Advanced MRI features of intraventricular inflammatory myofibroblastic tumor: a case report.

BACKGROUND: Inflammatory myofibroblastic tumor (IMT) is a rare central nervous system (CNS) tumor. We first report a rare case of IMT in the lateral ventricle and describe the magnetic resonance imaging (MRI) findings of the tumor with an emphasis on the advanced MRI features. CASE PRESENTATION: A 49-year-old female patient with headaches and blurred vision for 2 months. Brain MRI revealed a well-circumscribed, lobulated mass occupying the left lateral ventricle trigone, with marked perilesional brain edema. The tumor showed heterogeneous significant hyperintensity on T2-weighted imaging (T2WI) and hypointensity on T1-weighted imaging (T1WI). After the administration of gadolinium, the mass exhibited marked contrast enhancement and the halo sign was observed. On advanced MRI, the lesion showed decreased perfusion on perfusion MRI and reduced diffusion on diffusion-weighted imaging (DWI). On susceptibility-weighted imaging (SWI), there was a punctate low signal intensity in the tumor. The patient underwent surgical resection of the mass and a pathological examination confirmed the lesion to be an inflammatory myofibroblastic tumor with negative expression of anaplastic lymphoma kinase (ALK). This patient had remained healthy without evidence of recurrence during a 20-month follow-up. CONCLUSIONS: On MRI, marked perilesional brain edema, significant hyperintensity on T2WI, hypoperfusion on perfusion MRI but with an obvious enhancement, no diffusion restriction on DWI, and halo sign may be the characteristic findings of intraventricular IMT. The advanced MRI characteristics could provide abundant information to reflect the histological features and physiological metabolic characteristics of the tumor.

Genome-wide analysis of WRKY transcription factors in Aquilaria sinensis (Lour.) Gilg.

The WRKY proteins are a superfamily of transcription factor that regulate diverse developmental and physiological processes in plants. Completion of the whole-genome sequencing of Aquilaria sinensis allowed us to perform a genome-wide investigation for WRKY proteins. Here, we predicted 70 WRKY genes from the A. sinensis genome and undertaken a comprehensive bioinformatic analysis. Due to their diverse structural features, the 70 AsWRKY genes are classified into three main groups (group I-III), with five subgroups (IIa-IIe) in group II, except two belong to none of them. Distinct expression profiles of AsWRKYs with RNA sequencing data revealed their diverse expression patterns among different tissues and in the process of whole-tree-inducing agarwood formation. Based on the expression characteristics, we predict some AsWRKYs are pseudogenes, and some may be involved in the biosynthesis of agarwood sesquiterpenes as activators or repressors. Among the tested genes treated with MeJA and H2O2, most of them are induced by H2O2, but downregulated by MeJA, implying the complexity of their involvement in signal transduction regulation. Our results not only provide a basic platform for functional identification of WRKYs in A. sinensis but important clues for further analysis their regulation role in agarwood formation.

WRKY44 represses expression of the wound-induced sesquiterpene biosynthetic gene ASS1 in Aquilaria sinensis.

Agarwood is derived from wounds in Aquilaria trees and is widely used in traditional medicine, incense, and perfume. Sesquiterpenes are one of the main active components in agarwood and are known to be induced by wounding or injury; However, the molecular mechanisms by which wounding leads to sesquiterpene formation remain largely unknown. Agarwood sesquiterpene synthase 1 (ASS1) is one of key enzymes responsible for the biosynthesis of sesquiterpenes and is a crucial jasmonate (JA)-responsive wound-inducible synthase. However, it is not known why ASS1 is not expressed in healthy trees and how its expression is induced as a result of wounding. Here, we report that ASS1 is a wound-induced gene with a promoter in which a 242-bp region (-973 to -731bp) is identified as the core sequence for responding to wound signals. AsWRKY44 binds directly to this region and represses ASS1 promoter activity. Down-regulation or disruption of AsWRKY44 can relieve the inhibition and activate ASS1 expression. In addition, AsWRKY44 is degraded and the expression of ASS1 is significantly up-regulated in response to exogenous application of methyl jasmonate. Thus, AsWRKY44 is a crucial negative regulator of wound-induced ASS1 transcription, and is central to the mechanism of sesquiterpene biosynthesis in agarwood.

[Expression and Regulatory Effect of miR-30b on Dynamin in Cochlear Hair Cells].

OBJECTIVE: To determine the expression of miR-30b in hair cells of mice,and its regulatory effect on the target gene DNM1 and expression of Dynamin,the key protein of synaptic endocytosis in inner hair cells. METHODS: The basilar membrane of cochlear in adult C57 mice was obtained. The expression of miR-30b in the hair cells was detected by in situ hybridization. Luciferase vector was constructed and transfected into 293T cells with miR-30b. Changes in luciferase activity were measured to verify whether DNM1 was the target gene of miR-30b. Adeno-associated virus carrying miR-30b were micro-injected into cochlear via the round window membrane. mRNA expressions of DNM1 and miR-30b were detected by RT-PCR 14 days later. The expression of Dynamin was detected by Western blot. RESULTS: miR-30b expressed in the inner and outer hair cells scattered in the region of the nucleus and cytoplasm. miR-30b reduced luciferase activity from the reporter vector containing DNM11 (P<0.05),but not in its mutants. Increased expressions of miR-30b and decreased mRNA expressions of DNM1 and Dynamin were observed following transfection of AAV-miR-30b. CONCLUSION: miR-30b expresses in inner and outer hair cells,which is consistent with the morphological orientation of dynamin. miR-30b inhibits the expression of Dynamin by targeting DNM11 gene.

Preparation of poly(L-lactic acid) nanofiber scaffolds with a rough surface by phase inversion using supercritical carbon dioxide.

Phase inversion using supercritical carbon dioxide (SC-CO2) has been widely used in the development of tissue engineering scaffolds, and particular attention has been given to obtaining desired morphology without additional post-treatments. However, the main challenge of this technique is the difficulty in generating a three-dimensional (3D) nanofiber structure with a rough surface in one step. Here, a poly(L-lactic acid) (PLLA) 3D nanofiber scaffold with a rough surface is obtained via phase inversion using SC-CO2 by carefully choosing fabrication conditions and porogens. It is found that this method can effectively modulate the structure morphology, promote the crystallization process of semicrystalline polymer, and induce the formation of rough structures on the surface of nanofibers. Meanwhile, the porogen of ammonium bicarbonate (AB) can produce a 3D structure with large pores, and porogen of menthol can improve the interconnectivity between the micropores of nanofibers. A significant increase in the fiber diameter is observed as the menthol content increases. Furthermore, the menthol may affect the mutual transition between the α' and α crystals of PLLA during the phase separation process. In addition, the results of protein adsorption, cell adhesion, and proliferation assays indicate that cells tend to have higher viability on the nanofiber scaffold. This process combines the characteristic properties of SC-CO2 and the solubility of menthol to tailor the morphology of polymeric scaffolds, which may have potential applications in tissue engineering.

ZBP-89 reduces histone deacetylase 3 by degrading IkappaB in the presence of Pin1.

BACKGROUND: Histone deacetylase 3 (HDAC3) is overexpressed in cancers and its inhibition enhances anti-tumor chemotherapy. ZBP-89, a transcription factor, can induce pro-apoptotic Bak and reduce HDAC3 but the mechanism is unknown. Pin1, a molecular switch that determines the fate of phosphoproteins, is known to interact with HDAC3. The aim of this study was to investigate the mechanism how ZBP-89 downregulated HDAC3. METHODS: In this study, liver cells, Pin1-knockout Pin1(-/-) and Pin1 wild-typed Pin(+/+) cells were used to explore how ZBP-89 reduced HDAC3. The overexpression of ZBP-89 was achieved by infecting cells with Ad-ZBP-89, an adenoviral construct containing ZBP-89 gene. The role of NF-κB was determined using CAY10576, MG132 and SN50, the former two being inhibitors of IκB degradation and SN50 being an inhibitor of p65/p50 translocation. A xenograft tumor model was used to confirm the in vitro data. RESULTS: ZBP-89 reduced HDAC3, and it could form a complex with IκB and induce IκB phosphorylation to inhibit IκB. Furthermore, ZBP-89-mediated HDAC3 reduction was suppressed by IκB degradation inhibitors CAY10576 and MG132 but not by p65/p50 translocation inhibitor SN50, indicating that IκB decrease rather than the elevated activity of NF-κB contributed to HDAC3 reduction. ZBP-89-mediated HDAC3 or IκB reduction was significantly less obvious in Pin1(-/-) cells compared with Pin1(+/+) cells. In Ad-ZBP-89-infected Pin1(+/+) cancer cells, Pin1 siRNA increased HDAC3 but decreased Bak, compared with cells without ZBP-89 infection. These findings indicate that Pin1 participates in ZBP-89-mediated HDAC3 downregulation and Bak upregulation. The cell culture result was confirmed by in vivo mouse tumor model experiments. CONCLUSIONS: ZBP-89 attenuates HDAC3 by increasing IκB degradation. Such attenuation is independent of NF-κB activity but partially depends on Pin1. The novel pathway identified may help generate new anti-cancer strategy by targeting HDAC3 and its related molecules.

Bone mineral density change in systemic lupus erythematosus: a 5-year followup study.

OBJECTIVE: To determine changes of bone mineral density (BMD) over a 5-year period in a cohort of female patients with systemic lupus erythematosus (SLE) and to identify factors predictive of BMD loss. METHODS: Our longitudinal study involved 125 female patients with SLE with a mean (SD) age of 46.5 years (10.1) and a median disease duration of 10.4 years. Demographics and clinical data were collected and BMD at the femoral neck, total hip, and lumbar spine (L1-4) was performed by using dual-energy x-ray absorptiometry at baseline and followup. RESULTS: Average percentage changes of BMD over a mean followup of 5 years were -2.41% at the femoral neck, -1.63% at the total hip, and -0.62% at the lumbar spine, with significant changes at both the femoral neck (p < 0.0001) and total hip (p < 0.0005), but not at the lumbar spine (p = 0.128). Disease flare, new organ damage, and use of glucocorticoids during followup were significantly associated with larger decreases in BMD. BMD loss was arrested at the femoral neck and BMD increased at the total hip and lumbar spine in patients receiving antiosteoporosis therapy. In multivariate analyses, use of antiosteoporosis therapy was independently associated with increased BMD at any site and new organ damage was an independent predictor of BMD loss at the femoral neck. CONCLUSION: Significant BMD loss at the hip over a period of 5 years was found in patients with SLE. Disease activity, disease damage, and use of glucocorticoids are the disease-specific variables that contribute to bone loss in SLE.

[Computer aided design and computer aided manufacture of template for crown lengthening].

PURPOSE: To introduce a method applied in computer aided design and computer aided manufacture (CAD-CAM) of template for crown lengthening. METHODS: Point cloud data of dental stone model of the patient in the plan of crown lengthening surgery was obtained by laser scanning. The following processes were carried out, constructing 3-D curve of the gingiva, drawing template outline on the triangle mesh model, shelling it for 3-D model of template, and transferring the data to rapid prototyping equipment for manufacture. RESULTS: 3-D model of the template was preliminarily accomplished. The resin template was manufactured with rapid prototyping equipment. The fitness between resin template and plaster model was good. CONCLUSIONS: This method, as an integrated procedure including data acquisition, 3-D computer modeling and fabrication by rapid prototyping, is feasible to implement CAD-CAM of template for crown lengthening.

Structure and strength of the distal radius in female patients with rheumatoid arthritis: a case-control study.

The purpose of this work was to investigate the volumetric bone mineral density (vBMD), bone microstructure, and mechanical indices of the distal radius in female patients with rheumatoid arthritis (RA). We report a cross-sectional study of 66 middle-aged female RA patients and 66 age-matched healthy females. Areal BMD (aBMD) of the hip, lumbar spine, and distal radius was measured by dual-energy X-ray absorptiometry (DXA). High-resolution peripheral quantitative computed tomography (HR-pQCT) was performed at the distal radius, yielding vBMD, bone microstructure, and mechanical indices. Cortical and trabecular vBMD were 3.5% and 10.7% lower, respectively, in RA patients than controls, despite comparable aBMD. Trabecular microstructural indices were -5.7% to -23.1% inferior, respectively, in RA patients compared to controls, with significant differences in trabecular bone volume fraction, separation, inhomogeneity, and structural model index. Cortical porosity volume and percentage were 128% and 93% higher, respectively, in RA patients, with stress being distributed more unevenly. Fourteen RA patients had exaggerated periosteal bone apposition primarily affecting the ulnovolar aspect of the distal radius. These particular patients were more likely to have chronic and severe disease and coexisting wrist deformity. The majority of the differences in density and microstructure between RA patients and controls did not depend on menstrual status. Recent exposure to glucocorticoids did not significantly affect bone density and microstructure. HR-pQCT provides new insight into inflammation-associated bone fragility in RA. It detects differences in vBMD, bone microstructure, and mechanical indices that are not captured by DXA. At the distal radius, deterioration in density and microstructure in RA patients involved both cortical and trabecular compartments. Excessive bone resorption appears to affect cortical more than trabecular bone at distal radius, particularly manifested as increased cortical porosity. Ulnovolar periosteal apposition of the distal radius is a feature of chronic, severe RA with wrist deformity.

Increased organ damage associated with deterioration in volumetric bone density and bone microarchitecture in patients with systemic lupus erythematosus on longterm glucocorticoid therapy.

OBJECTIVE: To evaluate bone quality in patients with systemic lupus erythematosus (SLE) who were undergoing longterm glucocorticoid (GC) therapy, and to focus on the correlation between bone quality and organ damage. METHODS: Seventy-eight female patients with SLE and organ damage taking longterm GC, and 72 age-matched SLE patients without damage taking longterm GC were recruited for study. Clinical variables of interest included disease activity, cumulative organ damage (by Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index; SDI), major organ involvement (musculoskeletal damage and neuropsychiatric damage, etc.), and use of medication. Areal bone mineral density (aBMD) was measured by dual-energy X-ray absorptiometry. Bone geometry, volumetric BMD (vBMD), microarchitecture, and biomechanical properties were measured by high-resolution peripheral quantitative computed tomography (HR-pQCT). RESULTS: Patients were mean age of 45 years (SD 10) and 54% were postmenopausal. The median SDI score of the cohort was 1 (interquartile range 1-2, range 1-5). Compared with patients without damage, the prevalence of osteopenia at either total hip or lumbar spine was significantly higher, and there were trends of deterioration of bone geometry, vBMD, microarchitecture, and biomechanical properties in patients with organ damage. Potential risk factors for bone quality in patients with damage were screened by univariate analysis. During multiple regression analysis, SDI was the only clinical variable consistently associated with deterioration of vBMD and microarchitecture. CONCLUSION: Cumulative organ damage consistently correlated with deterioration of vBMD and bone microarchitecture in SLE patients with damage on longterm GC therapy. HR-pQCT provides an insight into the underlying mechanism of bone loss in SLE.

[17-ß estradiol promotes the expression of interleukin-6 in human periodontal ligament cells infected with Porphyromonas gingivalis].

OBJECTIVE: To investigate the effects of 17-ß estradiol (E(2)) and Porphyromonas gingivalis (Pg) W83 on the expression of interleukin (IL)-6 and IL-8 in human periodontal ligament cells (hPDLC). METHODS: Primary cultures of hPDLC were established and the cells of passage four were treated with 10(-10) mol/L E(2), 10(-7) mol/L E(2) or PgW83 individually or E(2) combined with PgW83. The expression levels of IL-6 and IL-8 protein at 12 h and 24 h were measured with enzyme-linked immunosorbent assay and the levels of mRNA at 24 h were detected with real-time reverse transcriptase polymerase chain reaction. RESULTS: The expression level of IL-6 reached (2482.88 ± 26.53) ng/L in hPDLC treated with Pg at multiplicity of infection (MOI) of 100 for 24 h, which was significantly higher than that in hPDLC treated with Pg at MOI of 10:1 [(734.09 ± 87.90) ng/L, P = 0.000], the controls [(425.8 ± 77.25) ng/L, P = 0.000] and that in hPDLC treated with Pg at MOI of 100 for 12 h [(1157.50 ± 234.65) ng/L, P = 0.000]. The expression level of IL-8 reached (4965.81 ± 1072.55) ng/L in hPDLC treated with Pg at MOI of 100 for 24 h, which was significantly higher than that in hPDLC treated with Pg at MOI of 10 [(803.51 ± 162.08) ng/L, P = 0.007], the controls [(400.75 ± 2.27) ng/L, P = 0.005] and that in hPDLC treated with Pg at MOI of 100 for 12 h [(1431.12 ± 82.78) ng/L, P = 0.001]. E(2) did not show remarkable effect on the expressions of IL-6 and IL-8. E(2) combined with Pg (MOI = 100:1) significantly promoted the expression levels of IL-6 at 24 h while did not influence those of IL-8. The relative mRNA level of IL-6 in hPDLC treated with 10(-10) mol/L E(2) or 10(-7) mol/L E(2) combined with Pg were 0.49 ± 0.15 (P = 0.021)and 0.53 ± 0.16 (P = 0.036) individually, which were significantly higher than that treated with Pg alone, 0.19 ± 0.06. The protein level of IL-6 in hPDLC treated with 10(-10) mol/L E(2) or 10(-7) mol/L E(2) combined with Pg were (5512.66 ± 1022.07) ng/L (P = 0.012) and (6988.78 ± 2279.13) ng/L (P = 0.000) individually, which were significantly higher than that treated with Pg alone, (3138.46 ± 183.72) ng/L. CONCLUSIONS: PgW83 significantly increased the expression levels of IL-6 and IL-8 in hPDLC in a dose-and time-dependent manner. Without the infection of periodontal pathogens, estrogen may exert no effect on the expression of IL-6 and IL-8 while it may promote the expression of IL-6 in hPDLC when combined with Pg, which may in turn promote the process of periodontal inflammation.

[Effect of periodontal surgery on the subgingival periodontal microbiota in artificial class III furcation defects: an experimental study in monkeys].

OBJECTIVE: This study was to longitudinally evaluate the change of prevalence of five periodontal putative pathogens in the subgingival plaque of artificial class III furcation defects at the three time-points, including before the establishment of furcation defects, before and 6 months after periodontal surgery. METHODS: Eighteen chronic infected class III FI defects were created at the mandibular first molars, second molars and second premolars of three adult male Macaca fascicularis. The samples of subgingival plaque were obtained from the subgingival area of furcation defects in buccal and lingual sites before the establishment of furcation defects, before and 6 months after periodontal surgery. 36 samples were obtained at one time-points. Five periodontal putative pathogens, including Porphyromonas gingivalis (Pg), Tannerella forsythensis (Tf), Treponema dinticola (Td), Actinobacillus actinomycetemcomitans (Aa) and Fusobacterium nucleatum (Fn), were detected with 16SrRNA based PCR. RESULTS: 1. The prevalence of Pg, Tf, Td and Fn was gradually increased, from 58.3% to 69.4% to 88.9%, 47,2% to 69.4% to 83.3%, 13.9% to 36.1% to 61.1% (P<0.01), and 69.4% to 91.7% to 91.7% (P<0.05), respectively during the experimental period. The prevalence of Fn was higher than Pg, Tf and Td. The prevalence of Aa was the lowest and no obvious difference among the three samplings(from 25.9% to 13.9% to 33.3%)was detected. 2. The prevalence of more than 3 species simultaneously detected was increased from 38.9% to 61.1% to 83.3% (P <0.01). The red complex (Pg + Tf + Td) was detected from 8.3% to 27.8% to 44.4% (P<0.01) at the different time point. 3. The combined detection frequency of red complex in the inflammatory sites (87.5%), which were histologically defined as inflammatory cells infiltrated in furcation area 6 months post-surgery, and the same sites pre-surgery (62.5%) was more than that in pre-creation of furcation defects (P<0.01). But there were no significant differences compared to that in non inflammatory area (60.0%, 40.0%), respectively. CONCLUSION: The prevalence of periodontal pathogenic bacteria correlated with the severity of local inflammation. The increase of coexistent rate of red complex at the second and third sampling times suggests that the red complex play important role in the pathogenesis of periodontitis. Fn may be a resident bacteria in the subgingival plaque, play a bridge role on the biofilm formation and maturation. Aa may not be a major causative bacteria in the clinical periodontitis.

Study of the adsorption reactions of thiophene on Cu(I)/HY-Al2O3 by Fourier transform infrared and temperature-programmed desorption: adsorption, desorption, and sorbent regeneration mechanisms.

This work mainly involved the investigation of the adsorption of thiophene on Cu(I)-supported HY-Al(2)O(3). It demonstrated a high sulfur capacity of 10 mg sulfur/g sorbent when the HY/Al(2)O(3) mass ratio was 3, loaded with 12% copper, calcined at 550 °C, and tested at ambient temperature. In situ Fourier transform infrared (FTIR) and temperature-programmed desorption (TPD) results indicated that the adsorption mechanisms on Cu(I)/HY-Al(2)O(3) primarily were π-complexation and sulfur-adsorbent (S-M; σ) bonds. Pyridine-FTIR showed the total weak Lewis acid contribution to the Cu(I)/HY-Al(2)O(3) adsorption desulfurization performance.

[The inhibitory effect of calcitriol on the proliferation of hPDLCs populations of different vitamin D receptor Fok I genotypes].

PURPOSE: The study was designed to investigate the effect of calcitriol [1,25-(OH)(2)D(3),VD(3)] on the proliferation of human periodontal ligament cells (hPDLCs) and to analyze the influence of vitamin D receptor Fok I polymorphism (rs2228570, T/C) on the effect. METHODS: Primary cultures of hPDLCs populations from individual donors were established. Restriction fragment length polymorphism method was used to detect the genotypes of the Fok I site. The fourth passage hPDLCs populations of FF, Ff and ff genotypes were respectively treated with 0.1% ethyl alcohol (control) or 10(-14)-10(-8) mol/L VD3 for 1, 3, 5 and 7 days. MTT method was performed to evaluate the proliferation of hPDLCs. The data was analyzed by one-way ANOVA and LSD Post Hoc test using SPSS 11.5 software package. RESULTS: VD3 inhibited the proliferation of hPDLCs populations of three genotypes in a time- and concentration-dependent manner. 10(-8) mol/L VD(3) exerted the most powerful inhibition compared with controls and other concentrations and the highest inhibition occurred on day 5 or day 7. Compared with the corresponding controls, the lowest concentrations of VD3 which exerted statistically significant inhibitory effect on the proliferation of hPDLCs populations were listed as follows: 10(-12) mol/L for the hPDLCs population of FF genotype (P<0.01), 10(-10) for Ff genotype (P<0.01) and 10(-8) mol/L for ff genotype (P<0.01). The highest proliferation inhibitory rate exerted by 10(-8) mol/L VD(3) on the ff population was 16.93%,which was significantly lower than FF (31.20%, P<0.05) and Ff (29.68%, P<0.05). On day 3 and day 5, 10(-8) mol/L VD(3) exhibited the strongest suppression on the proliferation of hPDLCs population of FF genotype, followed by Ff and ff genotypes (P<0.01). CONCLUSION: VD(3) can significantly inhibit the proliferation of hPDLCs, which may be affected by VDR Fok I polymorphism.

[Correlation analysis between plasma levels of 25-hydroxy vitamin D3 and osteocalcin in patients with aggressive periodontitis].

OBJECTIVE: To explore the relationship between plasmatic 25-hydroxy vitamin D3 (25OHD3) level and plasmatic osteocalcin level in patients with aggressive periodontitis (AgP). METHODS: Thirty four AgP patients and 29 healthy controls were included in this study. 25OHD3 and osteocalcin levels in plasma were measured using commercially available radioimmunoassay kits. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured using a standard hospital analytical technique. RESULTS: Plasmatic 25OHD3 level was significantly higher in AgP patients than that in healthy controls (8.65 microg/L vs 3.10 microg/L; P<0.01). Osteocalcin level was also significantly higher in AgP patients than that in healthy subjects (1.0 microg/L vs 0.8 microg/L; P=0.028). AST level was significantly lower in AgP patients than that in healthy controls(20.0 U/L vs 23.0 U/L). No correlations between the plasmatic levels of 25OHD3 and osteocalcin were detected in AgP patients or in healthy controls (r=0.271, P=0.12; r=-0.356, P=0.58). CONCLUSION: Plasmatic 25OHD3 and osteocalcin concentrations were not correlated but might be influenced by AgP.

[Clinical and radiographic evaluation of class III furcation defects in the treatment using autogenous periodontal ligament cells with or without enamel matrix derivatives].

OBJECTIVE: To evaluate whether the use of periodontal ligament cells(PDLC) with or without enamel matrix derivatives(EMD) influences periodontal tissue repair in class III furcation defects. METHODS: Three adult male Macaca fascicularis monkeys were used. Class III furcation defects were created at the mandibular second pre-molars, first molars and second molars. The autogenous PDLC were cultured in vitro with Bio Oss Collagen. Six furcation defects in the 3 monkeys were divided as follows, Group A (one second molar): PDLC/Bio Oss Collagen+EMD; Group B (another second molar): Bio Oss Collagen+EMD; Group C (one first molar): PDLC/ BiojOss Collagen; Group D(another first molar): Bio Oss Collagen; Group E (one second pre-molar): EMD; Group F (another second pre-molar): the empty control. All sites (including buccal and lingual side) were covered with collagen membranes. The monkeys were euthanized at the end of 6 months. The periodontal depth (PD) and clinical attachment level (AL) at the buccal and lingual furcation defects were examined before and 6 months after the implantation. X-rays were also obtained at the same time points. RESULTS: PD and AL were decreased in most sites, the reductions in groups E and F (the second pre- molars) were the most significant, and then in turn were in groups A, C, B and D. The repaired alveolar bones were almost full of furcation area in the second pre-molars, and the relatively clear lamina dura was also found. The alveolar bones in the other sizes only had a little repair, and the obviously low density area still remained in the coronal of the defects. CONCLUSION: The results of this study indicate that class III furcation defects can not be predictably resolved even with the combination of autogenous PDLC and EMD, although they may increase the repair of periodontal tissue in the area of class III furcation defects separately. The sizes of furcation defects and the coverage of gingival flap would influence the outcome of the treatment of class III furcation defects.

[Effects of calcitriol on the expression of vitamin D receptor, RANKL and osteoprotegerin in human periodontal ligament cells].

OBJECTIVE: To study the effects of 1, 25-dihydroxyvitamin D(3) (VD(3)) on the expression of vitamin D receptor (VDR), receptor activator of nuclear factor-kappa B ligand (RANKL) and osteoprotegerin (OPG) in human periodontal ligament cells (hPDLC) populations and to analyze the potential mechanisms. METHODS: Twelve hPDLC populations were primarily established from 12 donors individually. Two samples of each hPDLC population of passage three were treated respectively with 10(-8) mol/L VD(3) (V D(3) group) or 0.1% absolute ethyl alcohol as controls (control group). Six days later, the mRNA expression levels of VDR, RANKL and OPG in the samples were determined with real-time quantitative RT-PCR. The DNA base sequences upstream to the transcription start site of RANKL gene were also analyzed. RESULTS: Compared with the control group, the mRNA expression level of VDR increased significantly in the VD(3) group (P = 0.003), averagely (3.04 +/- 1.06) times of that in the control group; the mRNA expression level of RANKL was also up-regulated by VD(3) (P = 0.001), 9.82 (0.75-119.18) times of that in the control group; the OPG expression level was (94.48 +/- 39.15)% of the controls (P = 0.136); OPG/RANKL ratio was down-regulated in the VD(3) group to averagely 10.36% (1.01%-138.00%) of the controls (P = 0.003). No mutation was found in the DNA fragments upstream to the transcription start site in the RANKL gene and the genotypes of the polymorphism at -1832 (rs7984870, C/G) were not shown to be significantly related to the RANKL mRNA expression level. CONCLUSIONS: In hPDLC, VD(3) can significantly increase the mRNA expression level of VDR; VD(3) can increase RANKL mRNA expression level to decrease OPG/RANKL ratio, but it has little effect on OPG mRNA expression. The big differences of the RANKL mRNA regulation in response to VD(3) treatment among hPDLC populations may not be associated with the DNA sequences upstream to the transcription start site in the RANKL gene.

[Effect of 17-beta estradiol on the expression of receptor activator of nuclear factor kappaB ligand and osteoprotegerin of human periodontal ligament cells during their osteogenic differentiation].

OBJECTIVE: To investigate the effect of 17-beta estradiol (E(2))on the expression of receptor activator of nuclear factor kappaB ligand(RANKL) and osteoprotegerin(OPG) in human periodontal ligament cells( hPDLCs) during their osteogenic differentiation. METHODS: hPDLCs of passage four were treated with 0.1% absolute ethyl alcohol (control), 10(-10) mol/L(physiological dose) or 10(-7) mol/L(high dose) E(2) in DMEM supplemented with 10% FBS, 10 mmol/L beta-glycerophosphate and 5 microg/mL ascorbic acid for 24 hours, 48 hours and 72 hours. Levels of RANKL and OPG mRNA were detected with real-time quantitative RT-PCR. RESULTS: High dose E(2) induced a time-dependent decrease of RANKL mRNA and expression levels were 31.0%, 23.6% and 15.4% of the controls at 24 h, 48 h and 72 h respectively while expression levels with physiological dose E(2) were 28.4%, 87.3% and 22.6% of the controls respectively. On the other hand, E(2) increased OPG mRNA expression levels of hPDLCs which reached the maximum at 72 h with physiological dose E(2), 210.1% of the controls and with high dose E(2), 171.3% of the controls. CONCLUSION: These results suggest that estrogen may exert anti-resorptive effects on alveolar bone by regulating the expression of RANKL and OPG in hPDLCs during their osteogenic differentiation.