To summarize and analyze the clinical features of blastic plasmacytoid dendritic cell neoplasm (BPDCN), so as to enhance the understanding of this disease.

Petroleum hydrocarbon pollutants in soil are challenging to biodegrade, negatively impacting plant growth as well as the metabolic activity and community structure of soil microorganisms. Microorganisms immobilized by seed carriers can synergistically contribute to the remediation of petroleum hydrocarbon-contaminated soil. We prepared a rape seed carrier with immobilized microorganism by seed coating (with a mixture of diatomaceous earth and bentonite as fillers) and microbial immobilization. A pot experiment was conducted with the following treatments: control (CK, neither seeds nor microorganisms added), bare rapeseed (T1), rapeseed coated with diatomaceous earth and bentonite (T2), free-living Pseudomonas aeruginosa added (T3), rapeseed coated with diatomaceous earth and bentonite plus free-living P. aeruginosa (T4), and rapeseed coated with diatomaceous earth and bentonite immobilized with P. aeruginosa (T5). We measured rape seed growth, rhizosphere microbial community structure, and petroleum hydrocarbon removal efficiency. The results showed that 1) There were no significant difference in seed germination rate among T1, T2, T4, and T5 treatments. Compared to T1, leaf length, root length, biomass, and soluble protein content of rape seed significantly increased in T4 and T5 treatments, while T2 treatment showed no significant effect. Leaf width, stem length, chlorophyll content, and superoxide dismutase activity of rape seed in T2, T4, and T5 treatments were significantly higher than T1, while malondialdehyde content was signi-ficantly lower. 2) Compared to CK, the removal rate of petroleum hydrocarbon in the T1, T2, T3, T4, and T5 treatments increased by 0.8, 1.6, 0.5, 1.8, and 2.2 times, respectively. The T5 treatment achieved the highest petro-leum hydrocarbon removal rate of 54.0%. Soil dehydrogenase activity in all treatments increased significantly, with a positive correlation with the petroleum hydrocarbon removal rate (r=0.893). 3) The T5 treatment had the highest soil microbial α diversity and the abundances of Chloroflexi and Acidobacteria. In conclusion, seed carriers with immobilized microorganisms could regulate plant growth, modify the structures of microbial communities, enhance the biological activity of soil enzymes, thereby improving petroleum hydrocarbon removal efficiency. This provides a novel environmentally friendly approach for the joint remediation of petroleum hydrocarbon-polluted soil by plants and microorganisms.

To observe the role of miR-139-5p and Notch1 signaling pathway in regulation of homing of bone mesenchymal stem cells (BMSCs) of asthmatic rats.

To observe the reparative effects of human umbilical cord mesenchymal stem cell (hUC-MSC) transplantation on white matter injury (WMI) in neonatal rats and explore its mechanism through the nuclear factor-kappa B (NF-κB) signaling pathway mediated by microglial cells.

In order to establish a stable in vitro culture platform for chicken small intestine three-dimensional (3D) organoids, in this study, crypt cells were collected from the small intestine of 18-day-old embryos of AA broilers. On the basis of the L-WRN conditioned medium, we optimized the culture conditions of chicken small intestinal organoids by adjusting the proportions of nicotinamide, N-acetylcysteine, LY2157299, CHIR99021, Jagged-1, FGF, and other cytokines to select the medium suitable for the long-term stable growth of the organoids. The optimization results showed that the addition of 1.5 µmol/L CHIR99021 significantly improved the organoid formation efficiency and organoid diameter. When 0.5 µmol/L Jagged-1 was added, a small amount of bud-like tissue appeared in organoids. After the addition of 50 ng/mL FGF-2, the rate of organoid germination was significantly increased. The 1.5 µmol/L CHIR99021, 0.5 µmol/L Jagged-1, and 50 ng/mL FGF-2 added in the medium can cooperate with each other to improve the formation and speed up the proliferation and differentiation of organoids, while improving the stemness maintenance of cells. The morphology, cell types, and culture characteristics of chicken small intestinal organoids were studied by HE staining, transmission electron microscopy, reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), indirect immunofluorescence, and immunohistochemistry. The results showed that the 3D organoids of the chicken small intestine cultured in vitro were morphologically consistent with the chicken intestinal tissue and contained differentiated epithelial cells. In summary, we successfully established an in vitro culture system for chicken small intestinal organoids, providing a new method for the subsequent research on chicken intestinal physiology, pathology, and host-pathogen interaction mechanism and the development of relevant drugs.

The purpose of this study is to construct a muscle-specific synthetic promoter library, screen out muscle-specific promoters with high activity, analyze the relationship between element composition and activity of highly active promoters, and provide a theoretical basis for artificial synthesis of promoters. In this study, 19 promoter fragments derived from muscle-specific elements, conserved elements, and viral regulatory sequences were selected and randomLy connected to construct a muscle-specific synthetic promoter library. The luciferase plasmids pCMV-Luc and pSPs-Luc were constructed and transfected into the myoblast cell line C2C12. The activities of the synthesized promoters were evaluated by the luciferase activity assay. Two non-muscle-derived cell lines HeLa and 3T3 were used to verify the muscle specificity of the highly active promoters. The sequences of promoters with high activity, good muscle specificity, and correct sequences were analyzed to explore the relationship between the element composition and activity of promoters. We successfully constructed a muscle-specific promoter library and screened out 321 effective synthetic promoter plasmids. Among them, the activity of SP-301 promoter was 5.63 times that of CMV. The 15 promoters with high activity were muscle-specific. In the promoters with high activity and correct sequences, there was a relationship between their element composition and activity. Muscle-specific elements accounted for a high proportion in the promoters, while they had weak correlations with the promoter activity, being tissue-specific determinants. Viral elements accounted for no less than 20% in highly active promoters, which may be the key elements for the promoter activity. The content of conserved elements was proportional to the promoter activity. This study lays a theoretical foundation for the synthesis of tissue-specific efficient promoters and provides a new idea for the construction and application of in-situ gene delivery systems.

This study aims to investigate the effects of Bushen Huoxue Decoction regulating adipose-derived stem cells(ADSCs)-exosomes(Exos) on the apoptosis of intervertebral disc nucleus pulposus cells(NPCs) and extracellular signal-regulated kinase(ERK) signaling pathway. Tert-butyl hydrogen peroxide(TBHP)-induced NPCs were divided into control, model, drug-containing serum, blank Exos, normal serum Exos, and drug-containing serum Exos groups. Cell viability and proliferation were examined by the CCK-8 assay and EdU staining, respectively. The cell cycle and apoptosis were evaluated by flow cytometry. Enzyme-linked immunosorbent assay was employed to measure the levels of interleukin(IL)-1β, tumor necrosis factor(TNF)-α, and IL-6. The mRNA levels of aggrecan, collagen type Ⅱ alpha 1 chain(COL2A1), and ERK were determined by qRT-PCR, and the protein levels of aggrecan, COL2A1, and p-ERK were determined by Western blot. The results showed that compared with the model group, the treatments with drug-containing serum, blank Exos, and normal serum Exos enhanced the viability and proliferation of NPCs, decreased the proportion of cells in the G_0/G_1 phase, increased the proportion of cells in the S phase, reduced apoptosis, lowered the levels of IL-1β, TNF-α, and IL-6, up-regulated the mRNA and protein levels of aggrecan and COL2A1, and down-regulated the mRNA level of ERK and the protein level of p-ERK. Compared with the drug-containing serum, blank Exos, and normal serum Exos groups, the treatment with drug-containing serum Exos enhanced the viability and proliferation of NPCs, decreased the proportion of cells in the G_0/G_1 phase, increased the cells in the S phase, reduced apoptosis, lowered the levels of IL-1β, TNF-α, and IL-6, up-regulated the mRNA and protein levels of aggrecan and COL2A1, and down-regulated the mRNA level of ERK and the protein level of p-ERK. The results confirmed that the Exos secreted by ADSCs after treatment with Bushen Huoxue Decoction-containing serum promoted the proliferation of degenerated NPCs, inhibited apoptosis and the expression of inflammatory mediators, and promoted the production of proteoglycans and collagen, thus delaying the progression of intervertebral disc degeneration, the mechanism of which was related to the regulation of the ERK signaling pathway.

This study explores the reparative effect of Qixiong Zuogui Compound Prescription(QXZG) intervention on the blood-brain barrier(BBB) in the aging brain with middle cerebral artery occlusion(MCAO) in rats mediated by bone marrow stem cells(BMSCs)-derived exosomes, as well as its anti-aging mechanism. An aging MCAO composite model was established using D-galactose-induced aging combined with line embolism. Rats were divided into young sham surgery group, aging sham surgery group, model group, exosome group, and exosome with traditional Chinese medicine(TCM) intervention group. SA-β-gal staining combined with TTC staining were used to evaluate the model establishment. Exosomes group or exosomes with TCM intervention group were injected via the tail vein 2 h post-surgery and 48 and 96 h post-ischemia reperfusion. Neurological deficit scores were assessed at 2 h, 3 d, and 7 d post-modeling. Cognitive function was evaluated using novel object recognition tests on the 7th day, and brain infarct volume was observed via MRI. Evans Blue staining was used to evaluate BBB permeability. Western blot was performed to detect the expression of tight junction proteins ZO-1 and Occludin in brain tissue. ELISA was used to measure serum matrix metalloproteinase(MMP)-9 and MMP-2 levels. SA-β-gal staining was used to assess cellular senescence. RT-qPCR was performed to detect p21 mRNA expression, and qPCR was used to measure relative telomere length. The results showed that QXZG intervention-derived BMSCs exosomes significantly reduced brain infarct volume in aging MCAO rats, improved neurological deficit scores, and enhanced cognitive function. These exosomes suppressed MMP-2 and MMP-9 expression, promoted he expression of tight junction proteins ZO-1 and Occludin, reduced BBB permeability, lowered p21 mRNA expression, elongated telomeres, and delayed cell senescence. In conclusion, this study confirms that QXZG intervention-derived BMSC exosomes may repair the BBB by reducing MMP-2 and MMP-9 levels and increasing ZO-1 and Occludin expression. Moreover, its anti-aging mechanism may involve reducing p21 mRNA expression and limiting telomere shor-tening.

The establishment of modern biliary surgery system, alongside pivotal scientific paradigm shifts, has heralded a new era featured by precision, personalization, life-cycle care, and multidisciplinary management in the treatment of both benign and malignant biliary diseases. However, two formidable challenges persist in haunting the treatment of biliary diseases: (1) The refinement of surgical techniques has reached a plateau in reducing the disability associated with benign biliary conditions and in improving survival outcomes in biliary tract cancers; (2) Traditional evidence-based clinical studies have shown limited power in addressing complex dilemmas, such as determining whether to excise or preserve pathological gallbladders or selecting the optimal biliary drainage strategy. Consequently, the authors propose the conceptual framework of "biliary ecosystem". In this model, diverse and abundant cholangiocytes represent forest, while blood vessels, nerves, and lymphatic vessels serve as nurturing soil, biliary stem cells function as seeds, bile flows like river network, and hepatocytes mark the river's origins. Both benign and malignant biliary diseases exhibit significant spatiotemporal dynamics. The bile ducts form the "macro" environment, bile constitutes the "sub-macro" environment, and diverse cellular niches create the microenvironment. Specific pathological biliary conditions are shaped by intricate regulatory mechanisms that operate across these three tiers. Within the biliary ecosystem, cellular subpopulations exist remarkable diversity with states of homeostasis, oscillation, perturbation, or imbalance, underpinned by complex signaling networks. This holistic approach allows us to reframe and critically examine the pressing scientific issues confronting biliary tract diseases. Based on this framework, the authors distill key scientific questions and offer preliminary recommendations for embracing the paradigm shift. The authors anticipate that this conceptual model will promote interdisciplinary integration and accelerate clinical and translational researches.

Recurrence and metastasis remain the leading cause of death in breast cancer patients due to the lack of effective treatment. A microenvironment suitable for cancer cell growth, referred to as pre-metastatic niche (PMN), is formed in distant organs before metastasis occurs. Myeloid-derived suppressor cells (MDSCs) are a heterogenous population of immature myeloid cells with immunosuppressive effects. They can expand in large numbers in breast cancer patients and participate in the formation of PMN. MDSCs can remodel the extracellular matrix of pulmonary vascular endothelial cells and recruit cancer stem cells to promote the lung metastasis of breast cancer. Furthermore, MDSCs facilitate immune evasion of breast cancer cells to impact the efficacy of immunotherapy. It is proposed that MDSCs represent a potential therapeutic target for the inhibition of recurrence and metastasis in breast cancer. Therapeutic strategies targeting MDSCs have shown promising efficacy in preclinical studies and clinical trials. This review presents a summary of the principal factors involved in the recruitment and activation of MDSCs during the formation of PMN, and outlines MDSCs functions such as immunosuppression and the current targeted therapies against MDSCs, aiming to provide new ideas for the treatment of distant metastases in breast cancer.

Objective: To investigate the role of RNA m6A reader YTHDF2 in neurodegeneration of the aged mice. Methods: Eighteen 18-month-old control C57BL/6 mice and 22 Ythdf2 conditional knockout (cKO) mice of the same age (that exhibited significant aging characteristics) were used. Five pairs of mice were used for morphological analysis. Thirteen control mice and 17 cKO mice were used for behavioral experiments. Immunofluorescence analysis was performed to detect the expression of YTHDF2 in the brains of 18-month-old C57BL/6 mice. After establishing the neural progenitor cell-specific knockout mice of Ythdf2, their phenotypes were analyzed through comparison of body weight, brain weight and H&E staining. Subsequently, immunohistochemistry and immunofluorescence analyses were used to detect the expression of various neural cell-specific markers in the aged control mice and Ythdf2 cKO mice. Finally, behavioral tests, including the open field test, new object recognition, and water maze, were used to assess the levels of anxiety, depression, learning and memory abilities. Results: Immunofluorescence staining showed that YTHDF2 was mainly expressed in the neurons. Compared with the age-matched control mice, there was no significant change in the body weight of the Ythdf2 cKO mice, but the brain weight decreased significantly (P<0.05). The immunostaining showed that Ythdf2 cKO mice had fewer neurons, fewer astrocytes with defective morphology, more microglia and activation of microglia (P<0.05). Behavioral tests showed that the aged Ythdf2 cKO mice exhibited impaired movement, learning and memory abilities (P<0.05). Conclusions: YTHDF2 is mainly expressed in the neurons of the aged brain. Conditional knockout of Ythdf2 causes quantitative and structural abnormalities in hippocampal neuronal cells, and impairs motor ability and learning and memory of the aged mice, suggesting that YTHDF2 plays an important role in neurodegeneration of the aged mice.

Objective: To investigate the relationship between the expression patterns of SOX2 and FOXG1 and the differentiation/development level of neural components in immature teratoma and to determine the clinical significance and potential application of this correlation in a clinical setting. Methods: We conducted a comprehensive whole transcriptome sequencing analysis to identify differentially expressed genes (DEGs) across various subtypes of ovarian germ cell tumors. Additionally, immunohistochemical staining of paraffin-embedded tissue sections was employed to assess the nuclear staining pattern of SOX2 and FOXG1 proteins within the tumor tissues. Results: The transcriptome sequencing data showed that transcription factors SOX2 and FOXG1 exhibited high levels of expression typically in immature teratoma and occupied a pivotal position within the protein-protein interaction network. Immunohistochemical staining revealed the absence of both SOX2 and FOXG1 protein expression in dysgerminoma and yolk sac tumor samples. In immature teratoma, immunohistochemical staining demonstrated diffuse expression of SOX2 and FOXG1 proteins within the inner layer of densely-arranged primitive neuroepithelial tubules. This pattern of expression suggested the presence of stem cell-like properties within these tumor cells. In the sparsely peripheral neurogliocytes, FOXG1 maintained a diffuse nuclear staining pattern resembling that of neuroepithelial cells, while SOX2 exhibited a scattered pattern of positive staining, hinting at a neural lineage differentiation potential. This spatial differential expression pattern of SOX2 and FOXG1 proteins in immature teratoma suggested that primitive neural components within these tumors often recapitulated the trajectory of neural formation and cortical development that was typically observed during embryogenesis. The primitive neural tube acted as the center that constantly moved from inside to outside, with a dynamic shift from the interior to the exterior, paralleled by the sequential differentiation of cell lineages from primitive neuroepithelial stem cells to radial glia, intermediate progenitor cells, and ultimately to precursor glia. Conclusions: This spatial expression pattern of SOX2 and FOXG1 proteins observed in immature teratoma mirrors the lineage differentiation and migration trajectories of primitive neuroepithelial components typically seen in embryonic neurogenesis and cortical development. In daily practice, the combined application of SOX2 and FOXG1 SOX2 and FOXG1 helps identify the primitive neuroepithelial components in immature teratoma, avoid misjudgment of similar morphologies, and thereby assist in the histological grading and clinical decision-making.

Objective: To investigate the interventional effect of bone marrow mesenchymal stem cell (BMMSC) transplantation with different doses of X-ray irradiation induced hepatic injury in mice. Methods: Eighteen female C57BL/6J mice were randomly divided into 0, 2, and 3 Gy irradiation groups and 0, 2, and 3 Gy transplantation groups. The irradiation group was used as the control and injected with an equal volume of culture medium. The mice in the transplantation group were irradiated with different doses of X-ray irradiation, and BMMSCs were intravenously infused into the bone marrow. The mice were sacrificed for sampling at the end of the 21st day. Mice body weight changes were recorded daily. The changes in the content of peripheral blood lymphocytes, red blood cells, platelets, and hemoglobin were detected by an automatic blood tester. The morphological changes in mice liver tissues were observed by hematoxylin-eosin staining. The serum activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected by a biochemical analyzer. The reduced glutathione contents in liver tissue were detected by the microplate method. The malondialdehyde content in liver tissue was detected by thiobarbituric acid. The content of total superoxide dismutase (T-SOD) in liver tissue was detected by the hydroxylamine method. The expression of the F4/80 protein in liver tissue was detected by the immunohistochemistry method. The protein expression of nuclear transcription factor erythroid 2 related factor 2 (Nrf2) and heme oxygenase 1 (HO-1) in liver tissue was determined by the western blotting method. The mRNA expression of NLRP3, IL-6, and Nrf2 in liver tissue was detected by a real-time quantitative polymerase chain reaction. The multiple-group comparisons were analyzed by factorial analysis of variance. The inter-group comparisons were analyzed by the LSD method for statistical analysis. Results: The contents of peripheral blood lymphocytes, erythrocytes, platelets, and hemoglobin were significantly decreased in the 3 Gy irradiation group than the 0 Gy irradiation group (P<0.05), while the activities of serum ALT and AST were significantly increased (P<0.05). The malondialdehyde content, F4/80 protein expression level, nucleotide-binding domain and leucine-rich repeats, nucleotide oligomerization domain-like receptor family, pyrin domain containing 3 (NLRP3), and interleukin 6 mRNA expression levels were significantly increased in liver tissue, while the contents of T-SOD and glutathione, Nrf2 and HO-1 protein expression levels, and Nrf2 mRNA expression level in liver tissue were significantly decreased (P<0.05). The contents of peripheral blood lymphocytes, red blood cells, platelets, and hemoglobin were significantly increased in the 3 Gy transplantation group than the 3 Gy irradiation group (P<0.05), while the activities of serum ALT and AST were significantly decreased (P<0.05). The malondialdehyde content, F4/80 protein expression level, NLRP3 and interleukin-6 mRNA expression levels in liver tissue were significantly decreased (P<0.05), while the content of T-SOD and glutathione, Nrf2 and HO-1 protein expression levels, and Nrf2 mRNA expression level in liver tissue were significantly increased (P<0.05). Conclusion: X-ray irradiation at a dose of 3 Gy can induce liver oxidative damage in mice. BMMSC transplantation can improve X-ray irradiation-induced liver oxidative damage in mice, and its mechanism of action may be related to the regulation of the Nrf2/HO-1 pathway.

Objective: To explore the therapeutic efficacies of three different doses of human umbilical cord mesenchymal stem cell exosomes (hucMSC-EXO) on the injury of intestinal barrier structure and dysfunction in severely burned rats, and to identify the optimal dose of hucMSC-EXO for the repair of intestinal barrier injury. Methods: The hucMSC-EXO was isolated and identified by using an exosome extraction and purification kit. A total of 30 specific pathogen free (SPF) male Wistar rats (aged 6-8 weeks) were selected, and were randomly divided into five groups (n=6) using a random number table: sham group, burn group, burn+100 μg hucMSC-EXO group (Burn+EXO100), burn+200 μg hucMSC-EXO group (Burn+EXO200), and burn+400 μg hucMSC-EXO group (Burn+EXO400). The rats were immersed in 94 ℃ water, with the dorsal area exposed for 12 seconds and the ventral area for 6 seconds, to establish a 50% total body surface area (TBSA) third-degree burn model. The sham group rats were subjected under the same condition but with a 37 ℃ water bath. On day 1, 3, and 5 post-burn, the rats in sham group and burn group received an intraperitoneal injection of 0.5 ml phosphate buffered solution, and those in Burn+EXO100, Burn+EXO200, and Burn+EXO400 groups received intraperitoneal injections of 100, 200, and 400 μg/0.5 ml hucMSC-EXO, respectively. The activity of rats was observed and the weight was recorded daily. On day 7, the small intestine tissues and serum of the rats were collected. Hematoxylin-Eosin (HE) staining was used to observe the pathological changes of the small intestinal tissues, and the levels of inflammatory factors, such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-8, IL-10 and lipopolysaccharide (LPS) of small intestine tissues were detected by enzyme-linked immunosorbent assay (ELISA). The levels of diamine oxidase, D-lactic acid and bacterial endotoxin in serum were detected by intestinal barrier function biochemical analysis system. Results: The morphology of hucMSC-EXO was observed to be round or oval, with uniform size and a peak diameter of approximately 100 nm, expressing positive markers CD63 and TSG101. In the sham injury group, the rats' body weight increased by approximately (6.3±1.2) g/day, whereas in the burn group, the body weight significantly decreased on the first day post-injury and then gradually increased at a rate of (1.6±0.5) g daily. In contrast, the body weight of the Burn+EXO100, 200, and 400 groups increased at a rate of (2.9±1.1) g daily. By day 7 post-injury, the body weight in the Burn+EXO200 and Burn+EXO400 groups were significantly higher than those in the burn group and the Burn+EXO100 group (all P<0.05). HE staining showed that the villus height in the small intestine (duodenum, jejunum, ileum) of the burn group [(711±35), (526±25), (418±33) μm] was significantly reduced with severe structural damage, while the small intestine structure and villus height in the EXO-treated groups showed varying degrees of recovery. The villus height in the Burn+EXO200 group [(1 050±40), (798±30), (609±29)μm, respectively] and burn+EXO400 group [(1 102±46), (830±28), (625±33)μm, respectively] recovered significantly (all P<0.05). ELISA results indicated that the levels of pro-inflammatory cytokines TNF-α, IL-1β, IL-6, IL-8, and LPS in the burn group [(29.3±1.7), (81.2±2.5), (582.4±36.9), (22.1±0.6), (366.8±15.9)ng/L, respectively] were elevated, while those were significantly reduced in the EXO-treated groups, with Burn+EXO200 group [(17.9±1.0), (58.2±2.3), (206.6±38.7), (8.9±1.0), (94.9±7.3)ng/L, respectively] showing the most significant reduction (all P<0.05). The level of anti-inflammatory cytokine IL-10 was elevated in the burn group [(293.4±16.0) ng/L], and it was significantly increased in the burn+EXO100, 200, and 400 groups [(591.8±40.7), (672.5±53.7), (712.5±36.2)ng/L, respectively] (all P<0.05). Assessment of intestinal barrier function showed that serum diamine oxidase in the burn group decreased, while D-lactate and bacterial endotoxin increased. The EXO-treated groups demonstrated significant improvement in diamine oxidase, D-lactate, and bacterial endotoxin levels, with Burn+EXO200 group showing the most significant effect (all P<0.05). Conclusions: In this study, hucMSC-EXO was successfully isolated, extracted, and identified. An intraperitoneal injection of 200 μg of hucMSC-EXO demonstrates the most effective repair of intestinal structure and function in rats with severe burn injuries.

To explore ferroptosis-related genes in osteoporosis through bioinformatic analysis and in vitro study.

Objective: Investigating the changes of phenotype and moleculars associated with aging with the increase of passage times of human dental pulp stem cells (hDPSC), to explore the role of WW-containing transcriptional regulator 1 (WWTR1) in the aging mechanism. Methods: hDPSCs were cultured by tissue block method, and were divided into 4 groups according to the age, algebra, cell knockdown and overexpression of WWTR1 in hDPSCs. Group Ⅰ: hDPSCs from human teeth were further divided into youth group (15-25 years old) and group middle-aged group (40-50 years old) according to different ages. Group Ⅱ: according to different passage, hDPSCs were divided into young cells group (hDPSCs were transmitted to P3 generation), and old cells group (hDPSCs were transmitted to P10 generation). Group Ⅲ: hDPSCs were knocked down of WWTR1, which were further divided into knockdown group and knockdown carrier group. Group Ⅳ: hDPSCs were overexpressed of WWTR1, which were further divided into overexpression group and overexpression carrier group. Real-time fluorescence quantitative PCR (RT-qPCR) was used to detect the changes of WWTR1 expression in groups Ⅰ and Ⅱ, and cell counting kit-8 (CCK-8) was used for groups Ⅱ, Ⅲ, and Ⅳ. Cell proliferation capacity was detected by CCK-8 assay. The ability of osteogenic differentiation was detected by alizarin red staining. Cell senescence positive rate was detected by age-related β-galactosidase staining. The expression levels of age-related genes p53 and p21 were detected by RT-qPCR. Results: The proportion of senescent cells increased gradually with continuous culture. The proliferation and osteogenic differentiation of hDPSCs in the old group were significantly lower than those in the young group (P<0.001). The expression levels of senescence related genes p53 (2.09±0.24) and p21 (4.91±0.54) in old cell group were higher than those in young cell group respectively [p53: (1.08±0.09) and p21: (1.09±0.08)] (P<0.01, P<0.001). The WWTR1 expression levels of hDPSCs in middle-aged group and old cells group were both decreased compared with those in young group and young cells group (P<0.01). The proportion of senescent cells in knockdown group (44.50±2.42) was higher than that in knockdown carrier group (22.27±0.56) (P<0.001). After knocking down WWTR1 in hDPSCs, the expression levels of age-related genes p53 and p21 were up-regulated (P<0.001), and the abilities of proliferation and osteogenic differentiation in the knockdown group were lower than those in the knockdown carrier group (P<0.001). The proportion of senescent cells in overexpression empty carrier group (20.40±0.79) was higher than that in overexpression group (10.07±0.61) (P<0.001). After WWTR1 overexpression ins hDPSCs, the expression levels of age-related genes p53 and p21 were down-regulated, and the proliferation and osteogenic differentiation ability in overexpression group were higher than those in overexpression carrier group (P<0.001). Conclusions: WWTR1 can inhibit the expression levels of age-related genes p53 and p21, thus delaying the aging process as well as promoting the proliferation and osteogenic differentiation of hDPSCs.

Objective To investigate the molecular mechanism of IL-6 regulating the expression of secretory phosphoprotein 1 (SPP1) in human placental-derived mesenchymal stem cells (hfPMSCs) and influencing the polarization of macrophages. Methods hfPMSCs were prepared by enzymatic digestion and cultured in a defined, serum-free medium. The morphology of hfPMSCs was observed under a microscope, and the expression of surface markers CD14, CD34, CD45, CD73, CD90, CD105, and human leukocyte antigen DR (HLA-DR) was detected by flow cytometry. hfPMSCs were treated with IL-6 at the final dosage of 100 ng/mL for 24 hours. ELISA, Western blot, and real-time quantitative PCR were used to detect the expression of SPP1 at the protein and mRNA levels; after being treated with IL-6, hfPMSCs infected with SPP1 interference lentivirus were co-cultured with activated macrophage THP-1 cells induced by 100 ng/mL lipopolysaccharide (LPS) and 80 ng/mL phorbol 12-myristate 13-acetate (PMA), and flow cytometry was used to detect the proportion of CD11c and CD206 positive cells in THP-1 cells; hfPMSCs were treated with IL-6 and/or specific inhibitors of nuclear factor kappa B p65 (NF-κB p65), SC75741, and Western blot was used to detect the expression of genes in the NF-κB signaling pathway and SPP1. Results IL-6 significantly upregulates the expression of SPP1 in protein and mRNA levels in hfPMSCs; interference of SPP1 significantly downregulates the expression of SPP1 in hfPMSCs and significantly reduces the positive cell ratio of CD206 in co-cultured macrophages; IL-6 significantly upregulates the expression of p-p65 in hfPMSCs, activating the NF-κB signaling pathway; SC75741 significantly downregulates the expression of p-p65 and SPP1 in hfPMSCs treated with IL-6. Conclusion IL-6 upregulates the expression of SPP1 through the NF-κB signaling pathway in hfPMSCs, which enhances the capability to induce macrophages to polarize towards the M2 phenotype.

To review the research progress on bone repair biomaterials with the function of recruiting endogenous mesenchymal stem cells (MSCs).

To investigate the effects of R-spondin 2 (Rspo2) on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and bone mineral content in ovariectomized mice.

To culture and expand primary rat aortic vascular stem cells in vitro and evaluate their characteristics as mesenchymal stem cells.