ObjectiveThis paper aims to conduct a secondary analysis of a randomized controlled trial on the treatment of myelodysplastic syndrome （MDS） with deficiency of both the spleen and kidney and blockage of toxin and blood stasis with an arsenic-containing traditional Chinese medicine compound， by applying the mixed model for repeated measure （MMRM） and the method of stratified composite outcome with win ratio. The analysis includes the assessment of hematological efficacy and the composite outcome evaluation of adverse reactions， so as to more comprehensively assess the therapy of this regimen. MethodsThe MMRM and win ratio methods were used to evaluate the efficacy of a prospective，multi-center，double-blind，randomized controlled study. The blood routine （hemoglobin concentration，neutrophil count， and platelet count） and biochemical indexes （aspartate aminotransferase，alanine aminotransferase，serum creatinine，and serum ferritin） of the patients were detected at the time of enrollment and at the end of each course of treatment in the laboratory department of Xiyuan Hospital. The patients' syndromes at the time of enrollment and after treatment were recorded and scored according to the therapy standard of traditional Chinese medicine for diseases and syndromes. MMRM was used to analyze the blood routine indexes of the experimental group and the control group. This method has the advantages of high data reliability and dynamic efficacy under intervention and time. The win ratio method was used to evaluate the composite outcome of traditional Chinese medicine syndrome scores and biochemical indexes according to the priority and to verify the clinical safety of arsenic-containing traditional Chinese medicine compound. ResultsThe results of MMRM analysis showed that the hemoglobin concentration of patients in the group with arsenic-containing traditional Chinese medicine compound increased significantly compared with that before treatment in the group，while that in the placebo group decreased significantly （P<0.01）. When compared with that after treatment in the placebo group，the hemoglobin concentration of patients in the group with arsenic-containing traditional Chinese medicine compound increased significantly，and the mean difference of least squares （LS） was statistically significant （P<0.01）. When compared with those before treatment in the group，there were no statistically significant differences in the neutrophil count and platelet count in both groups. After treatment，there were no statistically significant differences in the neutrophil count， platelet count， and the mean difference of LS between the two groups. The analysis results of win ratio showed that the group with arsenic-containing traditional Chinese medicine compound had a significant advantage in the comparison of composite outcomes，with a win ratio （95% CI） of 2.01 （1.24-3.27） （P<0.01），and that the possibility of "winning" in terms of safety was 2.01 times that of the placebo group. The safety advantage of the group with arsenic-containing traditional Chinese medicine compound mainly came from the traditional Chinese medicine syndrome scores，renal function indexes， and iron reserve capacity indexes，and the number of winning times was less than that of losing times in the comparison of liver function outcomes. ConclusionThe MMRM analysis proves that the arsenic-containing traditional Chinese medicine compound can significantly improve the hemoglobin concentration of patients with myelodysplastic syndrome with refractory cytopenia and multilineage dysplasia （MDS-RCMD） of the type of deficiency of both the spleen and kidney and blockage of toxin and blood stasis. This conclusion is not interfered with by time trends and individual relationships and methodologically improves the credibility of the therapy of the arsenic-containing traditional Chinese medicine compound in treating MDS. Four outcomes are evaluated by the win ratio method，namely traditional Chinese medicine syndromes，liver function，renal function， and iron reserve capacity，proving that the arsenic-containing traditional Chinese medicine compound has the comprehensive advantages of improving the survival quality of the patients and reducing adverse reactions. The win ratio outcome provides clear comparative indexes for the evaluation of adverse reactions，making it easier for regulatory authorities，medical staff， and patients to understand the safety of the arsenic-containing traditional Chinese medicine compound in clinical application.

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”.

Type 1 diabetes mellitus is a chronic autoimmune disease caused by the destruction of pancreatic islet β cells. Pancreatic islet transplantation provides a treatment method for patients with type 1 diabetes mellitus to restore endogenous insulin secretion. However, some problems limit the widespread application of islet transplantation, such as the shortage of donors and post-transplantation rejection damage. Mesenchymal stem cell-derived exosome (MSC-Exo) has become a potential tool for islet transplantation therapy due to their immunomodulatory and tissue repair capabilities. MSC-Exo shows great promise for application, because of low immunogenicity, easily being stored and transported, and the potential as drug delivery vehicles. However, challenges such as preparation, purification, standardization and safety verification need to be overcome before converting MSC-Exo into clinical practice. Therefore, this article reviews the application and potential advantages of MSC-Exo in islet transplantation, aiming to providing more effective and safer treatment options for patients with type 1 diabetes mellitus.

To understand the background of Escherichia coli(E.coli)carried by artificially bred sika deer and the biological characteristics of the isolated strains,such as drug resistance and pathogenicity,in April 2024,we collected 184 fresh deer fecal samples from four deer farms in Luxiang Township,Shuangyang District,Changchun City,Jilin Province,for isolation and cultivation of E.coli.The isolates were tested for drug resistance and biochemical identification with a BD PhoenixTM-100 Automated Microbiology System.The virulence genes were detected with PCR,and the strains were molecularly typed with ERIC-PCR.A total of 165 E.coli strains were isolated from 184 samples of deer feces,with an isolation rate of 89.67%.Twenty strains had a drug resistance phenotype,and the drug resistance rate was 12.12%;these strains included 15 strains of multi-drug resistant bacteria and 11 strains of ESBL-producing bacteria.Virulence gene detection indicated that the sika deer isolates carried multiple diarrhea-associated virulence genes,such as EAST-1(12.12%),eae(1.21%),stx1(7.88%),stx2(7.27%),and STa(1.82%).ERIC-PCR demonstrated that the isolates showed high polymorphism.The ESBL-producing E.coli carried by sika deer are likely to spread drug resistance in the community and livestock population.Some isolates carried multiple diarrhea-associated virulence genes,thus posing a human transmission risk.Therefore,monitoring of drug resistance and virulence genes must be strengthened,and antibiotics must be used reasonably during the breeding process to avoid excessive use and misuse.

Objective The clinical characteristics of 21 cases of nocardiosis were reviewed and antimicrobial resistance of Nocardia strains was analyzed in order to improve the accuracy of clinical diagnosis and treatment of nocardiosis.Methods Clinical data of patients diagnosed with nocardiosis in Zhoukou Central Hospital from 2019-2023 and the corresponding results of antimicrobial susceptibility testing were retrospectively analyzed to summarize the clinical characteristics and outcomes of patients.Results Overall,the 21 cases of nocardiosis included 9 males and 12 females,aged 2-91 years.Underlying disease was reported in 15 patients.Most common type of nocardiosis was pulmonary nocardiosis in 15 cases,followed by skin and soft tissue infection,pleurisy,lymphadenitis,and disseminated nocardiosis.Laboratory tests showed increased levels of WBC,neutrophils percentage,erythrocyte sedimentation rate,C-reactive protein,and procalcitonin.The 21 strains of Nocardia included 4 strains of Nocardia cyriacigeorgica,2 strains each of Nocardia brasiliensis,Nocardia abscessus,Nocardia asiatica,Nocardia otitidiscaviarum and Nocardia beijingensis,and 1 strain each of Nocardia puris,Nocardia asteroides,Nocardia farcinica,Nocardia pneumoniae,Nocardia amamiensis,and 2 strains of unclassified Nocardia.All of the Nocardia strains(100％)were susceptible to linezolid,amikacin,and trimethoprim-sulfamethoxazole,followed by various levels of susceptibility to cefotaxime,moxifloxacin,imipenem and ceftriaxone,and lower susceptibility rate to cefepime,minocycline,ciprofloxacin and clarithromycin.Antimicrobial susceptibility of Nocardia strains varied with different Nocardia species.Of the 21 patients,two were referred to other hospitals,another two died,two patients received unknown treatment,and the remaining 15 patients were improved after antibiotic treatment,including sulfonamides combined with other antibiotics in 11 cases,other antibiotics in 4 cases.Conclusions Immunocompromised patients or those with underlying diseases are more susceptible to nocardiosis.The clinical features are complex and diverse.Antimicrobial susceptibility of Nocardia strains varied with different Nocardia species.Accurate identification and antimicrobial susceptibility test are essential for prescribing effective antibiotic treatment.

Aim To infer novel therapeutic and phar-macological targets related to lung cancer treatment through multiomics approaches,so as to provide new directions for developing more personalized and effec-tive treatment strategies.Methods Genome-wide as-sociation study(GWAS)data analysis,pan-cancer,single-cell,transcriptomics,and protein-protein interac-tion analysis were employed in this study.Results We analyzed biomarkers and therapeutic targets associ-ated with lung cancer.The study identified key bio-markers closely related to lung cancer progression and explored the interrelationships between these biomark-ers and viral infections.According to KEGG pathway annotation,the number of genes related to metabolic processes increased significantly.In particular,metab-olites such as alanine and isoleucine emerged as pivotal factors in therapeutic interventions.The IgD+CD24+and IgD+CD24-B cell subsets were identified as cen-tral elements in immune evasion and treatment re-sponse.Concurrently,the Lachnospiraceae and Prevo-tella were shown to modulate host immune responses and the tumor microenvironment by regulating short-chain fatty acid levels,thereby opening novel avenues for cancer research.Conclusions Through mul-tiomics analysis combined with transcriptomics and pro-teomics analysis,we identify several potential therapeu-tic targets for lung cancer,providing key insights for developing novel treatment strategies.

OBJECTIVE To investigate the distribution,drug resistance and molecular characteristics of the Staph-ylococcus aureus strains isolated from intensive care unit(ICU)environment in some hospitals of Shanghai Pudong New Area so as to provide bases for prevention and control of infections.METHODS The specimens were collected from ICU environment of 8 medical institutions in Pudong District in 2019 and 2023,the S.aureus strains were i-solated and identified.The drug susceptibility testing was performed by microbroth dilution method.The drug re-sistance genes and virulence genes were predicted based on virulence factors of bacterial pathogens,the compre-hensive antibiotic resistance database(mcmaster.ca)and PubMLST,multilocus sequence typing(MLST)and phylogenetic analysis were performed.RESULTS Totally 3408 samples were collected,and the isolated S.aureus strains accounted for 1.29％(44/3408);the isolated strains form both the shared articles and the articles that health care workers directly contact accounted for 20.45％.The drug resistance rate to penicillin was highest(72.73％,32/44);the strains were completely susceptible to vancomycin,rifampicin,sulfamethoxazole-trime-thoprim,daptomycin and nitrofurantoin.The methicillin-resistant Staphylococcus aureus(MRSA)strains accoun-ted for 25.00％.The multi-drug resistance rate was 40.91％.The strains carried 26 types of drug resistance genes in total,covering 9 categories;the carrying rate of fluroquinolones resistance gene nor A was 100.00％.The strains carried 12 types of enterotoxin genes,the carrying rate of sea gene was highest(50.00％,22/44),some of the strains carried lukS-PV,lukF-PV and tsst gene.There were 14 types of sequence type(ST),dominated by ST7(13/44,29.55％),and novel ST type ST9342 was discovered.The phylogenetic analysis showed that all of the strains were divided into 2 major branches,and the strains with the same ST type were phylogenetically close on the evolutionary tree.CONCLUSIONS The S.aureus strains are prevalent in ICU environment and are highly re-sistant to antibiotics.The strains exhibit various ST types,which,to a certain degree,is associated with the path-ogenicity.

In view of the characteristics of H5N6 subtype avian influenza virus(AIV)that it has both high pathogenicity and the risk of cross-species transmission,posing a serious threat to the poultry farming industry and public health security,in order to effectively prevent and control the spread of H5N6 avian influenza,a rapid,sensitive and specific detection technolo-gy was established in this study.The specific monoclonal antibodies against the neuraminidase N6 protein of avian influenza A virus subtype H5N6 were obtained through hybridoma and monoclonal antibody technology.These antibodies were coupled and labeled with carboxyl-functionalized fluorescent quantum dots,along with previously prepared specific antibodies against the hemagglutinin H5 protein.A rapid fluorescence immunochromatographic detection method for the H5N6 subtype of avian influ-enza virus was established according to the principle of double-antibody sandwich immunochromatography.This method a-chieved a detection sensitivity of 1 ng/mL for recombinant hemagglutinin H5 subtype protein and 0.1 ng/mL for recombinant neuraminidase N6 subtype protein.Moreover,the method exhibited no cross-reactivity with other influenza subtypes or patho-gens,such as Newcastle disease(ND),infectious bronchitis(IB),and infectious laryngotracheitis(ILT),thus demonstrating good specificity.The method effectively identified the highly pathogenic avian influenza virus H5 subtype and directly distin-guished the H5N6 subtype with good accuracy.The fluorescent quantum dot immunochromatographic typing detection method established herein met the sensitivity,specificity,and accuracy requirements for H5N6 subtype detection,and can be further used for rapid detection of the H5 and H5N6 subtypes of avian influenza virus.

Objective·To investigate the spatial epigenetic characteristics of spontaneous colon tumors in Apcmin/+mice.Methods·A spatial assay for transposase-accessible chromatin with high-throughput sequencing(ATAC-seq)technology platform was established using an eight-month-old male Apcmin/+mouse model with spontaneous colon tumors.One tumor from a mouse was harvested and embedded in OCT compound for serial cryosectioning;one tissue section was stained with hematoxylin-eosin(H-E)to observe its histological characteristics,while an adjacent section was processed using spatial ATAC-seq technology to generate spatially resolved DNA libraries,followed by sequencing to obtain spatial chromatin accessibility data.Another tumor from the same mouse was digested into a single-cell suspension,in which viable single cells were sorted by flow cytometry and processed for single-cell RNA sequencing.The results were integrated with spatial chromatin accessibility data to jointly analyze the epigenetic characteristics of the colon tumor microenvironment.Results·A stable spatial ATAC-seq platform was successfully established,dividing the tumor into malignant,non-malignant,and malignant-non-malignant boundary regions.Transcription factors enriched in malignant regions included NK2 homeobox 5(NKX2-5)and transcription factor 3(TCF3).Analysis of transcription factor enrichment in the 3 regions revealed two distinct expression trends:one showing a gradual decrease from malignant to boundary to non-malignant regions,and the other exhibiting high expression in malignant and boundary regions but low expression in non-malignant regions.Gene analysis across regions revealed significant upregulation of hypoxia response,transforming growth factor(TGF),and Kirsten rat sarcoma viral oncogene homolog(KRAS)signaling pathways in malignant regions,with cell cycle-related functions markedly enhanced.Analysis of cell-cell interactions in the tumor microenvironment revealed significant differences in interaction strength:strong interactions within non-malignant regions,moderate interactions between boundary and non-malignant regions,and weak interactions between malignant and boundary regions as well as between malignant and non-malignant regions.Conclusion·Colon tumors in Apcmin/+mice exhibit high spatial heterogeneity;malignant regions were enriched with transcription factors including TCF3,and cell interactions between malignant regions and boundary/non-malignant regions were relatively weak.