To investigate the effects and mechanisms of Schwann cells-derived extracellular vesicles （SCs-EVs） on pulp regeneration.
Methods
Human dental pulp stem cells（hDPSCs） were isolated and cultured in vitro， and treated with SCs-EVs， SCs-EVs combined with Wnt/β-catenin signaling pathway inhibitor IWR-1 （SCs-EVs group， SCs-EVs+IWR-1 group）， and a blank treatment group （Control group） was set up. The cell proliferative activity and cell clonogenesis ability were detected by CCK-8 method and plate cloning method respectively. The formation ability of calcium nodules was detected by alizarin red staining method. The lipid formation capacity in cells was detected by oil red O staining method. The mRNA expression levels of self-renewal marker genes （Oct4， Nanog， Sox2）， osteogenic differentiation marker genes （ALP， Runx2， COL-1），lipogenic differentiation marker genes （ADPN， FABP4， LPL） and neurogenic differentiation marker genes （Gfap， Nestin， Tubb3） were detected by RT-qPCR. The protein expression level of Runx2，ADPN， Nestin and Wnt/β-catenin signaling pathway related proteinswere detected by Western blot.The fluorescence intensity of β-catenin was detected by immunofluorescence. Independent sample t test or corrected t test was used for comparison between two groups， one-way ANOVA was used for comparison between multiple groups， and LSD-t test was used for further pair-to-group comparison（between groups with different SCs-EVs concentrations； SCs-EVs group and Control group；SCs- EVs+IWR-1 group and SCs-EVs grou）.
Results
Compared with the Control group， the cell proliferation activity in SCs-EVs group was enhanced （P ＜ 0.05）， together with an increasing of the number of clones （909.17 ± 52.86 vs 415.38 ± 25.61）， calcium nodule formation ability （0.79 ±0.05 vs 0.24 ± 0.02）， lipid formation ability （0.52 ± 0.03 vs 0.14 ± 0.01）， Oct4， Nanog， Sox2， ALP，Runx2， COL-1， ADPN， FABP4， LPL， Gfap， Nestin， Tubb3 mRNA expression levels， Runx2 （1.05 ±0.16 vs 0.18 ± 0.05）， ADPN （0.91 ± 0.09 vs 0.14 ± 0.04）， Nestin （0.96 ± 0.13 vs 0.15 ± 0.03），β-catenin （1.01 ± 0.12 vs 0.19 ± 0.03） protein expression levels and β-catenin fluorescence intensity（4.27 ± 0.31 vs 1.07 ± 0.12） （all P＜0.05）. Compared with SCs-EVs group， cell proliferation activity in SCs-EVs+IWR-1 group was decreased （P＜0.05）， together with a decreasing of the number of clones （482.55 ± 29.32 vs 909.17 ± 52.86）， calcium nodule formation ability （0.32 ± 0.03 vs 0.79 ±0.05）， lipid formation ability （0.20 ± 0.01 vs 0.52 ± 0.03）， Oct4， Nanog， Sox2， ALP， Runx2， COL- 1，ADPN， FABP4， LPL， Gfap， Nestin， Tubb3 mRNA expression levels， Runx2 （0.21 ± 0.05 vs 1.05 ±0.16）， ADPN （0.37 ± 0.06 vs 0.91 ± 0.09）， Nestin （0.34 ± 0.09 vs 0.96 ± 0.13）， β-catenin （0.31 ±0.05 vs 1.01 ± 0.12） protein expression levels and and β-catenin fluorescence intensity （1.29 ± 0.15 vs 4.27 ± 0.31） （all P ＜ 0.05）.
Conclusion
SCs-EVs promotes pulp regeneration by activating the Wnt/β-catenin signaling pathway to promote the proliferation， self-renewal and pluripotency of hDPSCs.
This study aims to analyze immune cell infiltration and key genes of in BKV nephropathy after renal transplantation based on transcriptomics.
Methods
We obtained microarray chip data from 17 renal tissue biopsies after renal transplantation from the GEO database.Through PCA dimensionality reduction analysis， differential expression analysis， enrichment analysis，PPI network construction and key gene screening， as well as cell infiltration analysis， combined with statistical methods such as the Wilcoxon ran-sum test and Spearman correlation analysis， we explored the patterns of immune cell infiltration and key genes in BKV nephropathy. The findings were subsequently validated through the external dataset GSE75693.
Results
The study found that biopsy samples from BKV nephropathy showed a distinct separation in gene expression patterns compared to normal renal transplant biopsy samples and BKV viremia renal tissue biopsy samples. Immune cell infiltration analysis revealed immune cell characteristics in BKV-related nephropathy， with significantly reduced proportions of plasma cells， resting mast cells， regulatory T cells， and activated NK cells （P ＜ 0.05）， while the proportions of resting CD4 memory T cells， memory B cells， activated CD4 memory T cells， and resting NK cells were significantly increased （P ＜ 0.05）. Identification of differentially expressed genes， GO and KEGG analyses， and PPI network construction with hub gene screening revealed that key genes such as KIF11， DLGAP5， CDK1， CDC20， BUB1， ASPM，KIF20A， BUB1B， TOP2A， and NUSAP1 showed varying degrees of correlation with the infiltration proportions of specific immune cell types （|r| ≥ 0.483， P ＜ 0.05）. The expression of these key genes was upregulated （|log2 fold change| ≥ 1， P ＜ 0.001）. Validation results from the external dataset showed that TOP2A， CDC20， and KIF20A were significantly upregulated in BKV nephropathy biopsy samples （P ＜ 0.05）.
Conclusion
This study reveals the immune cell infiltration patterns and molecular characteristics of BKV nephropathy， providing new insights into the pathogenesis of BKV nephropathy and potential targets for future therapeutic strategies.
To investigate the biological toxic effect and mechanisms of gold nanorods （AuNRs） on A549 cells.
Methods
Normal cultured A549 cells were treated with different concentrations of AuNRs for 6， 12 or 24 hours for subsequent detection. CCK-8 test and LDH test were used to observe cell viability and cell membrane damage. The distribution and morphology of AuNRs in cells were observed by transmission electron microscope. Western blot was used to detect the expression of autophagy related proteins ATG16， ATG4， Beclin1， LC3-Ⅱ and P62. ROS was detected by laser scanning confocal microscope. The levels of GSH/GSSG and T-AOC were detected by kits. One way ANOVA was performed after the homogeneity test of data variance and Student- Newman-Keuls test was used for pairwise comparison between different groups.
Results
CCK-8 results showed that compared with the 0 μg/mL， the cell viability of the 4 μg/ mL AuNRs [（80.05 ± 3.45）% vs （100 ± 2.47）%］ was decreased at 6 h after treated with AuNRs （P ＜0.01）. The cell viability of 2 μg/mL and 4 μg/mL AuNRs [（71.36 ± 3.87）% and （39.47 ± 4.16）%vs （100 ± 3.12）%］ were decreased （P ＜ 0.01） at 12 h after treated with AuNRs； The cell viability of 1 μg/mL， 2 μg/mL and 4 μg/mL AuNRs [（91.83 ± 1.98）%， （56.53 ± 3.57）% and （28.65 ± 3.09）%vs （100 ± 2.34）%］ were decreased （P ＜ 0.01） at 24 h after treated with AuNRs. LDH test showed that the leakage of LDH was increased significantly （P ＜ 0.01）. Transmission electron microscopy showed that AuNRs could enter A549 cells and exist in the cytoplasm and some membrane vesicles in the form of single particles or aggregates 6 h after treatment with 4 μg/mL AuNRs. Western blot showed that after treated with different concentrations of AuNRs （0， 0.5， 1， 2， 4 μg/mL）for 6 h， the expression levels of ATG16， ATG4， Beclin1 and LC3- Ⅱ proteins were significantly increased， while the expression level of autophagy substrate protein P62 was significantly decreased in A549 cells（P ＜ 0.01）. After treated with 100 μmol/L MnTBAP or 10 mmol/L NAC， the increased expression of ROS and LC3 induced by AuNRs could be reversed， and the content of T-AOC [（92.37 ± 8.49）%vs （68.46 ± 6.38）%， （110.49 ± 7.39）% vs （51.26 ± 7.39）%］ and GSH/GSSG [（92.56 ± 5.52）%vs （62.48 ± 5.52）% ， （104.49 ± 4.84）% vs （57.39 ± 4.84）%］ were increased （all P ＜ 0.05）.
Conclusion
AuNRs can enter A549 cells and induce cytotoxicity effect， which in a dose-dependent and time-dependent manner. The mechanism may be related to the reduction of antioxidative stress ability of A549 cells caused by AuNRs.
To understand the publication status， research hotspots， and trend analysis of hair follicle stem cells （HFSCs） in the field of hair loss， and to provide reference for future research on hair loss prevention and treatment.
Method
The visual analysis method was used to analyze the related literature of HFSCs in the field of hair loss based on CiteSpace （6.2.R4）software， which were searched in the database of CNKI， CBM and WOS.
Results
A total of 1 091 articles were included， including 235 Chinese articles and 856 English articles. The country with the highest number of English literature publications is the United States （337）， followed by China（154）， the United Kingdom （98）， Germany （97）， and Japan （89）. The foreign research institution with the highest number of publications is the University of California system （44 articles）； The domestic research institution is Southern Medical University （9 articles）. Professor Paus Ralf from the University of Lubeck in Germany （34 articles） is the most prolific English author. Professor Diao Bo from the Basic Medicine Laboratory of the Central Theater Command General Hospital（7 articles） is the most prolific Chinese author. The attention in this field is constantly increasing worldwide， and there is close cooperation between countries and institutions. China has an important international status. High frequency keywords include "hair follicle"， "expression"， "differentiation"，etc. The research hotspots focus on signal pathway mediated hair follicle regeneration， application of bioprinting technology， maintenance of hair follicle immune microenvironment homeostasis， and traditional physical stimulation and drug combination.
Conclusion
Through visual analysis， the research status and hot trends of hair follicle stem cells in the field of hair loss were obtained， which provided a useful reference for clinical research in the future.
To investigate the role and mechanisms of LncRNA NEAT1 （NEAT1）on lipopolysaccharide （LPS）-induced renal tubular epithelial cell damage. Methods Human renal tubular epithelial cells HK-2 were cultured in vitro， LPS was used to treat HK-2 cells under different concentrations. Flow cytometry was used to determine the apoptosis rate of cells treated with different concentrations of LPS. The expression of NEAT1， miR-129-5p， phosphorylated （p-） β-Catenin and β-Catenin were detected by RT-qPCR and Western blot. Experimental groups-1 were mimic NC group， miR-129-5p mimic group； miR-NC group， miR-129-5p inhibitor group. The above four groups were co-transfected with NEAT1-WT or NEAT1-MUT. Dual luciferase reporter gene experiment was used to analyze and verify the direct sequence interactions and negative regulatory relationships between NEAT1 and miR-129-5p in the above groups of cells. Experimental group-2 were the blank control group， si-NC group and si-NEAT1 group. si-NC and si-NEAT1 were transfected，respectively. The relative expression levels of NEAT1 and miR-129-5p were detected in the above three groups of cells. Cells were treated with 1 μg/mL LPS after transfection， and co- treated for 24 h. Cells were divided into LPS group （1 μg/mL LPS）， si-NC+miR-NC group （co- transfected with si- NC+miR- NC）， si- NEAT1+miR-NC group （co-transfected with si-NEAT1+miR- NC），si-NC+miR-129-5p inhibitor group （co-transfected with si-NC+miR-129-5p inhibitor），si- NEAT1+miR-129-5p inhibitor group （co-transfected with si-NEAT1+miR-129-5p inhibitor）.CCK-8 and flow cytometry experiment were used to detect cell viability， cell cycle and apoptosis rate； the expression of Bcl-2， Bax， Cleaved caspase-3， β-catenin， p-β-catenin， cyclin D1 and c-myc proteins in cells were detected by Western bot. One-way ANOVA analysis of variances was used for multi-group comparisons， and pairwised comparisons between groups were performed by post hoc Turkey's test. P ＜ 0.05 was considered statistically significant.
Results
Compared with LPS group（0 μg/ mL）， the cell viability [（83.70 ± 1.40）% vs （100.00 ± 1.70）%］， the relative expression levels of miR-129-5p （0.81 ± 0.07 vs 1.00 ± 0.13）， and phosphorylated （p-） β-catenin levels（0.78 ± 0.05 vs 1.00 ± 0.14） of HK-2 cells treated by 1μg/mL LPS was decreased， while the relative expression levels of NEAT1 （1.33 ± 0.12 vs 1.00 ± 0.08） and the apoptosis rate [（32.80 ± 1.30）%vs （5.20 ± 0.20） % ］ were increased. Compared with mimic NC group， the expression levels of miR-129-5p was increased （2.34 ± 0.22 vs 1.00 ± 0.06）， the luciferase activity of NEAT1-WT was decreased （0.36 ± 0.03 vs 1.00 ± 0.08）， the luciferase activity of NEAT1-MUT was no significant difference in the miR-129-5p mimic group. Moreover， compared with the si-NC group， the expression of miR-129-5p in the si-NEAT1 group was higher （2.18 ± 0.24 vs 1.04 ± 0.13）. After LPS treatment，compared with si-NEAT1+miR-NC group， cell viability [（67.30 ± 2.40）% vs （147.80 ± 5.60）%］，the proportion of S-phase cells [（19.50 ± 1.70）% vs （40.60 ± 1.60）%］， the proportion of G2/M phase cells [（8.30 ± 0.40）% vs （20.70 ± 0.07）%］， the Bcl-2 protein expression levels （0.21 ± 0.04 vs 1.38 ± 0.14）， the ratio of p-β- catenin/ β-catenin expression levels （0.23 ± 0.03 vs 1.51 ± 1.10），the cyclin D1 protein expression levels （0.17 ± 0.02 vs 0.64 ± 0.08）， the c-myc protein expression levels （0.18 ± 0.02 vs 0.65 ± 0.07） were decreased in si-NC+miR-129-5p inhibitor group，whilethe proportion of G0/G1 phase cells [（71.20 ± 1.30）% vs （43.30 ± 1.90）%］， the cell apoptosis rate[（39.50 ± 2.60）% vs （7.90 ± 1.10）%］， the Bax protein expression levels（1.21 ± 0.11 vs 0.35 ±0.03）and the Cleaved caspase-3 protein expression levels were increased（1.51 ± 0.11 vs 0.28 ± 0.02）.
Conclusion
LPS- induced renal tubular epithelial cell injury may be achieved by up-regulating the NEAT1/ miR- 129-5p axis to inhibit the Wnt/β-catenin pathway.
To investigate the clinical efficacy of polymyxin B combined with other antibiotics in the treatment of severe infection after hematopoietic stem cell transplantation.
Methods
Clinical data of 58 patients with severe infection after hematopoietic stem cell transplantation treated with polymyxin B combined with other antibiotics were retrospectively analyzed. Patients were divided into 2 groups based on the duration of polymyxin B treatment[≤7 d group （27 cases） ， ＞ 7 d group （31 cases）］. The total bacterial clearance rate and effective rate were compared between two groups. The quantitative data between the two groups were compared by independent sample t test or Mann-Whitney U test， and the qualitative data between the two groups were compared by χ2 test or Fisher exact test. Survival data between the two groups were compared using Kaplan-Meier survival curve method.
Results
Compared with the treatment duration ≤7 days group， the total bacterial clearance rate （48.39% vs 22.22%） and treatment effectiveness（70.97%vs 44.44%） were increased in the treatment duration ＞ 7 days group.
Conclusion
For patients with severe infection after hematopoietic stem cell transplantation， polymyxin B should be combined with other anti-infective drugs for a full course of treatment to benefit patients.
Mesenchymal stem cell derived nanovesicles （MSCNVs） represent an innovative cellular therapy alternative， characterized by high stability and reduced size， with therapeutic potential validated across various fields， including the nervous system disease， cardiovascular health，inflammatory diseases， as well as tissue regeneration and repair. Studies indicate that nanovesicles（NVs） exert positive effects on disease treatment and tissue repair through mechanisms such as promoting tissue regeneration， alleviating inflammatory responses， and modulating immune responses. However， MSCNVs still confront challenges regarding efficacy and long-term safety in current disease applications， necessitating further research and optimization. This review integrates the latest research findings and experimental data， providing a systematic analysis of the applications and therapeutic efficacy of MSCNVs in the treatment of various diseases， offering a comprehensive and updated perspective on the potential value of MSCNVs in disease therapy.
