyxin B combined with novel β-lactam/β-lac-tamase inhibitor combinations on biofilm of polymyxin B-resistant Pseudomonas aeruginosa and Klebsiella pneumo-niae.Methods The minimum inhibitory concentration(MIC),minimum biofilm inhibitory concentration(MBIC),and minimum biofilm eradication concentration(MBEC)of all antimicrobial agents were determined by micro-broth dilution method and MBECTM assay.The crystal violet staining method was adopted to evaluate the effect of poly-myxin B combined with cefepime/avibactam,ceftazidime/avibactam,meropenem/avibactam,aztreonam/avibactam,meropenem/vaborbactam,and imipenem/relebactam at sub-MIC doses on inhibition of biofilm formation and eradi-cation of mature biofilm.The best combination scheme for anti-biofilm activity was screened out,and anti-biofilm mechanism of this combination scheme was preliminarily explored with phenol-sulfuric acid method,bacterial motili-ty test,and quorum sensing inhibition test.Results The MBIC and MBEC of all antimicrobial agents were higher than MIC.The combination regimen based on polymyxin B could inhibit the formation of biofilms and eradicate ma-ture biofilm in Pseudomonas aeruginosa and Klebsiella pneumoniae.The combination of polymyxin B with cefepime/avibactam had the highest inhibition and eradication rates,ranging 67.99％-90.16％and 58.26％-63.86％,respectively.The combination of polymyxin B and cefepime/avibactam could inhibit the extracellular poly-saccharides of Klebsiella pneumoniae,with inhibition rates of 34.04％-61.10％,this combination could also re-duce the swimming and twitching motility diameters of bacteria.Cefepime/avibactam monotherapy on quorum sen-sing signaling molecules presented concentration dependent inhibitory effect,and when combined with polymyxin B,the inhibitory effect was consistent with that of monotherapy.Conclusion Polymyxin B and cefepime/avibactam may be potential scheme for clinical treatment for severe biofilm-associated infection caused by polymyxin B-resistant strains,and their mechanisms may be related to the inhibition of bacterial extracellular polysaccharides and motility.

Objective:To investigate the effects of moxibustion at Tianshu(ST25)acupoint on 5-fluorouracil(5-FU)-induced intestinal mucositis(IM)and its underlying mechanisms.Methods:Eighteen C57BL/6 male mice were randomly divided into four groups:normal control(NC),IM model(IM),moxibustion 15 min(MO 15 min),and moxibustion 30 min(MO 30 min).The IM model was established via intraperitoneal injection of 5-FU.Pathological changes in colon tissues were observed using hematoxylin and eosin(HE)staining.Protein expression levels of peroxisome proliferator-activated receptor alpha(PPARα),nuclear factor kappa-B(NF-κB),phosphorylated NF-κB(p-NF-κB),NOD-like receptor thermal protein domain associated protein 3(NLRP3),caspase-1,interleukin-1β(IL-1β),and interleukin-18(IL-18)were analyzed via Western blot,ELISA,and immunohistochemistry.Results:Compared with the NC group,the IM group showed significantly shortened colon length(P＜0.05),exhibited mucosal damage,inflammatory cell infiltration,and glandular disorder,along with upregulated protein expression of p-NF-κB,NLRP3,IL-1β,IL-18,and caspase-1(P＜0.05),and downregulated PPARα expression(P＜0.05).After moxibustion intervention,the MO 15 min group demonstrated increased intestinal length(P＜0.05),reduced pathological scores(P＜0.05),significantly downregulated expression of NLRP3,p-NF-κB,IL-1β,and IL-18(P＜0.05),and elevated PPARα expression(P＜0.05),while total NF-κB protein levels remained unchanged.Conclusion:Moxibustion at Tianshu(ST25)acupoint may alleviate 5-FU-induced intestinal mucosal inflammatory responses by activating PPARα to suppress the NF-κB/NLRP3 inflammasome signaling pathway.

Objective To investigate the effects of joint function screening and correction on intervention efficacy of prevention and assessment score of training injuries in new recruits.Methods A randomized controlled study was conducted on 265 new recruits subjected from two organizational units of an army unit with cluster sampling.Based on entire organizational unit,the participants were randomly divided into a control group(n=132)and an experimental group(n=133).The experimental group received joint function screening and corrective movement training,which was subsequently applied in the new recruit training,while the control group underwent training according to conventional methods.Joint function were collected before and after training.The demographic data,assessment score of training,and incidence of training injuries were collected through the participant's own organizational unit.Receiver operating characteristic(ROC)curve was plotted to evaluate the efficacy of joint function screening in predicting training injuries,and binary logistic regression and general linear regression analyses were applied to verify the correlation of joint function screening score with training injuries and assessment score of training.Results After new training,the score of joint function screening was significantly higher in the experimental group than the control group(16.62±1.87 vs 14.92±2.58,P<0.001).And the score was obviously increased in the experimental group(16.62±1.87 vs 12.82±1.98,P<0.001)and the control group(14.92±2.58 vs 12.95±1.81,P<0.001)when compared with the corresponding score before training.The area under the ROC curve(AUC)of joint function screening in predicting training injuries was 0.762(95%CI:0.694～0.830),indicating good predictive efficacy.During the new training process,the incidence of training injuries in the experimental group(13.53%)was significantly lower than that in the control group(24.24%,Chi-square=4.963,P=0.026).Binary logistic regression analysis showed that the pre-training assessment score of joint function screening was an important influencing factor for training injuries in new recruits(OR=0.552,95%CI:0.413～0.660,P<0.001).The experimental group obtained notably higher mean assessment score than the control group[733.00(716.00,752.75)vs 728.79(710.46,744.28),P=0.027].Linear regression analysis revealed a correlation between post-training score of joint function screening and the assessment score of newly trained personnel(P<0.001).Conclusion Joint function screening and correction for newly trained personnel can effectively prevent training-related injuries during the new training period,and correcting joint function through training can effectively improve the assessment score of newly trained personnel.

Objective To investigate the neuroprotective effects and mechanisms of abscisic acid(ABA)in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced Parkinson's disease(PD)mouse models.Methods Eight-week-old C57BL/6J mice were randomly divided into three groups,control group(Ctrl),MPTP group,and MPTP+ABA group,12 mice in each group.Except for the control group,mice in the other groups were intraperitoneally injected with MPTP 25 mg/kg daily for 8 consecutive days to establish a subacute PD model.The MPTP+ABA group received intraperitoneal injections of ABA 25 mg/kg daily for 11 consecutive days,starting 3 days prior to MPTP administration.Behavioral tests were performed 24 hours after the last administration.On day 3,the expression of tyrosine hydroxylase(TH)and glial fibrillary acidic protein(GFAP)in the substantia nigra pars compacta(SNc)and striatum(STR)was analyzed by Western blotting,and mRNA levels of inflammatory factors were measured by Real-time PCR.Immunofluorescent staining was used to detect the expression of TH,GFAP,and ionized calcium-binding adapter molecule 1(Iba1).Results Compared with the control group,MPTP-treated mice exhibited impaired motor function,a reduced number of TH-positive dopaminergic neurons in the SNc,down-regulated TH protein expression in both the SNc and striatum,up-regulated GFAP protein expression,increased numbers of GFAP-and Iba1-positive cells,and elevated levels of pro-inflammatory factors.In contrast,the MPTP+ABA group showed improved motor function,increased TH-positive neurons in the SNc,up-regulated TH protein expression,down-regulated GFAP protein expression,reduced numbers of GFAP-and Iba1-positive cells,and decreased pro-inflammatory factor levels compared to the MPTP group.Conclusion ABA ameliorates motor dysfunction in MPTP-induced PD model mice,reduces degeneration and death of dopaminergic neurons in the SNc,suppresses the proliferation and activation of astrocytes and microglia in the SNc and striatum,and alleviates neuroinflammation.These results suggest that ABA exerts neuroprotective effects in MPTP-induced PD model mice.