ObjectiveTo observe the clinical effectiveness and safety of Xiaozhong Zhitong Mixture （消肿止痛合剂） combined with antibiotic bone cement in the treatment of diabetic foot ulcer （DFU） with damp-heat obstructing syndrome. MethodsA total of 72 DFU patients with damp-heat obstructing syndrome were randomly assigned to treatment group （36 cases） and the control group （36 cases）. Both groups received standard treatment and topical antibiotic bone cement for ulcer wounds， while the treatment group received oral Xiaozhong Zhitong Mixture （50 ml per time， three times daily） in additionally. Both groups underwent daily wound dressing changes for 21 consecutive days. Ulcer healing rate， serum levels of tumor necrosis factor-alpha （TNF-α）， interleukin-1 beta （IL-1β）， malondialdehyde （MDA）， superoxide dismutase （SOD）， C-reactive protein （CRP）， and white blood cell （WBC） count were observed before and after treatment， and visual analog scale （VAS） scores for wound pain， traditional Chinese medicine （TCM） syndrome scores， and the DFU Healing Scale （DMIST scale） were also compared. Liver and kidney function were evaluated before and after treatment， and adverse events such as allergic reactions， worsening ulcer pain were recorded. ResultsTotally 35 patients in the treatment group and 33 in the control group were included in the final analysis. The ulcer healing rate in the treatment group was （87.93±9.34）%， significantly higher than （81.82±12.02）% in the control group （P = 0.035）. Compared to pre-treatment levels， both groups showed significant reductions in serum CRP， WBC， MDA， IL-1β， and TNF-α levels， with an increase in SOD level （P＜0.05）. TCM syndrome scores， VAS， and DMIST scores also significantly decreased in both groups （P＜0.05）， with greater improvements in the treatment group （P＜0.05）. No significant adverse reactions were observed in either group during treatment. ConclusionXiaozhong Zhitong Mixture combined with antibiotic bone cement has significant advantages in promoting DFU healing， reducing inflammatory response， and alleviating oxidative stress in DFU patients with damp-heat obstructing syndrome， with good safety for DFU patients with damp-heat obstructing syndrome.

BACKGROUND:Flap transplantation technique is a commonly used surgical procedure for the treatment of severe tissue defects,but postoperative flap necrosis is easily triggered by ischemia-reperfusion injury.Therefore,it is still an important research topic to improve the survival rate of transplanted flaps. OBJECTIVE:To review the pathogenesis and latest treatment progress of flap ischemia-reperfusion injury. METHODS:CNKI,WanFang Database and PubMed database were searched for relevant literature published from 2014 to 2024.The search terms used were"flap,ischemia-reperfusion injury,inflammatory response,oxidative stress,Ca2+overload,apoptosis,mesenchymal stem cells,platelet-rich plasma,signaling pathways,shock wave,pretreatment"in Chinese and English.After elimination of irrelevant literature,poor quality and obsolete literature,77 documents were finally included for review. RESULTS AND CONCLUSION:Flap ischemia/reperfusion injury may be related to pathological factors such as inflammatory response,oxidative stress response,Ca2+overload,and apoptosis,which can cause apoptosis of vascular endothelial cells,vascular damage and microcirculation disorders in the flap,and eventually lead to flap necrosis.Studies have found that mesenchymal stem cell transplantation,platelet-rich plasma,signaling pathway modulators,shock waves,and pretreatment can alleviate flap ischemia/reperfusion injuries from different aspects and to varying degrees,and reduce the necrosis rate and necrosis area of the grafted flap.Although there are many therapeutic methods for skin flap ischemia/reperfusion injury,a unified and effective therapeutic method has not yet been developed in the clinic,and the advantages and disadvantages of various therapeutic methods have not yet been compared.Most of the studies remain in the stage of animal experiments,rarely involving clinical observations.Therefore,a lot of research is required in the future to gradually move from animal experiments to the clinic in order to better serve the clinic.

BACKGROUND:The osteogenic differentiation ability of exosomes derived from osteogenic mesenchymal stem cells is well established.However,their impact on the osteogenic differentiation of human periodontal ligament stem cells in an inflammatory microenvironment remains unclear. OBJECTIVE:To examine the impact of exosomes derived from osteogenesis-induced human periodontal ligament stem cells on the osteogenic differentiation of human periodontal ligament stem cells within an inflammatory microenvironment. METHODS:Human periodontal ligament stem cells were isolated and cultured.After 3 days of osteogenic induction,exosomes were extracted.Human periodontal ligament stem cells were divided into four groups.Control group was treated with osteogenesis-induced medium.The exosome group was treated with osteogenesis-induced medium containing 5 μg/mL exosomes.Inflammatory model and inflammatory model+exosome groups were treated with 1 μg/mL lipopolysaccharide for 24 hours to construct a cellular inflammatory microenvironment.The inflammatory model group was treated with osteogenesis-induced medium after lipopolysaccharide intervention.The inflammatory model+exosome group was treated with osteogenesis-induced medium containing 5 μg/mL exosome.The osteogenic differentiation ability of human periodontal ligament stem cells was assessed using alkaline phosphatase staining and alizarin red staining.The expressions of Runt-related transcription factor 2,osteopontin,osteoblast-specific transcription factor Osterix(OSX)and wnt pathway-related protein β-catenin were detected by real-time fluorescence quantitative PCR and western blot assay. RESULTS AND CONCLUSION:(1)Compared with the control group,the relative area stained by alkaline phosphatase,the relative area stained by mineralized nodules and the expression levels of Runx2,osteopontin,and OSX were significantly decreased in the inflammatory model group(P<0.05).(2)Compared with the inflammatory model group,the expression of Runx2,osteopontin,and OSX in the inflammatory model+exosome group was significantly increased in the relative area of alkaline phosphatase staining,the relative area of mineralized nodules staining(P<0.05).(3)Compared with the control group,the expression of wnt pathway-related protein β-catenin was significantly increased in the inflammatory model group(P<0.05).Compared with the inflammatory model group,the expression of β-catenin in the inflammatory model+exosome group was significantly decreased(P<0.05).These findings indicate that exosomes derived from human periodontal ligament stem cells induced by bone formation can enhance the osteogenic differentiation of human periodontal ligament stem cells within an inflammatory microenvironment,and the mechanism may be related to wnt/β-catenin signaling pathway.

OBJECTIVE To optimize the forming process of Yindan huoxue tongyu granules， and evaluate the quality of the granules. METHODS Taking forming rate， angle of repose， moisture， moisture absorption rate and dissolution rate as indexes， single factor experiment combined with Plackett-Burman design was adopted to screen key process parameters； analytic hierarchy process combined with entropy weight method and Box-Behnken response surface method were used to optimize the molding process of Yindan huoxue tongyu granules， and the forming process was verified. The relative homogeneity index， bulk density， vibration density， Hausner ratio， angle of repose， moisture and hygroscopicity were used as secondary physical indexes to establish the physical fingerprints of 10 batches of Yindan huoxue tongyu granules to evaluate particle quality consistency. RESULTS The optimal molding process of Yindan huoxue tongyu granules was as follows： mannitol as the fixed excipient， the drug-assisted ratio was 1∶1（m/m） and the drying time was 1 h； 90% ethanol was used as wetting agent and the amount of it was 32%， the drying temperature was 70 ℃. The results of validation tests showed that the average comprehensive score was 97.45， which was close to the predicted value of 97.18. The similarities between the physical fingerprints of 10 batches of Yindan huoxue tongyu granules prepared by the optimal molding process and the reference physical fingerprint were all higher than 0.99. CONCLUSIONS The molding process is stable and feasible， and the quality of Yindan huoxue tongyu granules produced is stable and controllable.

Diabetic ulcer （DU） wound is one of the chronic and serious complications of diabetes characterized by prolonged wound healing， and it is more common in foot and lower extremity ulcers. DU has brought great economic and psychological pressure to patients and seriously affected the quality of life of patients because of its great difficulty in treatment， long treatment process， and high morbidity and mortality. Therefore， how to help the rapid healing of DU wounds， reduce the disability rate and mortality rate， protect limb function， and improve the quality of life is an important topic and hot spot in the field of medical research. The pathogenesis of DU is complex， mainly including microcirculation disorder， peripheral neuropathy， inflammation and infection， and excessive apoptosis of cells， involving physiological processes such as wound inflammation， granulation tissue hyperplasia and re-epithelialization. A large number of previous studies have found that Chinese medicine can regulate phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt）， Wnt/β-catenin， nuclear factor-κB （NF-κB）， Notch， nuclear factor E₂-related factor 2 （Nrf2）， transforming growth factor-β （TGF-β）/Smad， and other signaling pathways， regulate abnormal glucose metabolism， improve microcirculation， inhibit inflammation and oxidative stress， regulate cell proliferation and excessive apoptosis， and promote wound tissue growth to promote the rapid healing of DU wounds under the guidance of treatment based on traditional Chinese medicine （TCM） syndrome differentiation and internal and external treatment. Therefore， this paper reviewed Chinese medicinal monomers or Chinese medicinal compounds in recent years in regulating the above signaling pathways and the expression of key protein molecules and promoting the rapid healing of DU wounds， aiming to provide ideas and a theoretical basis for the in-depth study and clinical application of Chinese medicine in promoting the healing of DU wounds.

Objective: To analyze the relationship between microorganisms and endodontic disease by using 16 S rDNA sequencing to compare the composition of the microbial community in the root canals of teeth with pulpitis and apical periodontitis. Methods: Clinical samples were collected from teeth requiring root canal treatment.The total DNA of the bacteria in the samples and the gene fragments of the V3-V4 highly variable region on the 16S rDNA fragments were amplified through PCR.After sequencing by NovaSeq,statistical and bioinformatic analysis,including phylogenetic analysis,diversity analysis and analysis of group differences,were performed. Results: In total,6 teeth with pulpitis and 7 teeth with apical periodontitis were collected,and a total of 8 510 OTUs were obtained after next-generation sequencing,and the analysis of bacterial diversity showed that the difference between pulpitis and apical periodontitis in terms of the composition of the bacterial flora was statistically significant(P<0.05).In particular,the relative abundance of Proteobacteria,Acidobacteriota and Actinobacteriota phylum was significantly higher in the roots of teeth affected by pulpitis than apical periodontitis.The relative abundance of Bacteroidota phylum and Synergistota phylum was significantly higher in the root canals of teeth with apical periodontitis. Conclusion There is a complex diversity of infecting microorganisms in the root canals of teeth affected by endodontic diseases.The microbial communities in the infected root canals of pulpitis and apical periodontitis show some differences,and changes in the microbial composition of the root canals may be associated with the development of endodontic diseases.

Objective:To investigate the effects of Porphyromonas gingivalis derived outer membrane vesicles (Pg OMV) on osteoclast differentiation of macrophages and its underlying mechanisms. Methods:The morphology and the size distribution of Pg OMV were analyzed by transmission electron microscopy and nanoparticle tracing analysis, respectively. The osteoclast precursors were treated with 1, 3 and 10 mg/L Pg OMV (1, 3 and 10 mg/L OMV treatment group) or phosphate buffer solution (PBS)(control group). The formation of osteoclasts was analyzed by tartrate-resistant acid phosphase (TRAP) staining and F-actin staining and real-time quantitative PCR (RT-qPCR) were used to detect the expression of Fos and matrix metallopeptidase 9 (MMP9). Polymyxin B (PMB) was used to block lipopolysaccharide (LPS) and then Pg OMV was used to treat osteoclast precursor (PMB-OMV treatment group), and OMV treatment group was used as control. TRAP and F-actin staining were used to observe the formation of osteoclasts and actin rings. The effect of Pg OMV on the expression of Toll-like receptor (TLR) 2 and TLR4 in preosteoclasts was detected by Western blotting. The osteoclast precursors were pretreated with 10, 50, 100 and 200 μmol/L C29, an inhibitor of TLR2, and then treated with Pg OMV(OMV+10, 50, 100 and 200 μmol/L C29 treatment group) and OMV treatment group without C29 pretreatment was control. TRAP and F-actin staining were used to observe the formation of osteoclasts and actin rings. The osteoclast precursor cells were treated with OMV (OMV treatment group) and OMV incubated with PMB (PMB-OMV treatment group) and the expression of TLR2 in osteoclast precursor was detected by Western blotting.Results:Pg OMV showed classical vesicular structures, and the average particle size of Pg OMV were 179.2 nm. A large number of actin rings were observed in the 3 and 10 mg/L OMV treatment groups. The percentages of TRAP-positive osteoclast area in 3 mg/L OMV treatment group [(22.6±2.1)%] and 10 mg/L OMV treatment group [(32.0±2.3)%] were significantly increased compared with control group [(4.9±0.5)%] ( P<0.001). Compared with the control group (1.000±0.029), the mRNA relative expression of Fos in 3 mg/L OMV treatment group (1.491±0.114) and 10 mg/L OMV treatment group (1.726±0.254) was significantly increased ( P=0.013, P=0.001). Compared with the control group (1.007±0.148), the mRNA relative expression of MMP9 in the group of 10 mg/L OMV (2.232±0.097) was significantly increased ( P<0.001). Actin ring formation was less in PMB-OMV treatment groups than in OMV treatment groups. The proportion of TRAP-positive osteoclasts area [(14.8±3.8)%] in PMB-OMV treatment group was significantly lower than OMV treatment group [(31.5±6.7) %] ( P=0.004). The relative expression of TLR2 in OMV treatment group (1.359±0.134) was significantly higher than that in the control group (1.000±0.000) ( t=4.62, P=0.044). Compared with the OMV treatment group [(29.4±1.7)%], 50, 100 and 200 μmol/L C29 significantly decreased the formation of osteoclasts [(24.0±1.7)%, (18.5±2.1)%, (9.1±1.3) %] ( P=0.026, P<0.001, P<0.001). TLR2 protein expression in PMB-OMV group (0.780±0.046) was significantly lower than that in OMV group (1.000±0.000)( t=8.32, P=0.001). Conclusions:Pg OMV can promote osteoclast differentiation by carrying LPS, TLR2 plays an important role in Pg OMV mediated osteoclast differentiation.

The oral cavity is the second largest reservoir of microorganisms in the human body, containing more than 700 species. Periodontal microorganisms are an important part of oral microorganisms. Plaque biofilm, the initiator of periodontal disease, contains an abundance of oral microorganisms. The special complex anatomy of the periodontium allows for a higher abundance of the periodontal microbiota. There are growing evidences show that the periodontal microbiota is not only closely associated with oral diseases, but also plays an important role in mouth-brain interactions. Dysbiosis of the periodontal microbiota may facilitate the progression of neurodegenerative diseases including Alzheimer disease, Parkinson disease, and multiple sclerosis. Here, this paper reviews the bidirectional role of periodontal microbiota between the oral cavity and the brain, that is, the bridge effect of periodontal microbiota involved in the interaction between the two diseases, enumerates the epidemiological and biological evidences that dysregulation of the periodontal microbiota induces and exacerbates neurodegenerative diseases, and analyzes their possible mechanisms. The positive implications of periodontal microbial homeostasis in the prevention and treatment of neurodegenerative diseases are described with the aim of providing new ideas and insights into the pathogenesis and therapeutic approaches for neurodegenerative diseases.

[Objective]To investigate the protective effect and mechanism of Ganoderma lucidum extract(GLE)on liver cirrhosis in mice.[Methods]Ten male C57BL/6 mice were randomly selected as control group,the remaining forty mice were intraperitoneally injected with carbon tetrachloride olive oil suspension to induce liver cirrhosis model.They were randomly divided into model group and GLE low(50 mg/kg·d),medium(100 mg/kg·d)and high(200 mg/kg·d)dose groups,while the control group and model group received 0.9％sodium chloride solution gastric irrigation,and the control group mice were given the same volume of olive oil solution twice a week.Liver index was calculated.The activities of alanine aminotransferase(ALT),aspartate transaminase(AST)and the levels of total cholesterol(TC),total bilirubin(TB)and creatinine(Cr)in serum of mice were detected by automatic biochemical analyzer.Hematoxylin eosin(HE)staining was used to observe the histopathological changes of liver,and Masson staining was used to observe the degree of liver fibrosis.Terminal-deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL)staining was used to observe the apoptosis of hepatocytes.The levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-6,malondialdehyde(MDA)and activity of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in serum were detected by enzyme linked immunosorbent assay(ELISA).The relative expression of nuclear factor E2 related factor 2(Nrf2)and nuclear Nrf2,heme oxygenase-1(HO-1)and NAD(P)H:quinone oxidoreductase 1(NQO1),α-smooth muscle actin(α-SMA),Collagen Ⅰ and E-cadherin protein in liver tissue were detected by Western blot.[Results]Compared with control group,the liver had significant damage,the liver index,serum ALT,AST activities,TC,TB and Cr levels,liver fibrosis degree,hepatocyte apoptosis index,the levels of serum TNF-α,IL-1 β,IL-6 and MDA,the relative expression of α-SMA and Collagen I protein increased(P＜0.05),while the activity levels of serum SOD and GSH-Px,and the relative expression of total Nrf2 and nuclear Nrf2,HO-1,NQO1 and E-cadherin protein in liver tissue decreased in model group(P＜0.05).Compared with model group,liver injury gradually reduced,the liver index,serum ALT,AST activities,TC,TB and Cr levels,liver fibrosis degree,hepatocyte apoptosis index,the levels of serum TNF-α,IL-1β,IL-6,MDA and the relative expression of α-SMA,Collagen I protein decreased(P＜0.05),while the activity levels of serum SOD and GSH-Px,and the relative expression of Nrf2 and nuclear Nrf2,HO-1,NQO1 and E-cadherin protein in liver tissue increased in GLE low,medium and high dose groups(P＜0.05).[Conclusion]GLE can alleviate the histopathological damage and improve liver function in cirrhotic mice.This may be related to the decreased level of oxidative stress and inflammatory reaction after activation of Nrf2/ARE signaling pathway,which may interfere with liver fibrosis.

Exosomes(EXOs)are important mediators of intercellular communication that contain a variety of sub-stances,including miRNA,mRNA,DNA,and protein molecules,which can act on target cells and have broad medical prospects as"cell-free therapy".The inclusion of EXOs varies with the type and state of the donor cell,thus EXOs from different cell types may exhibit different biological effects.Dental mesenchymal stem cell(DMSC)-derived EXOs(DMSC-EXOs)have gained increasing research attention in the fields of tissue regenerative medicine and immune regulation.Current research on EXOs is focused on the homeostasis between proinflammatory(M1)/anti-inflammatory(M2)macro-phages and T-helper 17(Th17)/regulatory T(Treg)cells during periodontal immune regulation.Studies have shown that DMSC-EXOs can promote the transformation of macrophages and T cells and that this function may be dependent on the surrounding microenvironment and the tissue origin of stem cells.For instance,miR-1246 in dental pulp stem cell-de-rived EXOs promotes M2 macrophage polarization by inhibiting nuclear factor kappa-B(NF-κB)p65.Meanwhile,EXOs derived from stem cells from apical papilla promote DNA demethylase Tet2-mediated demethylation of FoxP3,maintain stable FoxP3 expression,and promote Treg cell transformation,thus alleviating local inflammation in periodontitis.In addition,the immunomodulatory activities of DMSC-EXOs can be affected by inflammatory factors.For example,EXOs derived from lipopolysaccharide-preconditioned dental follicle stem cells can reduce the receptor activator of NF-κB li-gand/osteoprotegerin ratio through the reactive oxygen species(ROS)/c-Jun N-terminal kinase signaling pathway and pro-mote M2 macrophage polarization through the ROS/extracellular signal-regulated kinase signaling pathway.Additional-ly,EXOs derived from gingiva-derived mesenchymal stem cells pretreated with tumor necrosis factor-α and interferon-α proinflammatory cytokines can promote M2 macrophage polarization through high expression of CD73 and CD5L,while EXOs derived from inflammatory periodontal ligament stem cells can promote M1 macrophage polarization.This article reviews the research progress on the immunoregulation and effects of DMSC-EXOs on the homeostasis of M1/M2 macro-phages and Th17/Treg cells during periodontal immune regulation and provides a reference for the treatment of peri-odontitis using DMSC-EXOs.