[Comfortable initial periodontal therapy: step by step].

With the transition of the medical model from the traditional biomedical model to the biopsychosocial one, there is a growing trend and requirement for oral operations that prioritize comfort, pain management, minimally invasive techniques, and visualization. Consequently, demands for comfortable dental treatments among individuals are increasing. However, initial periodontal therapy is often accompanied by pain, and patients' reactions to pain range from nervousness to dental fear, such as irritability, hyperventilation, even nausea, vomiting, and refusal to cooperate, which make the implementation of initial periodontal therapy difficult or even impossible. This article will focus on three key steps: firstly, the preparation of the clinic, the acquisition of patients' trust and the implementation of preventive sedation before treatment; secondly, the use of comfort operation and nursing, psychological intervention measures, local anesthesia, and sedation techniques during treatment; thirdly, the health education and follow-up after treatment. By addressing these aspects, we aim to clarify how to perform comfortable initial periodontal therapy step by step.

[Inhibitory effect of downregulating G protein-coupled receptor class C group 5 member A expression on lipopolysaccharide-induced inflammatory response in human gingival fibroblasts].

Objective: To clarify the effect and the mechanism of G protein-coupled receptor class C group 5 member A (GPRC5A) on lipopolysaccharide (LPS)-induced inflammatory response in human gingival fibroblasts (GFs), thus to provide a foundation for delving into the role of G protein coupled receptor (GPCR) in periodontitis. Methods: Gingival tissue samples were collected from 3 individuals periodontally healthy (health group) and 3 patients with periodontitis (periodontitis group) in Shandong Stomatological Hospital from December 2022 to February 2023. The expressions of GPRC5A of the two groups were detected by immunohistochemistry staining. GFs used in this study were isolated from a portion of gingiva for the extraction of impacted teeth in School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University from December 2022 to February 2023. GFs were isolated with enzymic digestion and transfected with 30, 50 and 80 µmol/L small interfering RNA-GPRC5A (siGPRC5A) or small interfering RNA-negative control (siNC), regarded as the experimental group and the negative control one, respectively. The silencing efficiency of siGPRC5A was evaluated by real-time fluorescence quantitative PCR (RT-qPCR). Experiments were then conducted using these cells which were divided into four groups of negative control (NC), LPS, siGPRC5A+LPS and siGPRC5A. The mRNA and protein levels of GPRC5A in GFs under 1 mg/L LPS-induced GFs inflammatory state were evaluated by RT-qPCR and Western blotting analysis after GPRC5A knockdown. RT-qPCR was used to detect the gene expression levels of the inflammatory cytokines in GFs induced by LPS, namely, interleukin (IL)-1ß, IL-6, IL-8, tumor necrosis factor (TNF)-α, prostaglandin endoperoxide synthase 2 (PTGS2) after GPRC5A knockdown. Western blotting analysis and immunofluorescence staining were used to further investigate the activation of nuclear factor-kappa B (NF-κB) signaling pathway. Results: Immunohistochemistry staining showed that the expression of GPRC5A in gingival tissues of periodontitis group (0.132±0.006) increased compared with that in periodontally healthy group (0.036±0.019) (t=8.24, P=0.001). Meanwhile, RT-qPCR results showed that the gene expression levels of GPRC5A at different time point (2, 6, 12, 24 h) in LPS-induced GFs (0.026±0.002, 0.042±0.005, 0.004±0.000, 0.016±0.000) were upregulated compared with those in the NC group (0.004±0.000, 0.004±0.000, 0.002±0.000, 0.007±0.000) (all P<0.001), respectively, and peaked at 6 h. The 50 µmol/L group displayed the most significant decrease in siGPRC5A expression (31.16±3.29) compared with that of the siNC group (100.00±4.88) (F=297.98, P<0.001). The results of RT-qPCR and Western blotting analysis showed that siGPRC5A (0.27±0.03, 0.71±0.00) suppressed the expressions of GPRC5A at both gene and protein levels, while LPS (1.30±0.10, 1.43±0.03) was able to promote the expressions of GPRC5A compared with those of the NC group (1.00±0.01, 1.00±0.00)(all P<0.001). The siGPRC5A+LPS group (0.39±0.03, 1.06±0.16) also inhibited the increase of GPRC5A at both gene and protein levels induced by LPS (1.30±0.10, 1.43±0.03) (F=208.38, P<0.001; F=42.04, P<0.001). RT-qPCR results showed that the expressions of IL-8, IL-1ß, IL-6, TNF-α, and PTGS2 at the gene level in LPS group were highly increased compared with those in the NC group (all P<0.001). siGPRC5A significantly suppressed LPS-induced expressions of these inflammatory cytokines in GFs (all P<0.001). Western blotting analysis showed that the levels of p65 and IκBα protein phosphorylation in the LPS group were highly increased compared with those in the NC group, and siGPRC5A could effectively suppressed LPS-induced protein phosphorylation (all P<0.01). Furthermore, immunofluorescence staining showed that NF-κB p65 in the control group was mainly concentrated in the cytoplasm, and partially translocated to the nucleus under the stimulation of LPS. siGPRC5A was able to inhibit LPS-induced intranuclear translocation of p65 to a certain extent. Conclusions: GPRC5A expression was upregulated in periodontitis, and GPRC5A knockdown inhibited LPS-induced inflammation. Moreover, GPRC5A played a role in inflammation regulation by interacting with NF-κB signaling pathway.

[Review on the development of periodontology in China].

Chinese Journal of Stomatology has gone through 70 years of ups and downs, witnessing the development of periodontics in China from a faltering start to twists and turns, and finally innovative development. This article aims to review the periodontology-related papers published in the Chinese Journal of Stomatology. Based on the characteristics of the times, they are summarized into five stages: staggering start, forced stagnation, vigorous development, standardized innovation, and disciplinary integration. Researches on periodontal diseases in China initially focused on learning and reference, gradually caught up with the international level, and finally created in-depth insights and innovations. Eventually, Chinese periodontology has formed a research system with Chinese characteristics and achieved substantial achievements in clinical diagnosis and treatment, basic research, periodontal medicine, and disciplinary integration. Although the current status of Chinese periodontology still lags behind that of developed countries, these representative studies demonstrate the unremitting efforts and hard work of periodontists for generations, laying a solid foundation for the innovation and development of periodontology in our country.

[The efficacy of chemotherapy re-challenge in third-line setting for metastatic colorectal cancer patients: a real-world study].

Objective: To explore the efficacy of chemotherapy re-challenge in the third-line setting for patients with metastatic colorectal cancer (mCRC) in the real world. Methods: The clinicopathological data, treatment information, recent treatment efficacy, adverse events and survival data of mCRC patients who had disease progression after treatment with oxaliplatin-based and/or irinotecan-based chemotherapy and received third-line chemotherapy re-challenge from January 2013 to December 2020 at Tianjin Medical University Cancer Institute and Hospital were retrospectively collected. Survival curves were plotted with the Kaplan-Meier method, and the Cox proportional hazard model was used to analyze the prognostic factors. Results: A total of 95 mCRC patients were included. Among them, 32 patients (33.7%) received chemotherapy alone and 63 patients (66.3%) received chemotherapy combined with targeted drugs. Eighty-three patients were treated with dual-drug chemotherapy (87.4%), including oxaliplatin re-challenge in 35 patients and irinotecan re-challenge in 48 patients. The remaining 12 patients were treated with triplet chemotherapy regimens (12.6%). Among them, as 5 patients had sequential application of oxaliplatin and irinotecan in front-line treatments, their third-line therapy re-challenged both oxaliplatin and irinotecan; 7 patients only had oxaliplatin prescription before, and these patients re-challenged oxaliplatin in the third-line treatment. The overall response rate (ORR) and disease control rate (DCR) reached 8.6% (8/93) and 61.3% (57/93), respectively. The median progression free survival (mPFS) and median overall survival (mOS) were 4.9 months and 13.0 months, respectively. The most common adverse events were leukopenia (34.7%) and neutropenia (34.7%), followed by gastrointestinal adverse reactions such as nausea (32.6%) and vomiting (31.6%). Grade 3-4 adverse events were mostly hematological toxicity. Cox multivariate analysis showed that gender (HR=1.609, 95% CI: 1.016-2.548) and the PFS of front-line treatments (HR=0.598, 95% CI: 0.378-0.947) were independent prognostic factors. Conclusion: The results suggested that it is safe and effective for mCRC patients to choose third-line chemotherapy re-challenge, especially for patients with a PFS of more than one year in front-line treatments.

[Long-term quality of life in patients with Brownâ ¡ maxillary defect repaired by different methods].

Objective: To analyze the long-term quality of life of patients with Brown Ⅱ maxillary defect repaired by tissue flap or prosthesis. Methods: Patients who underwent surgery for maxillary malignant tumors in the First Affiliated Hospital of Bengbu Medical College from 2014 to 2017 were selected to investigate the postoperative long-term (>5 years) quality of life using the fourth edition of the University of Washington quality of life questionnaire (UW-QOL). Mann Whitney U test was used to examine the differences between two groups. Results: In this study, 4 cases were lost to follow-up, 9 died, and a total of 46 valid questionnaires were collected, including 24 males and 22 females, aged 19-86 years. There were 26 cases of class Ⅱb/c and 20 cases of class Ⅱd. Tissue flap reconstruction was performed in 29 cases (tissue flap group) and prosthesis restoration in 17 cases (prosthesis group). The score of chewing QOL in the prosthesis group was higher than that in the tissue flap reconstruction group (Z=-2.787, P=0.005), but the scores of entertainment, swallowing, speech and emotion QOL in the former group were respectively lower than those in the latter group (Z=-3.185, -2.091, -2.556 and -1.996, respectively, all P values<0.05). In patients with Brown Ⅱb/c defect, the prosthesis repair could improve the chewing QOL score (Z=-2.830, P=0.005), but no statistically significant differences in other QOL scores between two groups. In patients with Brown Ⅱd defect, the tissue flap reconstruction could improve the scores of pain, entertainment, swallowing and speech QOL (Z=-2.741, -2.517, -2.320 and -2.843, respectively, all P values<0.05), and the average QOL score in tissue flap reconstruction group was also higher than that of the prosthesis group (Z=-2.276, P=0.023). Conclusion: For postoperative long-term quality of life, both prosthesis and tissue flap reconstruction can offer satisfactory results in patients with Brown Ⅱb/c defect, and patients with Brown Ⅱd defect repaired by tissue flap reconstruction have better speech and swallowing functions. Tissue flap reconstruction may bring more entertainment and emotional benefits.

[Research progress and application prospect of stem cells in inflamed dental tissues on periodontal bone regeneration].

Tissue engineering has become a research hotspot regarding periodontal bone regeneration in recent years. Generally, stem cells used in periodontal tissue engineering are derived from healthy dental tissues, while restricted due to the strict indication of tooth extraction and limited sources. Stem cells in inflamed dental tissues mainly derive from inflamed pulp, periapical and periodontal tissues. Stem cells in inflamed dental tissues are abundant and retain most of the basic characteristics of stem cell compared with the ones derived from healthy dental tissues, which can be a promising source of stem cells for periodontal bone regeneration. In this review, we summarize the current application and prospect of stem cells in inflamed dental tissues on periodontal bone regeneration, and then discuss their feasibility as seed cells, in order to provide a reference for future research and clinical application of stem cells in inflamed dental tissues.

[Advances in the periodontal surgical flaps techniques to preserve gingival papilla].

Papilla preservation in periodontal surgery is not only beneficial to maintain postoperative aesthetics and good oral hygiene but also contributes to obtaining good periodontal regeneration outcomes. Various periodontal flaps have been designed to preserve the gingival papilla, which constitutes the clinical basis for periodontal open flap debridement and periodontal regeneration surgery. A comprehensive understanding of their design purpose, indications, and technical key points will help clinicians to choose the optimal surgical plan, and thus improve the clinician's treatment levels, and obtain good clinical outcomes. Therefore, this article aims to introduce the design background, indications, and technical key points of various surgical flaps, such as papilla preservation technique, modified papilla preservation technique, simplified papilla preservation flap, etc.

[High expression of death-associated protein 5 promotes glucose metabolism in gastric cancer cells and correlates with poor survival outcomes].

OBJECTIVE: To investigate the prognostic value of death-associated protein 5 (DAP5) in gastric cancer (GC) and its regulatory effect on aerobic glycolysis in GC cells. METHODS: We analyzed DAP5 expression levels in GC and adjacent tissues and its association with survival outcomes of GC patients using public databases. We collected paired samples of GC and adjacent tissues from 102 patients undergoing radical resection of GC in our hospital from June, 2012 to July, 2017, and analyzed the correlation of DAP5 expression level detected immunohistochemically with the clinicopathological parameters of the patients. Cox regression analysis, Kaplan-Meier analysis, and ROC curves were used to explore the independent risk factors and the predictive value of DAP5 expression for 5-year survival of the patients. In the cell experiments, we observed the changes in aerobic glycolysis in MGC-803 cells following lentivirus-mediated DAP5 knockdown or overexpression by measuring glucose uptake and cellular lactate level and using qRT-PCR and Western blotting. RESULTS: Analysis using the public databases showed that DAP5 was highly expressed in GC and correlated with tumor progression and poor survival outcomes of the patients (P < 0.05). In the clinical samples, DAP5 expression was significantly higher in GC than in the adjacent tissues (3.19±0.60 vs 1.00±0.12; t=36.863, P < 0.01), and a high expression of DAP5 was associated with a reduced 5-year survival rate of the patients (17.6% vs 72.5%; χ2=29.921, P < 0.05). A high DAP5 expression, T3-4, N2-3, and CEA≥5 ng/mL were identified as independent risk factors affecting 5-year survival outcomes of GC (P < 0.05), for which DAP5 expression showed a prediction sensitivity, specificity and accuracy of 73.2%, 80.4% and 79.0%, respectively. In MGC-803 cells, DAP5 knockdown significantly reduced glucose uptake, lactate level and the expressions of GLUT1, HK2 and LDHA, and DAP5 overexpression produced the opposite effects (P < 0.05). CONCLUSION: A high expression of DAP5 in GC, which enhances cellular aerobic glycolysis to promote cancer progression, is correlated with a poor survival outcome and may serve as a biomarker for evaluating long-term prognosis of GC patients.

[Pachymic acid protects against Crohn's disease-like intestinal barrier injury and colitis in miceby suppressingintestinal epithelial cell apoptosis via inhibiting PI3K/AKT signaling].

OBJECTIVE: To investigate the effect of pachymic acid (PA) against TNBS-induced Crohn's disease (CD)-like colitis in mice and explore the possible mechanism. METHODS: Twenty-four C57BL/6J mice were randomized equally into control group, TNBS-induced colitis model group and PA treatment group. PA treatment was administered via intraperitoneal injection at the daily dose of 5 mg/kg for 7 days, and the mice in the control and model groups were treated with saline. After the treatments, the mice were euthanized for examination of the disease activity index (DAI) of colitis, body weight changes, colon length, intestinal inflammation, intestinal barrier function and apoptosis of intestinal epithelial cells, and the expressions of TNF-α, IL-6 and IL-1ß in the colonic mucosa were detected using ELISA. The possible treatment targets of PA in CD were predicted by network pharmacology. String platform and Cytoscape 3.7.2 software were used to construct the protein-protein interaction (PPI) network. David database was used to analyze the GO function and KEGG pathway; The phosphorylation of PI3K/AKT in the colonic mucosal was detected with Western blotting. RESULTS: PA significantly alleviated colitis in TNBS-treated mice as shown by improvements in the DAI, body weight loss, colon length, and histological inflammation score and lowered levels of TNF-α, IL-6 and IL-1ß. PA treatment also significantly improved FITC-dextran permeability, serum I-FABP level and colonic transepithelial electrical resistance, and inhibited apoptosis of the intestinal epithelial cells in TNBS-treated mice. A total of 248 intersection targets were identified between PA and CD, and the core targets included EGFR, HRAS, SRC, MMP9, STAT3, AKT1, CASP3, ALB, HSP90AA1 and HIF1A. GO and KEGG analysis showed that PA negatively regulated apoptosis in close relation with PI3K/AKT signaling. Molecular docking showed that PA had a strong binding ability with AKT1, ALB, EGFR, HSP90AA1, SRC and STAT3. In TNBS-treated mice, PA significantly decreased p-PI3K and p-AKT expressions in the colonic mucosa. CONCLUSION: PA ameliorates TNBS-induced intestinal barrier injury in mice by antagonizing apoptosis of intestinal epithelial cells possibly by inhibiting PI3K/AKT signaling.

[Analysis of efficacy and factors influencing sequential combination therapy with tenofovir alafenamide fumarate after treatment with entecavir in chronic hepatitis B patients with low-level viremia].

Objective: To observe the efficacy and factors influencing sequential or combined tenofovir alafenamide fumarate (TAF) after treatment with entecavir (ETV) in patients with chronic hepatitis B (CHB) with low-level viremia (LLV). Methods: 126 CHB cases treated with ETV antiviral therapy in the Department of Infectious Diseases of the First Affiliated Hospital of Nanchang University from January 2020-September 2022 were retrospectively collected. Patients were divided into a complete virologic response (CVR) group (n = 84) and a low-level viremia (LLV) group (n = 42) according to the HBV DNA level during treatment. Clinical characteristics and laboratory indicators of the two groups at baseline and 48 weeks were analyzed by univariate analysis. Patients in the LLV group were divided into three groups according to their continued antiviral treatment regimen until 96 weeks: continued use of ETV as a control group; replacement of TAF as a sequential group; and combination of ETV and TAF as a combined group. The data of the three groups of patients were analyzed by one-way analysis of variance for 48 weeks. HBV DNA negative conversion rate, HBeAg negative conversion rate, alanine aminotransferase (ALT), creatinine (Cr), and liver stiffness test (LSM) were compared among the three groups after 96 weeks of antiviral treatment. Multivariate logistic regression was used to analyze the independent factors influencing the occurrence of HBV DNA non-negative conversion in LLV patients at 96 weeks. Receiver operating characteristic curve (ROC) was used to evaluate the effectiveness of predicting the occurrence of HBV DNA non-negative conversion in LLV patients at 96 weeks. Kaplan-Meier was used to analyze the cumulative negative rate of DNA in LLV patients, and the Log-Rank test was used for comparison. HBV DNA and HBV DNA negative conversion rates during treatment were observed dynamically. Results: Univariate analysis showed statistically significant differences in age, BMI, HBeAg positivity rate, HBV DNA, HBsAg, ALT, AST, and LSM at baseline between the CVR group and the LLV group (P < 0.05). Univariate analysis of variance revealed no statistically significant difference among the three groups of LLV patients at 48 weeks (P > 0.05). HBV-DNA negative conversion rate in the sequential group and the combination group was significantly higher than that in the control group after 96 weeks of treatment (88.89% vs. 41.18%, 85.71% vs. 41.18%, χ (2) = 10.404, P = 0.006). HBeAg negative conversion rate was higher than that of the control group, with no statistically significant difference (P > 0.05).Compared with the control group, ALT, Cr, and LSM in the sequential group and the combined group were equally improved to varying degrees, with a statistically significant difference (P < 0.05). Subsequent use of ETV and HBV DNA at 48 weeks were independent risk factors for HBV DNA positivity at 96 weeks in LLV patients (P < 0.05). The AUC of HBV DNA at 48 weeks was 0.735 (95%CI: 0.578 ~ 0.891), the cut-off value was 2.63 log(10) IU/ml, and the sensitivity and specificity were 76.90% and 72.40%, respectively. DNA conversion rate was significantly lower in LLV patients receiving 48-week ETV and 48-week HBV DNA≥2.63 log10 IU/mL than in patients receiving sequential or combined TAF and 48-week HBV DNA < 2.63 log(10) IU/mL. HBV DNA negative conversion rates in the sequential group and combined group at 72 weeks, 84 weeks, and 96 weeks were higher than those in the control group during the period from 48 weeks to 96 weeks of continuous treatment, and the differences were statistically significant (P < 0.05). Conclusion: Sequential or combined TAF antiviral therapy could more effectively improve the 96-week CVR rate, as well as hepatic and renal function, and alleviate the degree of hepatic fibrosis in CHB patients with LLV following ETV treatment. Subsequent use of ETV and HBV DNA load at 48 weeks were independent predictors of HBV DNA positivity at 96 weeks in LLV patients.

[Clinical efficacy of femoral neck screw system combined with supported hollow screw in treatment of femoral neck fractures with posterometral comminution in young and middle-aged patients].

Objective: To investigate the clinical efficacy of dynamic cross screw system (FNS) for femoral neck fractures in young and middle-aged patients with posterior medial comminution. Methods: A retrospective cohort study. Clinical data of 197 young and middle-aged patients with femoral neck fractures accompanied by posterior medial comminution treated with closed reduction FNS and internal fixation with anti-rotation cannulated screws in Beijing Luhe Hospital, Beijing Jishuitan Hospital and Beijing Tongren Hospital from October 2019 to October 2021 were analyzed retrospectively. According to different surgical methods, the patients were divided into two groups. There were 102 patients in the FNS group, included 55 males and 47 females with a mean age of (40.49±19.79) years; and there were 95 patients in the FNS plus anti-rotation hollow screw group (combined group), included 51 males and 44 females with an average age of (40.03±18.82) years. All patients were followed-up for at least 1 year after surgery. The general clinical data, surgical conditions and Harris score of the hip joint at the last follow-up of the two groups were compared. And the clinical efficacy of the two surgical schemes were evaluated and compared. After surgery, routine X-ray and CT examinations were performed to evaluate the fracture reduction and internal fixation, and the shortening of the femoral neck on the affected side was compared to that of healthy side according to the Zlowodzki method. Results: At the last follow-up, the incidence of fracture reduction loss, screw resection and coxa vara in the combined group were all significantly lower than those in the FNS group [10 (10.5%) vs 28 (27.4%), 1 (1.0%) vs 7 (6.8%) and 9 (9.4%) vs 21 (20.5%), respectively, all P<0.05]. The incidence of nonunion and necrosis of the femoral head in the combined group were both lower than those in the FNS group, but there was no significant difference between two groups (both P>0.05). The postoperative mild, moderate and severe femoral neck shortening in the combined group were all lower than those in the FNS group, and the difference were not statistically significant (all P>0.05). At the last follow-up, the Harris score in the combined group was 84.60±2.08, and it was higher than that in the FNS group (79.57±4.31), but the difference was not statistically significant (P=0.403). Conclusion: FNS plus supporting hollow screw has a good clinical effect on femoral neck fractures in young and middle-aged adults with posterior medial comminution.

[Diosmetin regulates intestinal immune balance by inhibiting PI3K/AKT signaling to relieve 2, 4, 6-trinitrobenzene sulfonic acid-induced Crohn's disease-like colitis in mice].

OBJECTIVE: To investigate the therapeutic mechanism of diosmetin on 2, 4, 6-trinitrobenzene sulfonic acid (TNBS)-induced Crohn's disease (CD)-like colitis in mice. METHODS: Wild-type C57BL/6 mice were randomized into control group, TNBS-induced CD-like colitis group (TNBS group) and 50 mg·kg-1·d-1 diosmetin-treated group (n=8). Disease activity (DAI) scores, body weight changes, histological scores, colon lengths and colon mucosal levels of TNF-α, IFN-γ, and IL-17A were measured to evaluate the severity of colitis. The changes of T lymphocyte subsets (Th1/Th2 and Th17/Treg) in the mesenteric lymph nodes were analyzed by flow cytometry. Network pharmacology and molecular docking were used to analyze the effect of diosmetin on PI3K/AKT pathway. RESULTS: Compared with TNBS group, diosmetin treatment significantly lowered DAI scores, histological scores, body weight loss and colon mucosal levels of TNF-α, IFN-γ, and IL-17A (P < 0.05) and increased the colon length of the rat models, but these improvements did not reach the control levels (P < 0.05). Diosmetin significantly lowered the percentages of Th1/Th17 cells in the mesenteric lymph nodes in TNBS-treated mice, which remained higher than the control levels (P < 0.05); The percentages of Th2/Treg cells were significantly higher in diosmetin group than in TNBS group (P < 0.05) and the control group (P < 0.05). Network pharmacologic analysis identified 46 intersection targets of diosmetin and CD, and among them AKT1, EGFR, SRC, ESR1, MMP9 and PTGS2 were the top 6 core targets. GO and KEGG analyses showed that the PI3K/AKT signaling pathway was closely related with the therapeutic effect of diosmetin on CD-like colitis. Molecular docking suggested strong binding of diosmetin to the key core targets. Diosmetin significantly reduced the levels of p-PI3K and p-AKT in the colon mucosa in TNBS-treated mice (P < 0.05), but their levels remained higher than those in the control group (P < 0.05). CONCLUSION: Diosmetin ameliorates TNBS-induced CDPlike colitis in mice possibly by regulating Th1/Th2 and Th17/Treg balance to improve intestinal immune disorder through inhibition of PI3K/AKT signaling.

Porphyromonas gingivalis Evades Immune Clearance by Regulating Lysosome Efflux.

Porphyromonas gingivalis, a major periodontal pathogen, invades autophagosomes of cells, including gingival epithelial cells, endothelial cells, gingival fibroblasts, macrophages, and dendritic cells, to escape antimicrobial autophagy and lysosome fusion. However, it is not known how P. gingivalis resists autophagic immunity, survives within cells, and induces inflammation. Thus, we investigated whether P. gingivalis could escape antimicrobial autophagy by promoting lysosome efflux to block autophagic maturation, leading to intracellular survival, and whether the growth of P. gingivalis within cells results in cellular oxidative stress, causing mitochondrial damage and inflammatory responses. P. gingivalis invaded human immortalized oral epithelial cells in vitro and mouse oral epithelial cells of gingival tissues in vivo. The production of reactive oxygen species (ROS) increased upon bacterial invasion, as well as mitochondrial dysfunction-related parameters with downregulated mitochondrial membrane potential and intracellular adenosine triphosphate (ATP), upregulated mitochondrial membrane permeability, intracellular Ca2+ influx, mitochondrial DNA expression, and extracellular ATP. Lysosome excretion was elevated, the number of intracellular lysosomes was diminished, and lysosomal-associated membrane protein 2 was downregulated. Expression of autophagy-related proteins, microtubule-associated protein light chain 3, sequestosome-1, the NLRP3 inflammasome, and interleukin-1ß increased with P. gingivalis infection. P. gingivalis may survive in vivo by promoting lysosome efflux, blocking autophagosome-lysosome fusion, and destroying autophagic flux. As a result, ROS and damaged mitochondria accumulated and activated the NLRP3 inflammasome, which recruited the adaptor protein ASC and caspase 1, leading to the production of proinflammatory factor interleukin-1ß and inflammation.

A Germline-Targeting Chimpanzee SIV Envelope Glycoprotein Elicits a New Class of V2-Apex Directed Cross-Neutralizing Antibodies.

HIV-1 and its SIV precursors share a broadly neutralizing antibody (bNAb) epitope in variable loop 2 (V2) at the envelope glycoprotein (Env) trimer apex. Here, we tested the immunogenicity of germ line-targeting versions of a chimpanzee SIV (SIVcpz) Env in human V2-apex bNAb heavy-chain precursor-expressing knock-in mice and as chimeric simian-chimpanzee immunodeficiency viruses (SCIVs) in rhesus macaques (RMs). Trimer immunization of knock-in mice induced V2-directed NAbs, indicating activation of V2-apex bNAb precursor-expressing mouse B cells. SCIV infection of RMs elicited high-titer viremia, potent autologous tier 2 neutralizing antibodies, and rapid sequence escape in the canonical V2-apex epitope. Six of seven animals also developed low-titer heterologous plasma breadth that mapped to the V2-apex. Antibody cloning from two of these animals identified multiple expanded lineages with long heavy chain third complementarity determining regions that cross-neutralized as many as 7 of 19 primary HIV-1 strains, but with low potency. Negative stain electron microscopy (NSEM) of members of the two most cross-reactive lineages confirmed V2 targeting but identified an angle of approach distinct from prototypical V2-apex bNAbs, with antibody binding either requiring or inducing an occluded-open trimer. Probing with conformation-sensitive, nonneutralizing antibodies revealed that SCIV-expressed, but not wild-type SIVcpz Envs, as well as a subset of primary HIV-1 Envs, preferentially adopted a more open trimeric state. These results reveal the existence of a cryptic V2 epitope that is exposed in occluded-open SIVcpz and HIV-1 Env trimers and elicits cross-neutralizing responses of limited breadth and potency. IMPORTANCE An effective HIV-1 vaccination strategy will need to stimulate rare precursor B cells of multiple bNAb lineages and affinity mature them along desired pathways. Here, we searched for V2-apex germ line-targeting Envs among a large set of diverse primate lentiviruses and identified minimally modified versions of one chimpanzee SIV Env that bound several human V2-apex bNAb precursors and stimulated one of these in a V2-apex bNAb precursor-expressing knock-in mouse. We also generated chimeric simian-chimpanzee immunodeficiency viruses and showed that they elicit low-titer V2-directed heterologous plasma breadth in six of seven infected rhesus macaques. Characterization of this antibody response identified a new class of weakly cross-reactive neutralizing antibodies that target the V2-apex, but only in occluded-open Env trimers. The existence of this cryptic epitope, which in some Env backgrounds is immunodominant, needs to be considered in immunogen design.

Airway Phenotypes and Nocturnal Wearing of Dentures in Elders with Sleep Apnea.

The objective of this study was to examine to what extent the anatomic characteristics of the upper airway can influence the effect of nocturnal wearing of dentures on the sleep of edentulous elders with untreated sleep apnea. This study used the data from a randomized crossover clinical trial and an exploratory approach to address its objectives. Cone beam computed tomography scans of 65 edentulous individuals (female, n = 37; male, n = 28; mean ± SD age, 74.54 ± 6.42 y) with untreated obstructive sleep apnea (OSA) were used to identify anatomic variables. Polysomnography data were collected by means of one portable overnight recording. The respiratory variable values, including apnea-hypopnea index (AHI), with and without denture worn during sleep were used to calculate the change. Statistical analyses included multiple linear regressions, cluster analysis, and binary logistic regressions. A receiver operator characteristic curve was used to illustrate the accuracy of the statistical model. The regression model explained 15.8% (R2) of AHI change. An increase in the lateral dimension of the minimum cross-sectional area was associated with a decrease in AHI, oxygen desaturation index, and respiratory arousal index changes (P ≤ 0.041). Furthermore, an increase in the length of the hypopharynx was associated with an increase in AHI and oxygen desaturation index changes (P ≤ 0.027). An increase in the lateral dimension of the minimum cross-sectional area of the upper airway was associated with a decreased likelihood of being in the group having a worsened AHI (odds ratio = 0.85; 95% CI, 0.76 to 0.95; P = 0.006). An increase in the length of the oropharynx was associated with an increased likelihood of having increased AHI (odds ratio = 1.10; 95% CI, 1.01 to 1.20; P = 0.026). The nocturnal aggravation of respiratory variables in edentulous individuals with OSA who wear dentures at night can be linked to certain anatomic characteristics of the upper airway. Replication of these findings may open novel avenues for personalized advice regarding nocturnal wearing of dentures in edentulous individuals with OSA (ClinicalTrials.gov: NCT01868295).

[Introduction and interpretation of the European Federation of Periodontology S3 level clinical practice guideline for treatment of periodontitis].

2018 international classification of periodontal and implant diseases relates the classifications with the approaches of prevention and treatment based on the stages and grades of disease. European Federation of Periodontology (EFP) evaluated the available evidences following the methodological guidance of the Association of Scientific Medical Societies in Germany and the Grading of Recommendations Assessment, Development and Evaluation (GRADE), and published the EFP S3 level clinical practice guideline for the treatment of stage Ⅰ-Ⅲ and Ⅳ periodontitis in 2020 and 2022, respectively. The present manuscript gives introduction and interpretation based on the EFP S3 level clinical practice guideline and Chinese national conditions. On the base of the diagnostic key points of staging and grading, it introduces in detail the step treatment procedures of stageⅠ-Ⅲ periodontitis as well as the multi-disciplinary treatment procedures of stage Ⅳ periodontitis, compares the similarities and differences between the step and phase procedures, and then proposes a strategy for determining the recall interval more suitable for Chinese clinicians. The present manuscript aims to help dentists to learn and grasp the key points more quickly and accurately on the clinical application of the guideline and to assist them in making the optimal treatment plans after judging and evaluating the specific clinical circumstances, so as to maximize the chances of favorable outcome.

[Centromere protein U is highly expressed in colorectal cancer and associated with a poor long-term prognosis].

OBJECTIVE: To analyze the expression of centromere protein U (CENPU) in colorectal cancer and its predictive value for long-term prognosis of the patients. METHODS: We retrospectively analyzed the data of 102 patients with colorectal cancer undergoing radical resection in our hospital between January, 2005 and December, 2011. The expression level of CENPU in colorectal cancer tissue was detected immunohistochemically, and its association with clinicopathological characteristics of the patients were analyzed. The patients were divided into low expression group (n=51) and high expression group (n=51) based on the median CENPU expression level for analysis the value of CENPU for predicting long-term prognosis of the patients after radical resection of the tumors. In the in vitro study, we constructed colorectal cancer cell lines with CENPU interference and CENPU overexpression by lentiviral transfection and assessed the changes in the proliferation, migration and invasion of the cells using CCK-8 assay and Transwell assay. RESULTS: The protein expression level of CENPU was significantly higher in colorectal cancer tissues than in the adjacent tissues (P < 0.05) and was positively correlated with the expressions levels of Ki67 (r=0.569, P < 0.05) and VEGF-C (r=0.629, P < 0.05). CENPU expression level in colorectal cancer tissue was closely related with tumor progression and clinicopathological stage of the tumor (P < 0.05). Kaplan-Meier survival analysis showed that the patients with high CENPU expression had significantly decreased postoperative overall survival (χ2=11.155, P < 0.05); Cox multivariate regression analysis suggested that CENPU expression level was an independent risk factor affecting the overall survival of the patients after radical resection (HR=1.848, P < 0.05). The results of cell experiments demonstrated that high CENPU expression significantly promoted the proliferation, migration and invasion of the tumor cells. CONCLUSION: CENPU is highly expressed in colorectal cancer tissues in closely correlation with tumor progression and may serve as a potential biomarker for evaluating the long-term prognosis of colorectal cancer patients.