To explore the application value of the adherent culture method of large tissue blocks in the in vitro isolation, culture and large-scale preparation of human umbilical cord mesenchymal stem cells (hUC-MSCs).

The large tissue block method was used to isolate and culture hUC-MSCs. Cell morphology and growth status were observed by an inverted phase-contrast microscope. Cell proliferation was detected using the CCK-8 assay. Multidirectional differentiation potential was assessed through osteogenic/chondrogenic induction experiments. Cell surface markers were identified using flow cytometry.

Compared with the traditional adherent culture method, the harvest of hUC-MSCs was higher by the large tissue block method (1.5 × 10⁶ cells/dish vs 1.0 × 10⁶ cells/dish), with a statistically significant difference (P < 0.05). The proliferative activity of cells obtained by both methods was comparable, and there was no statistically significant difference in doubling time (P > 0.05). Cell migration was observed after 5 days of culture, fibroblast-like cells migrated out from 6 to 10 days, and the fusion rate exceeded 80 % after 2 weeks. The proliferation activity of P3 generation cells was excellent, the surface markers met international standards, and they had the ability to differentiate into osteoblasts and chondrocytes.

The large tissue block method is simple to operate, highly efficient and low-loss, could obtain high-purity and high-activity hUC-MSCs, with good promotion value.

To investigate the mechanism of tetramethylpyrazine (TMP) in alleviating CCl₄-induced liver fibrosis in mice by regulating ferroptosis through the SLC7A11/GPX4 signaling axis.

Thirty male C57BL/6 mice were randomly divided into control group, CCl₄ model group, and low-, medium-, high-dose TMP groups (50, 100, and 150 mg/kg). Except for the control group, mice were intraperitoneally injected with a mixture of CCl₄ and olive oil (1 : 10, 5 mL/kg) twice weekly for 8 weeks. From week 5, mice in the TMP groups were administered TMP by gavage once daily for 4 weeks. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured to evaluate liver function. Histopathological changes were assessed by hematoxylin-eosin (HE), Masson's trichrome, and Sirius red staining. The expression of α-SMA, COL1A1, Fibronectin, SLC7A11, and GPX4 was detected by immunohistochemistry and Western blot, and their mRNA levels were determined by qPCR. Fe²⁺, malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) levels were measured using commercial assay kits. One-way analysis of variance (ANOVA) was used for comparison among multiple groups, and Tukey's post-hoc test was used for pairwise comparison between groups. If the data did not conform to normal distribution or homogeneity of variance, Kruskal-Wallis H test was adopted, and Dunn's test was used for pairwise comparison between groups.

Compared with the model group, the levels of ALT [(251.70 ± 10.80) vs (450.00 ± 14.49) U/L] and AST [(236.70 ± 10.80) vs (427.50 ± 12.14) U/L], the degree of collagen fiber deposition in liver tissue [Masson: (1.10 ± 0.07)% vs (1.72 ± 0.17) %, Sirius Red: (0.17 ± 0.02) % vs (0.45 ± 0.04) %], the protein levels of α-SMA (1.17 ± 0.27 vs 8.08 ± 0.48), COL1A1 (4.30 ± 0.73 vs 18.78 ± 0.93) and Fibronectin (1.60 ± 0.32 vs 7.75 ± 0.60), the contents of Fe²⁺ [(2.60 ± 0.14) vs (3.63 ± 0.19) μmol/g protein] and MDA [(0.72 ± 0.03) vs (1.12 ± 0.07) nmol/mg protein] were decreased in the high-dose TMP group, while the protein levels of SLC7A11 (0.98 ± 0.05 vs 0.29 ± 0.03) and GPX4 (0.73 ± 0.08 vs 0.14 ± 0.03), the activity of SOD [(1 413.00 ± 47.19) vs (1 147.00 ± 42.74) U/mg protein] and the content of GSH [(1.60 ± 0.07) vs (1.15 ± 0.07) μmol/g protein], the mRNA levels of SLC7A11 (0.80 ± 0.07 vs 0.10 ± 0.03) and Gpx4 (0.75 ± 0.07 vs 0.37 ± 0.05) were increased, all these differences were statistically significant (all P < 0.05).

TMP may alleviate CCl₄-induced liver fibrosis in mice by regulating the SLC7A11/GPX4 signaling axis to inhibit ferroptosis, which has potential anti-fibrotic application value.

To prepare the effects of rat liver decellularized extracellular matrix scaffold on pancreatic islet cell viability and secretory function, providing a novel biomaterial for pancreatic islet transplantation in type 1 diabetes treatment.

Rat livers were processed using perfusion decellularization. The scaffold structure and core components were characterized through histological staining, immunofluorescence staining, and scanning electron microscopy. The retention of bioactive factors such as platelet-derived growth factor and fibroblast growth factor in the scaffold was detected using ELISA kits. Biocompatibility was assessed via DNA content quantification, DAPI staining, and CCK-8 assay using L929 cells. Functional modification of the decellularized scaffold was achieved by perfusion with platelet-rich plasma (PRP). The effects of the PRP-modified scaffold on pancreatic islet cell viability and function were verified using immunofluorescence staining and cell viability staining. Comparisons between two groups were performed using independent samples t-test, comparisons among three groups were conducted using one-way analysis of variance (ANOVA), followed by LSD-t test for pairwise comparisons. For the two-factor design, a two-way repeated-measures ANOVA was performed, with Bonferroni correction applied for multiple comparisons.

The liver decellularized scaffold prepared by perfusion appeared translucent white macroscopically, retained the natural multi-level vascular structure, and exhibited a porous network morphology microscopically. Various staining methods confirmed significant decellularization efficiency with preservation of core matrix proteins such as type I collagen and fibronectin, as well as multiple bioactive factors. Specifically, the relative retention rates of fibroblast growth factor and glycosaminoglycans reached (88.05 ± 9.87)% and (90.77 ± 3.32)%, respectively. The scaffold demonstrated good biocompatibility, being acellular with residual DNA content as low as (5.25 ± 2.18) ng/mg, and significantly promoted L929 cell proliferation. The scaffold enhanced the insulin secretory function of pancreatic islet cells, the stimulation index in the glucose-stimulated insulin secretion assay for the scaffold group was significantly higher than that of the control group (2.78 ± 0.08 vs 2.33 ± 0.14), with statistical significance (P < 0.01). PRP modification significantly increased the content of growth factors, such as platelet-derived growth factor, in the scaffold, showing statistical significance (P < 0.05). The PRP-modified scaffold positively promoted pancreatic islet cell viability and function.

The preparation process for the rat liver decellularized scaffold is reliable, yielding a scaffold with both structural integrity and bioactivity. The PRP-modified scaffold effectively supports the maintenance of islet cell viability and function, presenting a promising novel scaffold material and technical strategy for islet transplantation.

To investigate whether the mechanism by which high glucose weaken the protective effect of sevoflurane pretreatment on rat cardiomyocytes is related to the HIF-1α/LDHA signaling pathway.

H9c2 cardiomyocytes were randomly divided into 8 groups using a random number table, and named control group (N+Con group), hypoxia-reoxygenation group (N+H/R group), sevoflurane preconditioning group (N+SPC group), high glucose control group (H+Con group), high glucose-hypoxia-reoxygenation group (H+H/R group), high glucose sevoflurane preconditioning group (H+SPC group), high glucose sevoflurane preconditioning + HIF-1α agonist group (H+SPC+D group), and high glucose sevoflurane preconditioning + HIF-1α agonist + LDHA inhibitor group (H+SPC+D+A group). Cell viability of cardiomyocytes was assessed using the CCK-8 assay. Lactate dehydrogenase (LDH) levels in the culture medium were measured via the microplate method. Lactate levels were determined using the WST-8 assay. ATP levels were detected by chemiluminescence assay. The levels of key glycolytic enzymes were quantified using ELISA. Apoptosis rate was evaluated by flow cytometry, and mitochondrial membrane potential was measured using a fluorescent probe. The expression levels of HIF-1α and LDHA were detected by western blotting. Comparisons among multiple groups were performed using one-way ANOVA, and pairwise comparisons between groups were conducted using Tukey's multiple comparison test.

Compared with the N+SPC group, the H+SPC group showed decreases in cell viability [(70.12 ± 3.81)%vs (86.66 ± 5.48)%], lactate level [(5.36 ± 0.18) vs (6.94 ± 0.45) mmol/L], ATP level [(88.90 ± 17.33) vs (172.88 ± 26.83) μmol/L], mitochondrial membrane potential (0.55 ± 0.11 vs 0.78 ± 0.08), the activities of HK, PFK1, and PK [(72.43 ± 8.77) vs (134.23 ± 8.84) U/mL, (104.62 ± 10.65) vs (213.30 ± 12.36) U/mL, (92.87 ± 5.14) vs (251.67 ± 14.45) U/mL], the expression of HIF-1α (0.54 ± 0.01 vs 0.94 ± 0.04) and LDHA (0.69 ± 0.02 vs 1.07 ± 0.04) were decreased in the H+SPC group, while the LDH level in culture medium [(315.12 ± 25.22) vs (122.87 ± 8.03) U/L ] and apoptosis rate were increased [(21.42 ± 2.11)% vs (10.92 ± 0.69)%]. All differences were statistically significant (all P < 0.05). Compared with the H+SPC group, the cell viability [(83.72 ± 4.55)%vs (70.12 ± 3.81)%], lactate level [(6.40 ± 0.42) vs (5.36 ± 0.18) mmol/L], ATP level [(165.62 ± 24.28) vs (88.90 ± 17.33) μmol/L], mitochondrial membrane potential (0.82 ± 0.05 vs 0.55 ± 0.11), the activities of HK, PFK1, and PK [(122.34 ± 8.74 vs 72.43 ± 8.77) U/mL, (192.45 ± 20.98) vs (104.62 ± 10.65) U/mL, (208.76 ± 32.65) vs (92.87 ± 5.14) U/mL], the expression of HIF-1α (0.80 ± 0.03 vs 0.54 ± 0.01) and LDHA (0.87 ± 0.04 vs 0.69 ± 0.02) were increased in the H+SPC+D group, whereas culture medium LDH level [(154.33 ± 26.85) vs (315.12 ± 25.22) U/L ] and apoptosis rate [(15.43 ± 1.07)% vs (21.42 ± 2.11)%] were decreased. All differences were statistically significant (all P < 0.05). Compared with the H+SPC+D group, the cell viability [(68.43 ± 3.20)% vs (83.72 ± 4.55)%], lactate level [(5.03 ± 0.12) vs (6.40 ± 0.42) mmol/L], ATP level [(79.54 ± 19.22) vs (165.62 ± 24.28) μmol/L], mitochondrial membrane potential (0.57 ± 0.05 vs 0.82 ± 0.05), the activities of HK, PFK1, and PK [(66.83 ± 6.91) vs (122.34 ± 8.74) U/mL, (77.83 ± 5.94) vs (192.45 ± 20.98) U/mL, (132.80 ± 3.53) vs (208.76 ± 32.65) U/mL], the expression of HIF-1α (0.35 ± 0.02 vs 0.80 ± 0.03) and LDHA (0.45 ± 0.02 vs 0.87 ± 0.04) were decreased in the H+SPC+D+A group, while culture medium LDH level [(264.96 ± 13.46) vs (154.33 ± 26.85)U/L] and apoptosis rate [(19.83 ± 0.23)%vs (15.43 ± 1.07)%] were increased. All differences were statistically significant (all P < 0.05).

The mechanism by which high glucose attenuates the protective effect of sevoflurane preconditioning on rat cardiomyocytes may be related to the inhibition of HIF-1α/LDHA signaling pathway activity.

To explore the key genes, core cell subsets, and cell communication mechanisms underlying neural injury repair after intracerebral hemorrhage.

The rat intracerebral hemorrhage transcriptome dataset GSE288102, mouse intracerebral hemorrhage microglia transcriptome dataset GSE266602, and mouse intracerebral hemorrhage brain tissue single-cell RNA sequencing dataset GSE167593 were downloaded from the GEO database. The limma package was used to screen differentially expressed genes (DEGs), and the intersection of DEGs from the two transcriptome datasets was obtained to identify common DEGs. GO and KEGG enrichment analyses were performed using the DAVID database, and a protein-protein interaction (PPI) network was constructed via the STRING database. Key genes were screened using Cytoscape. Quality control, dimensionality reduction, clustering, and cell annotation of single-cell data were completed with the Seurat package, and intercellular communication pathways were analyzed using CellChat.

A total of 96 common DEGs were identified. GO terms was mainly enriched in immune cell activation, leukocyte activation, and injury repair-related processes, while KEGG enrichment was found in the ECM-receptor interaction pathway and the PI3K-Akt signaling pathway. Ten hub genes were screened, including LGALS3, ITGB2, TLR2, TIMP1, FN1, CD44, ICAM1, CCL2, TGFB1, and ITGAM. Single-cell RNA analysis identified five core cell populations: neurons, oligodendrocytes, oligodendrocyte precursor cells, microglia, and astrocytes. The hub genes showed specific expression in each cell subset: LGALS3 was downregulated and TGFB1 was upregulated in neurons; TGFB1, CCL2, and ICAM1 were significantly upregulated in astrocytes; and FN1 and TGFB1 were upregulated in microglia. Cell communication analysis revealed that SPP1- (ITGA5+ITGB1) and SPP1- (ITGAV+ITGB1) were the core signaling pathways for neural repair after intracerebral hemorrhage.

This study identified 10 core genes, 5 key cell populations, and 2 core cell communication pathways involved in neural injury repair after intracerebral hemorrhage, providing a reliable basis for investigating the mechanisms of neural repair and screening potential therapeutic targets for intracerebral hemorrhage.

Inherited primary arrhythmia syndromes (IPAS) encompass a group of disorders stemming from genetic anomalies in cardiac ion channels or their regulatory proteins, frequently associated with an elevated risk of sudden cardiac death. These diseases manifest a broad spectrum of clinical presentations, ranging from asymptomatic states to syncope, arrhythmias, and conduction disturbances, potentially leading to unexplained sudden cardiac death in phenotypically healthy individuals. Induced pluripotent stem cells (iPSCs) and induced pluripotent stem cells-derived cardiomyocytes (iPSC-CMs) offer robust in vitro disease models. It not only helps to analyze the mechanism of patient-specific diseases, but also avoids species differences in animal models and restrictions on the acquisition of human tissue samples. This article briefly introduces the current research status of iPSCs and iPSC-CMs, and summarizes the progress in the construction of patient-specific iPSC-CMs models, drug screening, and mechanistic studies for long QT syndrome, Brugada syndrome, early repolarization syndrome, and catecholaminergic polymorphic ventricular tachycardia.

Adipose-derived mesenchymal stem cells (ADSCs) have emerged as promising seed cells for bone tissue repair in regenerative medicine, owing to their distinct advantages including abundant sources, minimally invasive isolation, low immunogenicity, and multidirectional differentiation potential. The directional homing of ADSCs to the medullary cavity represents a core prerequisite for their therapeutic efficacy in bone regeneration. However, the specific regulatory details underlying this homing process remain poorly elucidated, which poses a critical bottleneck restricting their clinical translation. Focusing on the unique microenvironment of the medullary cavity, this review systematically summarizes the exclusive molecular mechanisms governing ADSCs homing to the medullary cavity, clarifies the bone marrow-targeted guidance characteristics of chemokine signaling axes, the patterns of integrin-mediated transendothelial migration across bone marrow vascular endothelium, as well as the synergistic regulatory network of intracellular signaling pathways. It further analyzes the specific effects of intrinsic cellular properties, medullary cavity microenvironment, and exogenous interventions on homing efficiency, and elucidates the bone marrow-specific mechanisms by which ADSCs promote bone tissue repair after successful homing. This review aims to provide a targeted theoretical basis and research insights for optimizing the medullary cavity homing efficiency of ADSCs and facilitating their clinical translation in the field of bone repair.

The prevalence of diabetes continues to rise, and traditional treatments struggle to reverse the decline in pancreatic islet β cell function. Cell therapy offers a new pathway for functional cure by restoring or replacing endogenous insulin secretion. This article systematically reviews the clinical progress of four major strategies: islet transplantation, stem cell-derived islets, mesenchymal stem cells (MSCs), and immune cell therapy. Islet transplantation has been clinically applied but is limited by the shortage of donors; stem cell-derived islets have made breakthroughs in providing a limitless cell source; MSCs improve glycemic control through immunomodulation; and immune cell therapy aims to restore immune tolerance. Each approach faces challenges such as immune rejection, cell survival, and functional maturation. Emerging technologies like gene editing, cell encapsulation, and 3D bioprinting offer novel solutions to these bottlenecks. Future research should focus on the development of universal cell products, precise immunological interventions, and multi-mechanism synergistic therapies to propel diabetes treatment from "glucose control" towards "functional reconstruction".