To investigate the antitumor effects and underlying molecular mechanisms of the p53 reactivator APR-246 in bladder cancer cells.

Human and murine bladder cancer cell lines with different p53 statuses were treated with APR-246 to assess drug sensitivity. p53-knockdown mutant cell models were established to evaluate the dependency of APR-246 on p53. For knockdown experiments, cells transduced with lentiviral shScramble were named as Scramble group, whereas cells transduced with shp53 were named as shp53 group. For overexpression experiments, cells transduced with empty pLKO-NC lentivirus were served as the control (NC) , and cells transduced with lentivirus overexpressing p53R248S were named as the OE-p53R248S group. Cell viability assay, ROS staining, and Western blot were performed to analyze cell death patterns and related protein expression. Cells were treated with DMSO (control) , 20 μmol/L APR-246, or 20 μmol/L APR-246 combined with 5 mmol/L NAC. Flow cytometry and a subcutaneous xenograft model in C3H mice were used to validate the biological effects in vitro and in vivo. Differences between two groups were assessed using the independent-samples t test. Comparisons among multiple groups were conducted by one-way analysis of variance (ANOVA) , followed by Dunnett's t test for multiple comparisons between each treatment group and the control group.

APR-246 significantly induced cell death in bladder cancer cell lines (EJ: IC₅₀ = 14.09 μmol/L; T24: IC₅₀ = 13.56 μmol/L; UMUC3: IC₅₀ = 15.14 μmol/L; SW780: IC₅₀ = 29.71 μmol/L; MB49: IC₅₀ = 7.02 μmol/L; MBT2: IC₅₀ = 1.12 μmol/L; all P < 0.05) , whereas mutant p53 cells exhibited greater sensitivity to the compound. Knockdown of p53 attenuated cellular responses to APR-246 (UMUC3: 17.69 ± 0.21 vs 24.16 ± 0.24; EJ: 12.24 ± 0.17 vs 19.61 ± 0.29; T24:11.39 ± 0.26 vs 16.24 ± 0.35; all P < 0.05) .Overexpression of mutant p53 reduced drug responsiveness [UMUC3: (18.93 ± 0.57) vs (13.42 ± 0.45) μmol/L; EJ: (13.05 ± 0.32) vs (8.21 ± 0.27) μmol/L; T24: (13.23 ± 0.46) vs (9.21 ± 0.42) μmol/L; all P < 0.05]. Cell death induced by APR-246 was primarily mediated through ROS-dependent ferroptosis and apoptosis. Treatment with APR-246 could downregulate the expression of SLC7A11 and GPX4 (P < 0.05) and increased the expression level of cleaved PARP (P < 0.05) . Furthermore, in bladder cancer cells harboring p53 missense mutations, APR-246 upregulated the expression of p21 and induced cell cycle arrest at the G0/G1 phase [ (37.44 ± 0.65) % vs (53.32 ± 0.83) %; P < 0.05]. In vivo studies further confirmed that APR-246 markedly inhibited tumor growth (P < 0.05) .

APR-246 exerts potent antitumor activity in bladder cancer by reactivating mutant p53 and inducing ROS-dependent ferroptosis and cell cycle arrest, providing a promising molecular target and experimental basis for precision therapy of bladder cancer.

To explore the mechanismof forkhead box p1 (FoxP1) , a forkhead box transcription factor, mediates the autophagy pathway to regulate the process of transforming growth factor-β1 (TGF-β1) -induced endothelial to mesenchymal transition (EndMT) .

The human umbilical vein endothelial cells were divided into the following groups: control group (Blank) , TGF-β1-induced EndMT model group (TGF-β1) , small interfering RNA negative control group (TGF-β1+si-NC) , FoxP1 small interfering RNA group (TGF-β1+si-FoxP1) , overexpressing RNA negative control group (TGF-β1+oe-NC) , FoxP1 overexpressing group (TGF-β1+oe-FoxP1) , and FoxP1 overexpression combined with an autophagy inhibitor group (TGF-β1+oe-FoxP1+3-MA) . Western blot were used to detect the expression of endothelial、mesenchymal markers and collagen. The mCherry-EGFP-LC3 dual fluorescence system was used to detect autophagy status. Cell migration ability was assessed through scratch assay. The independent samples t-test was used for the comparisons between two groups, while for comparisons among multiple groups, one-way analysis of variance (ANOVA) was employed, and the LSD-t method was used for pairwise comparisons.

Western blot results showed that, compared to the control group, the expression of endothelial markers VE-cadherin and CD31 were decreased in the TGF-β1 group, while the expression of mesenchymal markers α-SMA,Vimentin and collagen proteins Collagen Ⅰ (3.08 ± 0.09 vs 1.00 ± 0.08) , Collagen Ⅲ (3.14 ± 0.10 vs 1.00 ± 0.05) were increased. In addition, the cell migration ability [ (75.20 ± 4.30) %vs (36.80 ± 2.60) %] and the expression of p62 (2.24 ± 0.07 vs 1.00 ± 0.07) were increased in TGF-β1 group when compared to control group, while the expression levels of Beclin-1 (0.49 ± 0.03比1.00 ± 0.02) and LC3 Ⅱ/Ⅰ (0.17 ± 0.01 vs 1.00 ± 0.08) were decreased (all P < 0.01) . Compared to the TGF-β1+oe-NC group, the expression of VE-cadherin, CD31 and Beclin-1 (0.82 ± 0.01 vs 0.49 ± 0.03) , LC3 Ⅱ/Ⅰ (0.55 ± 0.02 vs 0.20 ± 0.01) (all P < 0.01) were increased in the TGF-β1+oe-FoxP1 group, while the expression of α-SMA, Vimentin and collagen proteins Collagen Ⅰ (2.08 ± 0.10 vs 4.38 ± 0.15) , Collagen Ⅲ (1.86 ± 0.07 vs 3.60 ± 0.14) as well as p62 (1.77 ± 0.09 vs 2.24 ± 0.08) , were decreased, along with areduced cell migration ability [ (46.66 ± 5.15) %比(77.56 ± 7.30) %] (all P < 0.01) . Compared to the TGF-β1+oe-FoxP1 group, the expression of Beclin-1 (1.60 ± 0.03 vs 1.96 ± 0.02) , LC3 Ⅱ/Ⅰ (2.20 ± 0.04 vs 3.88 ± 0.16) and E-cadherin, CD31 were decreased in the TGF-β1+oe-FoxP1+3-MA group, while the expression of p62 (0.70 ± 0.02 vs 0.49 ± 0.05) , Collagen I (0.80 ± 0.02 vs 0.51 ± 0.03) , CollagenⅢ (0.70 ± 0.01 vs 0.29 ± 0.02) and α-SMA, Vimentin were increased, along with the enhanced cell migration ability [ (58.63 ± 6.19) %比(40.84 ± 5.27) %] (all P < 0.01) .

Overexpression of FoxP1 can activate the autophagy pathway to inhibit the TGF-β1-induced EndMT process,while could provide a novel cytological perspective for the mechanistic research and therapeutic approaches to myocardial fibrosis.

To establish an induced pluripotent stem cells (iPSCs) model derived from peripheral blood mononuclear cells (PBMCs) of Down syndrom (DS) patients, providing a new research tool for studying DS and its associated neurological disorders.

The PBMCs of two children with DS were reprogrammed into iPSCs by using the electric reprogramming technology. The obtained iPSCs were systematically characterized by morphological observation, karyotype analysis, flow cytometry, immunofluorescence staining, and three-germ-layer differentiation experiments. Flow cytometry data are expressed as the percentage of positive cells.

Two iPSCs lines, DS1-iPSCs and DS2-iPSCs, with typical morphology were successfully obtained. Karyotype analysis revealed a karyotype of 47,XX or XY,+21, confirming that both patients had Trisomy 21. Flow cytometry and immunofluorescence results showed that the iPSCs expressed pluripotency markers SSEA4, TRA-1-60, OCT4, and SOX2. Three-germ-layer differentiation experiments confirmed that the iPSCs could differentiate into endoderm (AFP, SOX17) , mesoderm (α-SMA, BRACHYURY) , and ectoderm (Tuj1, PAX6) cells, demonstrating their pluripotency.

This study successfully established iPSCs model derived from the peripheral blood of two DS children, which possess trisomy 21 karyotype, expression of pluripotency markers and the ability to differentiate from the three germ layers, offering a new cell model and research tool for investigating DS and its associated neurological disorders.

Medulloblastoma (MB) is the most common malignant brain tumor in children. Currently, the conventional clinical treatments mainly include surgical resection, radiotherapy and chemotherapy. However, the heterogeneity of tumor cells and their interaction with complex microenvironments significantly increase the difficulty of precise treatment, leading to limited efficacy of traditional intervention methods and great challenges for patient prognosis. In recent years, the breakthrough development of single-cell RNA sequencing (scRNA-seq) technology has provided high-resolution research tools for understanding the biological mechanism of MB and the selection of therapeutic targets. This technology reveals the heterogeneous lineage characteristics and the dynamic evolution patterns of the microenvironment of MB tumor cell subtypes at the single-cell level through single-cell gene expression profiling analysis. This article systematically reviews the latest research progress of scRNA-seq in the heterogeneity of MB and the characteristics of the microenvironment, and explores the significance of these findings for understanding the biological mechanism of MB and its potential application value in clinical treatment. Meanwhile, in view of the challenges faced by scRNA-seq, such as the capture efficiency of tumor sample heterogeneity and the integration analysis of spatial transcriptomes, this review proposes its future research directions and application potential for the precise diagnosis and treatment of MB.

The aging of hematopoietic stem cells (HSCs) is the result of the combined effects of intrinsic cellular senescence programs and changes in the hematopoietic microenvironment, which is characterized by diminished self-renewal capacity, myeloid differentiation bias, homing and colonization defects, and the reduction of clonal diversity, etc. These functional declines significantly increase the risk of malignant blood disorders and various aging-related diseases. The application of technologies such as single-cell RNA sequencing, epigenomic mapping, and gene editing has not only elucidated the core mechanisms of HSCs aging such as gene damage, epigenetic alterations and changes in the hematopoietic microenvironment, but also led to the proposal of multiple strategies for HSCs rejuvenation. This review systematically summarizes recent advances in HSCs aging research, aiming to identify key regulatory nodes and potential intervention targets, providing a solid theoretical foundation and technical prospects for the prevention and treatment of related aging diseases.

Postoperative cognitive dysfunction (POCD) is a common central nervous system complication in elderly patients following surgery, primarily characterized by cognitive impairments such as memory decline, inattention, and reduced executive function. Currently, there is a lack of effective clinical strategies for its prevention and treatment. Mesenchymal stem cells (MSCs) have garnered widespread attention in the intervention of neurodegenerative and injury-related neurological diseases due to their multi-lineage differentiation potential and significant paracrine effects. This review systematically elucidates the core mechanisms by which MSCs prevent and treat POCD, with a focus on summarizing their therapeutic potential through multiple pathways including immunomodulation, neurotrophic support, alleviation of oxidative stress, and repair of the blood-brain barrier. It also analyzes the selection of MSCs sources, administration strategies, and preclinical research evidence. Furthermore, the review outlines future directions, such as optimization through cellular engineering techniques, to advance MSCs as a novel and effective therapeutic approach for POCD prevention and treatment.

Psychiatric disorders are a class of diseases with a high prevalence and complex treatment. Traditional drug treatments are often accompanied by metabolic side effects, while psychological interventions are clearly ineffective in terms of efficacy, which make the need for innovative treatments for psychiatric disorders even more urgent. Due to the characteristics of self-renewal, multi-directional differentiation and paracrine regulation, stem cells have opened up new strategies for the treatment of a variety of diseases. This review summarizes the stem cell applications and mechanisms in schizophrenia, depression, and autism, further highlights current clinical progress to inform optimization of future therapeutic strategies.

The promulgation of the "Regulations on the Clinical Research and Clinical Translation Application of New Biomedical Technologies" (hereinafter referred to as the "Regulations") constitutes a milestone in the history of regulating new biomedical technologies in China, marking the formal entry of this field into a new stage of rule-of-law characterized by "lenient entry, strict exit, and strong supervision". Based on the author's long-term practice experience in the field of clinical research and clinical transformation of stem cells, this article provides a systematic analysis and deep thinking on the legislative background, core framework, and underlying governance logic of the "Regulations". The article points out that the "lenient entry" established by the "Regulations" is, in essence, a dialectical unity of simplified procedural thresholds and strengthened substantive responsibilities. The "strict exit" emphasized by the "Regulations" builds a solid quality and ethical defense line for clinical translation through a national-level "dual-evaluation" mechanism and whole-cycle dynamic supervision. The "strong supervision" it relies on constructs a whole-process, clear-responsibility chain and deterrent system by granting substantive enforcement powers, establishing graded legal liabilities, and implementing a "dual punishment" system for individuals. Particularly importantly, the "Regulations" thoroughly unblock the medical technology pathway within the "dual-track" system for stem cell therapy, and provide clear guidance for technological innovation and translation through the "three stages, dual reviews, dual evaluations" (i.e., "3+2+2") regulatory framework. However, as a framework legislation, the supporting rules and technical guidelines for key links such as the review mechanism, qualification of assessment bodies, clinical application access standards, and the classification criteria of technologies and products in the "Regulations" still need to be clarified, and its implementation still faces challenges from "text" to "practice". Based on this, from the multiple perspectives of medical institutions, enterprises, and regulatory coordination, this article proposes systematic and actionable implementation suggestions, emphasizing that all parties need to plan ahead, consolidate internal capabilities, and collaborate to build together, in order to translate policy opportunities into clinical value, and jointly promote the standardized, safe, and efficient development of China's new biomedical technology industry.