BACKGROUND: Several studies have demonstrated the occurrence of secondary tumors as a rare but significant complication of chimeric antigen receptor T (CAR-T) cell therapy, underscoring the need for a detailed investigation. Given the limited variety of secondary tumor types reported to date, a comprehensive characterization of the various secondary tumors arising after CAR-T therapy is essential to understand the associated risks and to define the role of the immune microenvironment in malignant transformation. This study aims to characterize the immune microenvironment of a newly identified secondary tumor post-CAR-T therapy, to clarify its pathogenesis and potential therapeutic targets. METHODS: In this study, the bone marrow (BM) samples were collected by aspiration from the primary and secondary tumors before and after CD19 CAR-T treatment. The CD45 + BM cells were enriched with human CD45 microbeads. The CD45 + cells were then sent for 10× genomics single-cell RNA sequencing (scRNA-seq) to identify cell populations. The Cell Ranger pipeline and CellChat were used for detailed analysis. RESULTS: In this study, a rare type of secondary chronic myelomonocytic leukemia (CMML) were reported in a patient with diffuse large B-cell lymphoma (DLBCL) who had previously received CD19 CAR-T therapy. The scRNA-seq analysis revealed increased inflammatory cytokines, chemokines, and an immunosuppressive state of monocytes/macrophages, which may impair cytotoxic activity in both T and natural killer (NK) cells in secondary CMML before treatment. In contrast, their cytotoxicity was restored in secondary CMML after treatment. CONCLUSIONS This finding delineates a previously unrecognized type of secondary tumor, CMML, after CAR-T therapy and provide a framework for defining the immune microenvironment of secondary tumor occurrence after CAR-T therapy. In addition, the results provide a rationale for targeting macrophages to improve treatment strategies for CMML treatment.
Humans ; Lymphoma, Large B-Cell, Diffuse/therapy* ; Tumor Microenvironment/genetics* ; Antigens, CD19/metabolism* ; Leukemia, Myelomonocytic, Chronic/genetics* ; Immunotherapy, Adoptive/adverse effects* ; Male ; Single-Cell Analysis/methods* ; Female ; Sequence Analysis, RNA/methods* ; Receptors, Chimeric Antigen ; Middle Aged

BACKGROUND: China has made significant progress in medical accessibility and quality over the past decades, and quality improvements in gastroenterology and digestive endoscopy have been consistent. The study aimed to describe the status quo of gastroenterology and digestive endoscopy in the Chinese mainland based on the data from the National Clinical Improvement System (NCIS) and the Hospital Quality Monitoring System (HQMS). METHODS: Data were extracted from the NCIS and the HQMS. Data analysis included general information from the Department of Gastroenterology and Endoscopy centers, management of inpatients and outpatients, and annual volume and quality indicators of digestive endoscopy. Acute pancreatitis, gastrointestinal bleeding, inflammatory bowel disease, and cirrhosis were identified as priority diseases and were subjected to detailed analysis. RESULTS: Data from 4620 and 7074 hospitals were extracted from the NCIS and HQMS, respectively. In 2023, 9.6 gastroenterologists, 6.7 endoscopists, and 37.3 gastroenterology beds per hospital nationwide were observed, achieving 19,252.4 outpatient visits, 1615.2 hospitalizations (97.0 for acute pancreatitis, 146.1 for gastrointestinal bleeding, 40.2 for inflammatory bowel disease, and 111.4 for cirrhosis), and 9432.7 digestive endoscopic procedures per hospital. Overall, the quality of practice improved significantly. The proportion of early cancer among gastrointestinal cancers increased from 11.1% in 2015 to 23.4% in 2023, and the adenoma detection rate during colonoscopy increased from 19.3% in 2019 to 26.9% in 2023. Regarding priority diseases, hospitalizations increased, and 31-day unplanned readmission rates decreased between 2019 and 2023. The median hospitalization costs and median proportion of medication costs decreased for acute pancreatitis, gastrointestinal bleeding, and cirrhosis. However, it increased for inflammatory bowel disease. CONCLUSION This report evaluates the status quo and development of gastroenterology and digestive endoscopy in the Chinese mainland, providing guidance for future quality improvements.
Humans ; China ; Gastroenterology/statistics & numerical data* ; Gastrointestinal Hemorrhage ; Endoscopy, Gastrointestinal/statistics & numerical data* ; Endoscopy, Digestive System/statistics & numerical data*

The innate immune sensor NLRP3 inflammasome overactivation is involved in the pathogenesis of ulcerative colitis. PGAM5 is a mitochondrial phosphatase involved in NLRP3 inflammasome activation in macrophages. However, the role of PGAM5 in ulcerative colitis and the mechanisms underlying PGAM5 regulating NLRP3 activity remain unknown. Here, we show that PGAM5 deficiency ameliorates dextran sodium sulfate (DSS)-induced colitis in mice via suppressing NLRP3 inflammasome activation. By combining APEX2-based proximity labeling focused on PGAM5 with quantitative proteomics, we identify NEK7 as the new binding partner of PGAM5 to promote NLRP3 inflammasome assembly and activation in a PGAM5 phosphatase activity-independent manner upon inflammasome induction. Interfering with PGAM5-NEK7 interaction by punicalagin inhibits the activation of the NLRP3 inflammasome in macrophages and ameliorates DSS-induced colitis in mice. Altogether, our data demonstrate the PGAM5-NEK7 interaction in macrophages for NLRP3 inflammasome activation and further provide a promising therapeutic strategy for ulcerative colitis by blocking the PGAM5-NEK7 interaction.

Benzylisoquinoline alkaloids (BIAs) are a structurally diverse group of plant metabolites renowned for their pharmacological properties. However, sustainable sources for these compounds remain limited. Consequently, researchers are focusing on elucidating BIA biosynthetic pathways and genes to explore alternative sources using synthetic biology approaches. CYP80B, a family of cytochrome P450 (CYP450) enzymes, plays a crucial role in BIA biosynthesis. Previously reported CYP80Bs are known to catalyze the 3'-hydroxylation of (S)-N-methylcoclaurine, with the N-methyl group essential for catalytic activity. In this study, we successfully cloned a full-length CYP80B gene (StCYP80B) from Stephania tetrandra (S. tetrandra) and identified its function using a yeast heterologous expression system. Both in vivo yeast feeding and in vitro enzyme analysis demonstrated that StCYP80B could catalyze N-methylcoclaurine and coclaurine into their respective 3'-hydroxylated products. Notably, StCYP80B exhibited an expanded substrate selectivity compared to previously reported wild-type CYP80Bs, as it did not require an N-methyl group for hydroxylase activity. Furthermore, StCYP80B displayed a clear preference for the (S)-configuration. Co-expression of StCYP80B with the CYP450 reductases (CPRs, StCPR1, and StCPR2), also cloned from S. tetrandra, significantly enhanced the catalytic activity towards (S)-coclaurine. Site-directed mutagenesis of StCYP80B revealed that the residue H205 is crucial for coclaurine catalysis. Additionally, StCYP80B exhibited tissue-specific expression in plants. This study provides new genetic resources for the biosynthesis of BIAs and further elucidates their synthetic pathway in natural plant systems.
Cytochrome P-450 Enzyme System/chemistry* ; Benzylisoquinolines/chemistry* ; Hydroxylation ; Plant Proteins/chemistry* ; Alkaloids/metabolism* ; Stephania tetrandra/genetics*

Objective We aimed to investigate the TCM syndrome characteristics and risk factors of early death in higher-risk patients with myelodysplastic syndromes with excess blasts (HR-MDS-EB). Methods The data of 57 HR-MDS-EB patients in the hematology ward of Xiyuan Hospital, China Academy of Chinese Medical Sciences admitted between January 2013 and August 2020 were analyzed. Patients were divided into an early death group and a non-early death group according to the survival time. The basic information, clinical characteristics, specialized examination, complete blood cell counts and laboratory examination result for first diagnosis, TCM syndrome characteristics, and other data of patients were collected and analyzed to screen the risk factors of death.Results There were statistically significant differences in Charlson comorbidity index (CCI) score, Barthel index score, platelet count at first diagnosis, blast ratio in peripheral blood, lactate dehydrogenase levels, blazing pathogenic heat pattern, and constipation between the early death group and the non-early death group (P<0.05). According to the result of the above univariate analysis, logistic multivariate regression analysis was performed for significant factors P<0.05. CCI score had statistically significant effects on the risk of death (P<0.05).Conclusion Higher CCI score is an independent risk factor for early death in HR-MDS-EB patients.

The application of artificial intelligence(AI)in medical diagnosis and treatment is becoming increasingly widespread,providing doctors and patients with more high-quality,efficient and personalized medical services.However,it also raised a series of ethical issues such as data security,algorithm transparency,responsibility definition,fairness and justice,doctor-patient relationships,and other aspects.Based on the combing of existing research results,this paper analyzed the research status of medical ethics in the application of AI in diagnosis and treatment,as well as expected that future medical ethics research can further explore the ethical issues of AI technology in medical treatment in greater depth,thus ensuring the rational application of AI in the medical field and maximizing the protection of patients'rights and interests.

Objective To observe the effects of probiotics and lactulose synergistic action on gastroin-testinal motility,mucosal function and disease progression in elderly patients with type 2 diabetes mellitus(T2DM)nephropathy constipation.Methods A total of 108 elderly patients with T2DM nephropathy consti-pation admitted and treated in this hospital from May 2021 to March 2023 were selected as the study subjects and divided into the observation group,control group 1 and control group 2 by the random number table meth-od,36 cases in each group.The control group 1 was treated with bifidobacterium tetrad tablet,the control group 2 adopted the lactulose treatment,and the observation group was treated with bifidobacterium tetrad tablet combined with lactulose.The clinical efficacy,constipation symptoms before treatment and in 1,4 weeks after treatment,gastrointestinal motility,serum cerebrointestinal peptide[substance P(SP),nitric oxide(NO),vasoactive intestinal peptide(VIP)],intestinal mucosal barrier function[D-lactic acid(DLC),diamine oxidase(DAO),lipopolysaccharide(LPS)],renal function[creatinine(Cr),urea nitrogen(BUN),estimated glomerular filtration rate(eGFR)]and Toll-like receptor 4(TLR4)/nuclear factor κB(NF-κB)mRNA ex-pression were compared between the groups.Results After 1-,4-week treatment,the total effective rate,bari-um discharge rate at different time points in the observation group were higher than those in the control group 1 and control group 2,and the scores of rectal,abdominal and fecal traits in the observation group were lower than those in the control group 1 and control group 2(P＜0.05),but there was no statistically significant difference between the control group 1 and control group 2(P＞0.05).The serum NO and VIP levels in the observation group were lower than those in the control group 1 and control group 2(P＜0.05),the SP level was higher than that in the control group 1 and control group 2(P＜0.05).After 4-week treatment,the levels of serum DLC,DAO and LPS in the observation group were lower than those in the control group 1 and con-trol group 2(P＜0.05);the Cr and BUN levels,TLR mRNA expression and NF-κB mRNA expression in the observation group and control group 1 were decreased compared with before treatment(P＜0.05),the eGFR level was increased compared with before treatment(P＜0.05),but the above indexes had no statistically sig-nificant difference between these two groups(P＞0.05).Conclusion Probiotics and lactulose in the treat-ment of elderly patients with T2DM nephropathy constipation could play a synergistic role,alleviate the con-stipation symptom,promote the gastrointestinal motility and restore the intestinal mucosal barrier function,which may be reach the anti-inflammation and anti-apoptosis goal by inhibiting TLR4/NF-κB,so as to inhibit the further development of diabetic nephropathy.

Objective:To understand the health needs of patients with non-dementia cognitive impairment after stroke, to provide reference for targeted interventions.Methods:Using the convergent mixed research method, convenience sampling was used to select post-stroke patients with non-dementia cognitive impairment in China-Japan Friendship Hospital and Beijing University of Chinese Medicine Third Affiliated Hospital, a cross-sectional survey was conducted on 191 patients with non-dementia cognitive impairment after stroke using the health needs questionnaire in March to August 2023. A descriptive study was used to conduct semi-structured interviews with 16 patients.Results:A total of 191 questionnaires were distributed and 191 valid questionnaires were collected, including 103 male and 88 female patients, aged from 34 to 90 years old. The items of the post-stroke health questionnaire were (3.47 ± 0.54), with the highest need for understanding the rehabilitation program (148/191); multiple linear regression analysis showed that gender and primary caregiver type were factors influencing their health needs ( t = 2.39, 2.73, both P<0.05). A total of 16 patients with non-dementia cognitive impairment after stroke, 10 males and 6 females, aged from 58 to 90 years old, were interviewed. Four themes were extracted, namely, information support and behavioral guidance needs, psychological care needs, social support needs, and pre-established medical care plan needs. Conclusions:The health needs of patients with non-dementia cognitive impairment after stroke are at an above medium level and have diversified characteristics. Medical staff should conduct systematic health management based on patients′specific conditions and actual needs to help patients recover or maintain cognitive function.

Objective To investigate the expression of Midkine(MDK)in cholangiocarcinoma(CCA)and its value in predicting the prognosis of CCA,as well as the potential mechanism of the effect of MDK on the progression of CCA.Methods The data of CCA samples were obtained from TCGA database to analyze the difference in the expression of MDK between cancer tissue and paracancerous tissue and its association with clinical features,and the data collected from GEO database and 11 CCA patients who underwent surgical resection in The Fifth Medical Center of Chinese PLA General Hospital from June 2018 to September 2021 were used for validation.STRING and Cytoscape were used to construct a protein-protein interaction network,and gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were used to investigate the biological functions and tumor-related pathways involving MDK-related genes.In addition,TIMER and TISIDB databases were used to analyze the correlation between MDK expression and immune cell infiltration in CCA tissue.The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups,and the Fisher's exact test was used for comparison of categorical data between two groups.The Kaplan-Meier method was used to plot survival curves,and the Log-rank test was used for comparison between groups.The Spearman correlation analysis was used to investigate the correlation between two variables.Results The expression level of MDK in cancer tissue and paracancerous tissue of CCA patients was compared based on TCGA database,and the results of the non-paired and paired analyses showed that the expression level of MDK in CCA tumor tissue was significantly higher than that in paracancerous tissue(P<0.001).Transcriptome sequencing was performed for the tumor tissue and its corresponding paracancerous tissue from 11 CCA patients,and the results showed that the expression level of MDK in CCA tumor tissue was significantly higher than that in corresponding paracancerous tissue(P<0.01).High expression of MDK was associated with lymph node metastasis(P=0.045)and vascular invasion(P=0.044).Survival analysis showed that compared with the CCA patients with low MDK expression,the CCA patients with high MDK expression had significantly shorter overall survival time(χ2=5.30,P=0.028)and disease-specific survival time(χ2=6.25,P=0.019).The GO and KEGG enrichment analyses showed that the 30 MDK-related genes were closely associated with ubiquitin-mediated proteolysis and affected the prognosis of CCA patients.The TIMER analysis showed that the expression level of MDK was positively correlated with the infiltration of B cells(r=0.356,P=0.035 6)and dendritic cells(r=0.409,P=0.014 7)in tumor microenvironment of CCA;the TISIDB analysis showed that the expression level of MDK was positively correlated with CXCL16(r=0.465,P=0.004 67)and was negatively correlated with CXCL12(r=-0.389,P=0.019 7)and CXCR5(r=-0.393,P=0.018 5),and it was also negatively correlated with the immune checkpoint regulators VTCN1(r=-0.393,P=0.018 3),LTA(r=-0.380,P=0.022 7),and PVR(r=-0.350,P=0.037 3).Conclusion High expression of MDK is associated with poor prognosis in CCA patients,and MDK has the potential of being used as a molecular marker for predicting the prognosis of CCA.MDK may promote the development and progression of CCA by regulating ubiquitin-mediated proteolysis and the infiltration of B cells and dendritic cells.