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

BACKGROUND:Rheumatoid arthritis is an inflammatory disease.Many studies have shown that wogonin has a good anti-inflammatory effect on rheumatoid arthritis,but its exact efficacy and specific mechanism of action remain to be clarified. OBJECTIVE:To investigate the mechanism of wogonin ameliorating joint inflammation by regulating endoplasmic reticulum stress pathway in rats with collagen-induced arthritis. METHODS:(1)At the animal level:Female Wistar rats were divided into healthy control group,arthritis model group and wogonin treatment group.Rat models of arthritis in the latter two groups were established by subcutaneous injection of bovine type Ⅱ collagen and adjuvant.In the wogonin group,wogonin was given by gavage for 28 consecutive days after modeling.During this period,the rats in each group were weighed,and arthritis score and ankle swelling were measured every 7 days.After the experiment,the pathological changes of the joint were observed,the mRNA and protein levels of endoplasmic reticulum stress pathway GRP78 and CHOP were detected by qRT-PCR,western blot,and immunohistochemistry.(2)At the cellular level,cell counting kit-8 was used to detect the cytotoxic effect of wogonin on fibroblast-like synoviocytes from rats with collagen-induced arthritis.The fibroblast-like synoviocytes induced by thapsigargin were treated with different concentrations of wogonin.The levels of interleukin-1β and tumor necrosis factor-α in the cell supernatant were detected by ELISA,and the intracellular reactive oxygen species in each group were determined by DCFH-DA probe method.The mRNA and protein levels of GRP78,IRE1α,XBP1s and CHOP were detected by qRT-PCR and western blot,respectively. RESULTS AND CONCLUSION:Compared with the healthy control group,arthritis index score and ankle swelling degree in the arthritis model group were increased(P<0.01),synovial hyperplasia,inflammatory cell infiltration,cartilage destruction and bone erosion were observed in pathological sections,and the mRNA and protein expressions of GRP78 and CHOP in the ankle were significantly increased(P<0.01),which were mainly located in synovial tissue and articular surface.Compared with the arthritis model group,the arthritis index score and ankle swelling degree in the wogonin treatment group were decreased(P<0.05),synovial hyperplasia and the number of inflammatory cells were decreased,cartilage destruction and bone erosion were alleviated,the mRNA and protein expression levels of GRP78 and CHOP in the ankle were decreased(P<0.05),particularly in synovial tissue and on the articular surface.There was no significant difference in body mass among the three groups(P>0.05).In the cell experiment,200 μmol/L wogonin significantly reduced the survival rate of fibroblast-like synoviocytes(P<0.01).Compared with the blank control group,the levels of interleukin-1β,tumor necrosis factor-α,content of reactive oxygen species,and mRNA and protein expression of GRP78,IRE1α,XBP1s,and CHOP in the thapsigargin group were significantly increased(P<0.05);compared with the thapsigargin group,50 and 100 μmol/L wogonin significantly reduced the levels of interleukin-1β and tumor necrosis factor-α in the cell supernatant(P<0.05,P<0.01),and 100 μmol/L wogonin significantly reduced the content of reactive oxygen species(P<0.01)and down-regulated the mRNA and protein expression levels of GRP78,IRE1α,XBP1s and CHOP(all P<0.05).These results suggest that wogonin can effectively alleviate joint inflammatory responses in rats with collagen-induced arthritis,and the endoplasmic reticulum stress pathway may be the key target of its intervention.

ObjectiveTo investigate the effects of Congrong Zonggan capsules （CRZG） on cognitive impairment in the Alzheimer's disease （AD） model of mice and its related mechanisms. MethodsSPF grade 4-week-old 5×FAD mice were divided into a model group， low-dose CRZG （0.819 g·kg^-1） and high-dose CRZG （1.638 g·kg^-1） groups， and Donepezilepezil hydrochloride group （2 mg·kg^-1）， with eight mice in each group. Eight C57 mice with the same background were set as the normal group. After one week of adaptive feeding， mice were orally administered continuously for six months. On the 5th month of drug administration， Y maze， new object recognition， and Morris water maze tests were conducted separately. After administration， mouse brain tissue was taken， and the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in brain tissue were detected by enzyme-linked immunosorbent assay （ELISA）. Immunofluorescence （IF） was used to detect the expression of small glial cell markers Iba1， astrocyte markers GFAP， and amyloid protein 1-42 （Aβ_1-42） in the hippocampus of the brain tissue. The hematoxylin-eosin （HE） staining was used to detect pathological changes in the hippocampus of brain tissue. Western blot was used to detect the expression of nuclear factor-κB （NF-κB） p65， NOD-like receptor protein 3 （NLRP3）， cleaved Caspase-1， apoptosis-associated speck-like protein containing a CARD （ASC）， and other proteins in the brain tissue. ResultsCompared with those in the normal group， the mice in the model group had obvious cognitive impairment. The spontaneous alternation rate of the Y maze was decreased， and the discrimination index of novel object recognition was decreased significantly （P<0.01）. The escape latency in the water maze was shortened significantly （P<0.01）. The contents of IL-6 and TNF-α in brain tissue were increased. The fluorescence levels of Iba1 and Aβ_1-42 in the hippocampus were significantly increased （P<0.01）. There was a significant increase in neuronal lesions， neuronal atrophy， loose arrangement of tissue structure， and abnormal erythrocyte aggregation in the hippocampus. The protein expressions of p-NF-κB p65/NF-κB p65， cleaved Caspase-1， ASC， IL-6， and IL-1β were significantly increased （P<0.05， P<0.01）. Compared with the model group， the spontaneous alternation rate and discrimination index of the high-dose CRZG group were increased significantly （P<0.01）， and the escape latency was shortened significantly （P<0.05， P<0.01）. The content of IL-6 decreased in the brain， and that of TNF-α dropped significantly （P<0.01）. The expression of Iba1 protein and Aβ_1-42 in the hippocampus decreased significantly （P<0.05， P<0.01）. The hippocampal neurons were densely arranged， and the pyramidal nuclei were clear and centered. The abnormal aggregation of red blood cells was alleviated. The value of p-NF-κB/NF-κB proteins and the expression of ASC， cleaved Caspase-1， IL-6， and IL-1β were significantly decreased （P<0.05， P<0.01）. ConclusionCRZG can effectively improve cognitive impairment in 5×FAD mice with Alzheimer's disease， and its mechanism may be related to the regulation of the NF-κB/NLRP3 pathway to reduce the abnormal activation of microglia and inhibit neuroinflammation.

OBJECTIVES: To investigate the mechanism mediating the regulatory effect of miR-155-5p on proliferation of human submandibular gland epithelial cells (HSGECs) in primary Sjogren's syndrome (pSS). METHODS: Dual luciferase reporter assay was used to verify the targeting relationship between miR-155-5p and the PI3K/AKT pathway. In a HSGEC model of pSS induced by simulation with TRAIL and INF-γ, the effects of miR-155-inhibitor-NC or miR-155 inhibitor on cell viability, cell cycle, apoptosis and proliferation were evaluated using CKK8 assay, flow cytometry and colony formation assay. ELISA and RT-PCR were used to detect the expressions of inflammatory cytokines and miR-155-5p mRNA in the cells; Western blotting was performed to detect the expressions of proteins in the PI3K/AKT signaling pathway. RESULTS: Dual luciferase assay showed that miR-155-5p targets the PI3K/AKT pathway via PIK3R1 mRNA. The HSGEC model of pSS showed significantly decreased cell viability, cell clone formation ability and expressions IL-10 and IL-4 and increased cell apoptosis, cell percentage in G2 phase, expressions of TNF‑α, IL-6, miR-155-5p and PIK3R1 mRNA, p-PI3K/PI3K ratio, p-Akt/AKT ratio, and PIK3R1 protein expression. Treatment of the cell models with miR-155 inhibitor significantly increased the cell viability, G1 phase cell percentage, colony formation ability, and expressions of IL-10 and IL-4 levels, and obviously reduced cell apoptosis rate, G2 phase cell percentage, expressions of TNF-α, IL-6, miR-155-5p and PIK3R1 mRNA, p-PI3K/PI3K ratio, p-AKT/AKT ratio, and PIK3R1 protein expression. CONCLUSIONS In HSGEC model of pSS, inhibition of miR-155-5p can promote cell proliferation and reduced cell apoptosis by targeting PI3K1 mRNA to negatively regulate the overexpression of PI3K/AKT signaling pathway.
Humans ; MicroRNAs/genetics* ; Cell Proliferation ; Signal Transduction ; Proto-Oncogene Proteins c-akt/metabolism* ; Sjogren's Syndrome/pathology* ; Epithelial Cells/cytology* ; Submandibular Gland/cytology* ; Phosphatidylinositol 3-Kinases/metabolism* ; Apoptosis ; Class Ia Phosphatidylinositol 3-Kinase ; Cells, Cultured

Objective:To investigate associations between cardiac biomarkers with stroke severity and short-term outcome in patients with acute ischemic stroke (AIS).Methods:Patients with AIS admitted to the Affiliated Hospital of Qingdao University from June 2018 to February 2024 whose etiological classification was large artery atherosclerosis (LAA), small vessel occlusion (SVO) or cardioembolism (CE) were included retrospectively. According to the National Institutes of Health Stroke Scale score at admission, patients were divided into mild stroke group (≤8) and moderate to severe stroke group (>8). According to the modified Rankin Scale score at discharge, patients were divided into good outcome group (≤2) and poor outcome group (>2). Multivariate logistic regression analysis was used to determine the independent correlation between cardiac biomarkers and short-term outcome. The predictive value of cardiac biomarkers for poor outcome in patients with AIS and different stroke etiology subtypes were evaluated using receiver operating characteristic (ROC) curves. Results:A total of 2 151 patients with AIS were enrolled, including 1 256 males (58.4%), aged 67.40±11.34 years. 1 079 patents were LAA type (50.2%), 679 were SVO type (31.6%), and 393 were CE type (18.3%); 1 223 were mild stroke (56.86%) and 928 (43.14%) were moderate to severe stroke; 1 357 patients (63.09%) had good short-term outcome, and 794 (36.91%) had poor short-term outcome. Multivariate logistic regression analysis showed that N-terminal pro-B type natriuretic peptide (NT-proBNP), NT-proBNP/creatine kinase (CK) isoenzyme MB (CK-MB) ratio, and CK-MB/CK ratio were independent risk factors for poor short-term outcome. ROC curve analysis shows that the CK-MB/CK ratio had a higher predictive value for short-term poor outcome in patients with AIS (the area under the curve, 0.859, 95% confidence interval 0.839-0.879). Various cardiac biomarkers had a higher predictive value for short-term outcome of CE type and LAA type, but the predictive value for short-term outcome of SVO type was lower. Conclusions:Cardiac biomarkers are associated with the severity and poor outcome of AIS. NT-proBNP/CK-MB and CK-MB/CK ratios have higher predictive value for short-term poor outcome of AIS, especially in patients with CE type.

Objective:To evaluate the safety and effectiveness of laparoscopic ventral mesh rectopexy (LVMR) +anal sphincter plasty for complete rectal prolapse.Methods:From Jan 1, 2018 to Dec 31, 2022, 45 patients with complete rectal prolapse diagnosed in Beijing Erlong Road Hospital received laparoscopic ventral mesh rectopexy +anal sphincter plasty were included in this study.Result:There was no mortality rate associated with LVMR in this study cohort. The follow-up period was 8-76 months, with a median follow-up period of 59 months. There were 2 cases of recurrence, with a recurrence rate of 4%. Patients with concomitant fecal incontinence symptoms had a preoperative Jorge Wexner fecal incontinence score of 13.8±2.0, and postoperative Jorge Wexner fecal incontinence scores of 7.5±1.5, 5.3±1.3, 4.3±1.1, 2.8±1.0, and 1.8±0.5 at 3, 6, 12, 36, and 60 months, respectively, all P<0.001. Patients with concomitant constipation had a preoperative Wexner constipation score of 15.7 ± 1.5, and a postoperative Wexner constipation score of 9.0±1.8, 6.8±1.5, 5.2±1.4, 4.1±0.7, 2.0±0.0 at 3, 6, 12, 36, and 60 months, respectively, all P<0.001. Conclusions:LVMR +anal Sphincter plasty is safe and effective for the treatment of complete rectal prolapse, and there are few complications related to biological patches. Anal sphincter plasty can effectively improve anal function.

Objective To investigate the clinical effect of air moxibustion combined with pushpin-like needle acupuncture,which is formulated based on the"suppressing wood-supporting earth"theory in the doctrine of the five elements of Traditional Chinese Medicine(TCM),in the treatment of pain,nausea and vomiting after transcatheter arterial chemoembolization(TACE).Methods A total of 160 patients with hepatocellular carcinoma,who were admitted to the First Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine of China to receive initial TACE between July 2020 and June 2023,were enrolled in this study.The patients were randomly divided into 4 groups with 40 patients in each group.The patients of the control group received TACE with conventional nursing care,the patients of the pushpin-like needle group received pushpin-like needle acupuncture on the basis of the control group,the patients of the air moxibustion group received air moxibustion treatment on the basis of the control group,and the patients of the combination group received air moxibustion combined with pushpin-like needle acupuncture on the basis of the control group.The post-TACE pain degree(VAS scoring)and nausea/vomiting extent(INVR scoring)were compared between each other among the four groups.Results The differences in the VAS score and INVR score(including main effect and interaction effect)between each other among the four groups were statistically significant(all P＜0.05).Statistically significant differences in the VAS score and INVR score existed between the control group and the other three groups(all P＜0.05).The differences in the VAS score and INVR score between combination group and pushpin-like needle group,as well as between combination group and air moxibustion group were statistically significant(both P＜0.05).Conclusion Air moxibustion combined with pushpin-like needle acupuncture can effectively and remarkably relieve post-TACE pain,nausea and vomiting,which is superior to simple use of air moxibustion or pushpin-like needle acupuncture.Therefor,the combination use of air moxibustion and pushpin-like needle acupuncture should be popularized in clinical practice.

Objective:To establish the myocardial ischemia-reperfusion injury (MIRI) model by ligation of the left ventricular branch of coronary artery in rabbits.Methods:Totally 36 New Zealand white rabbits were randomly divided into 3 groups according to the random number table method, including the sham group ( n=6), the model group ( n=15), and the sustained ischemia group ( n=15). The rabbits were placed under general anesthesia, then endotracheal intubation and common carotid artery intubation were performed. The intercostal muscle between the 3rd and 4th ribs of the left thorax was divided with the aid of a ventilator. The left ventricular branch was located and processed according to the different groups. After successful modeling, the gross morphological change of the heart was observed by naked eye, and the survival rate and modeling success rate of rabbits in each group were calculated respectively. Record the waveform of limb lead Ⅱ electrocardiogram before left ventricular branch ligation (N), 45 minutes of ischemia (I45), 15 minutes of reperfusion (R15), 30 minutes of reperfusion (R30), and 60 minutes of reperfusion (R60). Serum cardiac troponin Ⅰ (cTnⅠ) level was detected by enzyme-linked immunosorbent assay (ELISA), myocardial infarct size ratio was detected by 2, 3, 5-triphenyltetrazolium chloride (TTC) staining, and the change of myocardial tissue changes were detected by hematoxylin-eosin (HE) staining. Results:The survival rate of the rabbits was 93.33% (14/15) in the model group, and the success rate of MIRI modeling was 86.67% (13/15). In the model group, the ST segment (5.00±0.71) mm at I45 was higher than that before the ligation (1.20±0.27) mm, the difference was statistically significant ( P<0.01), and it was stable throughout the whole ischemia period. After ligation removal, the ST segment decreased to (3.00±0.61) mm at R15, (2.20±0.45) mm at R30, and (1.30±0.27) mm at R60. There were statistically significant differences between I45 and N, R15 and I45, and R60 and R15 (all P<0.01). The pathological Q-wave was (1.60±0.55) mm at R15, and decreased to (3.60±0.22 and 5.10±0.22) mm at R30 and R60, respectively. There were statistically significant differences between R30 and R15, R60 and R30, and R60 and R15 (all P<0.01). ST segment (6.10±0.42, 5.80±0.45, 5.60±0.22, and 5.30±0.27) mm at I45, I60, I75 and I105 respectively were higher than the ST segment (1.10±0.22) mm at N in the sustained ischemia group, and the differences were statistically significant (all P<0.01). The serum cTnⅠlevels in the model group at N, I45, R30 and R60 were (62.74±1.60, 97.60±6.36, 159.30±17.64, and 166.40±18.56) ng/L, respectively, and the difference between I45 and N was statistically significant ( P<0.05). There were statistically significant differences between R30 and I45, R60 and I45, and R30 and N (all P<0.01). The serum cTnⅠlevels in the sustained ischemia group were (69.00±4.85, 107.90±7.12, 140.60±10.96, 171.00±15.40) ng/L at N, I45, I75 and I105, respectively. There were statistically significant differences between I45 and N, I75 and I45, and I105 and I75 (all P<0.01). The infarct size ratios of the model group and the sustained ischemia group were 39.93% and (52.16±0.06) %, respectively, and the difference was statistically significant ( P<0.05). Conclusions:The method of establishing a MIRI model by ligating the left ventricular branch of the coronary artery in rabbits is simple to operate, with stable and reliable, and the success rate of modeling is high.

Objective:To analyze the effect of perioperative SEPT9 level in peripheral blood on long-term prognosis of patients with colorectal tumors. Methods:Retrospectively analyzed the data of 334 patients with colorectal cancer admitted to the Department of Anus & Intestine Surgery from January 2017 to December 2022, including 197 male patients and 137 female patients, aged 29 to 83 (62.8±10.7) years. Positive group was consisted of 241 patients with positive SEPT9 before surgery, while negative group was consisted of 93 patients with negative SEPT9 before surgery. Among the positive group, 169 cases turned negative for SEPT9 on the one week after surgery (transnegative group), and another 72 cases did not turn negative (non negative group). Univariate and multivariate analysis of clinical general data were carried out to screen out the risk factors affecting the long-term prognosis of colorectal cancer patients after surgery. The survival curve was calculated by Kaplan-Meier method, and the Log-rank test was used to compare the difference in survival rate between groups. Results:All patients′ overall median survival time was 67 months, and the 1, 3 and 5 years overall survival rate was 91.9%, 70.9% and 57.1%. The results of multi-factor analysis showed that whether the tumor had lymph node metastasis, TNM stage, and preoperative SEPT9 methylation status were independent risk factors affecting the long-term prognosis of colorectal cancer ( P=0.004, <0.001, 0.041), while for patients with preoperative SEPT9 positive, TNM stage of tumor and whether SEPT9 turned negative after surgery were independent risk factors for prognosis ( P=0.026, 0.001). The median survival time of patients in positive group and negative group was 63 months and 71 months, respectively. The 1, 3 and 5 year survival rates after surgery were 90.4%, 67.0%, 55.0% and 95.7%, 79.1% and 64.6%, respectively( P=0.007). The median survival time of the patients in the transnegative group and nonnegative group was 45 months and 62 months, respectively. The 1, 3 and 5-year survival rates were 83.2%, 60.5%, 48.1% and 93.5%, 72.9%, 63.5%( P<0.001). Conclusions:Perioperative SEPT9 level is correlated with long-term prognosis of CRC patients, and patients with negative SEPT9 before surgery have better prognosis than those with positive SEPT9. Preoperative positive patients who do not turn negative after surgery often indicate poor prognosis of tumor.

Electroconvulsive therapy (ECT) is a physical therapy method with rapid onset and remarkable effect for schizophrenia(SZ). It is widely used in patients with agitated impulses, stupor and refractory patients with poor drug treatment effect. The neurophysiological mechanism of ECT effect has not been elucidated. In the past decade, the magnetic resonance imaging (MRI) studies of ECT for SZ have been gradually reported in domestic and international journal, involving magnetic resonance spectroscopy, structural state and resting state analysis, preliminary findings suggest that there may be a correlation between ECT efficacy and structural and functional brain changes, mainly involving morphological changes in brain regions such as the thalamus, hippocampus, insula and left medial prefrontal lobe, as well as functional changes in core resting state networks, particularly the dorsal frontoparietal network, default network and limbic network. More research and exploration are needed in the future to provide valuable clues to further explore the therapeutic mechanism of ECT treatment.