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

ObjectiveTo develop the Evidence Grading Scale for Ancient classical prescriptions in Traditional Chinese medicine， assess its reliability and validity， and apply it in practice to provide multi-source evidence for clinical practice guidelines development. MethodsLiterature retrieval was conducted to extract and screen existing evaluation dimensions， then the initial items were summarized using thematic analysis. Experts in the clinical medicine， medical history and literature participated in the Delphi questionnaire survey to evaluate and refine the items. An expert consensus meeting was conducted to finalize the included items， refine the method for items evaluation and evidence grading. The evidence quality rating scale for ancient classical traditional Chinese medicine （TCM） prescriptions was then established and tested for reliability and validity. ResultsThrough literature review， extraction， screening and summarization， a total of 3 dimensions and 12 initial items were formed. Questionnaires were sent to 69 experts to evaluate the initial items， with a questionnaire response rate of 100% and an expert authority coefficient of 0.92. All 12 items were retained for they had importance scores above 4. The Evidence Grading Scale on Ancient classical prescriptions in Traditional Chinese medicine includes 3 dimensions with 12 items. The 3 dimensions includes ancient evidence， inheritance status， and modern application. Each dimension contains 4 items， and each item has a full score of 5 points. The evidence was rated as high-level， moderate-level， and low-level according to the final scores. The content validity index （CVI） of the 12 items was ＞0.9， the average CVI of the scale was 0.98， and the intraclass correlation coefficient （ICC） was 0.90. ConclusionThe Evidence Grading Scale on Ancient classical prescriptions in Traditional Chinese medicine has good reliability and validity， which is practical for use in the development of TCM clinical guidelines and can better support clinical decision-making.

Objective To analyze the hearing screening effect of newborns in Ningbo City and provide references for further promoting neonatal hearing screening.Methods Otoacoustic emission was applied to screen the hearing of 835 323 newborns in Ningbo City from 2013 to 2023.Those who failed the initial screening were re-checked within 30 to 42 days.Those who failed the re-checking were referred to the municipal hearing diagnosis center for audiological diagnosis.The diagnosis data were enrolled,analyzed,and reported.Results Among a total of 835 323 newborns,828 043 of them actually underwent hearing screening,with an initial screening rate of 99.13％,761 369 newborns passed the initial screening with an initial screening pass rate of 91.95％.64 580 newborns were re-checked with a re-checking rate of 96.86％.There were 10 487 newborns re-checking failures,with a re-checking failure rate of 16.24％.A total of 10 164 cases were diagnosed with hearing loss,with a diagnosis rate of 96.92％.There were 323 cases of loss to follow-up with a loss to follow-up rate of 3.08％.A total of 1582 cases of hearing loss were diagnosed,with an incidence of hearing abnormalities of 1.91‰.Among the 1582 confirmed cases of hearing abnormalities,the diagnosis rate within 3 months was 12.01％,within 6 months was 57.90％,and within 12 months was 90.52％.The average diagnosed age was(7.21±6.55)months,while the median diagnosed age was 5.70 months.Conclusion The initial screening rate,initial screening pass rate,re-screening rate,re-checking recall rate,and diagnosis rate of Ningbo City have met the requirements of the guidelines and are at a leading level in China.The next step is to upgrade the information network to improve the diagnosis rate at 3 months of age,the intervention rate at 6 months of age,and the follow-up management of children with hearing loss are the key points in the future work of neonatal hearing screening.

Venetoclax (Ven) is now widely used for both acute myeloid leukaemia (AML) and high-risk myelodysplastic syndrome (MDS), yet there is no consensus on salvage regimens after Ven failure. This study retrospectively evaluated the efficacy and safety of selinexor combined with subcutaneous decitabine (DAC) in 10 patients with AML or MDS with excess blasts (MDS-EB1/2) who had experienced prior Ven treatment failure. A literature review was also performed. Among the 7 patients with AML, 1 achieved complete remission (CR), 2 achieved CR with incomplete hematologic recovery (CRi), 1 achieved partial remission (PR), 2 had no remission, and 1 experienced disease progression (PD). Among the 3 patients with MDS, 2 achieved marrow CR and 1 had stable disease (SD). The median duration of response among the 6 responding patients was 2 months (range, 0.5-6 months). All 10 patients experienced varying degrees of myelosuppression. Five patients had mild gastrointestinal reactions, all of which were manageable. The overall tolerability was good, and no treatment-related deaths occurred. These findings suggest that selinexor combined with subcutaneous decitabine offers a novel and well-tolerated therapeutic option for patients with myeloid malignancies who have previously failed venetoclax-based therapy.

Here we report the case of a CBF-AML patient with KIT p.D816V mutation who failed avapritinib induction therapy and subsequently underwent bridging allogeneic hematopoietic stem cell transplantation (allo-HSCT), along with a literature review. The patient was a 64-year-old male who did not achieve remission after induction therapy with the DA regimen (Daunorubicin + Cytarabine). After reinduction with avapritinib combined with the DCHG regimen (Decitabine + Homoharringtonine + Cytarabine + Granulocyte colony-stimulating factor), he attained complete remission (CR) and flow cytometry minimal residual disease (MRD) negativity, but the RUNX1::RUNX1T1 fusion gene remained positive. During consolidation therapy, flow MRD reappeared, and the fusion gene level progressively increased. The patient then underwent a 9/10 HLA-matched unrelated donor allo-HSCT. Post-transplant, the fusion gene persisted, prompting treatment with the "ITI" regimen (with dose adjustments, including sequential addition of interferon and interleukin-2, pomalidomide incorporation, and standard vs. escalated dosing of "ITI" regimen). Currently, MRD negativity has been maintained for over 5 months, with good treatment tolerance. This finding suggest that the "ITI" regimen may serve as a novel and well-tolerated therapeutic option for CBF-AML patients with persistent RUNX1::RUNX1T1 fusion gene positivity after allo-HSCT and KIT p.D816V mutation, particularly in cases of avapritinib treatment failure.

Objective:To analyze the treatment-free remission (TFR) outcomes after discontinuation of tyrosine kinase inhibitor (TKI) in children with chronic myeloid leukemia (CML).Methods:In this retrospective cohort study, clinical data of 14 chronic phase CML children aged <18 years who had achieved stable deep molecular response (DMR) for ≥ 2 years after standardized treatment with TKI and had a strong desire to discontinue TKI at Henan Cancer Hospital from September 30, 2016 to January 30, 2022 were collected retrospectively. According to the different TFR outcomes after discontinuation of TKI, patients were divided into loss of major molecular response (MMR) group and without loss of MMR group, differences in clinical characteristics between the two groups of children were analyzed using Mann-Whitney U test and Fisher exact test. Results:Out of 14 children with TKI discontinuation, 7 were male and 7 were female. The age at diagnosis was 14.0 (4.8, 17.0) years, and the age at TKI discontinuation was 22.0 (12.5, 27.0) years. Among them, 8 children were treated with imatinib prior to TKI discontinuation and 6 children were treated with second-line substitution of the second-generation TKI nilotinib or dasatinib prior to TKI discontinuation. The follow-up time was 37.0 (27.8, 47.5) months, and 7 cases lost MMR at the time of discontinuation of 3.0 (2.0, 11.0) months. Eight children gained TFR at 6 months, 7 children gained TFR at 12 and 24 months. Amongst the 6 children who received second-generation TKI prior to TKI discontinuation, 2 children lost MMR at 3 and 11 months and 4 children gained TFR, among the 8 children who discontinued imatinib, 5 children lost MMR at the time 3.0 (2.0, 9.0) months and 3 children gained TFR. The age at diagnosis and TKI discontinuation, the time from TKI treatment to the acquisition of DMR, the duration of TKI treatment before TKI discontinuation, the duration of DMR before TKI discontinuation, and the number of children treated with second-generation TKI were not statistically different between the 7 children in the group that did not lose the MMR and the 7 children in the group that lost the MMR (all P>0.05) . All the 7 children with confirmed loss of MMR immediately restarted TKI therapy, and all regained DMR after 2.0 (2.0, 11.0) months of therapy. None of the children had disease progression. After TKI discontinued, only 1 child had mild bone pain, which could be relieved by oral antipyretic analgesic drugs. Conclusions:Children with CML who have achieved a durable stable DMR for≥2 years on TKI therapy can discontinue the TKI and obtain TFR. Both the longer duration of TKI therapy, the longer duration of DMR and the use of second-generation TKI therapy before TKI discontinuation, may allow more children with CML who are expecting TKI discontinuation to have access to TFR.

BACKGROUND:Macrophage migration inhibitory factor(MIF)is a pleiotropic cytokine,which is secreted in different types of stem cells and can regulate the proliferation,differentiation and migration of various types of stem cells.Our previous research has confirmed that human embryonic stem cells secrete MIF and that its concentration in the culture medium is relatively stable.However,whether MIF is involved in the survival,proliferation and differentiation of human embryonic stem cells remains unclear. OBJECTIVE:To investigate the effects of MIF on survival,proliferation,and differentiation of human embryonic stem cells. METHODS:(1)Human embryonic stem cells H9 were cultured.The growth curve of cells was detected and plotted by CCK-8 assay.Enzyme-linked immunosorbent assay was used to determine the level of MIF in the medium.(2)To determine the effects of exogenous MIF on the survival and proliferation of human embryonic stem cells,different groups were established:the control group,which was cultured in stem cell medium without any modifications;the exogenous MIF group,which was treated with different concentrations(30,100,300 ng/mL)of MIF in the stem cell medium;the MIF inhibitor ISO-1 group,which was treated with different concentrations(2,7,21 μmol/L)of ISO-1 in the stem cell medium;and the MIF+ISO-1 group,which was treated with different concentrations of ISO-1 along with 100 ng/mL of MIF.Cell viability was assessed using the CCK-8 assay.(3)To further elucidate the effect of MIF gene on survival and proliferation of human embryonic stem cell,the MIF knockout H9 cell line was constructed by CRISPR-Cas 9 technology to observe the lineage establishment.(4)To determine the effect of high concentrations of MIF on human embryonic stem cell differentiation,100 ng/mL MIF and 100 ng/mL of CXCR4 neutralizing antibody were separately added to the normal stem cell culture medium.The expression levels of self-renewal factors(KLF4,c-MYC,NANOG,OCT4,and SOX2)and differentiation transcription factors(FOXA2,OTX2)were measured using real-time quantitative polymerase chain reaction,immunofluorescence staining,and western blot analysis. RESULTS AND CONCLUSION:(1)The logarithmic growth phase of H9 cells was between 3-6 days.Under normal growth conditions,human embryonic stem cells secreted MIF at a concentration of approximately 20 ng/mL,independent of cell quantity.(2)Compared to the control group,the addition of different concentrations of MIF had no effect on the proliferation of human embryonic stem cells(P>0.05).ISO-1 significantly inhibited the proliferation of human embryonic stem cells,with a stronger inhibition observed at higher concentrations of ISO-1(P<0.05).The addition of MIF in the presence of ISO-1 reduced the inhibitory effect of ISO-1(P<0.05).(3)Real-time quantitative polymerase chain reaction showed that knocking out 50%of the MIF gene resulted in a significant decrease in the growth vitality of human embryonic stem cells and failure to establish cell lines.(4)Adding 100 ng/mL exogenous MIF to the culture medium resulted in a decrease in the mRNA,protein,and fluorescence expression levels of the self-renewal transcription factor KLF4,while the mRNA,protein,and fluorescence expression levels of the differentiation factor FOXA2 increased.(5)When 100 ng/mL CXCR4 neutralizing antibody was added to the culture medium,the mRNA and protein expression levels of KLF4 increased,while the mRNA and protein expression levels of FOXA2 decreased,contrary to the expression trend observed in the MIF group.In conclusion,the endogenous secretion of MIF by human embryonic stem cells is essential for their survival.The addition of MIF to the culture medium does not promote the proliferation of human embryonic stem cells.However,it can lead to a decrease in the expression of the self-renewal factor KLF4 and an increase in the expression of the transcription factor FOXA2.This provides a clue for further investigation into the effects and mechanisms of MIF on the differentiation of human embryonic stem cells.The MIF-CXCR4 axis plays a regulatory role in this process.

OBJECTIVES: To propose a hypothesis that aloe-emodin may inhibit scar tissue fibrosis through thrombospondin-1(THBS1)-PI3K-Akt pathway. METHODS: By cultivating fibroblasts derived from scar tissue after cleft palate surgery in humans, aloe emodin of different concentrations (10, 20, 30, 40 and 50 μmol/L) was added to the cells which activity was detected. At the same time, transcriptome sequencing was performed on scar tissue and cells, and bioinformatics methods were used to explore potential targets and signaling pathways of scar tissue fibrosis. RESULTS: Aloe-emodin had a concentration dependent inhibitory effect on fibroblast proliferation,with the 40 μmol/L concentration group showing the most significant effect. The results of tissue and cell sequencing indicated that differentially expressed genes were significantly enriched in extracellular matrix-receptor interaction pathway, and shared a common differential gene which was THBS1. The ORA analysis results indicated that differentially expressed genes, including THBS1, were significantly enriched in the PI3K-Akt signaling pathway. CONCLUSIONS Aloe emodin may inhibit the PI3K-Akt pathway by downregulating THBS1, thereby reducing the proliferation activity of fibroblasts derived from postoperative palatal scar tissue.
Thrombospondin 1/genetics* ; Humans ; Signal Transduction/drug effects* ; Fibroblasts/cytology* ; Proto-Oncogene Proteins c-akt/metabolism* ; Fibrosis ; Phosphatidylinositol 3-Kinases/metabolism* ; Cicatrix/metabolism* ; Cell Proliferation/drug effects* ; Anthraquinones/pharmacology* ; Cells, Cultured

Objective To investigate the protective effects of Qiji Special Dietary Food on exercise-induced myocardial injury in mice.Methods An exercise-induced myocardial injury model was established using a treadmill running protocol,and at the same time,the modeled mice were administered Qiji Special Dietary Food via oral gavage,followed by a 4-week treadmill exhaustion test.Serum levels of cardiac troponin T(cTnT),creatine kinase(CK),and lactate dehydrogenase(LDH)were measured.Myocardial tissues were analyzed for superoxide dismutase(SOD)activity and malondialdehyde(MDA)content.Histopathological alterations and ultrastructural changes in myocardial tissue were evaluated using light microscopy and transmission electron microscopy(TEM).Results After 4 weeks of exhaustive training,compared to the control group,the model group exhibited significantly elevated serum cTnT,CK,and myocardial MDA levels(P<0.01),along with reduced myocardial SOD activity(P<0.01).Compared to the model group,high-,medium-,and low-dose treatment significantly attenuated the exhaustive exercise-induced increases in serum cTnT level and myocardial MDA content(P<0.01),restoring these indicators to the levels comparable to those of the normal control group(P<0.05).Additionally,all treatment groups had markedly increased myocardial SOD activity,with no significant difference from the normal group.Histopathological and ultrastructural analyses revealed markedly alleviated myocardial damage in the treatment groups,with the medium-dose group exhibiting the most pronounced protective effects.Conclusion Qiji Special Dietary Food demonstrates significant protective effects against exercise-induced myocardial injury in mice,which maybe associated with its antioxidant activity and mitigation of oxidative stress.

Objective To analyze the elevated blood pressure status and its influencing factors among manufacturing workers in Foshan City. Methods A total of 565 795 manufacturing workers who underwent occupational medical examinations in Foshan City from 2017 to 2023 were included. Data of workers were obtained from the Guangdong Provincial Key Occupational Disease Monitoring and Management Platform. The influencing factors of elevated blood pressure were analyzed using descriptive epidemiological methods. Results A total of 89 526 cases of elevated blood pressure were detected among the 565 795 workers, with a detection rate of 15.82%. From 2017 to 2023, the annual detection rate of elevated blood pressure was 14.11%, 15.00%, 14.69%, 15.94%, 17.00%, 16.90%, and 16.68%, respectively, showing an overall upward trend (P<0.01). The top three industries with the highest detection rates of elevated blood pressure were instrument and meter manufacturing; wood processing and the manufacture of wood, bamboo, rattan, palm, and straw products; as well as agricultural and sideline food processing. Binary logistic regression analysis showed that male workers had a higher risk of elevated blood pressure than female workers (P<0.01). The risk of elevated blood pressure in workers increased with age, years of occupational hazard exposure, and duration of dust exposure (all P<0.05). Workers exposed to noise for more than three years had a higher risk of elevated blood pressure than those without noise exposure (P<0.05). The risk of elevated blood pressure among workers in Nanhai, Shunde, and Gaoming districts was higher than that in Chancheng District (all P<0.01). Workers originating from Northeast and Northwest China had a higher risk of elevated blood pressure than those from North China (all P<0.05). Workers from smaller-scale enterprises had higher risk of elevated blood pressure (all P<0.01). Conclusion Targeted blood pressure health interventions in Foshan City should be strengthened for male manufacturing workers who are older, have longer working years, and originate from Northeast and Northwest China. The development of exemplary health enterprises should be promoted, particularly among micro- and small-sized enterprises and key industries. Enhanced occupational health management is especially needed for positions with high intensity noise exposure and high concentration dust exposure.