Gut microbiota regulation of T lymphocyte subsets during systemic lupus erythematosus.

BACKGROUND: Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by disturbance of pro-inflammatory and anti-inflammatory lymphocytes. Growing evidence shown that gut microbiota participated in the occurrence and development of SLE by affecting the differentiation and function of intestinal immune cells. The purpose of this study was to investigate the changes of gut microbiota in SLE and judge its associations with peripheral T lymphocytes. METHODS: A total of 19 SLE patients and 16 HCs were enrolled in this study. Flow cytometry was used to detect the number of peripheral T lymphocyte subsets, and 16 s rRNA was used to detect the relative abundance of gut microbiota. Analyzed the correlation between gut microbiota with SLEDAI, ESR, ds-DNA and complement. SPSS26.0 software was used to analyze the experimental data. Mann-Whitney U test was applied to compare T lymphocyte subsets. Spearman analysis was used for calculating correlation. RESULTS: Compared with HCs, the proportions of Tregs (P = 0.001), Tfh cells (P = 0.018) and Naïve CD4 + T cells (P = 0.004) significantly decreased in SLE patients, and proportions of Th17 cells (P = 0.020) and γδT cells (P = 0.018) increased in SLE. The diversity of SLE patients were significantly decreased. Addition, there were 11 species of flora were discovered to be distinctly different in SLE group (P < 0.05). In the correlation analysis of SLE, Tregs were positively correlated with Ruminococcus2 (P = 0.042), Th17 cells were positively correlated with Megamonas (P = 0.009), γδT cells were positively correlated with Megamonas (P = 0.003) and Streptococcus (P = 0.004), Tfh cells were positively correlated with Bacteroides (P = 0.040), and Th1 cells were negatively correlated with Bifidobacterium (P = 0.005). As for clinical indicators, the level of Tregs was negatively correlated with ESR (P = 0.031), but not with C3 and C4, and the remaining cells were not significantly correlated with ESR, C3 and C4. CONCLUSION: Gut microbiota and T lymphocyte subsets of SLE changed and related to each other, which may break the immune balance and affect the occurrence and development of SLE. Therefore, it is necessary to pay attention to the changes of gut microbiota and provide new ideas for the treatment of SLE.

Sema4D/PlexinB1 inhibition ameliorates blood-brain barrier damage and improves outcome after stroke in rats.

The inflammatory process in stroke is the major contributor to blood-brain barrier (BBB) breakdown. Previous studies indicated that semaphorin 4D (Sema4D), an axon guidance molecule, initiated inflammatory microglial activation and disrupted endothelial function in the CNS. However, whether Sema4D disrupts BBB integrity after stroke remains unclear. To study the effect of Sema4D on BBB disruption in stroke, rats were subjected to transient middle cerebral artery occlusion and targeted injection of lentivirus-mediated clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 gene disruption of PlexinB1. We found that Sema4D synchronously increased with BBB permeability and accumulated in the perivascular area after stroke. Suppressing Sema4D/PlexinB1 signaling in the periinfarct cortex significantly decreased BBB permeability as detected by MRI and fibrin deposition, and thereby improved stroke outcome. In vitro, we confirmed that Sema4D disrupted BBB integrity and endothelial tight junctions. Moreover, we found that Sema4D induced pericytes to acquire a CD11b-positive phenotype and express proinflammatory cytokines. In addition, Sema4D inhibited AUF1-induced proinflammatory mRNA decay effect. Taken together, our data provides evidence that Sema4D disrupts BBB integrity and promotes an inflammatory response by binding to PlexinB1 in pericytes after transient middle cerebral artery occlusion. Our study indicates that Sema4D may be a novel therapeutic target for treatment in the acute phase of stroke.-Zhou, Y.-F., Li, Y.-N., Jin, H.-J., Wu, J.-H., He, Q.-W., Wang, X.-X., Lei, H., Hu, B. Sema4D/PlexinB1 inhibition ameliorates blood-brain barrier damage and improves outcome after stroke in rats.

Report: Small cell neuroendocrine carcinoma (SCNEC) of the tongue: A case report.

Small cell neuroendocrine carcinoma (SCNEC) of the tongue is very rare. We here present a SCNEC impatient with distant metastasis. A 74-year-old Chinese male went to hospital to treat a tongue tumor, which was founded at a conventional physical examination in Weifang Stomatology Hospital. The check of positron emission tomography-computer tomography (PET-CT) by Weifang people's hospital revealed a tumor in the right root of tongue, and distant metastasis in the right submandibular area, neck, mediastinum, right hilar, abdominal, retroperitoneal multiple lymph nodes, left thyroid, right lower lung, right scapula and bilateral adrenal. The patient was diagnosed tongue SCNEC by the pathological analysis of the tissue section. Conforming to the diagnosis of tongue SCNEC, the patient received adjuvant chemotherapy for 6 cycles with etoposide and carboplatin, and is alive now 9 months after the diagnosis.

Assessing changes in quality of life using the Oral Health Impact Profile (OHIP) in patients with different classifications of malocclusion during comprehensive orthodontic treatment.

BACKGROUND: The objectives of this study were to investigated changes in OHRQoL among patients with different classifications of malocclusion during comprehensive orthodontic treatment. METHODS: Clinical data were collected from 81 patients (aged 15 to 24) who had undergone comprehensive orthodontic treatment. Participants were classified 3 groups: Class I (n = 35), II (n = 32) and III (n = 14) by Angle classification. OHRQoL was assessed using the Oral Health Impact Profile (OHIP-14). All subjects were examined and interviewed at baseline (T0), after alignment and leveling (T1), after correction of molar relationship and space closure (T2), after finishing (T3). Friedman 2-way analysis of variance (ANOVA) and Wilcoxon signed rank tests were used to compare the relative changes of OHRQoL among the different time points. A Bonferroni correction with P < 0.005 was used to declare significance. RESULTS: Significant reductions were observed in all seven OHIP-14 domains of three groups except for social disability (P > 0.005) in class I and class II, Handicap in class II and class III (P > 0.005). Class I patients showed significant changes for psychological disability and psychological discomfort domain at T1, functional limitation, physical pain at T2. Class III patients showed a significant benefit in all domains except physical pain and functional limitation. Class II patients showed significant changes in the physical pain, functional disability, and physical disability domains at T1. CONCLUSIONS: The impact of comprehensive orthodontic treatment on patients' OHRQoL do not follow the same pattern among patients with different malocclusion. Class II patients benefits the most from the stage of space closure, while class I patients benefits the first stage (alignment and leveling) of treatment in psychological disability and psychological discomfort domains.

[Analysis of pathogen spectrum of Encephalitis/Meningitis in northwestern area of China].

OBJECTIVE: To learn the characteristics of pathogen spectrum of Encephalitis /Meningitis in northwestern area of China. METHODS: Between January 1st 2009 and March 31st 2011, a total of 569 patients with clinical symptoms of Encephalitis/Meningitis were selected from the hospitals in Gansu, Qinghai,Inner Mongolia and Xinjiang province. 1514 samples of specimen were collected from the 515 patients, to detect the IgM of Japanese encephalitis virus (JEV), enterovirus (EV, including Coxsackie virus, ECHO virus and enterovirus 71), Mumps virus, Herpes simplex virus (HSV) in blood and cerebrospinal fluid. Meanwhile, Neisseria meningitis (Nm), Haemophilus influenzae Type B (Hib), Staphylococcus, Streptococcus pneumonia, Streptococcus Suis, E. Coli and Cryptococci were also identified. The detection results were analyzed by different region, time and age range. RESULTS: Pathogenic bacteria were identified in the specimen from 16 patients, with the rate at 3.65%, of which the dominant ones were Streptococcus pneumonia (7 patients, 43.75%). Virus were identified in the specimen from 132 patients, with the rate at 27.05%, of which the dominant types were EV and HSV, accounting for 33.33% (44 cases) and 31.82% (42 cases) respectively. The detection rate of virus showed a significant seasonal trend, with the peak appearing between June and November each year. The peak of EV detection was between July and September, with 24 cases detected out; the peak of HSV was between June and August (11 cases detected out); mumps virus was mainly found between July and December (25 cases). There was no significant time-distribution found in the detection of bacteria. The EV and HSV were mainly distributed in Gansu and Qinghai province (70 cases) ;most of mumps virus were found in Gansu province (24 cases);and JEV were only found in Gansu province (20 cases). The viral pathogen spectrum was identified in all ages, and the EV and mumps virus were mainly found in children aged 0-14 years old (42 and 17 cases respectively) ; JEV were identified in people over 15 years old, with 13 detected out of the 20 patients. CONCLUSION: The main pathogen of acute encephalitis and meningitis in northwestern area of China was virus, and the main pathogens of encephalitis and meningitis in children under 15 years were Herpes simplex virus and Mumps virus.

[Effect of collagen sponge on the healing process of alveolar fossa after tooth extraction in rats].

PURPOSE: To investigate the effect of collagen sponge on early bone healing process of alveolar fossa after tooth extraction in rats. METHODS: A total of 16 healthy female SD rats were selected. Animal models with tooth extraction were established. The right alveolar fossa inserted with collagen sponge was as the experimental group, and the left alveolar fossa was as the control group with treatment. The rats were sacrificed 1, 2, 4 and 8 weeks after tooth extraction, and the osteogenesis of alveolar fossa was observed. Real-time quantitative PCR (qt-PCR) was used to detect the changes of osteogenesis related gene expression. SPSS 19.0 software package was used for statistical analysis. RESULTS: After surgery, alveolar cavity healing was significantly better in the experimental group than in the control group. Osterix, Runx2 and Vegf genes were expressed in the experimental group and the control group, and the expression levels of related genes in the experimental group were significantly higher than the control group 1, 2, 4 and 8 weeks after surgery(P＜0.05). CONCLUSIONS: Collagen sponge could promote early alveolar bone healing, possibly related to the expression level of osteogenic genes regulated by collagen sponge.

[Analysis on epidemiology and the main clinical symptoms of viral encephalitis in Gansu, 2009-2011].

OBJECTIVE: To understand the epidemiological characteristics and the main clinical symptoms of viral encephalitis in Gansu. METHODS: A total of 322 viral encephalitis patients were recruited from province sentinel hospitals in Gansu province from 2009 to 2011, and their basic information were collected as well as their serum samples and cerebrospinal fluid samples. 296 out of the 322 cases were qualified for our study. Based on the patients' epidemiological characteristics and clinical features, we determined the detection of the virus types (at least one kind of virus detection was carried out for each case). ELISA was applied to test the IgM antibody of Japanese encephalitis (JE) virus (JEV), enterovirus (EV: including Coxsackie virus, echovirus, enterovirus 71), mumps virus and herpes simplex virus (HSV) in cerebrospinal fluid and serum specimen. The difference of positive detected rate between types of virus, among patients from different regions, time, or at different ages, as well as the different clinical symptoms between JE patients and other viral encephalitis patients, were analyzed and compared. RESULTS: The positive detected rate of virus in the 296 patients was 27.03% (80/296); the positive rate of JEV, EV, mumps virus, HSV detected was separately 7.53% (22/292), 8.75% (23/263), 13.84% (22/159) and 15.09% (40/265), and the difference was statistically significant (χ(2) = 10.849, P < 0.05). 90.91% (20/22) of the JEV positive cases were distributed in Tianshui, Longnan and Pingliang, and 95.45% (21/22) patients were infected from July to September. All the 23 EV detected positive patients were infected from April to December, while the ages of patients ranged from 1 to 44 years old. Mumps virus, HSV testing positive cases had onset every month. Logistic regression analysis showed that the patients who had the symptoms as disturbance of consciousness (OR = 15.487, 95%CI: 2.266 - 105.852), somnolence (OR = 11.659, 95%CI: 1.783 - 76.242), convulsions (OR = 11.062, 95%CI: 1.687 - 72.530) were more likely to infect JEV. CONCLUSION: HSV was the principal pathogen of viral encephalitis in Gansu. An obvious central tendency in the regional and time distribution was found in JEV infection; and the clinical symptoms of JE patients were more severe.

Lower lip recurrent keratoacanthoma: A case report.

BACKGROUND: This paper introduces a case of recurrent keratoacanthoma (KA). KA is a self-healing disease. Recurrence after surgical resection is rare. In this case, the local application of retinoic acid ointment after the second operation achieved a good prognosis after 2 years of follow-up. CASE SUMMARY: A 76-year-old male patient was admitted to the hospital for "lower lip rupture and scab for 3 mo". Treatment: A rectangular incision was made in the healthy tissue about 3 mm outside the periphery of the lower lip mass, and a modified Bernard sliding flap was designed to completely remove the mass. Pathology showed (lower lip) KA. When the patient returned 6 mo after surgery, the middle mucosa of the lower lip had a bulge with a diameter of about 0.5 cm. The boundary was still clear, the surface was ulcerated. A recurrence of lower lip KA was suspected and a fan-shaped incision was performed in the healthy tissue about 5 mm outside the lesion to completely resect. Pathological showed lower lip KA had recurred. Topical application of tretinoin cream was applied once a day for 3 mo. The lower lip wounds were clean at the 2-year postoperative follow-up and the mucosa was normal. CONCLUSION: Adjuvant retinoic acid treatment after KA surgical resection can achieve good results.

[Effect of ipriflavone on reconstruction of periodontal tissues during recurrence of orthodontic teeth in rats].

PURPOSE: To investigate the effect of ipriflavone on reconstruction of periodontal tissues during recurrence of orthodontic teeth. METHODS: Twenty-four male SD rats were randomized into 2 groups, ipriflavone group(IP group) and control group, there were 12 rats in each group. The model of recurrence after movement of orthodontic teeth in rats was established. After continuous loading for 10 days, the loading devices were removed. Rats in ipriflavone group (IP group) were given ipriflavone intragastrically for 10 mg/(kg·d) after the devices were removed, while rats in the control group were given an equivalent amount of normal saline after the devices were removed. On the 0th, 1st, 3rd, 7th, and 10th day of administration, the rat maxillary impression and plaster model of two groups were prepared under local anesthesia, the distance between maxillary first molar lingual sulcus point and third molar in lingual groove point was measured to evaluate the relapse distance. After drug infusion for 10 days, the collected tissue specimens were stained with H-E to observe periodontal reconstruction, and expression of bone morphogenetic protein-2(BMP-2) was detected by immunohistochemical staining. Software Image-Pro 6.0 was used to analyze the optical density values of the stained sections. The data were analyzed with SPSS 22.0 software package. RESULTS: After removing the orthopaedic devices for 10 days , there was a significant recurrence of the movement of the orthodontic teeth in both groups. The recurrent distance of IP group was significantly smaller than that of the control group, and still significantly smaller than that of the control group at 10 d. H-E staining and immunohistochemical staining results showed that the IP group had more new bone formation and more BMP-2 expression in the periodontal tissues compared to the control group in vivo. CONCLUSIONS: In the process of recurrence of orthodontic tooth movement, ipriflavone can promote the expression of BMP-2 in periodontal tissue, improve bone remodeling of periodontal tissue, and effectively reduce the recurrent rate of orthodontic tooth movement.

Isolation of high quality RNA and construction of a suppression subtractive hybridization library from ramie (Boehmeria nivea L. Gaud.).

Isolation of high quality RNA from ramie (Boehmeria nivea L. Gaud.) is difficult due to its high levels of polyphenols, polysaccharides, pectin, fat, wax and other secondary metabolites. A modified procedure based on guanidinium isothiocyanate for RNA preparation of ramie was developed in this study. High concentrations (5%, v/v) of guanidinium isothiocyanate, PVP-4000, sodium citrate and sodium lauryl sarcosinate and beta-mercaptoethanol were used in the extraction buffer, together with a low pH sodium acetate (pH 4.0) added to improve the RNA quality. The average yield was about 400 microg RNAg(-1) fresh leaves. One SSH library which was induced by ramie anthracnose was constructed by utilizing the RNA extracted through the present method. These results showed that our protocol was applicable for RNA isolation from recalcitrant ramie tissues.

Root shortening in patients treated with two-step and en masse space closure procedures with sliding mechanics.

OBJECTIVE: To investigate the comparative amount of root shortening between two-step and en masse space closure procedures. MATERIALS AND METHODS: Fifty-two patients were selected from a pool of patients satisfying the following inclusion criteria: no evidence of resorption on the pretreatment panoramic radiographs; no dental trauma; no dilacerations of incisor roots, anodontia, or impacted canines; complete root formation at the start of treatment; intact and caries-free incisors; no endodontically treated incisors; Angle Class I or II malocclusion; extraction of four first premolars; and space closure with moderate anchorage. Patients received treatment with either a two-step or an en masse procedure to close the extraction spaces after alignment and leveling with the same preadjusted appliances. Root shortening of the maxillary and mandibular incisors was evaluated on panoramic radiographs, taken before and after space closure, and measured in millimeters. Distortion of measurements caused by panoramic radiographs was corrected by using special metal rods ligated to brackets. Statistical comparisons of root shortening between space closure procedures were investigated with the two-sample t test. RESULTS: No difference was found in the amount of root shortening between space closure procedures. The average root shortening of maxillary central and lateral incisors was 0.43 +/- 0.12 mm and 0.58 +/- 0.10 mm, respectively, and that of mandibular central and lateral incisors was 0.23 +/- 0.07 mm and 0.22 +/- 0.06 mm, respectively. CONCLUSION: No difference should be expected in root resorption between two-step and en masse space closure procedure.

Inhibitory effects of resveratrol on orthodontic tooth movement and associated root resorption in rats.

OBJECTIVE: To investigate the effect of resveratrol (RSV) on orthodontic tooth movement (OTM) and orthodontic induced root resorption (OIRR) in rats. METHODS: Thirty-six male Wistar rats used in this study were randomly divided into three groups of 12 animals each. All test subjects underwent a 50 g orthodontic force each, generated from a nickel-titanium closed-coil spring. The control group were fed carboxymethylcellulose (CMC) while rats in other two groups were fed 5 mg/kg/d RSV or 10 mg/kg/d RSV (dissolved in CMC). After 14 days of OTM, all rats were sacrificed, after which each group was randomly divided into two subgroups (6 test subjects in each subgroup). One subgroup was used to measure the amount of OTM and assessed by hematoxylin and eosin (HE) staining, tartrate-resistant acid phosphatase (TRAP) staining, and immunohistochemistry staining of Receptor Activator of Nuclear Factor-κ B Ligand (RANKL), Osteoprotegerin (OPG), Runt-related transcription factor 2 (RUNX2), as well as Osteocalcin (OCN). The second subgroup was used to analyze OIRR via scanning electron microscopy. RESULTS: Compared with the control group, the RSV groups showed a significant decrease in the distance of OTM and the OIRR ratio (p＜0.05). The number of TRAP positive osteoclasts and the expression of RANKL in periodontal tissue of the RSV groups were significantly inhibited (p＜0.01) while the expression of OPG, RUNX2, and OCN were remarkably promoted (p＜0.05). The effect of 10 mg/kg/d RSV group was more obvious than that of 5 mg/kg/d RSV group (p＜0.05). CONCLUSIONS: RSV could reduce the extent of OTM and root resorption areas.

The effects of binge alcohol exposure on tooth movement and associated root resorption in rats.

BACKGROUND: Considering the increasing prevalence of alcohol use and the growing number of orthodontic patients, some orthodontic patients might engage in binge drinking during treatment. Nevertheless, little is known about the effect of alcohol use on orthodontic treatment. METHODS: Male Wistar rats were divided into ethanol and control groups (n = 32). The rats received a single daily intraperitoneal injection of 20% (vol/vol) ethanol/saline solution at a dose of 3 g/kg of ethanol or saline for three consecutive days, and no injection was given during the remaining four days each week. All rats received orthodontic appliances to draw the maxillary first molar mesially. The rats were sacrificed at days 14 and 28, respectively. The amount of tooth movement was measured. Root resorption area was evaluated by scanning electron microscope. Hematoxylin and eosin (H&E) staining and tartrate-resistant acid phosphatase (TRAP) staining were conducted. Immunohistochemistry staining was performed to evaluate the expressions of nuclear factor kappa B ligand (RANKL), osteoprotegerin (OPG), and inducible nitric oxide synthase (iNOS). RESULTS: There were no significant differences in tooth movement and root resorption between ethanol and control groups. The number of TRAP-positive cells was significantly higher in the ethanol group. The expression of RANKL was statistically increased in the ethanol group. In contrast, the expression of OPG was remarkably decreased in rats injected with ethanol. Moreover, the iNOS level was significantly up-regulated in the ethanol group. CONCLUSION: The tooth movement and root resorption in rats were not affected by binge alcohol exposure.

Synthesis and in vitro and in vivo evaluation of manganese(III) porphyrin-dextran as a novel MRI contrast agent.

5-(4-Aminophenyl)-10,15,20-tris(4-sulfonatophenyl) manganese(III) porphyrin conjugated with dextran was synthesized. Its potential of being used as a tumor-targeting magnetic resonance imaging contrast agent was evaluated in vitro and in vivo. The results demonstrated that the compound has a longitudinal relaxivity (R(1)) higher than Gd-DTPA, low cytotoxicity and binding specificity to tumor cell membrane.

Erythropoietin attenuates high glucose-induced oxidative stress and inhibition of osteogenic differentiation in periodontal ligament stem cell (PDLSCs).

Diabetes mellitus and periodontitis have long been considered to be biologically linked. Erythropoietin (EPO) has multiple biological functions, such as stimulating the proliferation and differentiation of erythroid progenitor cells and reducing glucose-induced oxidative stress via different mechanisms, acting as a direct antioxidant. The purposes of the study to examine the anti-oxidative effect of EPO on reducing high glucose-induced oxidative stress of hPDLSCs and provide a better understanding of the mechanism of these processes. PDLSCs were induced by highglucose (HG, 30 mM) in the presence or absence of EPO. Cell proliferation was measured by MTT assay. The reactiveoxygen species (ROS) level, malondialdehyde (MDA) level and superoxide dismutase (SOD) activity were detected to evaluate oxidative stress. qRT-PCR and western blot analysis were used to examine the expression of osteogenic related genes and protein (Runx2 and Osterix). Alizarin Red-S staining was used to detect mineralized nodule formation. The results showed that EPO promote the proliferation of PDLSCs, which was suppressed by high glucose (30 mM). Moreover, EPO attenuated high glucose (30 mM) induced oxidative stress by reducing the levels of ROS and MDA, and increasing the SOD activity. Furthermore,EPO alleviate high glucose(30 mM) induced suppression of osteogenic differentiation ability in PDLSCs, as evidenced by the up-regulated mRNA and protein expression of Runx2 and Osterix and increased ALP activity. In conclusion, EPO attenuates high glucose-induced oxidative stress, inhibitory effect of proliferation and inhibition of osteogenic differentiation in periodontal ligament stem cell (PDLSCs).

[Effect of psoralen on the stability after orthodontic tooth movement in rats].

PURPOSE: This study was designed to investigate the effect of psoralen on periodontal tissue reconstruction after orthodontic tooth movement(OTM) in rats. METHODS: Thirty-six male 6-week-old Wistar rats were randomly divided into 2 groups: the experimental group and the control group. The experimental group and the control group were all installed between the central incisor and the left maxillary first molars to pull the first molars away from the force device; after 21 days, the force was removed and the rats in 2 groups were given drug gavage. Rats in the experimental group were given a gavage of psoralen 8 mg/kg per day, while rats in the control group were given the same amount of 0.9% sodium chloride everyday. Maxillary casts were made every week during the experimental and were scanned by 3D Scanner to measure relapse distance, and histologic examination was conducted. After 28 days, the rats were sacrificed and rats' upper jaw was separated. The remaining sections were immunohistochemically stained with BMP2 and BMP4. SPSS 19.0 software package was used for statistical analysis. RESULTS：Both groups had relapse after the force device was removed. Significant decrease of relapse percentage was observed in the experimental group compared with the control group at day 7，day 14，day 21 and day 28（P<0.05）. The speed of relapse of both groups were fastest in the first week and slowed down in the second, third and fourth week gradually. The speed of relapse in the experimental group in the first week was significantly less than in the control group(P<0.05).The expression of BMP2 and BMP4 within periodontal membrane and alveolar bone was significantly higher in the experimental group than in the control group(P<0.05). CONCLUSIONS: Psoralen can accelerate the reconstruction of periodontal tissues of orthodontic tooth and reduce relapse.

Erythropoietin enhances osteogenic differentiation of human periodontal ligament stem cells via Wnt/ß-catenin signaling pathway.

OBJECTIVES: The aim of this study is to examine the roles of erythropoietin (EPO) in regulating proliferation and osteogenic differentiation of periodontal ligament stem cells (PDLSCs) and analyze the underlying signaling of these processes. MATERIALS AND METHODS: PDLSCs were isolated and characterized. The PDLSCs were transfected with ß-catenin shRNA. qRT-PCR and Western blot analysis were used to examine the osteogenic effects of EPO on the expression of osteogenic-related genes and protein (Runx2, OCN and Osterix) in PDLSCs. Alizarin Red-S staining was used to detect mineralized nodule formation. In addition, the relationship between the Wnt/ß-catenin pathway and the effect of EPO on the osteogenesis of PDLSCs was investigated. RESULTS: The results suggested that EPO exerts positive osteogenic effects on PDLSCs. The results showed that EPO decreased the growth of PDLSCs slightly and increased alkaline phosphatase activity and calcium deposition in a dose-dependent manner. The expression of Runx2, Osterix and OCN was increased after EPO administration. EPO increases ß-catenin and Cyclin D1 in PDLSCs. After transfected with ß-catenin shRNA, the osteogenic effect of EPO on PDLSCs was attenuated. CONCLUSION: EPO promotes osteogenic differentiation of PDLSCs. The underlying mechanism may be activating Wnt/ß-catenin signaling pathway.

Berberine improves advanced glycation end products­induced osteogenic differentiation responses in human periodontal ligament stem cells through the canonical Wnt/ß­catenin pathway.

The aim of the present study was to investigate the effects of advanced glycation end products (AGEs) and berberine hydrochloride (BBR) on the osteogenic differentiation ability of human periodontal ligament stem cells (hPDLSCs) in vitro, and their underlying mechanisms. hPDLSCs were subjected to osteogenic induction and were treated with AGEs or AGEs + BBR. Following varying numbers of days in culture, alkaline phosphatase (ALP) activity assays, ALP staining, alizarin red staining, ELISAs, and reverse transcription­quantitative polymerase chain reaction (RT­qPCR) and western blot analyses were performed to determine the osteogenic differentiation ability of hPDLSCs; RT­qPCR, western blot analysis, and immunofluorescence staining were conducted to investigate the underlying mechanisms. The canonical Wnt/ß­catenin pathway inhibitor XAV­939 and agonist CHIR­99021 were used to determine the contribution of the canonical Wnt/ß­catenin pathway to differentiation. Treatment with AGEs resulted in reduced ALP activity and Collagen I protein levels, decreased ALP staining, fewer mineralized nodules, and downregulated expression of osteogenic­specific genes [Runt­related transcription factor 2 (Runx2), Osterix, ALP, osteopontin (OPN), Collagen I and osteocalcin (OCN)] and proteins (Runx2, OPN, BSP and OCN); however, BBR partially rescued the AGE­induced decrease in the osteogenic potential of hPDLSCs. Furthermore, AGEs activated the canonical Wnt/ß­catenin signaling pathway and promoted the nuclear translocation of ß­catenin; BBR partially attenuated this effect. In addition, XAV­939 partially rescued the AGE­induced reduction in the osteogenic potential of hPDLSCs, whereas CHIR­99021 suppressed the BBR­induced increase in the osteogenic potential of hPDLSCs. The present study indicated that AGEs attenuated the osteogenic differentiation ability of hPDLSCs, in part by activating the canonical Wnt/ß­catenin pathway; however, BBR attenuated these effects by inhibiting the canonical Wnt/ß­catenin pathway. These findings suggest a role for BBR in periodontal regeneration induced by hPDLSCs in patients with diabetes mellitus.

Effects Of Triptolide On Tooth Movement And Root Resorption In Rats.

PURPOSE: The aim of this study was to investigate the effects of triptolide on the tooth movement and root resorption in rats during orthodontic treatment. MATERIAL AND METHODS: A total of 48 male Wistar rats were divided into three groups of 16 each. The right maxillary first molars of rats were drawn mesially by closed coil nickel-titanium spring with a force of 50 g. The two experimental groups received intraperitoneal injections of triptolide for 14 days at a dose of 15 µg/kg/day and 30 µg/kg/day, respectively. The control group received vehicle injections. After 14 days, the rats were humanely killed. The amount of tooth movement was measured. Eight rats from each group were randomly chosen for analysis of the percentage of root resorption area by scanning electron microscopy. For the remaining eight rats in each group, the H&E staining, tartrate-resistant acid phosphatase (TRAP) staining and immunohistochemistry analysis were performed. RESULTS: The amount of tooth movement and the ratio of root resorption area were significantly decreased in the triptolide-treated rats. The number of TRAP-positive cells was significantly lower in triptolide-treated groups. Moreover, the expression of nuclear factor kappa B ligand (RANKL) was reduced. In contrast, the expression of osteoprotegerin was significantly up-regulated. In the tension side, the expressions of runt-related transcription factor 2 and osteocalcin were significantly enhanced by triptolide injection. CONCLUSION: Triptolide injection could arrest orthodontic tooth movement and reduce root resorption in rats via inhibition of osteoclastogenesis. In addition, triptolide may exert a positive effect on osteoblastogenesis.

Muscone Promotes The Adipogenic Differentiation Of Human Gingival Mesenchymal Stem Cells By Inhibiting The Wnt/ß-Catenin Signaling Pathway.

OBJECTIVES: This study was performed to evaluate the effects of muscone on the proliferation, migration and differentiation of human gingival mesenchymal stem cells (GMSCs) and to explore the relevant mechanisms. MATERIALS AND METHODS: We performed studies to determine the effects and mechanisms of muscone on GMSC proliferation, migration and differentiation. We conducted CCK-8, colony formation, transwell chamber, scratch wound, alkaline phosphatase (ALP) staining and activity, and alizarin red and oil red O staining assays, as well as real-time quantitative polymerase chain reaction (qRT-PCR), to ascertain the effects of muscone on GMSC proliferation, migration and differentiation in vitro. The mechanism by which muscone influences the osteogenic and adipogenic differentiation of GMSCs was elucidated by qRT-PCR and Western blotting. RESULTS: We found that muscone significantly promoted GMSC proliferation, chemotaxis, wound healing and fat droplet formation and inhibited ALP activity and mineral deposition. Notably, we observed that the Wnt/ß-catenin pathway was closely related to the ability of muscone to inhibit the osteogenic differentiation and promote the adipogenic differentiation of GMSCs. The effect of muscone on the multidirectional differentiation capacity of GMSCs was significantly reversed by the agonist lithium chloride through the Wnt/ß-catenin signaling pathway. CONCLUSION: Muscone effectively increased the proliferation and migration, promoted the adipogenic differentiation and inhibited the osteogenic differentiation of GMSCs by inhibiting the Wnt/ß-catenin signaling pathway. These results may provide a theoretical basis for the application of GMSCs and muscone in tissue engineering and regenerative medicine.