Objective: To investigate the effect of concentrated growth factor (CGF) on the biological performance of human dental pulp stem cells (hDPSCs) under oxidative stress status induced by hydrogen peroxide (H2O2). Methods: The hDPSCs were isolated by using tissue block separation method from healthy permanent teeth extracted for orthodontic reason. hDPSCs surface markers CD34, CD45, CD90 and CD105 were detected by flow cytometry. Alkaline phosphatase (ALP), alizarin red S (ARS), oil red O staining and colony formation assay were used to identify hDPSCs. After the cell counting kit-8 (CCK-8) detection, the optimal H2O2 concentration was used to construct the hDPSCs oxidative stress model. CGF conditioned medium was prepared by repeated freeze-thaw methods. After CCK-8 detection, the optimum CGF concentration was chosen for the subsequent experiments. The hDPSCs were divided into control group, H2O2 (only H2O2 processing), H2O2+CGF group (H2O2 processing in combination with the CGF) and CGF group (only CGF processing). Subsequent experiments were performed according to these groups. The oxidative stress model was verified by reactive oxygen species, β-galactosidase staining and Western blotting. The effects of CGF on the proliferation and migration of hDPSCs under oxidative stress status were detected by CCK-8 and cell scratch assay, respectively. ALP activity and ARS staining were used to detect the effect of CGF on the osteogenic differentiation of hDPSCs under oxidative stress status. The mRNA expression levels of odontogenesis related genes were detected by real-time fluorescence quantitative PCR (RT-qPCR), and the expression levels of odontogenesis and osteogenesis related proteins were detected by Western blotting. Results: Isolated hDPSCs showed positive expression of mesenchymal stem cells surface markers of CD90, CD105, and negative expression of hematopoietic stem cells surface markers CD34, CD45. The hDPSCs were proved to have the capacity of osteogenic, adipogenic differentiation and clone formation. The optimal concentration to construct the oxidative stress model was 200 μmol/L H2O2. Twenty percent CGF was the optimal concentration for subsequent experiments. Compared with the control group, the expression of aging protein p53 was significantly up-regulated from (0.82±0.12) to (1.19±0.14) in H2O2 group (P<0.05), with deepened β-galactosidase staining and increased fluorescence intensity of reactive oxygen species. The proliferative capacity of cells in H2O2+CGF group on day 1, 3, 5 and 7 (0.23±0.01, 0.50±0.02, 1.60±0.07, 1.80±0.21) were all higher than in H2O2 group (0.15±0.01, 0.14±0.02, 0.50±0.03, 0.90±0.09) (P<0.05). Cell healing capacity of cells in H2O2+CGF group at 12 h and 24 h [(47±7)%, (58±44)%] also increased compared with the H2O2 group [(36±2)%, (44±2)%] (P<0.05), and similar results in the activity of ALP and the formation of mineralized nodules. On day 28, the mRNA expressions of dentin sialophosphoprotein (0.52±0.16) and dental matrix protein 1 (DMP-1) (0.39±0.13) in H2O2 group were all significantly lower than those in H2O2+CGF group (0.96±0.24, 0.83±0.30, respectively) and CGF group (1.12±0.18, 1.23±0.19, respectively) (P<0.05). On day 28, the expressions of odontogenesis related protein DMP-1 (0.27±0.04) and osteogenesis related protein Runt-related transcription factor-2 (0.42±0.15) in H2O2 group were all significantly lower than those in H2O2+CGF group (0.66±0.18, 0.68±0.04) and CGF group (1.15±0.13, 1.06±0.19, respectively) (P<0.05). Conclusions: H2O2 can induce oxidative stress in hDPSCs, while CGF can promote proliferation, migration, odontogenic and osteogenic differentiation of hDPSCs under oxidative stress status.

To summarize the latest research progress of graphene and its derivatives (GDs) in bone repair.

Hexavalent chromium Cr(VI), as a well-established carcinogen, contributes to tumorigenesis for many human cancers, especially respiratory and digestive tumors. However, the potential function and relevant mechanism of Cr(VI) on the initiation of esophageal carcinogenesis are largely unknown. Here, immortalized human esophageal epithelial cells (HEECs) were induced to be malignantly transformed cells, termed HEEC-Cr(VI) cells, via chronic exposure to Cr(VI), which simulates the progress of esophageal tumorigenesis. In vitro and in vivo experiments demonstrated that HEEC-Cr(VI) cells obtain the ability of anchorage-independent growth, greater proliferative capacity, cancer stem cell properties, and the capacity to form subcutaneous xenografts in BALB/c nude mice when compared to their parental cells, HEECs. Additionally, HEEC-Cr(VI) cells exhibited weakened cell motility and enhanced cell adhesion. Interestingly, HEECs with acute exposure to Cr(VI) failed to display those malignant phenotypes of HEEC-Cr(VI) cells, suggesting that Cr(VI)‍-induced malignant transformation, but not Cr(VI) itself, is the cause for the tumor characteristics of HEEC-Cr(VI) cells. Mechanistically, chronic exposure to Cr(VI) induced abnormal activation of Notch signaling, which is crucial to maintaining the capacity for malignant proliferation and stemness of HEEC-Cr(VI) cells. As expected, N-‍[N-‍(3,5-difluorophenacetyl)‍-L-alanyl]‍-S-phenylglycine t-butyl ester (DAPT), an inhibitor for the Notch pathway, drastically attenuated cancerous phenotypes of HEEC-Cr(VI) cells. In conclusion, our study clarified the molecular mechanism underlying Cr(VI)‍-induced esophageal tumorigenesis, which provides novel insights for further basic research and clinical therapeutic strategies about Cr(VI)‍-associated esophageal cancer.

It has been popular and challenging to undertake researches on the delay of acquired resistance of epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKI). As key cells for tumor initiation, cancer stem cells (CSC) play an important role in the process of resistance to EGFR-TKI. Although preliminary studies found that traditional Chinese medicine (TCM) could inhibit CSC properties and delay EGFR-TKI resistance, the specific molecular mechanism remains unclear. By summarizing the empirical syndrome treatment of EGFR-TKI resistance via TCM and combining recent researches on TCM intervention in CSC to delay EGFR-TKI resistance, this review discussed the potential molecular pathways and mechanisms of deceleration in resistance to EGFR-TKI by TCM via inhibiting CSC characteristics, in order to expand the research ideas of TCM in combination with targeted therapy.

End-stage liver disease includes liver failure and decompensated cirrhosis resulting from various etiologies and often leads to patient mortality due to complications and clinical symptoms such as severe jaundice, ascites, hepatic encephalopathy, coagulopathy, and hepatorenal syndrome. Liver transplantation is currently regarded as the most effective treatment, but its clinical application is limited by the shortage of donors, elevated expenses, and post-transplant rejection. Stem cells are a group of cells with multidirectional differentiation potential and self-renewal ability, which can improve the clinical indicator outcomes through mechanisms such as immunoregulation and promotion of tissue repair in patients with end-stage liver disease. Clinical trials of stem cell therapy have achieved a series of results for end-stage liver disease, proving the safety and effectiveness of stem cell therapy. This article reviews the clinical studies that have been registered and published at home and abroad and provides a reference for the clinical plan on stem cell therapy for end-stage liver disease.

Immunotherapy has led to groundbreaking advances in anti-tumor treatment, yet significant clinical challenges remain such as the low proportion of beneficiaries and the lack of effective platforms for predicting therapeutic response. Organoid technology provides a novel solution to these issues. Organoids are three-dimensional tissue cultures derived from adult stem cells or pluripotent stem cells that closely replicate the structural and biological characteristics of native organs, demonstrating particularly strong potential in modeling the tumor microenvironment (TME). Tumor organoids can simulate TME effectively by retaining endogenous matrix components, including various immune cells, or by adding immune cells, cancer-associated fibroblasts, and other components. This provides a novel platform for predicting immunotherapy outcomes, evaluating adoptive cell therapies, and selecting personalized treatment options for patients. Summarizing strategies for constructing tumor organoids that simulate the microenvironment and understanding their advancements in immunotherapy research and clinical application can provide new insights for the development of tumor immunotherapy.

Islet transplantation is one of the most promising curative methods for type 1 diabetes mellitus (T1DM), but early hypoxic death of the graft post-transplantation impedes successful treatment. To improve the efficacy of islet transplantation and enhance islet cell resistance to hypoxia, reducing hypoxic injury before revascularization is crucial. Mesenchymal stem cells (MSCs) are known to regulate immune responses and protect against hypoxic damage through paracrine mechanisms. This study aims to verify the protective effects of MSC-conditioned medium (CM) in enhancing islet cells' tolerance to hypoxic conditions and preserving islet graft function.

β-Nicotinamide mononucleotide (NMN), as the precursor of nicotinamide adenine dinucleotide (NAD), plays an important role in enhancing NAD levels. Intake of NMN can alter the composition and vitality of gut microbiota, restore mitochondrial function, inhibit inflammatory pathways, improve metabolism, counteract oxidative stress, and alleviate inflammation. NMN significantly improves recovery from aging-related diseases, such as diminished heart function, reduced fertility, memory decline, and diabetes. NMN demonstrates both efficacy and safety in anti-aging. The use of NMN in China has gradually gained acceptance, highlighting the importance of exploring the mechanism of NMN in anti-aging effects and improving the biosynthesis of NMN. In addition, NMN in combination with stem cells hold promise in the treatment of aging-related degenerative diseases and promote overall human and animal health.

The objective of the present study was to investigate the role and mechanism of bone marrow microenvironmental cells in regulating the mitochondrial mass of leukemia cells, and to uncover the mechanism of leukemia progression at the metabolic level. A mouse model of acute myeloid leukemia (AML) induced by the overexpression of the MLL-AF9 (MA9) fusion protein was established, and the bone marrow cells of AML mice were transplanted into mitochondrial fluorescence reporter mice expressing the Dendra2 protein (mito-Dendra2 mice). The proportion of Dendra2+ cells in bone marrow leukemia cells at different stages of AML was quantified by flow cytometry. The effects of transferred mitochondria on leukemia cells were studied by fluorescence-activated cell sorting (FACS), followed by functional experiments and bulk RNA sequencing. Finally, components within the bone marrow niche, such as mesenchymal stromal cells (MSCs) and endothelial cells (ECs), were co-cultured with leukemia cells in vitro. The proportion of leukemia cells that underwent mitochondrial transfer and the apoptosis level of leukemia cells were then detected by flow cytometry. The results showed that mitochondria from bone marrow cells were transferred to leukemia cells in the AML mouse model, and the proportion of mitochondrial transfer decreased with AML progression. The proportion of mitochondria transferred to leukemia stem cells (LSCs) was lower than that of mature AML cells. In AML cells receiving Dendra2+ mitochondria, there was a significant increase in the levels of intracellular reactive oxygen species (ROS) and apoptosis, while the levels of protein translation and their colony-forming capacities were decreased. The transplantation of Dendra2+ AML cells resulted in an extension of the survival of mice. RNA sequencing analysis demonstrated a significant downregulation of pathways related to translation, aerobic respiration and mitochondrial organization in AML cells that had received mitochondria. In vitro co-culture experiments indicated that MSCs within the bone marrow niche tended to transfer their mitochondria to leukemia cells and promoted the apoptosis of leukemia cells. These results indicate that in the MA9-induced AML mouse model, bone marrow niche cells can transfer mitochondria to leukemia cells, resulting in a reduction in the overall survival and function of the leukemia cells. Mitochondrial transfer in the bone marrow microenvironment may serve as a self-defensive mechanism of the host bone marrow niche cells, inhibiting the progression of AML.

Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal disorder of hematopoietic stem cells induced by PIG-A gene mutations. It is clinically manifested by hemolysis, bone marrow failure, and high-risk concurrent thrombosis, which are life-threatening in severe cases. Significant progress has been made in the pathogenesis research and clinical diagnosis and treatment of PNH in recent years. The Red Blood Cell Disease (Anemia) Group Chinese Society of Hematology, Chinese Medical Association, combined with the latest diagnosis and treatment progress of PNH, relevant foreign guidelines/consensus, and China's national conditions, jointly formulated the "Guidelines for the diagnosis and management of paroxysmal nocturnal hemoglobinuria (2024) " based on extensive solicitation of expert suggestions and opinions. This article discussed the key and difficult issues in PNH diagnosis and treatment, interpreted the updated part of the guidelines, and expanded the relevant recommendations of the guidelines according to the latest research progress at home and abroad, thereby providing more references for clinical practice.

Cancer immunotherapy including immune checkpoint inhibitors and adoptive cell therapy has gained revolutionary success in the treatment of hematologic tumors; however, it only gains limited success in solid tumors. For example, chimeric antigen receptor T (CAR-T) cell therapy has shown significant effects and potential for curing patients with B-cell malignancies. In contrast, it remains a challenge for CAR-T cell therapy to gain similar success in solid tumors. The anti-tumor effect of endogenous or adoptively transferred tumor-specific T cells depends largely on their differentiation status. T cells at early differentiation stage show better anti-tumor therapeutic effects than fully differentiated effector T cells. In cancer patients, the persistence of tumor-specific T cells with the stem cell memory or precursor phenotype is significantly associated with improved therapeutic outcomes; therefore, adoptively transfered CAR-T cells with stem cell memory and/or central memory is expected to gain better anti-tumor effects. Herein we focused on the in vitro optimized culture and expansion system to obtain CAR-T cells with stem cell memory or central memory phenotype for the review.

Objective: This study aimed to investigate the association between early immune reconstitution and Epstein-Barr virus (EBV) reactivation by analyzing changes in natural killer (NK), B, and T cells and their functional status in the peripheral blood during the early post-transplant period. Methods: This study included 23 patients who underwent haplo-hematopoietic stem cell transplantation (HSCT). The immune reconstitution of NK cells, T cells, and B cells as well as the expression levels of NK and T cell exhaustion markers (PD-1, TIM-3, and CTLA-4) and cytotoxic function at 1, 2, and 3 months post-transplantation were compared between patients with EBV activation (EBV+ group) and those without activation (EBV- group) post- transplantation. Results: EBV activation occurred in nine patients post-transplantation (EBV+ group), whereas 14 patients demonstrated no activation (EBV- group). All patients with EBV activation exhibited EBV viremia, and no EBV-associated diseases occurred. No significant differences in the clinical characteristics were found between the two groups of patients. The median proportion of CD3(+)CD8(+) T cells in the EBV+ group was significantly lower than that in the EBV- group at 1 month post-transplantation (P=0.033). The median proportion of the CD3(-)CD16(neg)CD56(bri) subset in the EBV+ group was significantly higher than that in the EBV- group at 2 months post-transplantation (P=0.046). No significant differences in the median proportions of CD3(-)CD19(+) B cells were observed between the two groups at 1, 2, and 3 months post-transplantation. The expression of CTLA-4 on CD3(-)CD16(bri)CD56(dim) NK cells in the EBV+ group was significantly higher than that in the EBV- group at 1 month post-transplantation (P=0.033). The expression of TIM-3 on CD3(+)CD8(+) T cells in the EBV+ group was significantly higher than that in the EBV- group (P=0.009). The expression level of TIM-3 on CD3(-)CD16(neg)CD56(dim) NK cells in the EBV+ group was significantly lower than that in the EBV- group at 2 months post-transplantation (P=0.023). The expression levels of TIM-3 on CD3(+)CD4(+) T cells in the EBV+ group than those in the EBV- group at 1 and 3 months post-transplantation (P=0.002, P=0.043). The median positive rate of Granzyme B expression in CD3(+)CD8(+) T cells and CD3(+)CD4(+) T cells in the EBV+ group was significantly lower than that in the EBV- group at 1-month post-transplantation (P=0.033, P=0.016). The median positive rate of Granzyme B expression in the CD3(-)CD16(bri)CD56(neg) cell subset in the EBV+ group was higher than that in the EBV- group at 2 months post-transplantation (P=0.012). The median positive rate of Granzyme B expression in CD3(+)CD4(+) T cells in the EBV+ group remained significantly lower than that in the EBV- group at 2 months post-transplantation (P=0.049). The median positive rate of perforin expression in the CD3(-)CD16(bri)CD56(dim) cell subset was significantly higher in the EBV+ group than in the EBV- group at 3 months post-transplantation (P=0.003). The median positive rate of IFN-γ expression in CD3(+)CD8(+) T cells in the EBV+ group was significantly lower than that in the EBV- group at 3 months post-transplantation (P=0.036) . Conclusion: Delayed NK cell and T lymphocyte reconstitution, high exhaustion marker expression, and weakened cytotoxic functions may be related to EBV reactivation after haploidentical HSCT.

Retinitis pigmentosa (RP) is a group of inherited retinal diseases characterized by progressive loss of photoreceptor cells and retinal pigment epithelium function. Its treatment has long been a focus and challenge in ophthalmic research. Despite advances in therapies such as stem cell transplantation, gene therapy, and retinal prosthetic implants, many difficulties remain. Optogenetic technology, an emerging biological technique combining optics and genetics, holds promise for treating RP by inducing the expression of light-sensitive proteins in degenerated retinal cells, thereby partially restoring patients' vision and light sensitivity. However, limitations such as difficulties in selecting target cells, potential risks of viral transfection, and uncertainties in visual restoration still exist. With the continuous improvement of light sensitivity of optogenetic proteins and the development of viral vectors with higher transfection efficiency, it is believed that optogenetic therapy will help more patients regain vision.

Objective: To investigate whether there is mitochondrial transfer in dental mesenchymal stem cells (MSCs) and its significance for the odontogenic differentiation. Methods: Flow cytometry and immunohistochemical staining were used to isolate dental mesenchymal stem cells. Immunofluorescence staining and live cell imaging were applied to determine whether there is mitochondrial transfer in dental MSCs. Transcriptome sequencing data re-analysis of human dental pulp stem cells (DPSCs) and bone marrow mesenchymal stem cells (BMSCs) from gene expression omnibus (GEO) data base demonstrated the importance of mitochondrial transfer in dental MSCs. Cells were managed with mitochondrial transfer inhibitor ML141 with dimethyl sulfoxide as the control. Immunofluorescence staining, senescence-associated β-galactosidase (SA-β-gal) staining, reactive oxygen species (ROS) assay, 5-ethynyl-2'-deoxyuridine(EdU) labelling, cell counting kit-8 (CCK-8) assay, Western blotting, live cell imaging and transmission electron microscope were used to investigate cell morphology, ROS level, cellular senescence, cell proliferation, MSCs marker paired related homeobox 1 (Prrx1) and Sp7 transcription factor (Sp7) expression, mitochondrial transfer and mitochondrial morphology, respectively. Further, after using ML141 during the induction of odontogenic differentiation, alkaline phosphatase (ALP) chromogenic kit was used to detect ALP activity and real-time fluorescence quantitative PCR (RT-qPCR) was used to detect the expression of odontogenic differentiation-related genes Alp, Sp7, dentin matrix protein 1 (Dmp1), and dentin salivary phosphoprotein (Dspp), which were applied to investigate the effect of mitochondrial transfer on odontogenic differentiation. Results: An ultrafine tunneling nanotubes (TNTs) structure labelled with F-actin existed between dental MSCs, and the presence of transferring mitochondria in this structure was also confirmed. Transcriptome sequencing data suggested that the gene expression profiles were significantly different between DPSCs and BMSCs. Genes related to mitochondrial transfer and mitochondrial dynamic were significantly increased in DPSCs compared to BMSCs. Compared with the control group, treatment with 1, 5, 10 μmol/L ML141, the mitochondrial transfer inhibitor, had little significant effects on the cell morphology, cytoskeleton and ROS level. SA-β-gal activity and the proportion of SA-β-gal positive cells in the ML141-treated groups [(3.93±0.21)%, (3.23±0.42)%, (4.06±0.84)%] had no significant differences with the control group [(3.83±0.28)%] (all P>0.05). In the cell proliferation assay, the proportion of EdU positive cells in the ML141-treated groups [(20.00±3.82)%, (19.48±1.96)%, (12.55±2.86)%] had no significant differences (all P>0.05) with the control group [(18.57±0.87)%], whereas the CCK-8 assay showed similar results in ML141-treated group of 1, 5 μmol/L (all P>0.05). Western blotting results showed that the protein expression levels of PRRX1 and SP7 in the ML141-treated group had no significant differences with the control group. Live cell imaging showed that compared with the control group [(31.42±4.01)%], the proportion of TNTs and mitochondrial transfer in the ML141-treated groups [(13.45±1.46)%, (10.36±3.47)%, (9.32±1.11)%] were significantly decreased in dental MSCs (all P<0.001). Scanning electron microscope showed that the mitochondrial morphology of dental MSCs in the ML141-treated group was similar to the control group, with globular and short-rod shape. After 7 days of odontogenic differentiation, the ALP staining intensity of the ML141-treated group was significantly lower than the control group. After 21 days of induction, RT-qPCR results showed that compared with control group, the relative mRNA expressions of Alp, Sp7, Dmp1 and Dspp were significantly decreased in the ML141-treated group (all P<0.05), indicating that the suppression of mitochondrial transfer in dental MSCs inhibited the odontogenic differentiation. Conclusions: Mitochondrial transfer exists between dental MSCs, and inhibition of mitochondrial transfer impairs the odontogenic differentiation.

Mesenchymal stem cells (MSC) possess unique immunomodulatory properties and have enormous potential in the treatment of graft-versus-host disease (GVHD). However, the low implantation and survival rates of MSC in vivo, coupled with their weak immunosuppressive functions, have resulted in unstable clinical efficacy in the treatment of GVHD. Preconditioning of MSC with hypoxia, active molecules and gene modification can enhance the function of MSC and improve the implantation rate, survival rate and therapeutic effect of MSC. This review summarized the strategies for enhancing the efficacy of MSC in the treatment of hematopoietic stem cell transplantation complicated with GVHD in recent years, aiming to provide new strategies for optimizing the application of MSC in the prevention and treatment of GVHD.

To harvest the primary Philadelphia chromosome-positive (Ph+) cells of B-acute lymphoblastic leukemia (B-ALL) and to establish the B-ALL mouse model.

To establish an efficient gene editing method of HLA-I gene to prepare HLA-I universal hematopoietic stem cells.

To prepare mesenchymal stem cells with antioxidant capacity (AO-MSC) from human umbilical cords and evaluate its cell biological properties.

To summarize the clinical characteristics of patients with combined pneumocystis jiroveci pneumonia (PJP) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

To investigate the correlation of the clinical characteristics, fever characteristics, serum biomarkers with cytokine release syndrome (CRS) in patients with relapsed/refractory multiple myeloma (R/R MM) treated with chimeric antigen receptor T cell (CAR-T) immunotherapy.