[Diagnostic performances of salivary gland ultrasonography for Sjögren's syndrome].

OBJECTIVE: To evaluate the diagnostic performances of salivary gland ultrasonography(SGUS)in Sjögren's syndrome(SS). METHODS: A total of 246 patients with dry mouth and/or eyes who were treated in the outpatient department and inpatient department of Rheumatology and Immunology Department of the Ninth People's Hospital, Shanghai Jiaotong University School of Medicine from December 2019 to January 2022 were collected. All patients received SGUS examination and scored by 2019 outcome measures in rheumatology clinical trial (OMERACT)ultrasonic scoring system.Their general information, unstimulated saliva flow rate(USFR), Schirmer test and serological test results were recorded. In the study, 193 cases had lip gland biopsy. The 2016 American College of Rheumatology(ACR)/ European League Against Rheumatism(EULAR)classification criteria were adopted as the diagnostic standard of SS. χ2 test was used to compare the difference of salivary gland ultrasonic scores between the two groups. The receiver operating characteristic(ROC) curve was used to evaluate the accuracy of SGUS in diagnosing SS, and the disease characteristics of SGUS positive group and negative group in the SS patients were compared. RESULTS: A total of 175 patients were SS group according to the ACR/EULAR classification, and the remaining 71 patients were non-SS group.There was no significant difference in age [(54.2±11.8) years vs. (53.4±14.9) years, P=0.705] and female (94.4% vs.93.1%, P=1.000) between SS and non-pSS groups. A total of 109 patients were SGUS positive (≥ 2 points), of whom 104 patients met the SS diagnosis and 5 patients did not meet the SS diagnosis. The positive rate of SGUS in SS group was significantly higher than that in non-SS group (59.4% vs. 7.0%, P < 0.001). The accuracy of 2019 OMERACT ultrasonic scoring system to predict ACR/EULAR classification was good, with an area under the curve of 0.762 (95%CI 0.701-0.823). The absolute agreement between the SGUS outcome and ACR-EULAR classification was 69.1%(170/246), with a sensiti-vity of 59.4%(104/175), specificity of 93%(66/71), positive predictive value of 95.4%(104/109) and negative predictive value of 48.2% (66/137). A total of 81 patients were positive SGUS combined with anti-SSA antibody, 100% (81/81) fulfilled the ACR-EULAR criteria, 85 patients were negative SGUS and anti SSA antibody, and 60 patients(70.6%, 60/85) did not fulfil the ACR-EULAR criteria. SGUS positive group had higher antinuclear antibody(ANA) positive rate(83.1% vs. 98.1%, P < 0.001) in the patients with SS. CONCLUSION: The OMERACT ultrasonic scoring system has high diagnostic value in SS. The combination of SGUS and anti-SSA antibody can improve the diagnostic value.

[Effects of primary preventive treatment under endoscope for esophageal and gastric varices on bleeding rate and its relevant factors].

Objective: To investigate the effects of primary preventive treatment under endoscope for esophageal and gastric varices on bleeding rate and its relevant factors. Methods: 127 cases with liver cirrhosis accompanied with esophageal and gastric varices without bleeding history were included in the endoscopic and non-endoscopic treatment group, respectively. Informed consent was obtained from both groups. Gastric varices (Lgf) and esophageal varices (Leg) were diagnosed according to LDRf classification criteria, and the corresponding treatment scheme was selected according to the recommended principle of this method.The incidence rate of bleeding from ruptured esophageal varices were observed at 3, 6 months, and 1, and 2 years in the treated and the untreated group, and the patients with different Child-Pugh scores were followed-up for 2 years. Gender, age, etiology, varicose degree, Child-Pugh grade, platelet count, prothrombin activity, portal vein thrombosis, collateral circulation, portal vein width and other factors affecting the bleeding rate were assessed. Measurement data were described as mean ± standard deviation (x¯±s), and qualitative data of categorical variables were expressed as percentage (%), and χ2 test was used. Results: 127 cases were followed up for 2 years. There were 55 cases in the endoscopic treatment group (18 cases underwent band ligation, 2 cases underwent band ligation combined with tissue adhesive embolization, 28 cases underwent sclerotherapy, and 7 cases underwent sclerotherapy combined with tissue adhesive embolization). Recurrent bleeding and hemorrhage was occurred in 5 (9.1%) and 28 cases (38.9%), respectively (P<0.05). In addition, there were 72 cases in the untreated group (P<0.05). Severe varicose veins proportions in treated and untreated group were 91.1% and 85.1%, respectively (P>0.05). There was no statistically significant difference in liver cirrhosis-related medication and ß-blocker therapy between the treated and untreated group (P>0.05). There was no statistically significant difference in the bleeding rate between the different treated groups (P>0.05). The bleeding rates at 3, 6 months, 1, and 2 years in endoscopic treated and untreated group were 2.00% vs. 2.59% (P>0.05), 2.30% vs. 5.88% (P>0.05), 3.10% vs. 7.55% (P>0.05) and 4.00% vs. 21.62% (P<0.05), respectively. All patients with Child-Pugh grade A, B and C in the treated and the untreated group were followed-up for 2 years, and the bleeding rates were 1.8% vs. 8.1% (P<0.05), 1.1% vs. 9.4% (P<0.05) and 9.1% vs. 10.1% (P>0.05), respectively. There were statistically significant differences in the rupture and bleeding of esophageal and gastric varices, varices degree, Child-Pugh grade and presence or absence of thrombosis formation in portal vein (P<0.05); however, no statistically significant differences in gender, age, etiology, platelet count, prothrombin activity, collateral circulation and portal vein width (P>0.05). There was no intraoperative bleeding and postoperative related serious complications in the treated group. Conclusion: The risk of initial episodes of bleeding from esophageal and gastric varices is significantly correlated with the varices degree, Child-Pugh grade, and portal vein thrombosis. Primary preventive treatment under endoscope is safe and effective for reducing the long-term variceal bleeding risk from esophageal and gastric varices.

[Risk factors and nutritional status analysis in patients with liver cirrhosis and concomitant chronic periodontitis].

Objective: To study and explore the prevalence, characteristics, preliminary risk factors, as well as their relationship with nutritional scores in liver cirrhotic patient with chronic periodontitis. Methods: 163 patients with liver cirrhosis who were hospitalized in the Hepatology Division, Department of Internal Medicine at Tianjin Third Central Hospital from June to September 2018 were enrolled as the case group, while the control group consisted 140 healthy individuals enrolled during the same period. Periodontal examination, biochemical examination and oral hygiene habits were investigated. The prevalence of periodontitis in the two groups was compared, and the risk factors of severe periodontitis were conducted by multivariate regression analysis. Results: The prevalence of chronic periodontitis was significantly higher in patients with liver cirrhosis than healthy control population, and the differences were statistically significant (P < 0.05). The prevalence of severe periodontitis and full edentulous jaws was significantly higher in patients with liver cirrhosis than healthy control group, and the differences were statistically significant (P < 0.05 and P < 0.001). Compared with the healthy control group, the depth of periodontal pocket and the degree of attachment loss were significantly increased in the liver cirrhosis group (P < 0.001). Multivariate regression analysis showed that liver cirrhosis was the independent risk factors for both groups of patients with severe periodontitis (χ (2) = 11.046, P < 0.001). Univariate and multivariate regression analysis showed that toothbrushing frequency, nutritional risk score, prealbumin level and Child-Pugh grade were independent risk factors for occurrence of severe periodontitis in liver cirrhotic patient (χ (2) = 5.252, P = 0.022; χ (2) = 24.162, P < 0.001; χ (2) = 4.159, P = 0.041; χ (2) = 9.249, P = 0.002). Conclusion: The prevalence of periodontitis is significantly higher in patients with liver cirrhosis than healthy individuals, and liver cirrhosis is an independent risk factor for the occurrence of severe periodontitis. Toothbrushing frequency, nutritional risk score, prealbumin level and Child-Pugh grade are risk factors for severe periodontitis in patients with liver cirrhosis.

Effects of transforming growth factor-ß1 on plasminogen activation in stem cells from the apical papilla: role of activating receptor-like kinase 5/Smad2 and mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) signalling.

AIM: To study the effects of TGF-ß1 on the plasminogen activation (PA) system of stem cells from the apical papilla (SCAP) and its signalling. METHODOLOGY: SCAP cells were isolated from the apical papilla of immature permanent teeth extracted for orthodontic reasons. They were exposed to various concentration of TGF-ß1 with/without pretreatment and coincubation by SB431542 (ALK/Smad2/3 inhibitor), or U0126 (MEK/ERK inhibitor). MTT assay, Western blotting and enzyme-linked immunosorbent assay (ELISA) were used to detect their effects on cell viability, and the protein expression of plasminogen activator inhibitor-1 (PAI-1), urokinase-type plasminogen activator (uPA), uPA receptor (uPAR) and their secretion. The paired Student's t-test was used for statistical analysis. RESULTS: TGF-ß1 significantly stimulated PAI-1 and soluble uPAR (suPAR) secretion of SCAP cells (P < 0.05), whereas uPA secretion was inhibited. Accordingly, TGF-ß1 induced both PAI-1 and uPAR protein expression of SCAP cells. SB431542 (an ALK5/Smad2/3 inhibitor) pretreatment and coincubation prevented the TGF-ß1-induced PAI-1 and uPAR of SCAP. U0126 attenuated the TGF-ß1-induced expression/secretion of uPAR, but not PAI-1 in SCAP. SB431542 reversed the TGF-ß1-induced decline of uPA. CONCLUSIONS: TGF-ß1 may affect the repair/regeneration activities of SCAP via differential increase or decrease of PAI-1, uPA and uPAR. These effects induced by TGF-ß1 are associated with ALK5/Smad2/3 and MEK/ERK activation. Elucidation the signalling pathways and effects of TGF-ß1 is useful for treatment of immature teeth with open apex by revascularization/revitalization procedures and tissue repair/regeneration.

[Relationship between the suppressor of cytokine signaling 3 expression and antiviral efficacy of nucleos(t)ide and interferon alpha therapy for chronic hepatitis B].

Objective: To investigate the molecular mechanism of poor response of nucleoside and interferon therapy in some patients with chronic hepatitis B (CHB) and the negative regulatory factor of suppressor of cytokine signaling 3 (SOCS3) expression in the interferon-signaling pathway. Also, study the clinical relationship between SOCS3 and antiviral efficacy of nucleoside and interferon. Methods: Peripheral blood and matched liver tissue samples from 54 CHB patients who participated in the OSST study were selected. HBsAg was measured at different time points (baseline and weeks 12, 24, 36, and 48) to observe the antiviral efficacy. Meanwhile, quantitative real-time PCR, and immunohistochemistry were used to detect the expression levels of SOCS3 mRNA in peripheral blood mononuclear cells (PBMCs) and matched liver tissues (baseline and 48 weeks). At the end of the 48-week treatment, patients with HBsAg negative or HBeAg seroconversion were defined as response group, and vice versa. Paired t-tests were used to compare normal distribution variables and the Mann-Whitney U test was used to compare the median differences between groups of non-normally distributed variables. Results: After 48 weeks of treatment, serum HBsAg levels in the Peg-IFN group continued to decline (average decrease of 1.14 log(10) IU / ml at week 48; P = 0.001 compared with baseline), while the entecavir group remained almost unchanged during treatment (average decrease was 0.05 log(10) IU / ml at week 48; compared with baseline P = 0.12). The expression of SOCS3 mRNA (Messenger RNA, mRNA) in peripheral blood and liver tissues of non-responder group was significantly higher than the response group in the course of Peg-IFNα2a treatment. The immunohistochemical results of liver tissue showed that the expression of SOCS3 in the non-responder group was significantly higher than that in the response group at baseline (P = 0.027). After 48 weeks of treatment with Peg-IFNα2a, the expression of SOCS3 in the non-responder group was significantly higher than that in the baseline and response groups (P = 0.003, P = 0.012, respectively). Conclusion: The expression of SOCS3 in peripheral blood mononuclear cells and liver tissues of non-responding CHB patients was significantly higher than that of responding CHB patients during interferon and nucleoside antiviral therapy. We speculated that SOCS3 might affect the antiviral efficacy through negative regulation of JAK-STAT signaling pathway, and partly expose the mechanism of interferon resistance.

TNF-α was involved in calcium hydroxide-promoted osteogenic differentiation of human DPSCs through NF-κB/p38MAPK/Wnt pathway.

It has been reported that calcium hydroxide can induce proliferation, migration, and mineralization in dental pulp stem cells (DPSCs), but the underlying molecular mechanisms are still unclear. In this study, we sought to explore the role of calcium hydroxide in the cell proliferation and directional differentiation of DPSCs and to study the regulatory effect of NF-κB, p38MAPK, and Wnt signaling on differentiation of DPSCs. CCK8 cell assay, Wound Healing Assay, and Alkaline Phosphatase Staining Assay were respectively used to determine the proliferation rate, migration and ALP expression of DPSCs. Alizarin Red Staining Assay was used to observe the mineralization of DPSCs. RT-PCR analysis and Western Blot Analysis displayed the expression of related fators at mRNA and protein level, respectively. In the present study, we found that NF-κB, p38MAPK, and Wnt signaling could abolish calcium hydroxide-induced proliferation of DPSCs. The inhibition of NF-κB, p38MAPK, and Wnt signaling suppressed the migration, ALP expression, and mineralization of DPSCs. NF-κB, p38MAPK, and Wnt signaling involved in directional differentiation of DPSCs. Moverover, calcium hydroxide could activate NF-κB, p38MAPK, and Wnt pathway by regulating TNF-α. Our study showed that NF-κB, p38MAPK, and Wnt signaling pathway were involved in calcium hydroxide-induced proliferation, migration, mineralization, and osteogenic differentiation in DPSCs. Calcium hydroxide affected NF-κB, p38MAPK, and Wnt pathway by regulating TNF-α.

[Comparison of vacuum bag fixation and Orfit rack with thermoplastic membrane fixation for image-guided cervical cancer radiotherapy].

Objective: To compare the difference of displacement between the vacuum bag fixation and the Orfit rack with thermoplastic membrane fixation of the cervical cancer patients, and to explore the individual fixation of the patients. Methods: We retrospectively analyzed the clinical data of 66 patients diagnosed as cervical cancer in Zhangzhou Municipal Hospital of Fujian Province from December 2014 to April 2016. Among them, 33 patients were fixed with vacuum bag, 33 patients were fixed with the Orfit rack with thermoplastic membrane. The cone-beam computed tomography (CBCT) images were acquired daily for the first three times of the radiotherapy, followed by once every other day for a total of 15 times. The CBCT scan images were matched with the CT scan images, and the matching results were recorded and analyzed. Results: The absolute value of the displacement in the left and right directions of the vacuum bag group was (0.28±0.30) cm, significantly lower than (0.38±0.46) cm in the Orfit rack with thermoplastic membrane group(P<0.001). The absolute value of the displacement in the anteroposterior direction of the vacuum bag group was (0.28±0.32) cm, with no significant difference of (0.27±0.23) cm in the Orfit rack with thermoplastic membrane group (P=0.580). The absolute value of the displacement in the up and down directions was (0.33±0.60) cm, with no statistically significant difference of (0.27±0.48) cm in the Orfit rack with thermoplastic membrane group (P=0.150). During the three times of CBCT scans, the differences of displacement in the left and right directions of the vacuum bag group were negligible, while apparently varied in the anteroposterior and up and down directions, however, the differences were not statistically significant (P>0.05). The change of the displacement in the three-dimensional direction in the Orfit rack with thermoplastic membrane group was marginal, and all of the differences were not significant(all P>0.05). Conclusions: Both the vacuum bag fixation and the Orfit rack with thermoplastic membrane fixation are suitable for the cone-beam CT image-guided radiotherapy of cervical cancer patients. However, the displacement in the left and right directions of the vacuum bag fixation is smaller than the Orfit rack with thermoplastic membrane fixation. During the period of treatment, the mean value of the difference of displacement in the anterior and posterior directions of the Orfit rack with thermoplastic membrane fixation is mild, which can be used individually by the patients with a flexible body and good tolerance.

[Comparison of MPure-12 Automatic Nucleic Acid Purification and Chelex-100 Method].

OBJECTIVES: To explore the forensic application value of MPure-12 automatic nucleic acid purification （MPure-12 Method） for DNA extraction by extracting and typing DNA from bloodstains and various kinds of biological samples with different DNA contents. METHODS: Nine types of biological samples, such as bloodstains, semen stains, and saliva were collected. DNA were extracted using MPure-12 method and Chelex-100 method, followed by PCR amplification and electrophoresis for obtaining STR-profiles. RESULTS: The samples such as hair root, chutty, butt, muscular tissue, saliva stain, bloodstain and semen stain were typed successfully by MPure-12 method. Partial alleles were lacked in the samples of saliva, and the genotyping of contact swabs was unsatisfactory. Additional, all of the bloodstains （20 µL, 15 µL, 10 µL, 5 µL, 1 µL） showed good typing results using Chelex-100 method. But the loss of alleles occurred in 1 µL blood volume by MPure-12 method. CONCLUSIONS: MPure-12 method is suitable for DNA extraction of a certain concentration blood samples．Chelex-100 method may be better for the extraction of trace blood samples．This instrument used in nucleic acid extraction has the advantages of simplicity of operator, rapidity, high extraction efficiency, high rate of reportable STR-profiles and lower man-made pollution.

Cartilage degradation in temporomandibular joint induced by unilateral anterior crossbite prosthesis.

OBJECTIVE: To investigate the pathological effects on temporomandibular joint cartilage of an experimentally created unilateral anterior crossbite prosthesis. METHODS: Six-week-old female Sprague-Dawley rats were divided into experimental and control groups and further divided into three subgroups for three time points (n = 9). In experimental groups, the unilateral anterior crossbite was created by bonding two metal tubes on left maxillary and mandibular incisors. 2, 4, or 8 weeks after operation, the histomorphological changes and the expression changes of PCNA, COL II, aggrecan, MMP-3, MMP-9, MMP-13, and TIMP-1 in condylar cartilage were investigated by hematoxylin-eosin and toluidine blue staining, real-time PCR, and immunohistochemistry. RESULTS: In experimental groups, decreasing in chondrocytes, cell-free areas, reduction in the thickness of hypertrophic layer, and loss of extracellular matrix were noticed. The expression level of PCNA was decreased at week 2 and week 4. The mRNA expression of COL II was decreased at 4 week and that of aggrecan was increased at 2 week but decreased at 4 and 8 week. The expression of MMP-9, MMP-13, and TIMP-1 was increased at 2-week and decreased at 4- and 8-week, while that of MMP-3 was increased at 4- and 8-week. CONCLUSIONS: The present unilateral anterior crossbite prosthesis led to degradation of mandibular condylar cartilage.

[Effects and techniques of soft tissue augmentation in immediate implant treatment].

Immediate implant placement can reduce the number of treatments and the time without teeth, but it carries a higher aesthetic risk. Soft tissue augmentation can reduce the risk of gingival recession to a certain extent, improve the predictability and long-term stability of immediate implant aesthetics, and is currently a hot research topic. A comprehensive understanding of the evidence-based medicine and surgical techniques using soft tissue augmentation in immediate implant surgery can assist in clinical diagnosis, treatment decisions and improve treatment outcomes. This article elucidates the changes in soft and hard tissues after immediate implant placement, aesthetic risks, and risk factors. It also discusses the advantages, timing, material selection, and commonly used clinical techniques of soft tissue transplantation in immediate implantation, aiming to provide reference for clinical doctors to improve the effectiveness of immediate implantation.

ENAM Mutations Can Cause Hypomaturation Amelogenesis Imperfecta.

Amelogenesis imperfecta (AI) is a diverse group of inherited diseases featured by various presentations of enamel malformations that are caused by disturbances at different stages of enamel formation. While hypoplastic AI suggests a thickness defect of enamel resulting from aberrations during the secretory stage of amelogenesis, hypomaturation AI indicates a deficiency of enamel mineralization and hardness established at the maturation stage. Mutations in ENAM, which encodes the largest enamel matrix protein, enamelin, have been demonstrated to cause generalized or local hypoplastic AI. Here, we characterized 2 AI families with disparate hypoplastic and hypomaturation enamel defects and identified 2 distinct indel mutations at the same location of ENAM, c588+1del and c.588+1dup. Minigene splicing assays demonstrated that they caused frameshifts and truncation of ENAM proteins, p.Asn197Ilefs*81 and p.Asn197Glufs*25, respectively. In situ hybridization of Enam on mouse mandibular incisors confirmed its restricted expression in secretory stage ameloblasts and suggested an indirect pathogenic mechanism underlying hypomaturation AI. In silico analyses indicated that these 2 truncated ENAMs might form amyloid structures and cause protein aggregation with themselves and with wild-type protein through the added aberrant region at their C-termini. Consistently, protein secretion assays demonstrated that the truncated proteins cannot be properly secreted and impede secretion of wild-type ENAM. Moreover, compared to the wild-type, overexpression of the mutant proteins significantly increased endoplasmic reticulum stress and upregulated the expression of unfolded protein response (UPR)-related genes and TNFRSF10B, a UPR-controlled proapoptotic gene. Caspase, terminal deoxynucleotidyl transferase UTP nick-end labeling (TUNEL), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assays further revealed that both truncated proteins, especially p.Asn197Ilefs*81, induced cell apoptosis and decreased cell survival, suggesting that the 2 ENAM mutations cause AI through ameloblast cell pathology and death rather than through a simple loss of function. This study demonstrates that an ENAM mutation can lead to generalized hypomaturation enamel defects and suggests proteinopathy as a potential pathogenesis for ENAM-associated AI.

Phosphorus removal in a membrane-assisted BNR process with focus on evolutions of PAOs and DPAOs.

A bench-scale UCT (University of Cape Town)-type membrane bioreactor (UCT-MBR) fed with low-strength synthetic wastewater was operated to investigate phosphorus removal with reference to poly-phosphate accumulating organisms (PAOs) and denitrifying poly-phosphate accumulating organisms (DPAOs). A series of kinetic assays of PAOs and DPAOs were also conducted to analyze the metabolic activities of PAOs and DPAOs. Results showed that 93% of chemical oxygen demand (COD) and 77% of total nitrogen could be removed at 0.08 kgCOD kg(-1) MLSS d(-1) and 0.015 kgN kg(-1) MLSS d(-1) loading (MLSS: mixed liquor suspended solids). Removal efficiencies of total phosphorus increased during the experimental phase, with an ultimate removal efficiency of 96.1%. Kano and Kaer increased from 1.95 and 6.29 mgPO4(3-)-P g(-1) MLSS h(-1) to 5.47 and 11.13 mgPO4(3-)-P g(-1) MLSS h(-1) for DPAOs and PAOs respectively, with the increased ratio of DPAOs to PAOs from 31 to 49% implying DPAO metabolic activity increased faster than that of PAOs during the DPAO accumulation phase. Pano-uptake increased by 6.6 mg L(-1) and the ratio of PTano-uptake to PTupt increased from 58.97 to 91.62%. The ratio of DPAOs to PAOs tended to stabilize at around 50% over time.

Ncor1 Deficiency Promotes Osteoclastogenesis and Exacerbates Periodontitis.

Nuclear receptor corepressor 1 (Ncor1) has been reported to regulate different transcription factors in different biological processes, including metabolism, inflammation, and circadian rhythms. However, the role of Ncor1 in periodontitis has not been elucidated. The aims of the present study were to investigate the role of Ncor1 in experimental periodontitis and to explore the underlying mechanisms through an experimental periodontitis model in myeloid cell-specific Ncor1-deficient mice. Myeloid cell-specific Ncor1 knockout (MNKO) mice were generated, and experimental periodontitis induced by ligation using 5-0 silk sutures was established. Ncor1 flox/flox mice were used as littermate controls (LC). Histological staining and micro-computed tomography scanning were used to evaluate osteoclastogenesis and alveolar bone resorption. Flow cytometry was conducted to observe the effect of Ncor1 on myeloid cells. RNA sequencing was used to explore the differentially targeted genes in osteoclastogenesis in the absence of Ncor1. Coimmunoprecipitation (Co-IP), chromatin immunoprecipitation (ChIP) experiments, and dual luciferase assays were performed to explore the relationship between NCoR1 and the targeted gene. Alveolar bone resorption in the MNKO mice was significantly greater than that in the LC mice after periodontitis induction and osteoclastogenesis in vitro. The percentage of CD11b+ cells, particularly CD11b+ Ly6G+ neutrophils, was substantially higher in gingival tissues in the MNKO mice than in the LC mice. Results of RNA sequencing demonstrated that CCAAT enhancer binding protein α (Cebpα) was one of the most differentially expressed genes between the MNKO and LC groups. Mechanistically, Co-IP assays, ChIP experiments, and dual luciferase assays revealed that NCOR1 interacted with peroxisome proliferator-activated receptor gamma (PPARγ) and cooperated with HDAC3 to control the transcription of Cebpα. In conclusion, Ncor1 deficiency promoted osteoclast and neutrophil formation in mice with experimental periodontitis. It regulated the transcription of Cebpα via PPARγ to promote osteoclast differentiation.

Effect of triethylene glycol dimethacrylate on the cytotoxicity, cyclooxygenase-2 expression and prostanoids production in human dental pulp cells.

AIM: To evaluate the effect of TEGDMA on cell cycle progression as well as alterations of cell cycle-related gene and protein expression. METHODOLOGY: Human dental pulp cells were exposed to 0-5 mmol L(-1) TEGDMA for 24 h. Cytotoxicity was evaluated by 3-(4, 5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. Cell cycle progression was analysed by propidium iodide (PI) flow cytometry. Cell death pathway was surveyed by annexin V/PI dual-staining flow cytometry. The mRNA expression of cell cycle-related genes (cdc2, cyclinB1 and p21) and COX-2 was evaluated by reverse transcriptase-polymerase chain reaction, and their protein expression was evaluated by Western blotting. The production of PGE(2) and PGF(2α) in the culture medium was determined by enzyme-linked immunosorbent assay. RESULTS: Triethylene glycol dimethacrylate inhibited cellular growth and induced cell cycle deregulation in dental pulp cells. High-dose exposure provoked both necrotic and apoptotic cell death. The gene and protein expression of cdc2, cyclin B1 and cdc25C declined obviously whilst cells treated with 2.5 mmol L(-1) TEGDMA concurrent with the elevated expression of p21. The mRNA and protein expression of COX-2, along with production of PGE(2) and PGF(2α), are drastically raised by 2.5-5 mmol L(-1) TEGDMA. CONCLUSIONS: Triethylene glycol dimethacrylate induced cytotoxicity, cell cycle arrest and apoptosis in dental pulp cells, which was associated with the decline of cdc2, cyclin B1, cdc25C expression and elevation of p21 expression. Concomitantly, COX-2 expression, PGE(2) and PGF(2α) production increased. These effects may contribute to explain the pulpal damage and inflammation induced by TEGDMA after operative procedures.

[Preliminary study on centrifugation time of liquid plasmatrix for soft tissue regeneration].

Objective: To provide reference for clinical application of liquid plasmatrix, and to investigate the optimal centrifugation time of liquid plasmatrix prepared by horizontal centrifugation for soft tissue regeneration from the aspects of mechanical properties, biological properties, and the effect of promoting soft tissue regeneration. Methods: Venous blood was collected from 6 healthy volunteers [3 males and 3 females, aged (26±2) years, with informed consent] who volunteered to donate blood at School of Stomatology, Wuhan University from September to November 2021. The collected venous blood was centrifuged at 500 ×g for 3, 5, 8 and 12 min to obtain liquid plasmatrix. The volume, weight, solidification time, and mechanical properties of liquid plasmatrix prepared at different centrifugation time were measured and recorded (the sample size at each time point was 3). The microstructure of different groups of liquid plasmatrix clot was observed by scanning electron microscope (SEM). The rheological properties of each group of liquid plasmatrix clot were measured by rheological test. The number and concentration of cells in the whole blood group and in each liquid plasmatrix group were measured using complete blood count test. The distribution of cells in the liquid plasmatrix clots was observed by hematoxylin-eosin staining. The effect of control group (Dulbecco's modified Eagle's medium containing 20% fetal bovine serum) and liquid plasmatrix clot exudates in 3, 5, 8, 12 min group (the sample size at each time point was 3) on gingival fibroblast migration was detected by cell migration method. Finally, the effects of control group and liquid plasmatrix clot exudates on the morphology of gingival fibroblasts were observed by fluorescence microscope. Results: The volume of liquid plasmatrix in 3, 5, 8 and 12 min group were approximately (2.47±0.12), (2.67±0.12), (3.53±0.12) and (3.73±0.12) ml, respectively. The weight of liquid plasmatrix in 3, 5, 8 and 12 min group were approximately (0.35±0.01), (0.46±0.02), (0.88±0.06) and (1.03±0.01) g, respectively. The maximum tensile force of liquid plasmatrix clots in 3, 5, 8 and 12 min group were (0.55±0.03), (0.56±0.03), (1.31±0.05) and (1.38±0.02) N, respectively. SEM results showed that the fibers inside the liquid plasmatrix clot became denser with increased centrifugation time. Compared with other groups, the concentrations of leukocytes, neutrophils, monocytes and lymphocytes in 8 min group were the highest, and the distribution of cell was more even. Compared with other groups, the efficiency of stimulating gingival fibroblast migration in 8 min group was the best (1.60±0.01). Fluorescence staining test showed that the liquid plasmatrix clot exudates could make gingival fibroblasts more stretched compared with control group. Conclusions: The present study shows that liquid plasmatrix prepared by centrifugation with 500 ×g centrifugal force for 8 min has higher concentration of viable cells and the ability to promote the migration of gingival fibroblasts.

Patterns and forces of occlusal contacts during lateral excursions recorded by the T-Scan II system in young Chinese adults with normal occlusions.

The objective was to characterise the patterns and forces of occlusal contacts during lateral mandibular excursions in a young Chinese adult population. Angle Class I normal occlusions in 85 dental students aged 20-29 were evaluated using the T-Scan II occlusal imaging and analysis system. The frequency of occlusal contacts was recorded for right and left lateral excursions at the position of lateral disclusion, at the canine-to-canine position and at the lateral maximal position. At the canine-to-canine position, the occlusal contact patterns were classified, and the relative forces were compared by multiple regression analysis, with α = 0·05. At the position of lateral disclusion, a high percentage of canine contacts (58·5%) and individual posterior tooth contacts on the working side was observed. At the canine-to-canine position, the canines on the working side contacted most frequently (68·9%), and the occlusal contact patterns could be classified into six groups. At the lateral maximal position, incisors (39·0%) and canines (32·3%) contacted most frequently. At the canine-to-canine position, subjects with occlusal contacts only on canines or on first premolars on the working side had the lowest relative occlusal forces (P < 0·001). Canine protection and group function do not identify all descriptors for lateral occlusal contact patterns in the natural dentition. Six occlusal contact patterns were found at the lateral canine-to-canine position.

Exogenous nucleation sites fail to induce detectable polymerization of actin in living cells.

Most nonmuscle cells are known to maintain a relatively high concentration of unpolymerized actin. To determine how the polymerization of actin is regulated, exogenous nucleation sites, prepared by sonicating fluorescein phalloidin-labeled actin filaments, were microinjected into living Swiss 3T3 and NRK cells. The nucleation sites remained as a cluster for over an hour after microinjection, and caused no detectable change in the phase morphology of the cell. As determined by immunofluorescence specific for endogenous actin and by staining cells with rhodamine phalloidin, the microinjection induced neither an extensive polymerization of endogenous actin off the nucleation sites, nor changes in the distribution of actin filaments. In addition, the extent of actin polymerization, as estimated by integrating the fluorescence intensities of bound rhodamine phalloidin, did not appear to be affected. To determine whether the nucleation sites remained active after microinjection, cells were first injected with nucleation sites and, following a 20-min incubation, microinjected with monomeric rhodamine-labeled actin. The rhodamine-labeled actin became extensively associated with the nucleation sites, suggesting that at least some of the nucleation activity was maintained, and that the endogenous actin behaved in a different manner from the exogenous actin subunits. Similarly, when cells containing nucleation sites were extracted and incubated with rhodamine-labeled actin, the rhodamine-labeled actin became associated with the nucleation sites in a cytochalasin-sensitive manner. These observations suggest that capping and inhibition of nucleation cannot account for the regulation of actin polymerization in living cells. However, the sequestration of monomers probably plays a crucial role.

Single particle tracking of surface receptor movement during cell division.

We have used fluorescent latex beads to label membrane receptors on cultured NRK cells. Movement of individual beads during cell division was recorded with digital imaging techniques. Surface-bound beads showed no organized movement during metaphase but started to migrate toward the equator approximately 1 min after anaphase onset, when chromosomes moved out of the equatorial region to create the interzone. The movement was most active in the central region of the cell near separating chromosomes, while beads located near the poles of the cell underwent primarily random motion. Most beads showed a surge in speed upon the passage of chromosomes, suggesting a possible link between chromosome separation and cortical reorganization. Furthermore, treatment of anaphase cells with cytochalasin D induced a rapid, simultaneous collapse of beads and cortical actin filaments into aggregates, indicating that the movement of beads was closely related to the reorganization of the actin cortex. In contrast to normal directional movement, cytochalasin-induced movement occurred in random directions and caused some beads in the equatorial region to move toward poles. Our results indicate that cytokinesis involves contractile activities, not only along the equator, but over a wide area of the actin-containing cortex. In addition, organized cortical activities appear to be temporally activated at anaphase onset, and spatially modulated by the spindle interzone or separating chromosomes.

Early growth of nano-sized calcium phosphate on phosphorylated bacterial cellulose nanofibers.

It is believed that studies on the early hydroxyapatite (HAp) deposition on nano-sized substrates may possibly allow us to understand the formation mechanisms of biominerals at the molecular level. In this study, bacterial cellulose (BC) nanofibers were phosphorylated and used as nano-sized templates for early mineralization of calcium phosphate (Ca-P). To initiate mineralization the BC nanofibers were immersed in 1.5 times simulated body fluids (1.5 SBF) at 37 degreees C for varying periods of time. The deposited minerals on the nanofiber surfaces were characterized using scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transformed infrared spectroscopy (FTIR). SEM observations confirmed that early growth (at 4 h) of the Ca-P minerals was heterogeneous, which was followed by extensive spread of the minerals on the entire surfaces of the nanofibers. XRD and FTIR analyses indicated that octacalcium phosphate (OCP) was the precursor of HAp formed on BC nanofibers. Furthermore, HAp deposited on BC nanofibers elongated along the c axis. Nucleation and growth of the Ca-P minerals were analyzed in this paper.

A new covered stent designed for intracranial vasculature: application in the management of pseudoaneurysms of the cranial internal carotid artery.

BACKGROUND AND PURPOSE: The management of intracranial pseudoaneurysms is controversial. The purpose of this study was to provide a preliminary evaluation of the clinical efficacy of a Willis covered stent specially designed for the intracranial vasculature in the management of a pseudoaneurysm of the cranial internal carotid artery (CICA). MATERIALS AND METHODS: Eight patients with pseudoaneurysms of the CICA were treated with use of the Willis covered stent. The flexibility of the entire stent system was gauged from the resistance met when reaching the target lesion and was categorized as no resistance, no apparent resistance, or resistance that could be overcome. The apposition of the Willis stent after deployment was scored as excellent with no endoleak, good with a small endoleak, or bad with an apparent endoleak. Follow-up angiography was performed 3 to 12 months after placement of the stent, and angiographic assessments were categorized as endoleak, stenosis of the covered segment of vessel, or occlusion of parent arteries. Follow-up clinical evaluations were also performed, and outcomes were graded as full recovery, improvement, unchanged, and aggravation. RESULTS: Endovascular treatment was technically successful in all aneurysms without procedural-related complications, and all of the stents were easily navigated to the targeted lesions in the CICA. Complete resolution of the pseudoaneurysm was observed in 6 patients immediately after the procedure, and a minimal endoleak into the aneurysm persisted in 2 patients. No morbidity or mortality and no technical adverse event occurred. A follow-up angiogram confirmed complete reconstruction of the internal carotid artery, with no recurrent aneurysmal filling and no occurrence of stenosis in the area of the stent. By the final follow-up visit, 4 patients had fully recovered, 3 had improved, and 1 patient's condition was unchanged. CONCLUSION: On the basis of our preliminary experience, the Willis covered stent specially designed for the intracranial vasculature can manage a CICA pseudoaneurysm safely and effectively, but longer follow-up and expanded clinical trials are needed.