Objective: To investigate the inhibitory effect of neoeriocitrin (Neo) on ligature-induced experimental periodontitis in mice and evaluate its biosafety, providing experimental evidence for novel candidate drugs in periodontitis treatment. Methods: This study has been approved by the Animal Welfare and Ethical Safety Committee. A periodontitis model was established in C57BL/6J mice using silk ligation. The mice were divided into control, periodontitis model, and three Neo treatment groups (5, 10, 20 mg/kg Neo). After 2 weeks of intervention, alveolar bone resorption was analyzed by micro-computed tomography (micro-CT); periodontal tissue pathological changes were observed via hematoxylin-eosin (HE) and Masson staining; osteoclasts were counted using tartrate-resistant acid phosphatase staining; serum inflammatory factor levels [tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), interleukin-10 (IL-10)] were detected by enzyme-linked immunosorbent assay; gut microbiota changes were analyzed by 16S rRNA sequencing; and biosafety was assessed through HE staining of the heart, liver, spleen, lung, kidney, and colon, as well as zonula occludens-1 (ZO-1) immunohistochemical staining of the colon. Results: Compared with the periodontitis model group, the 10 mg/kg Neo treatment group showed significantly alleviated alveolar bone resorption, manifested as reduced cementoenamel junction to alveolar bone crest distance, increased bone mineral density, bone volume fraction, and trabecular thickness. Additionally, reduced inflammatory cell infiltration, improved collagen fiber arrangement, and a significantly decreased number of osteoclasts were observed in periodontal tissues. Serum levels of pro-inflammatory factors TNF-α and IL-6 were also reduced. Furthermore, 10 mg/kg Neo intervention modulated the gut microbiota structure without causing significant multi-organ toxicity or impairing intestinal barrier function. Conclusion Neo can effectively mitigate the progression of experimental periodontitis by inhibiting alveolar bone resorption, reducing periodontal tissue inflammation, modulating systemic immunity, and improving gut microbiota. With good biosafety, Neo is a promising candidate drug for the treatment of periodontitis.

Objective To investigate the effects of neurotrophic factor Neuritin overexpression on the angiogenic effects of chicken embryonic allantoic membrane (CAM) and human umbilical vein endothelial cells (HUVECs),and to provide a new direction for the treatment of angiogenic diseases. Methods Thirty fresh yellow-skinned breeding eggs were selected to establish a CAM model,which were divided into three groups by randomized numerical table method:positive control group (bFGF),negative control group (NS) and experimental group (Neuritin),with 10 eggs in each group. The positive control group was loaded with 2500 U/mL of bFGF,the experimental group was loaded with 10 μg/mL of Neuritin protein,and the negative control group was loaded with NS. 10 μL loading volume was loaded into each group,and all CAMs were incubated at the same temperature,relative humidity,and time,and the vascular branching,number,and size of the CAMs in each group were recorded after 72 h of incubation. Fresh umbilical cords from healthy pregnant women were selected to produce primary HUVECs,which were divided into three groups:transfected with recombinant plasmid (HUVEC-neu group),transfected with empty vector (HUVEC-3.1 group),and untransfected (HUVEC group). Primary HUVECs in the HUVEC-neu group were transfected with the recombinant plasmid Neuritin,and those in the HUVEC-3.1 group were transfected with the empty vector. HUVEC-3.1 group was transfected with the empty vector plasmid,and HUVEC group was not given any special treatment,and all three groups received the same culture regimen. Western blot was used to detect the protein expression level of Neuritin in HUVEC-3.1 and HUVEC-neu groups. CCK-8 assay,cell scratch assay,Transwell assay,and tube formation assay were used to detect protein expression level of Neuritin in HUVEC-3.1 group and HUVEC-neu group,and HUVEC-neu groups for cell proliferation,migration and tube formation. Results (1) The number of CAM vessel branch points and microvessels in the experimental group was significantly increased compared with that in the negative control group (P<0.01),but there was no statistically significant difference in the number of large and medium-sized vessels between the two groups (P>0.05);(2) Neuritin was successfully overexpressed in HUVECs in the HUVEC-neu group. (3) Compared with the HUVEC-3.1 group,the proliferation vigor of cells in the HUVEC-neu group was decreased (P<0.05),but their migration and tube formation abilities were significantly enhanced (P<0.01). Conclusion Neuritin overexpression promotes angiogenesis and participates in the regulation of neovascularization by affecting cell prolif-eration,migration,and tube formation ability.

Objective: To examine the effect of intracellular vesicles (IVs) and extracellular vesicles (EVs) that originated from periodontal ligament stem cells (PDLSCs) on the osteogenic differentiation of PDLSCs within a lipopolysaccharide (LPS)-simulated inflammatory microenvironment, and to provide new insights for the application of IVs in the repair and regeneration of periodontal tissue in periodontitis. Methods: Ethical approval was obtained from the institution. Human-origin PDLSCs were extracted, and the IVs and EVs from PDLSCs at the 3rd-6th passages were gathered and identified using transmission electron microscopy, nano flow cytometry (Nano FCM) analysis, and Western Blot. The 3rd-6th generations of PDLSCs were categorized into the following groups: Control group, LPS group, LPS + 100 μg/mL EVs group (LPS+EVs group), and LPS + 100 μg/mL IVs group (LPS+IVs group). The effects of the IVs and EVs on the anti-inflammatory and osteogenic differentiation of PDLSCs in an inflammatory microenvironment were assessed by using a Cell Counting Kit-8 (CCK-8), enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qRT-PCR), Western Blot, alkaline phosphatase (ALP) staining, and alizarin red staining (ARS). Results: Under transmission electron microscopy, the IVs and EVs derived from PDLSCs displayed a double-layer membrane structure. NanoFCM analysis revealed that the average diameters of the IVs and EVs were 79.6 nm and 82.1 nm, respectively. Western Blot analysis indicated that the surface proteins CD9, CD63, and CD81 of the IVs and EVs were positively expressed, while calnexin was negatively expressed, indicating that IVs and EVs were successfully obtained. Compared with the Control group, the proliferation of PDLSCs in the LPS group was reduced, while the levels of inflammatory cytokine interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the cell supernatant were increased, the mRNA expressions of osteogenic differentiation-related genes, including osteoblast-related genes runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin (OCN) of PDLSCs were reduced, the protein expressions of RUNX2 and osteopontin (OPN) were also decreased (P<0.05); compared with the LPS group, the proliferation of PDLSCs in the LPS+EVs group and LPS+IVs group were significantly increased, while the levels of IL-6, TNF-α were significantly reduced, and the mRNA expressions of RUNX2, ALP, OCN were significantly increased, the protein expressions of RUNX2 and OPN were also significantly increased (P<0.05). Further, in the inflammatory microenvironment, Compared with EVs, IVs more significantly promote the proliferation of PDLSCs, inhibit TNF-α expression, enhance the expression of RUNX2 mRNA, upregulate the expression of RUNX2 and OPN proteins, increase ALP activity, and promote the formation of mineralized nodules (P<0.05). Conclusion IVs and EVs derived from PDLSCs can boost the proliferation of PDLSCs in an inflammatory microenvironment, inhibit the expression of inflammatory factors, and advance the osteogenic differentiation of PDLSCs. The anti-inflammatory and osteogenic effects of IVs are superior to those of EVs.

Objective To investigate the effects of neurotrophic factor Neuritin overexpression on the angiogenic effects of chicken embryonic allantoic membrane (CAM) and human umbilical vein endothelial cells (HUVECs),and to provide a new direction for the treatment of angiogenic diseases. Methods Thirty fresh yellow-skinned breeding eggs were selected to establish a CAM model,which were divided into three groups by randomized numerical table method:positive control group (bFGF),negative control group (NS) and experimental group (Neuritin),with 10 eggs in each group. The positive control group was loaded with 2500 U/mL of bFGF,the experimental group was loaded with 10 μg/mL of Neuritin protein,and the negative control group was loaded with NS. 10 μL loading volume was loaded into each group,and all CAMs were incubated at the same temperature,relative humidity,and time,and the vascular branching,number,and size of the CAMs in each group were recorded after 72 h of incubation. Fresh umbilical cords from healthy pregnant women were selected to produce primary HUVECs,which were divided into three groups:transfected with recombinant plasmid (HUVEC-neu group),transfected with empty vector (HUVEC-3.1 group),and untransfected (HUVEC group). Primary HUVECs in the HUVEC-neu group were transfected with the recombinant plasmid Neuritin,and those in the HUVEC-3.1 group were transfected with the empty vector. HUVEC-3.1 group was transfected with the empty vector plasmid,and HUVEC group was not given any special treatment,and all three groups received the same culture regimen. Western blot was used to detect the protein expression level of Neuritin in HUVEC-3.1 and HUVEC-neu groups. CCK-8 assay,cell scratch assay,Transwell assay,and tube formation assay were used to detect protein expression level of Neuritin in HUVEC-3.1 group and HUVEC-neu group,and HUVEC-neu groups for cell proliferation,migration and tube formation. Results (1) The number of CAM vessel branch points and microvessels in the experimental group was significantly increased compared with that in the negative control group (P<0.01),but there was no statistically significant difference in the number of large and medium-sized vessels between the two groups (P>0.05);(2) Neuritin was successfully overexpressed in HUVECs in the HUVEC-neu group. (3) Compared with the HUVEC-3.1 group,the proliferation vigor of cells in the HUVEC-neu group was decreased (P<0.05),but their migration and tube formation abilities were significantly enhanced (P<0.01). Conclusion Neuritin overexpression promotes angiogenesis and participates in the regulation of neovascularization by affecting cell prolif-eration,migration,and tube formation ability.

Objective:To explore the serum miRNA-144-5p (miR-144-5p) level in liver cancer patients and its relationship with clinical characteristics, as well as the target genes of miR-144-5p and the possible pathogenic mechanisms.Methods:The morning fasting serum samples were retrospectively collected from 100 newly diagnosed liver cancer patients (liver cancer group) before any treatment in Shanxi Province Cancer Hospital from June 2019 to June 2020, as well as 100 healthy individuals (healthy control group) during physical examinations in the same period. The real-time fluorescence quantitative polymerase chain reaction method was used to measure the transcription level expression of miR-144-5p in serum. The high and low expressions of miR-144-5p in patients were determined based on the median relative expression level of miR-144-5p; the distribution differences between patients with high and low expressions of miR-144-5p among different clinical pathological features were compared. The online tools miRDB, TargetScan 8.0 and miRWalk were used to predict potential target genes for miR-144-5p, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed through a bioinformatics platform, and complementary sequences between miR-144-5p and target genes were analyzed using RNA22 software.Results:The median age of the liver cancer group was 54 years old, ranging from 38 to 74 years old, including 80 males (80.0%) and 20 females (20.0%); the median age of the healthy control group was 46 years old, ranging from 34 to 66 years old, including 69 males (69.0%) and 31 females (31.0%); there was no statistically significant difference in age and gender composition between the two groups (both P > 0.05). The median transcription level relative expression of miR-144-5p in liver cancer patients was lower than that in the healthy control group [ M ( Q1, Q3)] [0.311 (0.066, 2.270) vs. 1.067 (0.263, 3.620)], and the difference was statistically significant ( Z = -4.16, P < 0.001). The proportion of patients with low expression of miR-144-5p was higher in the group with maximum diameter of tumor > 5 cm compared to the group with maximum diameter of tumor ≤5 cm [59.3% (35/59) vs. 36.6% (15/41)] and in the group with metastasis compared to the group without metastasis [60.4% (29/48) vs. 40.4% (21/52)], and the differences were statistically significant (both P < 0.05). There was no statistically significant difference in the distribution of patients with high and low expressions of miR-144-5p among different subgroups based on gender, age >55 years, presence of hepatitis, cirrhosis, alpha fetoprotein >400 μ g/L, and differentiation degree (all P > 0.05). According to predictions, there were 34 miR-144-5p target genes shared by the three bioinformatics online tools. GO and KEGG enrichment analyses showed that these target genes were associated with ubiquitination, primarily enriched in the ubiquitin-mediated proteolysis pathway (hsa04120) and the mTOR signaling pathway (hsa04150). RNA22 software analysis showed that miR-144-5p mainly had complementary sequences with 3' untranslated regions of UBR5 and UBE4A genes. Conclusions:The serum miR-144-5p level in liver cancer patients is relatively low, and its expression level may be related to tumor size and metastasis. miR-144-5p may affect the occurrence and development of liver cancer by regulating ubiquitination level through target genes such as UBR5 and UBE4A.

Cemental tear is a rare and indetectable condition unless obvious clinical signs present with the involvement of surrounding periodontal and periapical tissues. Due to its clinical manifestations similar to common dental issues, such as vertical root fracture, primary endodontic diseases, and periodontal diseases, as well as the low awareness of cemental tear for clinicians, misdiagnosis often occurs. The critical principle for cemental tear treatment is to remove torn fragments, and overlooking fragments leads to futile therapy, which could deteriorate the conditions of the affected teeth. Therefore, accurate diagnosis and subsequent appropriate interventions are vital for managing cemental tear. Novel diagnostic tools, including cone-beam computed tomography (CBCT), microscopes, and enamel matrix derivatives, have improved early detection and management, enhancing tooth retention. The implementation of standardized diagnostic criteria and treatment protocols, combined with improved clinical awareness among dental professionals, serves to mitigate risks of diagnostic errors and suboptimal therapeutic interventions. This expert consensus reviewed the epidemiology, pathogenesis, potential predisposing factors, clinical manifestations, diagnosis, differential diagnosis, treatment, and prognosis of cemental tear, aiming to provide a clinical guideline and facilitate clinicians to have a better understanding of cemental tear.
Humans ; Dental Cementum/injuries* ; Consensus ; Diagnosis, Differential ; Cone-Beam Computed Tomography ; Tooth Fractures/therapy*

With the growing emphasis on maternal and child oral health, the significance of managing oral health across preconception, pregnancy, and infancy stages has become increasingly apparent. Oral health challenges extend beyond affecting maternal well-being, exerting profound influences on fetal and neonatal oral development as well as immune system maturation. This expert consensus paper, developed using a modified Delphi method, reviews current research and provides recommendations on maternal and child oral health management. It underscores the critical role of comprehensive oral assessments prior to conception, diligent oral health management throughout pregnancy, and meticulous oral hygiene practices during infancy. Effective strategies should be seamlessly integrated across the life course, encompassing preconception oral assessments, systematic dental care during pregnancy, and routine infant oral hygiene. Collaborative efforts among pediatric dentists, maternal and child health workers, and obstetricians are crucial to improving outcomes and fostering clinical research, contributing to evidence-based health management strategies.
Humans ; Pregnancy ; Female ; Infant ; Consensus ; Mouth Diseases/therapy* ; Pregnancy Complications/therapy* ; Oral Health ; Infant, Newborn ; Delphi Technique ; Oral Hygiene

Objective:To explore the serum miRNA-144-5p (miR-144-5p) level in liver cancer patients and its relationship with clinical characteristics, as well as the target genes of miR-144-5p and the possible pathogenic mechanisms.Methods:The morning fasting serum samples were retrospectively collected from 100 newly diagnosed liver cancer patients (liver cancer group) before any treatment in Shanxi Province Cancer Hospital from June 2019 to June 2020, as well as 100 healthy individuals (healthy control group) during physical examinations in the same period. The real-time fluorescence quantitative polymerase chain reaction method was used to measure the transcription level expression of miR-144-5p in serum. The high and low expressions of miR-144-5p in patients were determined based on the median relative expression level of miR-144-5p; the distribution differences between patients with high and low expressions of miR-144-5p among different clinical pathological features were compared. The online tools miRDB, TargetScan 8.0 and miRWalk were used to predict potential target genes for miR-144-5p, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed through a bioinformatics platform, and complementary sequences between miR-144-5p and target genes were analyzed using RNA22 software.Results:The median age of the liver cancer group was 54 years old, ranging from 38 to 74 years old, including 80 males (80.0%) and 20 females (20.0%); the median age of the healthy control group was 46 years old, ranging from 34 to 66 years old, including 69 males (69.0%) and 31 females (31.0%); there was no statistically significant difference in age and gender composition between the two groups (both P > 0.05). The median transcription level relative expression of miR-144-5p in liver cancer patients was lower than that in the healthy control group [ M ( Q1, Q3)] [0.311 (0.066, 2.270) vs. 1.067 (0.263, 3.620)], and the difference was statistically significant ( Z = -4.16, P < 0.001). The proportion of patients with low expression of miR-144-5p was higher in the group with maximum diameter of tumor > 5 cm compared to the group with maximum diameter of tumor ≤5 cm [59.3% (35/59) vs. 36.6% (15/41)] and in the group with metastasis compared to the group without metastasis [60.4% (29/48) vs. 40.4% (21/52)], and the differences were statistically significant (both P < 0.05). There was no statistically significant difference in the distribution of patients with high and low expressions of miR-144-5p among different subgroups based on gender, age >55 years, presence of hepatitis, cirrhosis, alpha fetoprotein >400 μ g/L, and differentiation degree (all P > 0.05). According to predictions, there were 34 miR-144-5p target genes shared by the three bioinformatics online tools. GO and KEGG enrichment analyses showed that these target genes were associated with ubiquitination, primarily enriched in the ubiquitin-mediated proteolysis pathway (hsa04120) and the mTOR signaling pathway (hsa04150). RNA22 software analysis showed that miR-144-5p mainly had complementary sequences with 3' untranslated regions of UBR5 and UBE4A genes. Conclusions:The serum miR-144-5p level in liver cancer patients is relatively low, and its expression level may be related to tumor size and metastasis. miR-144-5p may affect the occurrence and development of liver cancer by regulating ubiquitination level through target genes such as UBR5 and UBE4A.

Recent clinical studies have shown that mutation of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene in cancer cells may be associated with immunosuppressive tumor microenvironment (TME) and poor response to immune checkpoint blockade (ICB) therapy. Therefore, efficiently restoring PTEN gene expression in cancer cells is critical to improving the responding rate to ICB therapy. Here, we screened an adeno-associated virus (AAV) capsid for efficient PTEN gene delivery into B16F10 tumor cells. We demonstrated that intratumorally injected AAV6-PTEN successfully restored the tumor cell PTEN gene expression and effectively inhibited tumor progression by inducing tumor cell immunogenic cell death (ICD) and increasing immune cell infiltration. Moreover, we developed an anti-PD-1 loaded phospholipid-based phase separation gel (PPSG), which formed an in situ depot and sustainably release anti-PD-1 drugs within 42 days in vivo. In order to effectively inhibit the recurrence of melanoma, we further applied a triple therapy based on AAV6-PTEN, PPSG^@anti-PD-1 and CpG, and showed that this triple therapy strategy enhanced the synergistic antitumor immune effect and also induced robust immune memory, which completely rejected tumor recurrence. We anticipate that this triple therapy could be used as a new tumor combination therapy with stronger immune activation capacity and tumor inhibition efficacy.

Odontogenic maxillary sinusitis(OMS)is a subtype of maxillary sinusitis(MS).It is actually inflammation of the maxillary sinus that secondary to adjacent infectious maxillary dental lesion.Due to the lack of unique clinical features,OMS is difficult to distinguish from other types of rhinosinusitis.Besides,the characteristic infectious pathogeny of OMS makes it is resistant to conventional therapies of rhinosinusitis.Its current diagnosis and treatment are thus facing great difficulties.The multi-disciplinary cooperation between otolaryngologists and dentists is absolutely urgent to settle these questions and to acquire standardized diagnostic and treatment regimen for OMS.However,this disease has actually received little attention and has been underrepresented by relatively low publication volume and quality.Based on systematically reviewed literature and practical experiences of expert members,our consensus focuses on characteristics,symptoms,classification and diagnosis of OMS,and further put forward multi-disciplinary treatment decisions for OMS,as well as the common treatment complications and relative managements.This consensus aims to increase attention to OMS,and optimize the clinical diagnosis and decision-making of OMS,which finally provides evidence-based options for OMS clinical management.