Objective: To investigate the clinical application value of single nucleotide polymorphism (SNP) haplotype analysis in the preimplantation genetic diagnosis (PGD) of monogenic diseases. Methods: The whole genome amplification products of biopsied trophectoderm cells were analyzed by SNP haplotype analysis and verified by Sanger sequencing. Results: A total of 205 embryos were performed SNP haplotype analysis and Sanger sequencing. Among them, Sanger sequencing failed in 14.63% (30/205) of embryos, and SNP haplotype analysis failed in 0.98% (2/205) of embryos. The failure rate of the latter was significantly lower than that of the former (P＜0.05). There were consistent results in 155 (75.61%) embryos, and inconsistent results in 18 (8.78%) embryos. Forty-five embryos in 41 cycles were performed embryo transplantation. The clinical pregnancy rate was 70.73% (29/41) and the implantation rate was 71.11% (32/45). The results of prenatal diagnosis of amniotic fluid during the second trimester of pregnancy were completely consistent with those of SNP haplotype analysis. Conclusion SNP haplotype analysis is accurate, and its failure rate is lower than that of Sanger sequencing. It can be effectively used in the PGD of clinical monogenic diseases.

Objective: To investigate the effects of electrochemically dealloying of Ti6Al4V abutments on human gingival fibroblasts (HGFs) and to provide experimental evidence for surface modification of implant abutments. Methods: The samples were divided into an NC group (negative control, no other treatment on a smooth surface), an NM-1 group (nanomesh-1, electrochemical dealloying treatment in 1 mol/L NaOH 1 h on 2 V voltage), and an NM-2 group (nanomesh-2, electrochemical dealloying treatment in 5 mol/L NaOH 1 h on 2 V voltage). The surface morphologies of the samples and the adhesion of HGFs on the sample surfaces were observed with scanning electron microscopy (SEM). The surface hydrophilicities of the samples were measured with a contact angle measuring instrument. The proliferation of HGFs on the different samples were evaluated with CCK-8, and the expression of adhesion-related genes, including collagen Ⅰ (COL1A1), collagen Ⅲ (COL3A1), fibronectin 1 (FN1), focal adhesion kinase (FAK), vinculin (VCL), integrin α2 (ITGA2), and integrin β1 (ITGB1), on the different samples was measured with qRT-PCR. The expression of vinculin on the surfaces of HGFs was observed via confocal laser scanning microscopy (CLSM) after immunofluorescent staining. Collagen fiber secretion and syntheses of HGFs from different samples were evaluated via Sirius red staining. Results: SEM revealed the formation of ordered and uniform three-dimensional mesh structures on the surfaces of the NM-1 and NM-2 groups, with grid diameters of approximately 30 nm for the NM-1 group and approximately 150 nm for the NM-2 group. Compared with that of the NC group, the water contact angles of the NM-1 group and NM-2 groups were significantly lower (P<0.000 1). Cell proliferation in the NM-1 group was significantly greater than that in the NC group (P<0.01). Moreover, there was no significant difference in the water contact angles or cell proliferation between the NM-1 group and the NM-2 group. SEM revealed that HGFs were adhered well to the surfaces of all samples, while the HGFs in the NM-1 and NM-2 groups showed more extended areas, longer morphologies, and more developed pseudopodia than did those in the NC group after 24 h. qRT-PCR revealed that the expression levels of the adhesion-related genes COL1A1, COL3A1, FN1, FAK and VCL in the NM-1 group were significantly greater than those in the NC and NM-2 groups (P<0.01). The expression of vinculin protein in the NM-1 group was the highest, and the number of focal adhesions was greatest in the NM-1 group (P<0.01). The results of Sirius red staining showed that the NM-1 group had the highest secretion and syntheses of collagen fibers (P<0.000 1). Conclusion The three-dimensional nanomechanical structure of Ti6Al4V modified by electrochemical dealloying promoted the adhesion, proliferation, collagen fiber secretion and syntheses of HGFs, and electrochemical dealloying of Ti6Al4V with a grid diameter of approximately 30 nm obviously promoted HGF formation.

Objective: To evaluate the bidirectional association between periodontitis and Sjögren's syndrome using the Mendelian randomization (MR) method. Methods: Genome-wide association study (GWAS) data of periodontitis (N = 45 563) and Sjögren's syndrome (N = 214 435) were selected to meet the requirements of the same ethnicity and different regions. Inverse variance-weighted (IVW), MR-Egger, and weighted median (WM) tests were used to evaluate the causal effect. Cochran's Q statistics, MR-Egger intercept, MR-PRESSO and leave-one-out analysis were used as sensitivity analyses to assess the stability and reliability of the results. Results: After screening, the GWAS data of Sjögren's syndrome were based on the Finnish region, and the periodontitis GWAS data were based on the UK region, both of which originated from European ancestry. Using IVW (OR = 1.017, 95% CI = 0.956-1.082), MR-Egger (OR = 0.985, 95% CI= 0.956-1.082), and WM (OR =1.021, 95% CI = 0.948-1.099), no causal effect of Sjögren's syndrome on periodontitis was found using any of the three methods. Conversely, no causal effect of periodontitis on Sjögren's syndrome was found (IVW, OR = 1.024, 95% CI = 0.852-1.230; MR-Egger, OR = 0.978, 95% CI = 0.789-1.212; WM, OR = 1.024, 95% CI = 0.846-1.260). The sensitivity analyses indicated that the results were stable and reliable. Cochran's Q test and MR-PRESSO revealed that there was no significant heterogeneity among the instrumental variables, which included single nucleotide polymorphisms (SNPs). The intercept of MR-Egger regression indicated no pleiotropy in the included SNPs. No individual SNP was found that significantly affected the results using the leave-one-out method. Conclusion This study does not support a bidirectional causal effect between periodontitis and Sjögren's syndrome.

Acute Disease ; Child ; Child, Preschool ; Cord Blood Stem Cell Transplantation ; Graft vs Host Disease ; therapy ; Humans ; Immunosuppressive Agents ; therapeutic use ; Infant ; Mesenchymal Stem Cell Transplantation

Objective: To investigate the changes of mitochondrial homeostasis of human gingival fibroblasts (HGFs) in periodontitis, and to provide a basis for exploring the mechanism of periodontitis. Methods: This study has been reviewed and approved by the Ethics Committee. Gingival tissue was collected from patients who underwent periodontal surgery at the Department of Periodontology, School of Stomatology, the Fourth Military Medical University, from June 1, 2023 to December 31, 2023. All of the subjects signed informed consent forms prior to surgery. Gingival connective tissues were collected from patients with chronic periodontitis (CP group) and healthy individuals (control group) undergoing flap surgery and crown lengthening surgery, respectively. There were 6 cases in each group. The primary HGFs obtained from healthy periodontal subjects were cultured and divided into the control group (cultured in complete medium for 24 h) and the Pg.LPS group (cultured in medium with 5μg/mL Pg.LPS for 24 h). The number, morphology, and structure of mitochondria in gingival connective tissue and HGFs were observed by transmission electron microscopy. The number, circumference, and surface area of mitochondria were quantitatively analyzed. MitoSOXTMRed, TMRM, and an ATP kit were used to determine the production levels of mitochondrial reactive oxygen species, mitochondrial membrane potential, and ATP in each group of HGFs. Results: Transmission electron microscopy showed that the morphology and structure of mitochondria were abnormal in the gingival connective tissues of the periodontitis group and HGFs, which were stimulated by Pg.LPS. The mitochondrial ridges were broken or were not visible in these groups. The number of mitochondria decreased significantly, and the surface area and circumference of the mitochondria increased significantly (P < 0.05). In addition, after stimulation by Pg.LPS, the reactive oxygen species level in HGFs was significantly higher than that in the control group (P < 0.05). The mitochondrial membrane potential and the level of ATP production was significantly lower than that of the control group (P < 0.05). Conclusion The number, morphological structure, and function of mitochondria in HGFs changed significantly in periodontitis. The mitochondrial homeostasis is closely related to the occurrence and development of periodontitis.

Objective: To analyze the trends and hotspots in research related to microbiomes and microbes of dental caries; in addition, the study seeks to provide a reference for caries research. Methods: We searched the Web of Science Core Collection (WOSCC) to extract relevant literature in the field of microbiomes and microbes of dental caries published from 2014 to 2023. We used bibliometric visualization evaluation methods such as CiteSpace to conduct visualized analysis of factors that include the number of publications, journals, countries, authors, institutions, co-cited references, and keywords. Results: A total of 3 192 references were extracted, including 2 664 articles and 528 reviews. The number of annual publications is increasing. The United States and China lead the number of publications, with the United States demonstrating a greater capacity for international collaboration. The top 10 journals in percentage of literature are mainly in the field of dentistry followed by the field of microbiology. The author cooperation networks with the highest number of publications include the network led by Zhou Xuedong of Sichuan University, and the network led by Xu Hockin H.K and Weir Michael D of the University of Maryland, Baltimore. The research on microbiomes and microbes of dental caries focuses on the cariogenic toxicity and interaction of microorganisms, oral microbiomes, and the relationship between dental caries and systemic diseases. The articles with high citation frequency mainly involve topics such as dental caries, oral biofilm, oral microbiota, and Streptococcus mutans. Keyword research showed that “dental caries,” “Streptococcus mutans,” “bacteria,” “dental plaque,” and “antibacterial activity” have been the primary focus of research in the last decade. The number of keywords, such as “health” and “oral health,” is on the rise. The latest emergence of “gut microbiome/microbiota” suggests that the oral gut microbiome axis is at the forefront of research in this field, and researchers’ focus is gradually shifting toward the connection between dental caries and systemic diseases. Conclusion Over the last decade, the number of publications in the field microbiomes and microbes of dental caries has increased annually. This research trend will be the multi-omics of the overall oral microbiome, and new research methods and techniques will contribute to the field of cariology.

T helper cells (Th cells) play an important role in periodontitis. During the progression of periodontitis, the levels of pro-inflammatory cytokines such as INF-γ and IL-17, which are produced by Th1 and Th17 cells, are elevated, while the levels of anti-inflammatory cytokines such as IL-4 and TGF-β, which are secreted by Th2 cells and regulatory T cells (Tregs), are diminished. Interventions using mesenchymal stem cells (MSCs) or their exosomes can alter the dynamics of helper T cell populations and their associated cytokine profiles, thereby mitigating the bone loss associated with periodontitis or even promoting bone regeneration. Mesenchymal stem cell-derived exosomes (MSC-exos) have been shown to directly modulate Th cell activity through the proteins and microRNAs they transport. Recent studies indicate that MSC-exos carry immune-suppressive protein molecules: PD-L1 and IDO contribute to regulating the balance between Th17 and Tregs; TGF-β inhibits the proliferation of T lymphocytes while facilitating differentiation into Tregs by sustaining forkhead box protein O3 (FOXP3) and Smad expression; and CD73 catalyzes the conversion of monophosphate adenosine into adenosine, which interacts with A2A receptors on Th1 cells to induce apoptosis in Th1 cells. In addition, microRNAs exhibit immunoregulatory functions: periodontal ligament stem cell-derived exosomes contain miRNA-155-5p, which targets sirtuin-1 to suppress Th17 cell differentiation. Furthermore, evidence in rat models of periodontitis suggests that these exosomes may also carry miR-205-5p targeting XBP1 to restore the balance between Th17 and Tregs. Dental pulp stem cell-derived exosomes reestablish this balance via the miR-1246/Nfat5 axis. Bone marrow mesenchymal stem cell-derived exosomes harbor miR-1246, which targets ACE2 to promote differentiation towards Tregs. Moreover, MSC-exos can indirectly enhance the differentiation of Tregs through interactions with other immune entities, such as antigen-presenting cells or macrophages. This article reviews the changes and roles of helper T cells in periodontitis, as well as the regulatory role of exosomes on helper T cells, hoping to provide new ideas for immunotherapy in the treatment of periodontitis.

Immediate implant placement in the aesthetic zone has become increasingly widespread and has gradually evolved into a conventional techniques for implant procedures in the aesthetic region. To achieve favorable aesthetic and long-term outcomes, clinicians must possess extensive clinical experience as well as proficient surgical and restorative skills. This study summarizes the key factors influencing the long-term success of immediate implants in the aesthetic zone: strict adherence to the indications for immediate implant placement; thorough preoperative assessment of the patient’s systemic and local conditions, along with comprehensive evaluation of aesthetic risks; minimally invasive tooth extraction while preserving the integrity of the labial bone plate; selection of appropriately designed implants and their placement in an ideal three-dimensional position based on the implant’s characteristics; utilization of suitable bone and soft tissue augmentation techniques according to the patient’s specific hard and soft tissue anatomy, extent of bone defects, and periodontal phenotype; dynamic shaping of soft tissues through continuous adjustments in the emergence profile of provisional restorations; design of definitive restorations from the perspectives of health, function, and aesthetics; and implementation of regular follow-up and maintenance protocols after implant treatment, with increased emphasis on peri-implant care for patients who smoke, have diabetes, or undergo anti-osteoporosis therapy. This study proposes a decision-making framework to achieve long-term stable clinical outcomes with immediate implants in the aesthetic zone, providing a reference for clinicians in their clinical decision-making and treatment planning: ① for patients assessed as low aesthetic risk (e.g., thick gingival biotype, absence of hard and soft tissue defects, intact labial bone plate with thickness >1 mm, no acute infection), immediate implant placement after minimally invasive extraction is recommended, with the implant positioned in an ideal three-dimensional location, along with bone grafting in the gap between the implant and the labial bone plate and consideration of connective tissue grafting when required; ② for patients assessed as moderate aesthetic risk (e.g., thin gingival biotype, absence of soft tissue defects, intact labial bone plate but with thickness <1 mm or mild to moderate bone defects involving less than 50% height loss, chronic infection present), immediate implant placement with optimal three-dimensional positioning is feasible, accompanied by bone grafting in the implant-labial bone gap or external bone grafting on the labial aspect, with simultaneous or staged connective tissue grafting, or alternatively, use of the socket shield technique for immediate implant placement; ③ for patients assessed as high aesthetic risk (e.g., thin gingival biotype, presence of soft tissue defects, vertical bone deficiency, severe labial bone loss involving >50% height loss, acute infection present), ridge preservation followed by delayed implant placement is advised. By adhering to these treatment principles, immediate implant placement in the aesthetic zone can achieve reliable success rates and excellent aesthetic outcomes.

Periodontitis is a chronic inflammatory disease that affects the tooth-supporting tissues, and it constitutes a major global public health concern. Methylation modifications, including DNA methylation, histone methylation, and RNA m6A modification, represent reversible processes coordinately regulated by methyltransferases, demethylases, and binding proteins. In periodontitis, aberrant methylation modifications suppress Toll-like receptor 2 expression, leading to oral microbial dysbiosis. These modifications further disrupt normal immune regulatory functions through C-C motif chemokine ligands, Fc-γ receptor-mediated phagocytosis, and NF-κB signaling pathways, resulting in localized immune-inflammatory imbalance in periodontal tissues. In addition, various methylation modifications regulate the expression of Runt-related transcription factor 2 (RUNX2), osteoblast-specific transcription factor Osterix (OSX), and receptor activator of nuclear factor-κB ligand (RANKL), thereby interfering with osteoclast and osteoblast differentiation, disrupting bone homeostasis, and ultimately driving alveolar bone resorption. Methylation-related biomarkers demonstrate promising potential for periodontitis screening and prognostic evaluation. While numerous abnormally methylated sites have been identified in periodontitis, the precise signaling pathways and comprehensive epigenetic regulatory networks remain to be fully elucidated. This review systematically summarizes the functional roles of DNA methylation modifications in the pathogenesis of periodontitis and explores their potential value in etiological studies, diagnostic biomarker discovery, and targeted therapeutic interventions, with the aim of providing novel perspectives for periodontitis prevention and treatment strategies.

Objective: To investigate the effects of ginsenoside Rg3 on the formation of vasculogenic mimicry (VM) in gastric cancer cell line SGC7901 and its molecular mechanism. Methods: MTT assay was used to detect the effect of different concentrations of Rg3 on the proliferation of SGC7901 cells. SGC7901 cells were grouped as follows: BML-284 group, XAV-939 group, Rg3 group, Rg3+ BML-284 group and blank group. Transwell chamber assay was used to detect cell invasion and migration; the formation of VM was observed by tube formation assay; the secretion of MMP-9 and MMP2 was detected by ELISA; the mRNA expressions of GSK-3β and Wnt2B were detected by qPCR; the expression of β-Catenin protein in cells was analyzed by WB; and nuclear entry of β-Catenin was examined by Immunofluorescence. Results: Ginsenoside Rg3 inhibited the proliferation of SGC7901 cells in a time- and concentrationdependent manner; compared with the blank group, 40 mg/L Rg3 significantly inhibited the invasion and migration of SGC7901 cells (both P<0.05) and VM formation (P<0.05); in the meanwhile, the expressions of intracellular GSK-3β, Wnt2B mRNA and β-catenin protein, as well as the nuclear entry of β-catenin were significantly inhibited (all P<0.05). The invasion, migration and VM formation of SGC7901 cells in Rg3+BML-284 group were not significantly different from those in the blank group (all P>0.05). Conclusion: Rg3 can inhibit cell invasion, migration and VM formation in SGC7901 cells by inhibiting the activation of Wnt/β-Catenin pathway.