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1.
BMC Oral Health ; 24(1): 632, 2024 May 29.
Article in English | MEDLINE | ID: mdl-38811912

ABSTRACT

BACKGROUND: Anxiety is common preceding tooth extraction; hence, it is crucial to identify patients with dental anxiety (DA) and to manage DA. This study assessed the level of DA and influencing factors in tooth extraction patients in a dental hospital in China and changes in their blood pressure (BP) and heart rate (HR) during the tooth-extraction procedure. METHODS: The study was a cohort study. The Dental Anxiety Scale (DAS) was used to assess the level of DA of 120 patients before tooth extraction. A Demographics and Oral Health Self-Assessment Form was used to assess factors influencing DA. The correlations of DAS scores with HR and BP were measured. The effects of local anesthesia and general anesthesia on HR and BP were also compared using a Datex-Ohmeda anesthesia monitor to detect HR and BP continuously before and after anesthesia. Independent sample t-tests, OLS multiple regression model and one-way analysis of variance were applied to analysis the results. RESULTS: Based on the DAS score, 12.5% of the participants were identified as suffering from DA. DA was related to age, gender, and the self-assessment of oral health. The DAS score was correlated with increased BP (P < 0.05). BP showed an overall upward trend after local anesthesia, while it was generally stable after general anesthesia. The systolic BP at 4 and 5 min and the HR at 2 and 4 min increased remarkably (P < 0.05) after local anesthesia compared with those before anesthesia. The HR and BP of patients under local anesthesia were generally higher than those of patients under general anesthesia were during the operation. CONCLUSIONS: The prevalence of DA in adults was 12.5% in this study population. DA was related to gender, age, and the self-assessment of oral health. The score of DAS was correlated with BP. Compare to local anesthesia, general anesthesia can make the vital signs of tooth extraction patients more stable.


Subject(s)
Anesthesia, Dental , Anesthesia, General , Blood Pressure , Dental Anxiety , Heart Rate , Tooth Extraction , Humans , Female , Male , Heart Rate/physiology , Anesthesia, Dental/methods , Adult , Blood Pressure/physiology , Middle Aged , Anesthesia, Local , Cohort Studies , Sex Factors , Age Factors , Young Adult , Vital Signs , Aged
2.
BMC Oral Health ; 24(1): 466, 2024 Apr 17.
Article in English | MEDLINE | ID: mdl-38632582

ABSTRACT

OBJECTIVE: To investigate the association of leisure-time physical activity and serum cotinine levels with the risk of periodontitis in the general population and to further analyze the interaction between leisure-time physical activity and serum cotinine levels on the risk of periodontitis. METHODS: This was a cross-sectional study, extracting data from 9605 (56.19%) participants in the National Health and Nutrition Examination Survey (NHANES) database from 2009 to 2014, and analyzing the relationship and interaction effects of serum cotinine level, leisure time physical activity, and risk of periodontitis by weighted univariate logistic modeling; Effect sizes were determined using ratio of ratios (OR), 95% confidence intervals (95% CI). RESULTS: 5,397 (56.19%) of 9,605 participants had periodontitis; an increased risk of periodontitis was found in those in the leisure time physical activity intensity < 750 MET × min/week group (OR = 1.44, 95% CI: 1.17-1.78). Serum cotinine levels ≥ 0.05 ng/ml were associated with an increased risk of periodontitis (OR = 1.99, 95% CI: 1.69-2.33). The group with low leisure physical activity and serum cotinine levels ≥ 0.05 ng/ml had an increased risk of periodontitis compared to the group with high leisure physical activity and serum cotinine levels < 0.05 ng/ml (OR = 2.48, 95% CI: 1.88-3.27). Interaction metrics RERI = 0.90 (95% CI: 0.44-1.36) and API = 0.36 (95% CI: 0.18-0.55); CI for SI = 2.55 (95% CI: 1.03-6.28). for API 0.36. CONCLUSION: Leisure time physical activity intensity interacted with smoking exposure on periodontitis risk and may provide the general population with the opportunity to Increasing leisure-time physical activity and smoking cessation may provide recommendations for the general population.


Subject(s)
Periodontitis , Tobacco Smoke Pollution , Humans , Cotinine/analysis , Smoking/adverse effects , Tobacco Smoke Pollution/adverse effects , Nutrition Surveys , Cross-Sectional Studies , Periodontitis/epidemiology , Exercise , Leisure Activities
3.
J Biophotonics ; 16(8): e202300031, 2023 08.
Article in English | MEDLINE | ID: mdl-37074336

ABSTRACT

To alleviate the ill-posed of the inverse problem in fluorescent molecular tomography (FMT), many regularization methods based on L2 or L1 norm have been proposed. Whereas, the quality of regularization parameters affects the performance of the reconstruction algorithm. Some classical parameter selection strategies usually need initialization of parameter range and high computing costs, which is not universal in the practical application of FMT. In this paper, an universally applicable adaptive parameter selection method based on maximizing the probability of data (MPD) strategy was proposed. This strategy used maximum a posteriori (MAP) estimation and maximum likelihood (ML) estimation to establish a regularization parameters model. The stable optimal regularization parameters can be determined by multiple iterative estimates. Numerical simulations and in vivo experiments show that MPD strategy can obtain stable regularization parameters for both regularization algorithms based on L2 or L1 norm and achieve good reconstruction performance.


Subject(s)
Image Processing, Computer-Assisted , Tomography , Fluorescence , Image Processing, Computer-Assisted/methods , Phantoms, Imaging , Tomography/methods , Algorithms , Coloring Agents
4.
Biomed Opt Express ; 14(3): 1159-1177, 2023 Mar 01.
Article in English | MEDLINE | ID: mdl-36950247

ABSTRACT

Fluorescence molecular tomography (FMT) is a promising molecular imaging technique for tumor detection in the early stage. High-precision multi-target reconstructions are necessary for quantitative analysis in practical FMT applications. The existing reconstruction methods perform well in retrieving a single fluorescent target but may fail in reconstructing a multi-target, which remains an obstacle to the wider application of FMT. In this paper, a novel multi-target reconstruction strategy based on blind source separation (BSS) of surface measurement signals was proposed, which transformed the multi-target reconstruction problem into multiple single-target reconstruction problems. Firstly, by multiple points excitation, multiple groups of superimposed measurement signals conforming to the conditions of BSS were constructed. Secondly, an efficient nonnegative least-correlated component analysis with iterative volume maximization (nLCA-IVM) algorithm was applied to construct the separation matrix, and the superimposed measurement signals were separated into the measurements of each target. Thirdly, the least squares fitting method was combined with BSS to determine the number of fluorophores indirectly. Lastly, each target was reconstructed based on the extracted surface measurement signals. Numerical simulations and in vivo experiments proved that it has the ability of multi-target resolution for FMT. The encouraging results demonstrate the significant effectiveness and potential of our method for practical FMT applications.

5.
Zhejiang Da Xue Xue Bao Yi Xue Ban ; 50(2): 171-178, 2021 04 25.
Article in English | MEDLINE | ID: mdl-34137222

ABSTRACT

: To assess the () recombinant gingivalis gingipain R2 (rRgpB)-induced Ca mobilization in human gingival fibroblast (HGF) mediated by protease-activated receptor (PAR) and its downstream signal transduction pathways. : Flow cytometry was used to detect the expression of PAR in HGF. The proliferation of HGF was measured by CCK-8. The dynamic changes of intracellular Ca concentration in HGF induced by rRgpB and the blocking effect of PAR-1 antagonist were observed by laser confocal microscopy. Western blot was performed to determine the phosphorylation levels of c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK) 1/2, p38 mitogen-activated protein kinase (p38 MAPK) and p65 in HGF. : PAR-1 and PAR-3 were expressed in HGF, and the rRgpB could promote the proliferation of HGF. rRgpB caused a transient increase in [Ca], which could be completely suppressed by vorapaxar, a PAR-1 antagonist. The phosphorylation levels of JNK, ERK1/2 and p65 were significantly up-regulated after the induction of rRgpB for and (all <0.05), which was completely inhibited by vorapaxar. However, the phosphorylation level of p38 MAPK had no significant change after rRgpB stimulation. : rRgpB causes an increase in [Ca] in HGF mediated by PAR-1. JNK, ERK1/2 and nuclear factor-κB may be involved in intracellular signal transduction after PAR-1 activation.


Subject(s)
Calcium/metabolism , Fibroblasts , Signal Transduction , Humans , JNK Mitogen-Activated Protein Kinases/metabolism , MAP Kinase Signaling System , Phosphorylation , p38 Mitogen-Activated Protein Kinases/metabolism
6.
J Periodontol ; 91(4): 462-472, 2020 04.
Article in English | MEDLINE | ID: mdl-31471902

ABSTRACT

BACKGROUND: The development of platelet concentrated biomaterials has gained increasing awareness for regenerative medicine. With different protocol, derivatives such as advanced platelet-rich fibrin (A-PRF), injected platelet-rich fibrin, and concentrated growth factor (CGF) have been demonstrated effectively in preclinical and clinical studies. The aim of this study was to compare the level of growth factors releasing from A-PRF and CGF, and their clinical efficacy in the regenerative management of intrabony defects (IBDs). METHODS: Thirty-two blood samples were collected from eight healthy donors and assessed for platelet-derived growth factor-αß, vascular endothelial growth factor, bone morphogenetic protein-2, and transforming growth factor-ß1 release at indicated times. In addition, the clinical records of 45 patients (15 per group) who had undergone guided tissue regeneration (GTR) with or without A-PRF/CGF were retrieved. The probing depth (PD) and clinical attachment level (CAL) were recorded preoperatively and 6 months postoperatively. Intrabony component (IC) depth, radiographic bone level (RBL), and bone defect filling were assessed radiographically. RESULTS: A-PRF had a looser fibrin network than the CGF but presented larger amounts of growth factors with a more sustained release period. Although there was no difference in PD reduction, CAL gain, RBL height change and defect filling (%) between A-PRF and CGF group, both achieved a more favorable clinical result in IC height reduction and defect filling (%) than the control. CONCLUSIONS: A-PRF and CGF have the ability to stimulate a continual and steady release of total growth factors over a 14-day period. A-PRF and CGF show a similar effectiveness in periodontal bone regeneration with a potential benefit of improving GTR outcomes in IBD treatment.


Subject(s)
Alveolar Bone Loss/surgery , Platelet-Rich Fibrin , Guided Tissue Regeneration, Periodontal , Humans , Treatment Outcome , Vascular Endothelial Growth Factor A
7.
ACS Omega ; 4(24): 20644-20653, 2019 Dec 10.
Article in English | MEDLINE | ID: mdl-31858050

ABSTRACT

Salbutamol (SAL), one of the prohibited veterinary drugs, has been proven to be harmful to animals, but very few studies reported the underlying mechanism of actions and the effects after SAL intake. In this study, Ba-Ma minipigs were used as the animal model to demonstrate the impacts of SAL residues on blood lipid and the lung bronchial structures and the regulation of gene expression and gut microorganism population. The results showed that (1) SAL decreased the indexes of serum lipid and organ, (2) SAL widely retained in various tissues and organs, (3) the lung bronchial expanded under the influence of SAL, (4) the gene expression of growth-related ghrelin has increased, and (5) the residues of SAL affected the composition of gut microorganism population, which could be associated with the mechanism of action of SAL on pig. The findings suggest that SAL could be harmful to minipigs by altering the blood lipid, bronchial morphology, gastric mucosal gene expression, and the gut microorganism population.

8.
Eur J Oral Sci ; 127(4): 287-293, 2019 08.
Article in English | MEDLINE | ID: mdl-31175838

ABSTRACT

Arginine-specific cysteine proteinases, such as Arg-gingipain B (RgpB), mediate inflammation by activating protease-activated receptors (PARs). Arg-gingipain B is produced by Porphyromonas gingivalis, and is implicated in the causation of periodontal disease. The purpose of the present study was to observe the influence of recombinant RgpB protein (rRgpB) on PAR activation by monitoring intracellular Ca2+ ion concentration ([Ca2+]i) and inositol-1,4,5-triphosphate (IP3) levels in human gingival fibroblasts (HGFs). Our findings showed that rRgpB could cause a transient increase in [Ca2+]i. This increase in [Ca2+]i was completely suppressed by vorapaxar, a PAR-1 antagonist. Recombinant Arg-gingipain B increased the concentration of IP3, reaching a maximum at 60 s after treatment; this was completely inhibited by vorapaxar. We therefore conclude that rRgpB-induced calcium signaling in HGFs is mainly caused by PAR-1 activation. This suggests that PAR-1 activation plays a significant role in chronic inflammatory periodontal disease induced by P. gingivalis RgpB.


Subject(s)
Calcium Signaling , Fibroblasts/metabolism , Gingipain Cysteine Endopeptidases/pharmacology , Porphyromonas gingivalis/enzymology , Receptor, PAR-1/metabolism , Bacterial Proteins/pharmacology , Cells, Cultured , Fibroblasts/drug effects , Humans , Inositol 1,4,5-Trisphosphate , Lactones/pharmacology , Pyridines/pharmacology , Receptor, PAR-1/antagonists & inhibitors , Recombinant Proteins/pharmacology
9.
Sensors (Basel) ; 16(7)2016 Jul 22.
Article in English | MEDLINE | ID: mdl-27455271

ABSTRACT

The main limitation of high-temperature piezoresistive pressure sensors is the variation of output voltage with operating temperature, which seriously reduces their measurement accuracy. This paper presents a passive resistor temperature compensation technique whose parameters are calculated using differential equations. Unlike traditional experiential arithmetic, the differential equations are independent of the parameter deviation among the piezoresistors of the microelectromechanical pressure sensor and the residual stress caused by the fabrication process or a mismatch in the thermal expansion coefficients. The differential equations are solved using calibration data from uncompensated high-temperature piezoresistive pressure sensors. Tests conducted on the calibrated equipment at various temperatures and pressures show that the passive resistor temperature compensation produces a remarkable effect. Additionally, a high-temperature signal-conditioning circuit is used to improve the output sensitivity of the sensor, which can be reduced by the temperature compensation. Compared to traditional experiential arithmetic, the proposed passive resistor temperature compensation technique exhibits less temperature drift and is expected to be highly applicable for pressure measurements in harsh environments with large temperature variations.

10.
BMC Oral Health ; 15(1): 128, 2015 Oct 17.
Article in English | MEDLINE | ID: mdl-26476532

ABSTRACT

BACKGROUND: Protease-activated receptors (PARs) are G-protein-coupled receptors with an active role in mediating inflammation, pain and other functions. The oral pathogen Porphyromonas gingivalis (P. gingivalis) secretes proteases that activate PARs. The aim of this study was to elucidate the role of PARs in the pathogenesis of chronic periodontitis by expression analysis of PARs in human gingival epithelial cells (GECs) before and after P. gingivalis supernatants treatment. METHODS: GECs were isolated from healthy human gingival tissue samples. The expression of PARs in GECs was determined by reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry. The effect of P. gingivalis proteases was investigated by quantitative real-time reverse transcription polymerase chain reaction (QRT-PCR) and flow cytometry. RESULTS: PAR-1, PAR-2, and PAR-3 were expressed in GECs. PAR-4 was not found by both RT-PCR and flow cytometry. Analysis of gene expression using QRT-PCR showed an up-regulation of PAR-2 mRNA in comparison to the untreated control cells (P < 0.05). In contrast, the mRNA expressions of PAR-1 and PAR-3 were significantly down-regulated (P > 0.05) in response to P. gingivalis supernatant compared to that in unstimulated control cells. This effect was abrogated by the protease inhibitor TLCK (P < 0.05). The results of flow cytometry indicated PARs protein levels consistent with mRNA levels in the results of QRT-PCR. CONCLUSIONS: Our study shows that PAR-1, PAR-2 and PAR-3 are expressed in GECs. P. gingivalis proteases play a role in the regulation of innate immune responses in GECs. GECs use PARs to recognize P. gingivalis and mediate cell responses involved in innate immunity.


Subject(s)
Porphyromonas gingivalis , Receptors, Thrombin , Epithelial Cells , Gingiva/metabolism , Gingiva/microbiology , Humans , Immunity, Innate , Receptors, Thrombin/metabolism , Up-Regulation
11.
Zhonghua Kou Qiang Yi Xue Za Zhi ; 49(1): 21-6, 2014 Jan.
Article in Chinese | MEDLINE | ID: mdl-24697883

ABSTRACT

OBJECTIVE: To investigate the expression types of protease-activated receptors(PAR) in human gingival fibroblasts(HGF) and the functions of PAR in periodontitis. METHODS: Primary HGF were cultured.Reverse transcription PCR(RT-PCR) was used to detect the expression of PAR in HGF. Recombinant gingipain R (rRgp) was applied to HGF. The change of PAR expression on the cell surface was analyzed by real-time quantitative RT-PCR, and enzyme-linked immunosorbent assay (ELISA) was used to detect the change of the interleukin (IL)-6 production from HGF. The results of RT-PCR and ELISA were statistically analyzed using the two independent samples t-test of SPSS10.0 software. RESULTS: HGF expressed PAR-1 and PAR-3. The expression of PAR-1 and PAR-3 changed after two rRgp treatment with HGF cells. The relative expression of PAR-1 was decreased from 1.04 ± 0.31 to 0.67 ± 0.11 and 0.31 ± 0.11. The relative expression of PAR-3 was decreased from 1.01 ± 0.44 to 0.79 ± 0.13 and 0.44 ± 0.12(P < 0.05). The level of IL-6 was increased after rRgp treatment for 8 h. The control group was (18.77 ± 4.09) µg/L, the rRgp treatment groups were (179.36 ± 15.81) and (320.56 ± 26.19) µg/L respectively. CONCLUSIONS: HGF expressed PAR-1 and PAR-3 and were involved in periodontal inflammation.


Subject(s)
Gingiva/metabolism , Periodontitis/metabolism , Receptors, Proteinase-Activated/biosynthesis , Adhesins, Bacterial , Cell Membrane , Cysteine Endopeptidases , Enzyme-Linked Immunosorbent Assay , Fibroblasts , Gingipain Cysteine Endopeptidases , Humans , Interleukin-6
12.
Arch Oral Biol ; 59(4): 393-9, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24509446

ABSTRACT

OBJECTIVE: Protease-activated receptors (PARs) are a unique class of receptors which are implicated in mediating inflammation, pain and other functions. The aim of this study was to elucidate the role of PARs in the pathogenesis of chronic periodontitis by differential expression analysis of PARs in the gingival tissues of chronic periodontitis patients compared with those of healthy control individuals. DESIGN: Gingival tissue specimens were collected from chronic periodontitis patients (n=20) and control individuals (n=20). The expression of PAR-1, -2, -3 and -4 was determined in these tissues by immunohistochemistry and differential expression between the two groups was investigated by quantitative real-time reverse transcription-polymerase chain reaction analysis. RESULTS: PAR-1, -2, -3 and -4 were expressed in all gingival tissues. A significant overexpression of PAR-3 was detected in chronic periodontitis-affected tissues compared to healthy gingival tissues. However, expression of PAR-2 was decreased in periodontal lesions. CONCLUSIONS: Our study shows that PAR-1, -2, -3 and -4 are expressed in both healthy and inflamed gingival tissues. Furthermore, PAR-2 and PAR-3 may contribute to the inflammatory responses associated with chronic periodontitis.


Subject(s)
Chronic Periodontitis/metabolism , Gingiva/metabolism , Receptors, Proteinase-Activated/metabolism , Adult , Aged , Case-Control Studies , Female , Humans , Immunohistochemistry , Male , Middle Aged , Reverse Transcriptase Polymerase Chain Reaction
13.
Tissue Eng Part A ; 19(19-20): 2226-32, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23617625

ABSTRACT

This study aimed to investigate the ability of osteoclasts during bone resorption activities to regulate the differentiation and calcification of osteoblast precursor cells. The bone resorption model was established using in vitro cortical bone slices and mouse RAW264.7 cells, which were differentiated into osteoclasts by stimulation with the receptor activator of nuclear factor-κB ligand and macrophage colony-stimulating factor. Tartrate-resistant acid phosphatase (TRAP) staining, reverse transcriptase-polymerase chain reaction (RT-PCR), and scanning electron microscopy (SEM) were used to detect osteoclast differentiation. The osteoblast precursor cell line MC3T3-E1 was cultured with the bone resorption supernatant (BRS). Involvement of the phosphatidylinositol 3-kinase (PI3K)/AKT pathway in osteogenesis was evaluated by Western blotting, RT-PCR, and ELISA analysis of markers of the early (runt-related transcription factor-2 and alkaline phosphatase) and late (osteocalcin [OCN]) stages of osteogenesis, and Alizarin Red S staining of matrix mineralization. TRAP staining, RT-PCR, and SEM analysis demonstrated the successful establishment of the bone resorption model. Osteoclast BRS effectively increased the differentiation and calcification of MC3T3-E1 cells. Western blot analysis indicated that the BRS enhanced AKT and p-AKT expression levels in MC3T3-E1 cells. Following AKT2 knockdown and treatment with the PI3K/AKT pathway inhibitor LY294002, the expression of OCN in MC3T3-E1 cells was decreased (p<0.05), as was the calcification area (p<0.05). The data obtained in this study indicated that the osteoclast bone resorption medium promoted the differentiation and calcification of MC3T3-E1 cells and that the PI3K/AKT pathway played a role in this process.


Subject(s)
Osteoblasts/cytology , Osteoblasts/metabolism , Osteoclasts/cytology , Osteoclasts/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Animals , Cell Line , Mice , Osteogenesis/genetics , Osteogenesis/physiology , Reverse Transcriptase Polymerase Chain Reaction
14.
Immunology ; 130(2): 262-72, 2010 Jun.
Article in English | MEDLINE | ID: mdl-20113368

ABSTRACT

Little is known of how Toll-like receptor (TLR) ligands are processed after recognition by TLRs. This study was therefore designed to investigate how the TLR2 ligand FSL-1 is processed in macrophages after recognition by TLR2. FSL-1 was internalized into the murine macrophage cell line, RAW264.7. Both chlorpromazine and methyl-beta-cyclodextrin, which inhibit clathrin-dependent endocytosis, reduced FSL-1 uptake by RAW264.7 cells in a dose-dependent manner but nystatin, which inhibits caveolae- and lipid raft-dependent endocytosis, did not. FSL-1 was co-localized with clathrin but not with TLR2 in the cytosol of RAW264.7 cells. These results suggest that internalization of FSL-1 is clathrin dependent. In addition, FSL-1 was internalized by peritoneal macrophages from TLR2-deficient mice. FSL-1 was internalized by human embryonic kidney 293 cells transfected with CD14 or CD36 but not by the non-transfected cells. Also, knockdown of CD14 or CD36 in the transfectants reduced FSL-1 uptake. In this study, we suggest that (i) FSL-1 is internalized into macrophages via a clathrin-dependent endocytic pathway, (ii) the FSL-1 uptake by macrophages occurs irrespective of the presence of TLR2, and (iii) CD14 and CD36 are responsible for the internalization of FSL-1.


Subject(s)
CD36 Antigens/immunology , Clathrin/immunology , Diglycerides/pharmacology , Endocytosis/immunology , Lipopolysaccharide Receptors/immunology , Macrophages, Peritoneal/immunology , Oligopeptides/pharmacology , Toll-Like Receptor 2/immunology , Animals , CD36 Antigens/genetics , Cell Line , Chlorpromazine/pharmacology , Clathrin/genetics , Clathrin/metabolism , Diglycerides/immunology , Diglycerides/metabolism , Dopamine Antagonists/pharmacology , Dose-Response Relationship, Drug , Endocytosis/drug effects , Endocytosis/genetics , Humans , Ionophores/pharmacology , Lipopolysaccharide Receptors/genetics , Macrophages, Peritoneal/metabolism , Membrane Microdomains/genetics , Membrane Microdomains/immunology , Membrane Microdomains/metabolism , Mice , Mice, Knockout , Nystatin/pharmacology , Oligopeptides/immunology , Oligopeptides/metabolism , Toll-Like Receptor 2/genetics , Toll-Like Receptor 2/metabolism
15.
Zhejiang Da Xue Xue Bao Yi Xue Ban ; 37(6): 622-8, 2008 11.
Article in Chinese | MEDLINE | ID: mdl-19084961

ABSTRACT

OBJECTIVE: To determine the effect of Porphyromonas gingivalis lipopolysaccharide (LPS) on induced secretion of inflammatory cytokines by different cell lines. METHODS: LPS of P. gingivalis strain ATCC33277 (Pg-LPS) was extracted with phenol-water method, and identified by Limulus test and infrared spectrum analysis. KB, HGF-1 and THP-1 cells were treated with Pg-LPS of different concentrations and time duration, a commercial LPS of E.coli strain O111:B4 (E-LPS) was used as the control. TNF-alpha, IL-1beta, IL-6 and IL-8 levels in culture supernatants were measured by quantitative ELISA. RESULT: The minimal dosages of both Pg-LPS and E-LPS to solidify Limulus agents were 15 ng/ml and their infrared spectrums were similar. With the treatment of Pg-LPS or E-LPS, the TNF-alpha and IL-1beta levels secreted by HGF-1 cells were remarkably increased with a single perk (P<0.01) while a continuous enhancement of secretion by THP-1 cells was observed (P<0.01). Either Pg-LPS or E-LPS stimulated HGF-1 or THP-1 cells to continuously increase the secretion of IL-6 (P<0.01). Both Pg-LPS and E-LPS induced IL-8 secretion by THP-1 cells (P<0.01), but only Pg-LPS showed the similar effect on HGF-1 cells (P<0.01). Neither Pg-LPS nor E-LPS induced KB cells to secrete inflammatory cytokines. CONCLUSION: Pg-LPS can promote target cells to increase their secretion of inflammatory cytokines. KB cells can not be used as the target cell to determine inflammation-causing effect of LPS.


Subject(s)
Interleukin-1beta/metabolism , Lipopolysaccharides/pharmacology , Porphyromonas gingivalis/chemistry , Tumor Necrosis Factor-alpha/metabolism , Cell Line , Epithelial Cells/cytology , Epithelial Cells/metabolism , Fibroblasts/cytology , Fibroblasts/metabolism , Humans , Interleukin-6/metabolism , Interleukin-8/metabolism
16.
Blood ; 112(3): 886-90, 2008 Aug 01.
Article in English | MEDLINE | ID: mdl-18535204

ABSTRACT

The adult erythron is maintained via dynamic modulation of erythroblast survival potentials. Toward identifying novel regulators of this process, murine splenic erythroblasts at 3 developmental stages were prepared, purified and profiled. Stage-to-stage modulated genes were then functionally categorized, with a focus on apoptotic factors. In parallel with BCL-X and NIX, death-associated protein kinase-2 (DAPK2) was substantially up-modulated during late erythropoiesis. Among hematopoietic lineages, DAPK2 was expressed predominantly in erythroid cells. In a Gata1-IE3.9int-DAPK2 transgenic mouse model, effects on steady-state reticulocyte and red blood cell (RBC) levels were limited. During hemolytic anemia, however, erythropoiesis was markedly deficient. Ex vivo ana-lyses revealed heightened apoptosis due to DAPK2 at a Kit(-)CD71(high)Ter119(-) stage, together with a subsequent multifold defect in late-stage Kit(-)CD71(high)Ter119(+) cell formation. In UT7epo cells, siRNA knock-down of DAPK2 enhanced survival due to cytokine withdrawal, and DAPK2's phosphorylation and kinase activity also were erythropoietin (EPO)-modulated. DAPK2 therefore comprises a new candidate attenuator of stress erythropoiesis.


Subject(s)
Apoptosis Regulatory Proteins/physiology , Calcium-Calmodulin-Dependent Protein Kinases/physiology , Erythroblasts/cytology , Erythropoiesis/genetics , Erythropoietin/pharmacology , Anemia, Hemolytic , Animals , Apoptosis Regulatory Proteins/analysis , Apoptosis Regulatory Proteins/genetics , Calcium-Calmodulin-Dependent Protein Kinases/analysis , Calcium-Calmodulin-Dependent Protein Kinases/genetics , Cell Lineage , Death-Associated Protein Kinases , Hemostasis , Mice , Mice, Transgenic , Phosphorylation/drug effects , Spleen/cytology , Up-Regulation/genetics
17.
Plant Mol Biol ; 58(3): 401-19, 2005 Jun.
Article in English | MEDLINE | ID: mdl-16021403

ABSTRACT

We have used oligonucleotide microarrays to detect Arabidopsis gene expression during early flower development. Among the 22,746 genes represented on the Affymetrix ATH1 chip, approximately 14,660 (approximately 64.5%) genes were expressed with signal intensity at or more than 50 in each of the six organs/structures examined, including young inflorescences (floral stages 1-9), stage-12 floral buds, developing siliques, leaves, stems, and roots. 17,583 genes were expressed with an intensity at or above 50 in at least one tissue, including 12,245 genes that were expressed in all the six tissues. Comparison of genes expressed between young inflorescence or stage-12 floral buds with other tissues suggests that relatively large numbers of genes are expressed at similar levels in tissues that are related morphologically and/or developmentally, as supported by a cluster analysis with data from two other studies. Further analysis of the genes preferentially expressed in floral tissues has uncovered new genes potentially involved in Arabidopsis flower development. One hundred and four genes were determined to be preferentially expressed in young inflorescences, including 22 genes encoding putative transcription factors. We also identified 105 genes that were preferentially expressed in three reproductive structures (the young inflorescences, stage-12 floral buds and developing siliques), when compared with the vegetative tissues. RT-PCR results of selected genes are consistent with the results from these microarrays and suggest that the relative signal intensities detected with the Affymetrix microarray are reliable estimates of gene expression.


Subject(s)
Arabidopsis/genetics , Flowers/genetics , Gene Expression Profiling/methods , Genome, Plant , Analysis of Variance , Arabidopsis/growth & development , Cluster Analysis , Flowers/growth & development , Gene Expression Regulation, Developmental , Gene Expression Regulation, Plant , Oligonucleotide Array Sequence Analysis , Reverse Transcriptase Polymerase Chain Reaction , Time Factors
18.
Genomics ; 85(1): 117-30, 2005 Jan.
Article in English | MEDLINE | ID: mdl-15607427

ABSTRACT

DYRKs are an emerging family of dual-specificity kinases that play key roles in cell proliferation, survival, and development. Up to seven mammalian DYRK isoforms have been reported, but only the DYRK1A gene (and its products) has been well characterized. Defined here are the genomic structures (and phylogenetics) of DYRK3, four additional murine and/or human DYRKs, and two related HIPK genes. For murine DYRK3, direct BAC sequences are provided, a basis for differential processing of murine versus human transcripts is defined, and tissue expression profiles plus a functional DYRK3 promoter are characterized. Unlike complex DYRKs 1A, 1B, and 4A, DYRK3 and DYRK2 possess simple 4-exon structures. For DYRK3, expression is strong in erythroid cells and testis, but is also detected in adult kidney and liver in situ. In addition, a 1930-bp DYRK3 promoter drives erythroid expression and transcript expression is inhibited sharply on GATA1-induced G1E cell differentiation.


Subject(s)
Erythropoiesis/physiology , Gene Expression Regulation/physiology , Protein Serine-Threonine Kinases/biosynthesis , Protein-Tyrosine Kinases/biosynthesis , Animals , Erythropoiesis/genetics , Exons/genetics , Gene Expression Regulation/genetics , Humans , Mice , Phylogeny , Protein Isoforms/biosynthesis , Protein Isoforms/genetics , Protein Serine-Threonine Kinases/genetics , Protein-Tyrosine Kinases/genetics , Sequence Homology, Nucleic Acid , Dyrk Kinases
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