Objective:Chimeric antigen receptor T-cell(CAR-T)immunotherapy has made major breakthroughs in the treatment of blood tu-mors.However,current CAR-T therapies face several limitations:they require autologous cells,involve a lengthy and costly production pro-cess,and use lentiviral transduction that carry risk of insertional carcinogenesis due to random integration.Therefore,there is an urgent need to develop a universal cost-effective cancer immunotherapy method generating CAR-T cells for in vivo cancer immunotherapy.Meth-ods:This study successfully established an exosome-mediated,T-cell targeted delivery system,demonstrating both precise design and func-tional efficacy for biomedical applications.To optimize CAR-T cell generation the transfection dose was adjusted,and the kinetics of CAR-T cell percentage were recorded.The cytotoxicity of the resulting CAR-T cells was evaluated in vitro by calcein-AM release.To test the tumor-killing in vivo of engineered exosomes,human PBMCs were injected into NPG mice via the tail vein to establish humanized mice,followed by intravenous injection of tumor cells to induce cancer.Results:To overcome the limitations of conditional autologous CAR-T cells,we de-veloped a T cell-targeted exosome system capable of specifically targeting human CD3+,CD4+,and CD8+T cells.CAR-T production was dose-dependent,with transfection efficiency reaching upto 97.8%at 106 particles/cell.Both in vitro cytotoxicity assays and in vivo animal experi-ments demonstrated that exosome-incubated CAR-T cells effectively eliminated CD19-positive Raji cells,highlighting their specificity and therapeutic potential in antigen-directed applications.Conclusions:We successfully established a CD8-targeting exosome delivery system for CAR-T cell production capable of transforming CD8+T cells into functional CAR-T cells,which showed significant tumor-killing ability in vitro and in mice.Compared with the traditional lentiviral vector for the preparation of CAR-T cells in vitro,in vivo-reprogrammed CAR-T cells us-ing our CD8-targeted exosome delivery system,with higher transfection efficiency,shorter production period,lower cost,and eliminated the risk of insertion carcinogenesis.This strategy promises to bring a new era of universal CAR-T medicine,which can improve cancer immuno-therapy and may hold promise as a therapeutic platform to treat various diseases.

Most cancers are currently incurable, partly due to abnormal post-translational modifications (PTMs). In this study, we initially used multiple myeloma (MM) as a working model and found that SUMOylation activating enzyme subunit 1 (SAE1) promotes the malignancy of MM. Through proteome microarray analysis, SAE1 was identified as a potential target for bioactive colcemid or its derivative colchicine. Elevated levels of SAE1 were associated with poor clinical survival and increased MM proliferation in vitro and in vivo. Additionally, SAE1 directly SUMOylated and upregulated the total protein expression of p27, leading to LLPS-mediated nuclear export of p27. Our study also demonstrated the involvement of SAE1 in other types of cancer cells, and provided the first monomer crystal structure of SAE1 and its key binding model with colchicine. Colchicine also showed promising results in the Patient-Derived Tumor Xenograft (PDX) model. Furthermore, a controlled clinical trial with 56 MM patients demonstrated the clinical efficacy of colchicine. Our findings reveal a novel mechanism by which tumor cells evade p27-induced cellular growth arrest through p27 SUMOylation-mediated nuclear export. SAE1 may serve as a promising therapeutic target, and colchicine may be a potential treatment option for multiple types of cancer in clinical settings.

Objective:Chimeric antigen receptor T-cell(CAR-T)immunotherapy has made major breakthroughs in the treatment of blood tu-mors.However,current CAR-T therapies face several limitations:they require autologous cells,involve a lengthy and costly production pro-cess,and use lentiviral transduction that carry risk of insertional carcinogenesis due to random integration.Therefore,there is an urgent need to develop a universal cost-effective cancer immunotherapy method generating CAR-T cells for in vivo cancer immunotherapy.Meth-ods:This study successfully established an exosome-mediated,T-cell targeted delivery system,demonstrating both precise design and func-tional efficacy for biomedical applications.To optimize CAR-T cell generation the transfection dose was adjusted,and the kinetics of CAR-T cell percentage were recorded.The cytotoxicity of the resulting CAR-T cells was evaluated in vitro by calcein-AM release.To test the tumor-killing in vivo of engineered exosomes,human PBMCs were injected into NPG mice via the tail vein to establish humanized mice,followed by intravenous injection of tumor cells to induce cancer.Results:To overcome the limitations of conditional autologous CAR-T cells,we de-veloped a T cell-targeted exosome system capable of specifically targeting human CD3+,CD4+,and CD8+T cells.CAR-T production was dose-dependent,with transfection efficiency reaching upto 97.8%at 106 particles/cell.Both in vitro cytotoxicity assays and in vivo animal experi-ments demonstrated that exosome-incubated CAR-T cells effectively eliminated CD19-positive Raji cells,highlighting their specificity and therapeutic potential in antigen-directed applications.Conclusions:We successfully established a CD8-targeting exosome delivery system for CAR-T cell production capable of transforming CD8+T cells into functional CAR-T cells,which showed significant tumor-killing ability in vitro and in mice.Compared with the traditional lentiviral vector for the preparation of CAR-T cells in vitro,in vivo-reprogrammed CAR-T cells us-ing our CD8-targeted exosome delivery system,with higher transfection efficiency,shorter production period,lower cost,and eliminated the risk of insertion carcinogenesis.This strategy promises to bring a new era of universal CAR-T medicine,which can improve cancer immuno-therapy and may hold promise as a therapeutic platform to treat various diseases.

Background: In the field of neuroscience research, executive functions (EFs) are of great significance. Patients with Parkinson’s disease (PD) often encounter the problem of EFs impairment, which severely affects their lives and health.However, currently, the methods for evaluating EFs in experimental mice are limited, and there is a lack of effective evaluation indicators for touchscreen behavioral tests in different disease model mice. This study aims to establish a paradigm process and an evaluation baseline for touchscreen behavioral analysis in PD model mice, deeply explore the mechanisms of EFs impairment in PD, and provide a crucial foundation for subsequent research and treatment. Methods: Thirty clean - grade SNCA*A53T transgenic mice and forty clean - grade C57BL/6J wild - type mice were selected. For A53T mice, genetic identification was carried out. Molecular biology techniques such as PCR were used to determine their genetic characteristics. Protein detection was conducted through methods like Western blot to clarify the expression of relevant proteins. In terms of behavioral tests, the five - choice serial reaction time task (5 - CSRT) was adopted, and various behavioral data of mice in this task were recorded. Four groups were set up: the control group, the MPTP group. Different groups of mice were given specific treatments. For example, the MPTP group of mice was injected with MPTP to construct a drug - induced PD model. Principal component analysis (PCA) and receiver operating characteristic (ROC) curves were used to analyze the obtained touchscreen behavioral baseline indicators of mice, and key indicators were screened and evaluated. Results: In the 5 - CSRT, the optimal stage for wild - type C57 mice to achieve an accuracy rate of ≥ 80% was from the 11th to the 13th cycle, while for A53T mice, it was the 11th cycle. The EFs of A53T mice were impaired, with abnormalities in the accuracy rate, trace number, and number of punished times in the 5 - CSRT. When identifying drug - induced PD models, the 13th cycle of the 5 - CSRT was more effective; when identifying transgenic PD models, the 11th cycle was more suitable. Interventions could be carried out after the baseline accuracy rate reached 80%. Through the “genetic - environmental” dual - axis drive, the “chronic - acute” time - dimension complementarity, and the “mechanism - transformation” multi - level verification, the A53T and MPTP dual models were used to comprehensively cover the pathological cycle of PD EFs impairment. Conclusions This study has successfully established a paradigm process and an evaluation baseline for touchscreen behavioral analysis in PD model mice. This provides an important basis for a deep understanding of the mechanisms of EFs impairment in PD patients, has potential guiding significance for the development of intervention strategies and treatment methods for EFs impairment in PD patients, and is expected to promote new progress in PD research in the field of neuroscience.

Objective: Heavy metal and harmful element contamination are frequently reported in Chinese herbal medicines (CHMs), and roots and rhizomes parts showed a higher content than other parts. To investigate the residue level and assess the potential human health risk of heavy metals and harmful elements in roots and rhizomes, 720 batches of the sample representing 20 species of herbs from different sources were collected. Methods: The content of Pb, Cd, As, Hg, and Cu in the digests was determined using ICP-MS. The chronic hazard index estimate based on non-cancer hazard quotient (HQ) was applied for potential health risk assessment of Pb, Cd, As, Hg, and Cu via consumption of CHMs. Results: Compared with the Chinese limit standard (Chinese Pharmacopoeia Commission, 2020 edition) of Pb, Cd, As, Hg, and Cu in Ginseng Radix et Rhizoma, the exceedance percentage of Pb in total samples was 14.1%, which were generally far higher than Cd, As, Hg, and Cu. Health risk assessment results based on hazard quotient calculating showed that total HQ of Cu, Pb, As, Cd, and Hg in Pulsatillae Radix and Clematidis Radix et Rhizoma exceeded 1, with the value of 1.543 and 1.235. Besides, Arsenic had the highest HQ value (0.957) in Pulsatillae Radix. Conclusion: Consuming raw materials of Pulsatillae Radix and Clematidis Radix et Rhizoma may pose a potential risk and Arsenic residues in Pulsatillae Radix deserved special attention.

Objective: To explore the efficacy and safety of chimeric antigen receptor T (CAR-T) cells in the treatment of central nervous system leukemia (CNSL). Methods: Two leukemia patients with CNSL were treated with CD19-CAR-T cells. The process and results of the entire treatment is reported and related literature review is conducted. Results: The patients were diagnosed as acute myeloid leukemia (AML)-M₂ with B lymphoid antigen expression and B cell acute lymphoblastic leukemia(B-ALL) by morphology and immunophenotype assay. The immunophenotype was consistent with the abnormal manifestations of AML-M₂ and B-ALL. Their clinical manifestations and laboratory tests met the diagnostic criteria of CNSL. The diagnosis was clear and the two patients were treated with CD19-CAR-T cell immunotherapy. Central nervous system symptoms were relieved. The imaging abnormalities of patient one has disappeared but cytokines release syndrome (CRS) occurred during the treatment. Cerebrospinal fluid of patient two was negative and no obvious CRS reaction was found. Conclusions CAR-T cell immunotherapy is likely to induce the remission of CNSL and improve the prognosis.

Objective To explore the value of the molecular probe USPIO-PEG-sLeX on nasopharyngeal carcinoma xenograft in nude mice.Methods The USPIO nanoparticles was synthesized by physical deposition method,and which was modified by PEG to synthesize USPIO-PEG-sLeX .The nude mice of nasopharyngeal carcinoma xenograft were divided into experimental and control groups.USPIO-PEG-sLeX and USPIO-PEG were injected into nude mice of experimental and control groups by caudal vein,respectively.MR T2 mapping imaging was scanned before and after the injection,and analyzed the changes of T2 values between experimental and control groups. Results USPIO-PEG-sLeX had a good representation.The non-enhanced T2 values between control and experimental group had no statistical significance (P >0.05).However,T2 values of the mice in two groups before and after injections were statistically significant (P <0.05);and T2 values of experimental group were much lower than that of the control group after the injection,additionally,the difference of enhanced rate between the two groups was statistically significant (P <0.05).Conclusion USPIO-PEG-sLeX magnetic nanoparticles is potential to be a targeted contrast agent to ELAM-1 expression of nasopharyngeal carcinoma,and can be valuable in non-invasive dynamic monitoring the expression of ELAM-1.