OBJECTIVES: This study aimed to clarify the effects of Foxp3 silencing on the expression of inflammatory cytokines in human periodontal ligament cells (hPDLFs) in an inflammatory environment and on cell proliferation and invasiveness, as well as to explore the role of Foxp3 gene in the development of periodontitis. METHODS: An small interfering RNA (siRNA) construct specific for Foxp3 was transfected into hPDLFs. Foxp3 silencing efficiency was verified by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, and the siRNA with the optimum silencing effect of Foxp3 gene was screened. Using lipopolysaccharide to simulate an inflammatory environment in vitro, CCK-8 detected the effect of silencing Foxp3 on hPDLFs proliferation under inflammatory conditions. Wound-healing experiments and transwell assays were conducted to detect the effect of silencing Foxp3 on hPDLF migration under inflammatory conditions. The expression of the inflammatory cytokines interleukin (IL)-6 and IL-8 was detected by RT-PCR and Western blotting under inflammatory conditions. RESULTS: After siRNA transfection, RT-PCR and Western blotting analyses showed that the expression of Foxp3 mRNA in the Foxp3-si3 group decreased significantly (t=21.03, P<0.000 1), and the protein expression of Foxp3 also decreased significantly (t=12.8, P<0.001). In the inflammatory environment, Foxp3 gene silencing had no significant effect on hPDLFs proliferation (P>0.05), and Foxp3 gene silencing promoted hPDLFs migration (P<0.05). Moreover, the expression of IL-6 and IL-8 increased (P<0.05). CONCLUSIONS In an inflammatory environment, Foxp3 gene silencing promoted hPDLFs migration but had no significant effect on hPDLFs proliferation. The expression of inflammatory factors expressed in hPDLFs increased after Foxp3 gene silencing, indicating that Foxp3 gene inhibited inflammation in periodontitis.
Humans ; Cell Proliferation/genetics* ; Cells, Cultured ; Cytokines/metabolism* ; Fibroblasts/metabolism* ; Forkhead Transcription Factors/metabolism* ; Gene Silencing ; Interleukin-6/metabolism* ; Interleukin-8/metabolism* ; Periodontal Ligament/metabolism* ; Periodontitis/metabolism* ; RNA, Small Interfering/metabolism* ; Transcription Factors/metabolism*

Objective : To investigate the effect of transforming growth factor β1 (TGF-β1) on human periodontal ligament fibroblasts (hPDLFs) stimulated by lipopolysaccharide from Porphyromonas gingivalis (Pg-LPS). Methods: hPDLFs were obtained and identified by immunohistochemistry. The stimulating concentration of Pg-LPS was determined by qRT-PCR and CCK-8. The hPDLFs were divided into 4 groups: blank control group, 100 μg/mL pure Pg-LPS; low concentration group, 1 ng/mL TGF-β1+100 μg/mL Pg-LPS; medium concentration group, 10 ng/mL TGF-β1+100 μg/mL Pg-LPS; and high concentration group 100 ng/mL TGF-β1+100 μg/mL Pg-LPS. Cell proliferation was measured by CCK-8 assay at 72 hours, cell migration was measured by scratch and Transwell chamber assays at 24 hours, and the cell cycle of the hPDLFs was measured by flow cytometry at 72 hours. The expression of Forkhead/winged helix transcription factor p3 (Foxp3), interleukin-6 (IL-6) and Epstein-Barr virus-induced gene 3 (EBI3) mRNA in hPDLFs at 72 hours was measured by qRT-PCR, and the expression of Foxp3, IL-6 and EBI3 proteins in hPDLFs at 72 hours was detected by western blot. Results : The immunohistochemistry results showed that anti-vimentin was positive and anti-keratin was negative. At a concentration of 100 μg/mL Pg-LPS, the expression of IL-6 mRNA in hPDLFs was increased (P<0.000 1), and the proliferation of hPDLFs was decreased (P<0.000 1). Therefore, 100 μg/mL PG-LPS was selected to simulate the inflammatory state. 10, 100 ng/mL TGF-β1 could improve the proliferation ability of hPDLFs in inflammatory state (P<0.000 1) ; 1, 10 and 100 ng/mL TGF-β1 could promote the migration ability of hPDLFs in inflammatory state (P<0.000 1). 1, 10 and 100 ng/mL TGF-β1 could accelerate the cell cycle of hPDLFs in inflammatory state (P<0.000 1). 1, 10 and 100 ng/mL TGF-β1 could inhibit the expression of IL-6 gene and protein in hPDLFs in inflammatory state (P<0.000 1), 1 and 10 ng/mL TGF-β1 could increase the expression of EBI3 gene and protein in hPDLFs in inflammatory state (P<0.000 1). 1, 10 ng/mL TGF-β1 could increase the expression level of Foxp3 gene in hPDLFs in inflammatory state, and 10 ng/mL TGF-β1 could increase the expression level of Foxp3 protein (P<0.05). Conclusion TGF-β1 can promote the proliferation and migration of hPDLFs under inflammatory conditions, upregulate EBI3 and inhibit inflammation, which may be related to the expression of the transcription factor Foxp3.

IL-6 and IL-17 levels in saliva and serum were measured by ELISA in 116 cases of chronic periodontitis(CP)in Mulei district of Xinjiang,including 62 cases of Han and 54 of Kazak.Health controls included 50 subjects of Han and 45 of Kazak.IL-6 and IL-17 levels in serum and saliva were higher in CP groups than in the controls(P <0.05),and correlated with the severity of CP(P <0.05),in Kazak CP group were higher than in Han CP group(P <0.05).

Microglia play a pivotal role in clearance of Aβ by degrading them in lysosomes, countering amyloid plaque pathogenesis in Alzheimer's disease (AD). Recent evidence suggests that lysosomal dysfunction leads to insufficient elimination of toxic protein aggregates. We tested whether enhancing lysosomal function with transcription factor EB (TFEB), an essential regulator modulating lysosomal pathways, would promote Aβ clearance in microglia. Here we show that microglial expression of TFEB facilitates fibrillar Aβ (fAβ) degradation and reduces deposited amyloid plaques, which are further enhanced by deacetylation of TFEB. Using mass spectrometry analysis, we firstly confirmed acetylation as a previously unreported modification of TFEB and found that SIRT1 directly interacted with and deacetylated TFEB at lysine residue 116. Subsequently, SIRT1 overexpression enhanced lysosomal function and fAβ degradation by upregulating transcriptional levels of TFEB downstream targets, which could be inhibited when TFEB was knocked down. Furthermore, overexpression of deacetylated TFEB at K116R mutant in microglia accelerated intracellular fAβ degradation by stimulating lysosomal biogenesis and greatly reduced the deposited amyloid plaques in the brain slices of APP/PS1 transgenic mice. Our findings reveal that deacetylation of TFEB could regulate lysosomal biogenesis and fAβ degradation, making microglial activation of TFEB a possible strategy for attenuating amyloid plaque deposition in AD.
Alzheimer Disease ; metabolism ; pathology ; Amyloid beta-Peptides ; metabolism ; Amyloid beta-Protein Precursor ; genetics ; metabolism ; Animals ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; chemistry ; genetics ; metabolism ; Brain ; metabolism ; Cells, Cultured ; Chloride Channels ; genetics ; metabolism ; Disease Models, Animal ; HEK293 Cells ; Humans ; Lysosomes ; genetics ; metabolism ; Mice ; Mice, Transgenic ; Microglia ; cytology ; metabolism ; Mutagenesis, Site-Directed ; Peptides ; analysis ; chemistry ; Protein Binding ; RNA Interference ; Sirtuin 1 ; antagonists & inhibitors ; genetics ; metabolism

Objective To investigate the change of peripheral blood lymphocyte immunophenotype in the patients with malignant carcinoma and its clinical significance .Methods The flow cytometry was adopted to detect the expression levels of peripheral blood T lymphocyte subpopulations ,CD19+ cells ,CD3-CD56+ cells ,CD3+CD56+ cells in 167 patients with malignant carcinoma (inclu‐ding respiratory tract cancers ,gastrointestinal cancers and reproductive system cancer) and 170 normal controls as the healthy con‐trol group .The contrastive analysis was performed .Furthermore the analysis was conducted according to whether having tumor me‐tastasis and chemotherapeutic cycles .Results The total absolute value of the lymphocytes and the proportion of CD3+CD4+ cells in the patients with malignant carcinoma were significantly lower than those in the healthy control group (P<0 .05) ,but the propor‐tions of CD3+CD8+ ,CD3+CD56+ and CD3-CD56+ cells were significantly higher than those in the healthy control group (P<0 .05) .The proportion of CD3+CD4+ cells in the patients with tumor metastasis were significantly lower than those in the ones without tumor metastasis ;but the proportions of CD3+ CD8+ ,CD3-CD56+ and CD3+ CD56+ cells were significantly higher than those in the ones without tumor metastasis ,the differences were statistically significant(P<0 .05) .In the chemotherapy treatment , the proportions of CD3+CD4+ ,CD3+CD56+ and CD3-CD56+ cells were gradually increased with the increase of chemotherapeutic cycles ,but the proportion of CD3+CD8+ cells was gradually reduced .Conclusion In the process of development ,progression and treatment of tumor ,the cellular immunity function plays a dominant role ,which could be used as one of indexes for clinically moni‐toring the cellular immune function in the patients with tumor .

Objective To increase the recognition of the clinicians and laboratorians to hereditary spherocytosis for reducing the misdiagnosis and missed diagnosis ,and improving the diagnostic level .Methods The data in 4 cases of definitely diagnosed heredi‐tary spherocytosis in our hospital were retrospectively analyzed ,and the misdiagnosis reasons were analyzed and summarized .Results In 4 cases ,2 cases were misdiagnosed as Mediterranean anemia ,1 case was misdiagnosed as hemolytic anemia and 1 case as hemo‐globin disease .Conclusion Correctly mastering the key points of the diagnosis ,strengthening the communication between the labo‐ratorians and clinicians ,paying attention to the red blood cells morphologic examination under microscope and high level diagnosis a‐bility of blood cell morphology are the important means to reduce the misdiagnosis and missed diagnosis and improve the diagnostic accuracy rate .

BACKGROUND:The biological function of human periodontal ligament stem cells is a hot area of research in the treatment of periodontal disease. Human periodontal ligament cells are one of the end cells derived from human periodontal ligament stem cells;meanwhile, it can also provide supports to the development of human periodontal ligament stem cells. However, few studies are reported about the difference of biological characteristics between human periodontal ligament stem cells and human periodontal ligament cells. OBJECTIVE:To compare the differences of biological characteristics between human periodontal ligament stem cells and human periodontal ligament cells. METHODS:The human periodontal ligament stem cells and human periodontal ligament cells were isolated and purified using tissue explant method and cellclone method, respectively, and then were observed under light microscope to compare the differences of morphology. cellproliferation curves of human periodontal ligament stem cells and human periodontal ligament cells were drawn respectively with cellcounting kit 8 assay. Flow cytometry analysis was used to detect their cellcircles and their surface markers expressions. The alkaline phosphatase gene, proliferating cellnuclear antigen gene and Scleraxis gene of human periodontal ligament stem cells and human periodontal ligament cells were detected by Real-time PCR assay.RESULTS AND CONCLUSION:The human periodontal ligament stem cells and human periodontal ligament cells showed a notable difference in morphology under the light microscope observation. During the first 5 days, the cellproliferation curve of human periodontal ligament stem cells was lower than that of human periodontal ligament cells, but 5 days later, the curve of human periodontal ligament stem cells was significantly higher than that of human periodontal ligament cells. The cellcircles of human periodontal ligament stem cells and human periodontal ligament cells were 41.1%and 23.9%, respectively. The surface markers of human periodontal ligament stem cells and human periodontal ligament cells were similar, but their expression rates had significant difference. The expressions of alkaline phosphatase gene, proliferating cellnuclear antigen gene and Scleraxis gene of human periodontal ligament stem cells were significantly higher than those of human periodontal ligament cells. The above results suggest that human periodontal ligament stem cells have much stronger potential ability than human periodontal ligament cells in osteogensis and cellproliferation.

Objective:To investigate the adjuvant effect of initial periodontal therapy by observing the eradiation rate of gastric helicobacter pylori( Hp) in the patients with gastric ulcer treated by the therapy. Methods:Totally 88 patients with stomach ulcer and chronic periodontitis were enrolled and randomly divided into two groups. The control group was treated by triple therapy,the experimental group was treated by initial periodontal therapy additionally with the treatment course of one month. The changes of Hp in stomach and periodontium were observed and the eradiation rate of Hp was compared between the two groups respectively after 1-, 3-, 6-and 12-month treatment. Results:After 3, 6 and 12 months, the eradiation rate in the experimental group was significantly higher than that in the other group (P < 0. 05). Conclusion: Initial periodontal therapy combined with triple therapy can obviously improve the eradication rate of gastric Hp in the patients with gastric ulcer and chronic periodontitis.

OBJECTIVEThis study aims to investigate the feasibility of tongue reconstruction by a rectus abdominis musculoperitoneal flap with neurovascular pedicled in a canine model.

METHODSTwelve Beagle dogs were enrolled to the experiment. The animals were randomly divided into thee groups, two of which (group A and B) had nerve anastomosis. The left sides were experimental sides, whereas the right sides were control sides. Twelve weeks after operation, electrophysiological test was performed to detect hypoglossal nerve latency amplitude and conduction velocity as well as to evaluate the reinnervation of the rectus abdominis musculoperitoneal flap.

RESULTSAmong the 12 Beagle dogs, nine animal tongue reconstruction models by rectus abdominis musculoperitoneal flap with neurovascular pedicled were successful, whereas one male Beagle dog died from ventral hemia 3 d after the operation, two female rectus peritoneal flaps were abandoned because their arterial anatomy differed from the male, which was not ideal. Hypoglossal nerve conduction velocity of group A and B were restored to the normal side of the 40%, 30%.

CONCLUSIONAnimal models of tongue reconstruction can be established by a rectus abdominis musculoperitoneal flap with neurovascular pedicled in Beagle dogs. Denervated rectus abdominis musculoperitoneal flap can regain hypoglossal nerve innervation. Hypoglossal nerve functions partly recover.

BACKGROUND:Periodontal tissue engineering technology provides new ideas and new ways for periodontitis-induced bone defect repair. OBJECTIVE:To develop a culture model for the periodontal ligament cells of miniature swine, which was constructed with hydroxyapatite, to investigate the biocompatibility with hydroxyapatite. METHODS:Periodontal ligament cells from miniature swine were harvested by using tissue explant method. Immunofluorescence was used to detect the expression of stromal cel antigen 1 in the periodontal ligament cells of miniature swine. The third passage cells were co-cultured with a three-dimension hydroxyapatite scaffold, and the biological characteristics of the cells were observed under a scanning electron microscope at days 1, 3, 7 of co-culture. RESULTS AND CONCLUSION:The pirmary miniature swine periodontal ligament cells grew wel , and they were positive for stromal cel antigen 1. Under the scanning electron microscope, the periodontal ligament cells of miniature swine grew wel on the hydroxyapatite scaffold at days 1, 3, 7 of co-culture. These prove that the miniature swine periodontal ligament cells, which can be separated using tissue explant method and cultured successful y in vitro, can grow wel on the hydroxyapatite scaffold.