Objective: To explore new therapeutic strategies for periodontitis, this study prepared extracellular vesicles (EVs) derived from human umbilical cord mesenchymal stem cells (hUC-MSCs) loaded with saikosaponin D (SSD) and evaluated their efficacy in a mouse model of periodontitis. Methods: This study was approved by the Medical Ethics Committee and the Laboratory Animal Ethics Committee of the unit. hUC-MSCs at logarithmic growth phase were seeded into 96 well plates and then treated with various concentrations of SSD for 24 hours. The CCK-8 assay was used to determine the SSD concentration that preserves hUC-MSC viability. Flow cytometry (FCM) assessed SSD-induced apoptosis in hUC-MSCs and determined drug loading and loading efficiency. Nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM) characterized the morphology and size of hUC-MSCs-EVs and hUC-MSCs-SSD-EVs. C57BL/6J mice were divided into five groups with six mice per group: control group, periodontitis model group (PD), PD+SSD group (2 mg/kg), PD+hUC-MSCs-EVs group (10 μg/μL) and PD+hUC-MSCs-SSD-EVs group (10 μg/μL). Administration was performed 3 times a week for 4 consecutive weeks. Micro-CT and hematoxylin-eosin (HE) staining evaluated alveolar bone resorption in periodontal tissues of a mouse model of periodontitis. RT-qPCR measured inflammatory cytokine expression in periodontal tissues of a mouse model of periodontitis. Immunofluorescence analyzed macrophage M1/M2 polarization and runt-related transcription factor 2 (RUNX2) protein expression in periodontal tissues of a mouse model of periodontitis. Results: The CCK-8 assay showed that 10 μmol/L SSD had little effect on hUC-MSCs viability (P>0.05). FCM confirmed the successful loading of SSD into EVs with a loading efficiency of 11.40%. TEM and NTA revealed that hUC-MSCs-SSD-EVs presented round or oval membranous structures with diameters ranging from 200 to 300 nm. Micro CT and HE staining showed that, compared with the PD group, the distance from cementoenamel junction to alveolar bone crest (CEJ-ABC) was markedly shortened (P<0.01）.The alveolar bone of periodontitis‑model bone volume fraction (BV/TV) and trabecular number (Tb.N) were significantly elevated in the PD+hUC-MSCs-SSD-EVs group (P<0.01). RT-qPCR and IF results showed that hUC-MSCs-SSD-EVs in periodontal tissues markedly downregulated the expression of pro-inflammatory cytokines including interleukin-1β, interleukin-6, tumor necrosis factor-α, interleukin-12 and interleukin-22 (P<0.01), upregulated the expression of the anti-inflammatory factor interleukin-10 (P<0.01), promoted the polarization of macrophages from the M1 phenotype to the M2 phenotype, and elevated the protein expression level of RUNX2 (P<0.01). Conclusion hUC-MSC-SSD-EVs effectively alleviated inflammatory responses in a mouse model of maxillary periodontitis and reduced alveolar bone loss, providing a new idea for the treatment of periodontitis.

Periodontal bone defects, primarily caused by periodontitis, are highly prevalent in clinical settings and manifest as bone fenestration, dehiscence, or attachment loss, presenting a significant challenge to oral health. In regenerative medicine, harnessing developmental principles for tissue repair offers promising therapeutic potential. Of particular interest is the condensation of progenitor cells, an essential event in organogenesis that has inspired clinically effective cell aggregation approaches in dental regeneration. However, the precise cellular coordination mechanisms during condensation and regeneration remain elusive. Here, taking the tooth as a model organ, we employed single-cell RNA sequencing to dissect the cellular composition and heterogeneity of human dental follicle and dental papilla, revealing a distinct Platelet-derived growth factor receptor alpha (PDGFRA) mesenchymal stem/stromal cell (MSC) population with remarkable odontogenic potential. Interestingly, a reciprocal paracrine interaction between PDGFRA⁺ dental follicle stem cells (DFSCs) and CD31⁺ Endomucin⁺ endothelial cells (ECs) was mediated by Vascular endothelial growth factor A (VEGFA) and Platelet-derived growth factor subunit BB (PDGFBB). This crosstalk not only maintains the functionality of PDGFRA⁺ DFSCs but also drives specialized angiogenesis. In vivo periodontal bone regeneration experiments further reveal that communication between PDGFRA⁺ DFSC aggregates and recipient ECs is essential for effective angiogenic-osteogenic coupling and rapid tissue repair. Collectively, our results unravel the importance of MSC-EC crosstalk mediated by the VEGFA and PDGFBB-PDGFRA reciprocal signaling in orchestrating angiogenesis and osteogenesis. These findings not only establish a framework for deciphering and promoting periodontal bone regeneration in potential clinical applications but also offer insights for future therapeutic strategies in dental or broader regenerative medicine.
Receptor, Platelet-Derived Growth Factor alpha/metabolism* ; Humans ; Neovascularization, Physiologic/physiology* ; Dental Sac/cytology* ; Single-Cell Analysis ; Transcriptome ; Mesenchymal Stem Cells/metabolism* ; Bone Regeneration ; Animals ; Dental Papilla/cytology* ; Periodontium/physiology* ; Stem Cells/metabolism* ; Regeneration ; Angiogenesis

Objective:To study the whole bone marrow cellular composition of jaw and long bones, and further analyze the heterogeneity of mesenchymal stem cells (MSCs) derived from these two tissue, aiming at exploring the differences in functional characteristics of bone MSCs from different lineage sources.Methods:The Seurat package of R language was used to analyze the mandibular and femur whole bone marrow single-cell RNA-sequencing (scRNA-seq) datasets in the literature, and the subpopulations were annotated by reference to the marker genes reported by previous studies. The differentially expressed genes between mandible-derived MSCs (M-MSCs) and femur-derived MSCs (F-MSCs) were calculated, and cell-cell communication analysis between M-MSCs or F-MSCs with other cell populations was performed. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on up-regulated and down-regulated differentially expressed genes of M-MSCs, and Gene Set Enrichment Analysis (GSEA) was performed on M-MSCs or F-MSCs.Results:cRNA-seq analysis showed that the mandible and femur had the same bone marrow cell composition, but there were differences in the proportion of specific cell populations. Also, there were significantly differentially expressed genes between M-MSCs and F-MSCs. In addition, cell-cell communication analysis revealed differences in numbers of ligand-receptor pairs between M-MSCs or F-MSCs with other cell populations. Furthermore, GO, KEGG and GSEA analysis showed that M-MSCs had higher extracellular matrix production potential than F-MSCs, but had lower ability to regulate other cells in the bone marrow, especially immune cells.Conclusions:M-MSCs and F-MSCs showed distinct differences in the gene expression pattern and up-regulated signaling pathways, which may be closely related to the developmental sources and functional characteristics of jaw and long bones.

Objective:To investigate the predictive values of baseline hematoma mean CT value and shape regularity (SR) for hematoma enlargement (HE) in patients with spontaneous intracerebral hemorrhage (ICH).Methods:Patients with ICH admitted to Yulin First Hospital from June 2018 to December 2021 were retrospectively included. The first head CT scan was performed within 24 h of onset, and the second head CT scan was performed within 72 h of the first scan. HE was defined as an increase in hematoma volume of at least 6 ml or 33% from the first CT. 3D Slicer software was used to reconstruct 3D images and SR was calculated. Multivariate logistic regression analysis was used to determine the independent factor for HE. Receiver operator characteristic (ROC) curve was used to evaluate the predictive value of baseline hematoma mean CT value for HE. Results:A total of 249 patients with ICH were enrolled, including 134 males (53.8%), and aged 62.2±12.1 years. The median baseline Glasgow Coma Scale score was 12, and the median time from onset to first CT scan was 3.1 h. The median baseline hematoma volume was 10.9 ml, and 58 patients (23.3%) showed HE. The baseline hematoma mean CT value in the HE group (58.5±3.2 HU vs. 60.3±3.3 HU; P<0.01) and baseline SR (0.615±0.146 vs. 0.688±0.100; P<0.001) were significantly lower in the non-HE group. Multivariate logistic regression analysis showed that the time from onset to first CT scan (odds ratio [ OR] 0.867, 95% confidence interval [ CI] 0.786-0.957; P=0.004), the baseline hematoma volume ( OR 1.050, 95% CI 1.028-1.073; P<0.001), and the baseline hematoma mean CT value ( OR 0.809, 95% CI 0.725-0.902; P<0.001) were the independent predictors of HE, while the baseline SR had no significant independent correlation with HE. ROC curve analysis showed that the area under the curve of baseline hematoma mean CT value for predicting HE was 0.652 (95% CI 0.573-0.731; P<0.001), with an optimal cutoff value of 57.97 HU. The sensitivity and specificity for predicting HE were 50% and 75.9%, respectively. Conclusion:The baseline hematoma mean CT value is an independent factor for HE in patients with ICH and has certain predictive value for HE.

Objective:To study the clinical effect of early postoperative enteral nutrition and parenteral nutrition after radical ex-cision of gastric cancer to provide a better way of treating gastric cancers. Methods:Retrospective analysis of 140 gastric cancer pa-tients who were admitted to the PLA General Hospital between February 2009 and February 2011 was conducted. These patients were randomized into two groups based on the clinical trial, i.e., 70 in the control group received an intravenous parenteral nutrition for the treatment, and for the other 70 in the observation group, jejunostomy was done 1 to 5 days after the radical surgery by using Supportan as the enteral nutritional agent with a dose of at TPF-T 500 mL/d to 1 000 mL/d. The postoperative long-term survival rate of the pa-tients, the serum albumin, hemoglobin, alanine aminotransferase, aspartate aminotransferase levels before and after the treatment, as well as the situation of IgA, IgG, IgM and CD4+cells, NK cells, and B lymphocytes in the blood at the first and the seventh day after surgery were observed in the patients. Results:After the implementation of early enteral nutrition in the observation group, the 1-and 3-year survival rates were 84.29% (59/70) and 61.43% (43/70) respectively, whereas in the control group, the survival rates were 64.29% (45/70) and 47.143% (33/70) respectively, with statistically significant differences between the two groups (P<0.05). At the first and seventh day after surgery, the serum albumin, hemoglobin, alanine aminotransferase, and aspartate aminotransferase levels were significantly better in the observation group than in the control group, with statistically significant differences between the two (P<0.05). Compared with the parameters in the observation group at the first day after surgery and those in the control groups at the eighth day after surgery, the levels of IgA , IgG, IgM and CD42+cells, NK cells, and B lymphocytes were significantly increased in the obser-vation group at the seventh day after surgery. The differences among them were statistically significant (P<0.05). Conclusion: Early postoperative enteral nutrition for the gastric cancer patients undergoing radical surgery can be effective in improving the nutrition level of the patients and in enhancing their long-term survival rate, Thus, the treatment has value in clinical application.

Glucose metabolism of cancer cells presents Warburg effect.In resent years,more and more experiments demonstrate that the therapeutics based on aerobic glycolysis pathway in cancer cells restrict energy and inhibit tumor proliferation through the inhibition of a variety of moleculars,genes and signal pathways.These can provide an opportunity for targeted cancer therapies and possess enormous potential advantages and broad prospects in clinical application.

Cancer stem cells are a subclass of stem-cell-like tumor cells.They have an apparently higher tumorigenicity than other cell populations within the same tumor.They play a Very important role in genesis,development and maintenance of cancer.The data from research in leukemia and solid timors suggest that in each cancer tissue,only a small fraction of the cells have the ability to initiate tumor,which are called cancer stem cells.In recent years,with in-depth research on stem cells and cancer stem cells,accumulating evidence suggests that cancer stem cells are the cause of malignant tumor metastasis and recurrence and are resistant current callcer drugs.Drugs targeting cancer stem cells are urgently needed. Flow cytometry and magnetic cell sorting using the identified tumor stem cell surface markers are mainly used to separate cancer stem cells or stem cell-like cells. In this article,the concept of cancer stem cells and the commonly used approaches to isolate and identify cancer stem cells,especially for gastric cancer stem cells will be reviewed.

Objective To investigate the adjuvant effect of dimo-thylidioctyl ammonium bromide (DDA) and/or DDA-BCG polysaccharide nucleic acid( BCG-PSN), which was combined with a Mycobacterium tuberculosis fusion protein AMM ( Ag 8 5 B - MPT64190-198 - Mtb8.4 ) to boost BCG primed immunization. Methods DDA with or without BCG PSN was mixed with the fusion protein AMM to construct the boosting vaccine. Mice were immunized with BCG and then boosted twice with AMM formulated with the adjuvant DDA with or without BCG-PSN. PBS or BCG vaccination without boosting was used as control. The humoral and cell-mediated immune responses were analyzed by ELISA and ELISPOT. Moreover, the protective efficacy of BCG prime-AMM subunit vaccine boosting against Mycobacterium tuberculosis infection was analyzed. Results With in vitro stimulation of Ag85B and PPD( purified protein derivative) antigen, the number of IFN-γ secreting cells from the mice boosted twice by AMM/DDA/BCG-PSN and AMM/DDA were higher than BCG and PBS group (P <0.05). The CFU in lungs of mice boosted with AMM/DDA/BCG-PSN was less than that of PBS group(P <0.05), while the CFU of AMM/DDA-boosted mice was less than that of BCG and PBS group(P < 0.05).However, fewer lesions were seen in lungs of mice immunized with BCG alone or BCG-prime-AMM/DDA/BCG-PSN boosting than the other groups. Conclusion DDA is an idea adjuvant for tuberculosis subunit vaccine;BCG-PSN might play a role in alleviating the immunity-mediated pathology.

Objective:To determine the diagnostic value of multislice CT urography (MSCTU) in patients with transitional cell carcinoma ( TCG) of upper urinary tract by comparing other imageology methods used. Methods: Two hundred and thirty four cases of transitional cell carcinoma of upper urinary tract, in which 82 cases were diagnosed pathologically with pelvic carcinoma and 152 cases with ureteral carcinoma, between June 2004 and September 2006 in our institute were enrolled in a retrospective study. Most of them underwent urological ultrasound, intravenous urogram (IVU) , retrograde pyelography and MSCTU. We compared the positive rate (PR) and diagnostic rate (DR) of these methods used by chi-square test. Results: Among the 234 cases, 215 patients underwent urologic ultrasound, in which 152 cases were detected to be abnormal, with the PR of 70.1% ;Meanwhile, 58 cased were diagnosed by this examination, with the DR of 27. 0%. IVU was performed in 193 patients and 132 cases were found to be abnormal, and the PR was 68. 4% , 65 cases were diagnosed by IVU and the DR was 33.7%. And 132 patients underwent retrograde pyelography, by which 115 cases of lesion were detected, with the PR of 87. 1% ; In the meantime, 93 cases were diagnosed, with the DR of 70. 5%.MSCTU was performed in 226 cases and 220 cases were found to be abnormal, and the PR was 97.3% ;214 cases were diagnosed by MSCTU, with the DR of 94. 7%. The DR of detecting TCC of retrograde pyelography had statistically significant difference with that of ultrasound and IVU(P＜0.001). As compared with retrograde pyelography, MSCTU had statistically significant superiority (P＜0.001). Conclusion: To shorten the diagnosis time and mitigate the sufferings, patients with hematuria supposed to be TCC of upper urinary tract should be recommended to undergo MSCTU first.

Objective To construct protective immunity to Mycobncterium tuberculosis latent infection, a novel fusion protein consisting of HspX, the 190 to 198 peptide of Mpt64 and Ag85B, which were confirmed to be the effective protective antigens mainly expressed in the dormancy and exponential phase of growth, was constructed and its immunogenicity was investigated. Methods Ag85B and Mpt64190-198-HspX sequences were amplified by PCR and cloned into plasmids pET-28a. The fusion protein, Ag85BMpt64190-198-HspX (AMH) was expressed in E. coli BL21 and purified with Ni-NTA resins. C57BL/6 mice were immunized three times at 2-week intervals subcutaneously with AMH formulated with the adjuvant composed of dimethyl-dioctyldecyl ammonium bromide (DDA) and BCG polysaccharide nucleic acid (BCGPSN). Humoral and cell-mediated immunity responses were analyzed at five weeks after the last injection. Results AMH was expressed stably in E. coli and could be purified well by Ni-NTA affinity chromatography. C57BL/6 mice immunized with AMH subunit vaccine generated specific cellular and humoral immunologic response to the stimulation of Ag85B, Mpt64190-198 and HspX. Conclusion It suggested that AMH was a promising candidate antigen of tuberculosis subunit vaccine.