中医治未病领域中免疫组库(TCR/BCR)作为一种新型“全息”生物标志物的应用与展望

中医“治未病”理论构成了中医药健康管理的主要内容,“治未病”思想为中医药健康管理体系赋予科学内涵,中医药健康管理体系也是“治未病”思想的外在体现,该体系的建立与完善离不开中医“治未病”思想的深度解析。“治未病”思想的深度解析需要从科学意义上深度思考三个关键问题:一是如何客观描述“未病”的生物学基础并阐明“未病”发生机制;二是如何阐释中医“治未病”科学内涵及中药复方的特殊药理效应机制;三是如何构建以中医“治未病”为内核的中医药精准健康管理新模式。本文从监管科学角度出发,首次提出“全息”生物标志物免疫组库(TCR/BCR)可作为新型“全息”生物标志物,用以客观表征中医治未病生物学基础和干预效果的科学评价体系,进而作为新的科学证据解释上述三个关键科学问题。

Early proactive monitoring of DNA-thioguanine in patients with Crohn’s disease predicts thiopurine-induced late leucopenia in NUDT15/TPMT normal metabolizers

BACKGROUND Thiopurine-induced leucopenia significantly hinders the wide application of thiopurines.Dose optimization guided by nudix hydrolase 15(NUDT15)has significantly reduced the early leucopenia rate,but there are no definitive biomarkers for late risk leucopenia prediction.AIM To determine the predictive value of early monitoring of DNA-thioguanine(DNATG)or 6-thioguanine nucleotides(6TGN)for late leucopenia under a NUDT15-guided thiopurine dosing strategy in patients with Crohn’s disease(CD).METHODS Blood samples were collected within two months after thiopurine initiation for detection of metabolite concentrations.Late leucopenia was defined as a leukocyte count<3.5×10^(9)/L over two months.RESULTS Of 148 patients studied,late leucopenia was observed in 15.6%(17/109)of NUDT15/thiopurine methyltransferase(TPMT)normal and 64.1%(25/39)of intermediate metabolizers.In patients suffering late leucopenia,early DNATG levels were significantly higher than in those who did not develop late leucopenia(P=4.9×10^(-13)).The DNATG threshold of 319.43 fmol/μg DNA could predict late leucopenia in the entire sample with an area under the curve(AUC)of 0.855(sensitivity 83%,specificity 81%),and in NUDT15/TPMT normal metabolizers,the predictive performance of a threshold of 315.72 fmol/μg DNA was much more remarkable with an AUC of 0.902(sensitivity 88%,specificity 85%).6TGN had a relatively poor correlation with late leucopenia whether in the entire sample(P=0.021)or NUDT15/TPMT normal or intermediate metabolizers(P=0.018,P=0.55,respectively).CONCLUSION Proactive therapeutic drug monitoring of DNATG could be an effective strategy to prevent late leucopenia in both NUDT15/TPMT normal and intermediate metabolizers with CD,especially the former.

Effectiveness and tolerability of programmed cell death protein-1 inhibitor+chemotherapy compared to chemotherapy for upper gastrointestinal tract cancers

BACKGROUND The combination of programmed cell death protein-1(PD-1)inhibitor and che-motherapy is approved as a standard first-or second-line treatment in patients with advanced oesophageal or gastric cancer.However,it is unclear whether this combination is superior to chemotherapy alone.AIM To assess the comparative effectiveness and tolerability of combining PD-1 inhibitors with chemotherapy vs chemotherapy alone in patients with advanced gastric cancer,gastroesophageal junction(GEJ)cancer,or oesophageal carcinoma.METHODS We searched the PubMed and Embase databases for studies that compared the efficacy and tolerance of PD-1 inhibitors in combination with chemotherapy vs chemotherapy alone in patients with advanced oesophageal or gastric cancer.We employed either random or fixed models to analyze the outcomes of each clinical trial,en-compassing data on overall survival(OS),progression-free survival(PFS),objective response rate,and adverse events(AEs).RESULTS Nine phase 3 clinical trials(7016 advanced oesophageal and gastric cancer patients)met the inclusion criteria.Our meta-analysis demonstrated that the pooled PD-1 inhibitor+chemotherapy group had a significantly longer OS than the chemotherapy-alone group[hazard ratio(HR)=0.76,95%confidence interval(CI):0.71-0.81];the pooled PFS result was consistent with that of OS(HR=0.67,95%CI:0.61-0.74).The count of patients achieving an objective response in the PD-1 inhibitor+chemotherapy group surpassed that of the chemotherapy-alone group[odds ratio(OR)=1.86,95%CI:1.59-2.18].AE incidence was also higher in the combination-therapy group than in the chemotherapy-alone group,regardless of whether≥grade 3 only(OR=1.30,95%CI:1.07-1.57)or all AE grades(OR=1.88,95%CI:1.39-2.54)were examined.We performed a subgroup analysis based on the programmed death-ligand 1(PD-L1)combined positive score(CPS)and noted extended OS and PFS durations within the CPS≥1,CPS≥5,and CPS≥10 subgroups of the PD-1 inhibitor+chemotherapy group.CONCLUSION In contrast to chemotherapy alone,the combination of PD-1 inhibitor and chemotherapy appears to present a more favorable option for initial or subsequent treatment in patients with gastric cancer,GEJ tumor,or oesophageal cancer.This holds true particularly for individuals with PD-L1 CPS scores of≥5 and≥10.

Higher intakes of lysine,threonine and valine are inversely associated with non-alcoholic fatty liver disease risk:a community-based case-control study in the Chinese elderly

The associations of individual amino acid with non-alcoholic fatty liver disease(NAFLD)risk remained unclear.The present study aimed to investigate the associations between the two in the Chinese elderly.Methods:A community-based health check-up program was conducted in Qingdao,China.NAFLD was diagnosed by ultrasonography accompanied by epidemiological investigation.The dietary intakes of amino acids were investigated with 3-day,24-h dietary records and calculated by Nutrition Calculator software.Restricted cubic spline model was used to evaluate a nonlinear relationship between amino acid intake and NAFLD risk.Results:400 NAFLD subjects were identified,and 400 participants were randomly selected as controls and matched by gender and age(±3 years)Dose-response analysis showed that 1000 mg increment of aromatic amino acids(AAAs)was associated with reduced 16%risk of NAFLD.Dietary increments of 750 mg/d threonine,950 mg/d valine,or 1700 mg/d lysine were associated with a 20%reduction in the NAFLD risk(all P for linearity<0.05).Conclusion:The present study demonstrated that the dietary increases in milk,eggs and deep-sea fish,which are rich in the amino acids,might contribute to protecting against NAFLD in the elderly.

拟微绿球藻发酵肽的抗衰老作用研究

探讨了拟微绿球藻发酵肽(FNPs)的提取、组成分析及其对果蝇体内抗衰老效果。与非发酵组相比,FNPs的多肽含量增加约100条,肽中的抗氧化位点显著增加。FNPs的最大吸收强度在258 nm处,且具有剂量依赖关系。在果蝇实验中,经口干预后,5μg/mL FNPs可使自然衰老的黑腹果蝇的平均寿命延长约13天。同时观察到,在70~90日龄时,0.5,5和10μg/mL的FNPs均可缓解因年龄引起的MDA、SOD和CAT等氧化应激指标的恶化,并将其提高到阳性组水平。结果表明,FNPs是潜在的缓解自然衰老引起的健康问题的生物活性成分,其资源丰富且易于获得,在作为新型抗氧化成分进行功能性食品开发或营养保健膳食制剂将有很好的应用前景。

Impacts of different pancreatic resection ranges on endocrine function in Suncus murinus

BACKGROUND Surgical intervention involving the pancreas can lead to impaired glucose tolerance and other types of endocrine dysfunction.The scope of pancreatectomy and whether it includes the ventral pancreas are the key factors in the development of postoperative diabetes.The ventral and dorsal pancreases are almost separated in Suncus murinus(S.murinus).AIM To investigate the effects of different extents of pancreatic resection on endocrine function in S.murinus.METHODS Eight-week-old male S.murinus shrews were randomly divided into three experimental groups according to different pancreatic resection ranges as follows:ventral pancreatectomy(VPx)group;partial pancreatectomy(PPx)group;subtotal pancreatectomy(SPx)group;and a sham-operated group.Postprandial serum insulin,glucagon-like peptide-1(GLP-1),pancreatic polypeptide(PP),and somatostatin(SST)levels,as well as food intake,weight,blood glucose,and glucose tolerance were regularly measured for each animal.RESULTS S.murinus treated with PPx and SPx suffered from varying degrees of impaired glucose tolerance,but only a small proportion of the SPx group developed diabetes.Only S.murinus in the SPx group showed a significant decrease in food intake accompanied by severe weight loss,as well as a significant increase in postprandial serum GLP-1 levels.Postprandial serum PP levels decreased in both the VPx and PPx groups,but not in the SPx group.Postprandial serum SST levels decreased in both VPx and PPx groups,but the decrease was marginal.CONCLUSION Severe weight loss after pancreatectomy may be related to loss of appetite caused by compensatory elevation of GLP-1.PP and GLP-1 may play a role in resisting blood glucose imbalance.

Simulation of Haloxylon Ammodendron Stand Basic Diameter Structure Based on Fuzzy Distribution Function

Based on the investigation data of 12 Haloxylon ammodendron plots in the south edge of Gurbantunggut Desert, Fuzzy distribution was introduced into the study of Haloxylon ammodendron base diameter structure fitting according to the consistency between the characteristics of Fuzzy distribution function and the distribution series of cumulative percentage of stand base diameter, and the fitting precision and effect of Fuzzy distribution function were discussed. The root mean square error RMSE and determination coefficient R2 values showed that Fuzzy-Γ1, Fuzzy-Γ2, Fuzzy-Γ3, Fuzzy-Γ4 had good fitting performance, among which Fuzzy-Γ1 had relatively high fitting precision, and its parameters were closely related to stand age and density, Fuzzy-Γ2 distribution function was the second, and Fuzzy-Γ4 distribution function had the worst fitting effect. By introducing a parameter c from the similarity of four distribution function formulas, a generalized Fuzzy distribution function Fuzzy-Γ5 is obtained. This function shows the highest fitting accuracy. Most of the values of parameter c are near 1 or 2, which shows that the diameter distribution is mainly approximate to Fuzzy-Γ1 and Fuzzy-Γ2.

Effects of different moxibustion durations on lumbar disk herniation:a clinical study

Objective:To explore the best duration of moxibustion on lumbar disk herniation(LDH).Methods:A total of 88 patients were randomly divided into control group and 15-min,30-min,and 45-min moxibustion groups.The control group was treated with conventional therapy,while the other 3 groups were intervened with different moxibustion durations.Low back pain,dysfunction,lumbar function,and effective rates were evaluated before,in the first week and second week of intervention.Results:In low back pain,compared with the control group,the score of the 15-min group had no significant difference,but it was significantly lower in the 30-min and 45-min groups.In dysfunction and lumbar function,compared with the control group,the scores of the 15-min,30-min,and 45-min groups were significantly higher,and the scores of the 30-min and 45-min groups were significantly higher,but there was no significant difference between the 30-min and 45-min group.In effective rates,there was no significant difference between the control group and 15-min group;the effective rates of the 30-min and 45-min groups were significantly higher than those of the control group.Conclusions:Moxibustion has good therapeutic effect on LDH with specific moxibustion time.

Research on Adaptive Cluster Sampling Method Based on PPS

This paper introduces the principle of PPS-based adaptive cluster sampling method and its modified HH estimator and HT estimator calculation method. It compares PPS-based adaptive cluster sampling method with SRS sampling and SRS-based adaptive group. The difference between the group sampling and the advantages and scope of the PPS adaptive cluster sampling method are analyzed. According to the case analysis, the relevant conclusions are drawn: 1) The adaptive cluster sampling method is more accurate than the SRS sampling;2) SRS adaptive The HT estimator of the cluster sampling is more stable than the HH estimator;3) The two estimators of the PPS adaptive cluster sampling method have little difference in the estimation of the population mean, but the HT estimator variance is smaller and more suitable;4) PPS The HH estimator of adaptive cluster sampling is the same as the HH estimator of SRS adaptive cluster sampling, but the variance is larger and unstable.

CoB and BN composites enabling integrated adsorption/catalysis to polysulfides for inhibiting shuttle-effect in Li-S batteries

Lithium-sulfur(Li-S)batteries are hampered by the infamous shuttle effect and slow redox kinetics,resulting in rapid capacity decay.Herein,a bifunctional catalysis CoB/BN@rGO with integrated structure and synergy effect between adsorption and catalysis is proposed to solve the above problems.The integrated CoB and BN are simultaneously and uniformly introduced on the rGO substrate through a one-step calcination strategy,applied to modify the cathode side of PP separator.The transition metal borides can catalyze the conversion of lithium polysulfides(Li_(2)Sn,n≥4),whereas the bond of B-S is too weak to absorb LPS.Thus BN introduced can effectively restrict the diffusion of polysulfides via strong chemisorption with LiSnLi+…N,while the rGO substrate ensures smooth electron transfer for redox reaction.Therefore,through the integrated adsorption/catalysis,the shuttle effect is suppressed,the kinetics of redox reaction is enhanced,and the capacity decay is reduced.Using CoB/BN@rGO modified PP separator,the Li-S batteries with high initial capacity(1450 mAh g^(-1)at 0.35 mA cm^(-2))and long-cycle stability(700 cycles at 1.74 mA cm^(-2)with a decay rate of 0.032%per cycle)are achieved.This work provides a novel insight for the preparation of bifunctional catalysis with integrated structure for long-life Li-S batteries.

Dissecting the genome of star fruit(Averrhoa carambola L.)

Averrhoa carambola is commonly known as star fruit because of its peculiar shape,and its fruit is a rich source of minerals and vitamins.It is also used in traditional medicines in countries such as India,China,the Philippines,and Brazil for treating various ailments,including fever,diarrhea,vomiting,and skin disease.Here,we present the first draft genome of the Oxalidaceae family,with an assembled genome size of 470.51 Mb.In total,24,726 protein-coding genes were identified,and 16,490 genes were annotated using various well-known databases.The phylogenomic analysis confirmed the evolutionary position of the Oxalidaceae family.Based on the gene functional annotations,we also identified enzymes that may be involved in important nutritional pathways in the star fruit genome.Overall,the data from this first sequenced genome in the Oxalidaceae family provide an essential resource for nutritional,medicinal,and cultivational studies of the economically important star-fruit plant.

Clinical Analysis of Primary Tracheobronchial Tumors in Children and Evaluation of the Predicting Models for Mucoepidermoid Carcinoma

Objective:To determine the clinical characteristics and prognosis of primary tracheobronchial tumors(PTTs)in children,and to explore the most common tumor identification methods.Methods:The medical records of children with PTTs who were hospitalized at the Children's Hospital of Chongqing Medical University from January 1995 to January 2020 were reviewed retrospectively.The clinical features,imaging,treatments,and outcomes of these patients were statistically analyzed.Machine learning techniques such as Gaussian na?ve Bayes,support vector machine(SVM)and decision tree models were used to identify mucoepidermoid carcinoma(ME).Results:A total of 16 children were hospitalized with PTTs during the study period.This included 5(31.3%)children with ME,3(18.8%)children with inflammatory myofibroblastic tumors(IMT),2 children(12.5%)with sarcomas,2(12.5%)children with papillomatosis and 1 child(6.3%)each with carcinoid carcinoma,adenoid cystic carcinoma(ACC),hemangioma,and schwannoma,respectively.ME was the most common tumor type and amongst the 3 ME recognition methods,the SVM model showed the best performance.The main clinical symptoms of PPTs were cough(81.3%),breathlessness(50%),wheezing(43.8%),progressive dyspnea(37.5%),hemoptysis(37.5%),and fever(25%).Of the 16 patients,7 were treated with surgery,8 underwent bronchoscopic tumor resection,and 1 child died.Of the 11 other children,3 experienced recurrence,and the last 8 remained disease-free.No deaths were observed during the follow-up period.Conclusion:PTT are very rare in children and the highest percentage of cases is due to ME.The SVM model was highly accurate in identifying ME.Chest CT and bronchoscopy can effectively diagnose PTTs.Surgery and bronchoscopic intervention can both achieve good clinical results and the prognosis of the 11 children that were followed up was good.

Management of regional citrate anticoagulation for continuous renal replacement therapy:guideline recommendations from Chinese emergency medical doctor consensus

Continuous renal replacement therapy(CRRT)is widely used for treating critically-ill patients in the emergency department in China.Anticoagulant therapy is needed to prevent clotting in the extracorporeal circulation during CRRT.Regional citrate anticoagulation(RCA)has been shown to potentially be safer and more effective,and is now recommended as the preferred anticoagulant method for CRRT.However,there is still a lack of unified standards for RCA management in the world,and there are many problems in using this method in clinical practice.The Emergency Medical Doctor Branch of the Chinese Medical Doctor Association(CMDA)organized a panel of domestic emergency medicine experts and international experts of CRRT to discuss RCA-related issues,including the advantages and disadvantages of RCA in CRRT anticoagulation,the principle of RCA,parameter settings for RCA,monitoring of RCA(mainly metabolic acid-base disorders),and special issues during RCA.Based on the latest available research evidence as well as the paneled experts'clinical experience,considering the generalizability,suitability,and potential resource utilization,while also balancing clinical advantages and disadvantages,a total of 16 guideline recommendations were formed from the experts'consensus.

Location and Capacity Determination Method of Electric Vehicle Charging Station Based on Simulated Annealing Immune Particle Swarm Optimization

As the number of electric vehicles(EVs)continues to grow and the demand for charging infrastructure is also increasing,how to improve the charging infrastructure has become a bottleneck restricting the development of EVs.In other words,reasonably planning the location and capacity of charging stations is important for development of the EV industry and the safe and stable operation of the power system.Considering the construction and maintenance of the charging station,the distribution network loss of the charging station,and the economic loss on the user side of the EV,this paper takes the node and capacity of charging station planning as control variables and the minimum cost of system comprehensive planning as objective function,and thus proposes a location and capacity planning model for the EV charging station.Based on the problems of low efficiency and insufficient global optimization ability of the current algorithm,the simulated annealing immune particle swarm optimization algorithm(SA-IPSO)is adopted in this paper.The simulated annealing algorithm is used in the global update of the particle swarm optimization(PSO),and the immune mechanism is introduced to participate in the iterative update of the particles,so as to improve the speed and efficiency of PSO.Voronoi diagram is used to divide service area of the charging station,and a joint solution process of Voronoi diagram and SA-IPSO is proposed.By example analysis,the results show that the optimal solution corresponding to the optimisation method proposed in this paper has a low overall cost,while the average charging waiting time is only 1.8 min and the charging pile utilisation rate is 75.5%.The simulation comparison verifies that the improved algorithm improves the operational efficiency by 18.1%and basically does not fall into local convergence.

T cells in pancreatic cancer stroma:Tryptophan metabolism plays an important role in immunoregulation

Several studies have shown that the immune system is highly regulated by tryptophan metabolism,which serves as an immunomodulatory factor.The indoleamine 2,3-dioxygenase 1(IDO1),as an intracellular enzyme that participates in metabolism of the essential amino acid tryptophan in the kynurenine pathway,is an independent prognostic marker for pancreatic cancer(PC).First,overexpression of IDO1 inhibits the maturation of dendritic cells and T-cell proliferation in the liver and spleen.Second,the high expression of kynurenine induces and activates the aryl hydrocarbon receptor,resulting in upregulated programmed cell death protein 1 expression.Third,the induction of IDO1 can lead to loss of the T helper 17 cell/regulatory T cell balance,mediated by the proximal tryptophan catabolite from IDO metabolism.In our study,we found that overexpression of IDO1 upregulated CD8+T cells and reduced natural killer T cells in pancreatic carcinoma in mice.Hence,it may be essential to pay more attention to tryptophan metabolism in patients,especially those who are tolerant to immunotherapy for PC.

Comparison and development of machine learning for thalidomideinduced peripheral neuropathy prediction of refractory Crohn’s disease in Chinese population

BACKGROUND Thalidomide is an effective treatment for refractory Crohn’s disease(CD).However,thalidomide-induced peripheral neuropathy(TiPN),which has a large individual variation,is a major cause of treatment failure.TiPN is rarely predictable and recognized,especially in CD.It is necessary to develop a risk model to predict TiPN occurrence.AIM To develop and compare a predictive model of TiPN using machine learning based on comprehensive clinical and genetic variables.METHODS A retrospective cohort of 164 CD patients from January 2016 to June 2022 was used to establish the model.The National Cancer Institute Common Toxicity Criteria Sensory Scale(version 4.0)was used to assess TiPN.With 18 clinical features and 150 genetic variables,five predictive models were established and evaluated by the confusion matrix receiver operating characteristic curve(AUROC),area under the precision-recall curve(AUPRC),specificity,sensitivity(recall rate),precision,accuracy,and F1 score.RESULTS The top-ranking five risk variables associated with TiPN were interleukin-12 rs1353248[P=0.0004,odds ratio(OR):8.983,95%confidence interval(CI):2.497-30.90],dose(mg/d,P=0.002),brainderived neurotrophic factor(BDNF)rs2030324(P=0.001,OR:3.164,95%CI:1.561-6.434),BDNF rs6265(P=0.001,OR:3.150,95%CI:1.546-6.073)and BDNF rs11030104(P=0.001,OR:3.091,95%CI:1.525-5.960).In the training set,gradient boosting decision tree(GBDT),extremely random trees(ET),random forest,logistic regression and extreme gradient boosting(XGBoost)obtained AUROC values>0.90 and AUPRC>0.87.Among these models,XGBoost and GBDT obtained the first two highest AUROC(0.90 and 1),AUPRC(0.98 and 1),accuracy(0.96 and 0.98),precision(0.90 and 0.95),F1 score(0.95 and 0.98),specificity(0.94 and 0.97),and sensitivity(1).In the validation set,XGBoost algorithm exhibited the best predictive performance with the highest specificity(0.857),accuracy(0.818),AUPRC(0.86)and AUROC(0.89).ET and GBDT obtained the highest sensitivity(1)and F1 score(0.8).Overall,compared with other state-of-the-art classifiers such as ET,GBDT and RF,XGBoost algorithm not only showed a more stable performance,but also yielded higher ROC-AUC and PRC-AUC scores,demonstrating its high accuracy in prediction of TiPN occurrence.CONCLUSION The powerful XGBoost algorithm accurately predicts TiPN using 18 clinical features and 14 genetic variables.With the ability to identify high-risk patients using single nucleotide polymorphisms,it offers a feasible option for improving thalidomide efficacy in CD patients.

High-efficiency preparation of multifunctional conjugated electrospun graphene doped PVDF/CF yarns for energy harvesting and human movement monitoring in TENG textile

Portable power is an effective solution to realize self-powered sensors for wearable devices,promoting future sustainable development.Membrane-based triboelectric nanogenerators(M-TENGs)have emerged as a promising technology for harvesting biomechanical energy from human motion owing to their advantages,such as simple structure,lightweight design,and efficient energy conversion.However,the poor durability,low adaptability,and un-washability of two-dimensional membrane materials have largely hindered their application in wearable electronics.In this study,we propose a sheath-core polyvinylidene fluoride(PVDF)/graphene(G)-carbon fiber(CF)yarn fabricated via conjugate electrospinning,comprising a commercial CF core and an electrospun graphene-doped PVDF sheath,which improves the fatigue resistance of electrospun nanofiber films under prolonged friction and keeps a high degree of freedom.The resulting electronic textile,woven with the large-scale electrospun PVDF/G-CF yarn,demonstrates a remarkable power density of 25.5 mW·m^(-2).The tight distribution of PVDF/G nanofibers on the textile surface ensures excellent softness,washability,and durability.Furthermore,the electrospun PVDF/G-CF textile exhibits significant potential in pressure sensing,self-powered operation,and motion detection,making it highly suitable for wearable electronics applications.

Suberoylanilide hydroxamic acid upregulates reticulophagy receptor expression and promotes cell death in hepatocellular carcinoma cells

BACKGROUND Hepatocellular carcinoma(HCC)is a common clinical condition with a poor prognosis and few effective treatment options.Potent anticancer agents for treating HCC must be identified.Epigenetics plays an essential role in HCC tumorigenesis.Suberoylanilide hydroxamic acid(SAHA),the most common histone deacetylase inhibitor agent,triggers many forms of cell death in HCC.However,the underlying mechanism of action remains unclear.Family with sequence similarity 134 member B(FAM134B)-induced reticulophagy,a selective autophagic pathway,participates in the decision of cell fate and exhibits anticancer activity.This study focused on the relationship between FAM134B-induced reticulophagy and SAHA-mediated cell death.AIM To elucidate potential roles and underlying molecular mechanisms of reticulophagy in SAHA-induced HCC cell death.METHODS The viability,apoptosis,cell cycle,migration,and invasion of SAHA-treated Huh7 and MHCC97L cells were measured.Proteins related to the reticulophagy pathway,mitochondria-endoplasmic reticulum(ER)contact sites,intrinsic mitochondrial apoptosis,and histone acetylation were quantified using western blotting.ER and lysosome colocalization,and mitochondrial Ca^(2+)levels were characterized via confocal microscopy.The level of cell death was evaluated through Hoechst 33342 staining and propidium iodide colocalization.Chromatin immunoprecipitation was used to verify histone H4 lysine-16 acetylation in the FAM134B promoter region.RESULTS After SAHA treatment,the proliferation of Huh7 and MHCC97L cells was significantly inhibited,and the migration and invasion abilities were greatly blocked in vitro.This promoted apoptosis and caused G1 phase cells to increase in a concentration-dependent manner.Following treatment with SAHA,ER-phagy was activated,thereby triggering autophagy-mediated cell death of HCC cells in vitro.Western blotting and chromatin immunoprecipitation assays confirmed that SAHA regulated FAM134B expression by enhancing the histone H4 lysine-16 acetylation in the FAM134B promoter region.Further,SAHA disturbed the Ca^(2+)homeostasis and upregulated the level of autocrine motility factor receptor and proteins related to mitochondria-endoplasmic reticulum contact sites in HCC cells.Additionally,SAHA decreased the mitochondrial membrane potential levels,thereby accelerating the activation of the reticulophagy-mediated mitochondrial apoptosis pathway and promoting HCC cell death in vitro.CONCLUSION SAHA stimulates FAM134B-mediated ER-phagy to synergistically enhance the mitochondrial apoptotic pathway,thereby enhancing HCC cell death.

Efficacy of Children Neuropsychological and Behavioral Scale in Screening for Autism Spectrum Disorders through a Combination of Developmental Surveillance

Objective This study aimed to explore the clinical value of Children Neuropsychological and Behavioral Scale-Revision 2016(CNBS-R2016)for Autism Spectrum Disorder(ASD)screening in the presence of developmental surveillance.Methods All participants were evaluated by the CNBS-R2016 and Gesell Developmental Schedules(GDS).Spearman’s correlation coefficients and Kappa values were obtained.Taking GDS as a reference assessment,the performance of the CNBS-R2016 for detecting the developmental delays of children with ASD was analyzed with receiver operating characteristic(ROC)curves.The efficacy of the CNBS-R2016 to screen for ASD was explored by comparing Communication Warning Behavior with Autism Diagnostic Observation Schedule,Second Edition(ADOS-2).Results In total,150 children aged 12–42 months with ASD were enrolled.The developmental quotients of the CNBS-R2016 were correlated with those of the GDS(r=0.62–0.94).The CNBS-R2016 and GDS had good diagnostic agreement for developmental delays(Kappa=0.73–0.89),except for Fine Motor.There was a significant difference between the proportions of Fine Motor,delays detected by the CNBS-R2016 and GDS(86.0%vs.77.3%).With GDS as a standard,the areas under the ROC curves of the CNBS-R2016 were above 0.95 for all the domains except Fine Motor,which was 0.70.In addition,the positive rate of ASD was 100.0%and 93.5%when the cut-off points of 7 and 12 in the Communication Warning Behavior subscale were used,respectively.Conclusion The CNBS-R2016 performed well in developmental assessment and screening for children with ASD,especially by Communication Warning Behaviors subscale.Therefore,the CNBS-R2016 is worthy of clinical application in children with ASD in China.

Vitamin A deficiency suppresses CEACAM1 to impair colonic epithelial barrier function via downregulating microbial-derived short-chain fatty acids

Vitamin A (VA) plays an essential role in modulating both the gut microbiota and gut barrier function.Short-chain fatty acids (SCFAs),as metabolites of the gut microbiota,protect the physiological intestinal barrier;however,they are compromised when VA is deficient.Thus,there is an urgent need to understand how and which SCFAs modulate colonic epithelial barrier integrity in VA deficiency (VAD).Herein,compared with normal VA rats (VAN),at the beginning of pregnancy,we confirmed that the colonic desmosome junction was impaired in the VAD group,and the amounts of acetate,propionate,and butyrate declined because of the decreased abundance of SCFA-producing bacteria (Romboutsia ,Collinsella ,and Allobaculum ).The differentially expressed genes correlated with the gut barrier and the histone deacetylase complex between the VAD and VAN groups were enriched by RNA sequencing.In the VAD group,the expression levels of colonic CEA cell adhesion molecule 1 (CEACAM1) were down-regulated,and the levels of histone deacetylase 1 (HDAC1) and HDAC3 were up-regulated.Intriguingly,the above changes in the VAD groups were rescued by VA supplementation in the early postnatal period.Further study indicated that in Caco-2 cells,butyrate treatment significantly repressed the enrichment of HDAC3 on the promoter of the CEACAM1 gene to induce its expression.Our findings support that butyrate intervention can alleviate the impairment of colonic barrier function caused by VAD,and timely postnatal VA intervention may reverse the damage caused by VAD on gut barrier integrity during pregnancy.