OBJECTIVEThe aim of this study was to observe the human hair follicle apoptosis status affected by fluorine and the antagonism effect by selenium in vitro.

METHODSThe single hair follicles were separated and cultured, then they were added in different concentrations of sodium fluoride and sodium selenite. Chosen the appropriate concentrations, they were divided into 7 groups. The TUNEL was used to investigate the apoptotic cells of different parts. The morphous of hair follicles was observed consecutively and electron microscope was used.

RESULTSWe found that in 1 mmol/L and 10 mmol/L sodium fluoride groups, when the human hair follicles in vitro were cultured on the 5th day, the apoptotic cells of outer root sheath (ORS), dermal sheath and hair papilla, hair bulb were obviously increased. But 0.01 mmol/L sodium selenite weakened the toxicity of 1 mmol/L sodium fluoride at the outer root sheath and hair bulb (P < 0.05).

CONCLUSIONSDifferent concentrations of sodium fluoride had different effect on the growth of human hair follicle in vitro which were cultured on 5th day. Sodium fluoride of certain concentration could accelerate the apoptosis of human hair follicle in vitro. Sodium selenite of certain concentration could act antagonism to the toxicity of sodium fluoride.

Adolescent ; Adult ; Apoptosis ; drug effects ; Hair Follicle ; drug effects ; Humans ; Middle Aged ; Sodium Fluoride ; pharmacology ; Sodium Selenite ; pharmacology ; Tissue Culture Techniques ; Young Adult

OBJECTIVETo investigate the effects of putative bacteriocin immunity proteins on the growth mode of Streptococcus mutans (Sm). To observe the differences of antimicrobial sensitivity in planktonic Sm wild-type strains and mutant strains caused by the inactivation of bacteriocin immunity proteins and their influence on the biofilm formation.

METHODSSm wild-type strains (WT) and its knockout mutants defective in immA and immB (ΔimmA(-) and ΔimmB(-) mutants) coding putative bacteriocin immunity proteins were cultured in brain heart infusion (BHI) and selected by erythromycin at the concentration of 10 mg/L. Optical density was detected by spectrophotometer every hour and growth curve was drawn. WT, ΔimmA(-) and ΔimmB(-) mutants were treated with ampicillin (0.04, 0.05, 0.06, 0.07, 0.08 mg/L), sodium fluoride (50, 100, 150, 200, 250 mg/L) and sodium hypochlorite (0.078%, 0.156%, 0.313%, 0.625%, 1.250%) for 24 hours. Optical density was detected by multifunctional micro plate reader. WT and the mutants were cultured in MBEC(TM) P&G Assay for 24 hours. The minimum biofilm eradication concentration (MBEC) of chlorhexidine against Sm was determined by serial dilution method. Confocal laser scanning microscopy (CLSM) was used to visualize the biofilm architecture, depth and ratio of live to dead bacteria.

RESULTSGrowth curve showed that it took about 3 hours to reach exponential phase and about 7 hours to stationary phase for WT, while 4 hours to exponential phase and 8 hours to stationary phase for mutants. Optical density of mutants were lower than WT in the presence of various antimicrobial agents (P < 0.01). In 0.06 mg/L ampicillin group, optical density value of WT, ΔimmA(-) and ΔimmB(-) mutants were 0.334 ± 0.016, 0.027 ± 0.016 and 0.047 ± 0.018. In 150 mg/L sodium fluoride group, optical density value of WT and mutants were 0.254 ± 0.018, 0.129 ± 0.011 and 0.167 ± 0.010. In 0.313% sodium hypochlorite group, optical density value of WT and mutants were 0.467 ± 0.008, 0.017 ± 0.006 and 0.050 ± 0.006. The MBEC of chlorhexidine against Sm WT, ΔimmA(-) and ΔimmB(-) mutants were 6.25, 1.57, and 3.13 mg/L. The results by CLSM showed a noticeable difference in biofilm architecture. The depth of WT biofilm was higher than the mutants biofilm (P < 0.01). The ratio of live to dead bacteria of WT biofilm was higher than ΔimmA(-) mutants in all layers (P < 0.05) and ΔimmB(-) mutants in the outer and intermedium layer (P < 0.01). There is no significant different between the inner layers of WT and ΔimmB(-) mutants (P = 0.191).

CONCLUSIONSPutative bacteriocin immunity proteins have influence on the growth mode of Sm. The antimicrobial sensitivity of planktonic Sm can be up-regulated by the inactivation of immA or immB. The MBEC of chlorhexidine against ΔimmA(-) and ΔimmB(-) mutants is lower than WT. The inactivation of immA or immB affects the biofilm formation.

Ampicillin ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Bacteriocins ; genetics ; immunology ; Biofilms ; drug effects ; growth & development ; Cariostatic Agents ; pharmacology ; Chlorhexidine ; pharmacology ; Disinfectants ; pharmacology ; Microbial Sensitivity Tests ; Mutation ; Plankton ; drug effects ; Sodium Fluoride ; pharmacology ; Sodium Hypochlorite ; pharmacology ; Streptococcus mutans ; drug effects ; genetics

BACKGROUND: Accurate measurement of blood glucose concentrations is essential for defining diabetes, and the minimization of ex vivo glycolysis has been recommended. Recent guidelines advocate two kinds of methods for sample collection and processing: either the sodium fluoride (NaF) method or immediate refrigeration using a serum separation tube (SST). We investigated the difference between the two methods in measuring subsequent glucose concentrations using blood specimens from participants recruited for the fourth Korean National Health and Nutrition Examination Survey. METHODS: Paired venous blood samples were collected in an SST and a NaF tube from 1,103 men and women. SST serum was separated within 30 min, including standing for 15 min, and then refrigerated. The NaF samples were refrigerated, but not separated until immediately before analysis. We compared the blood glucose concentrations between the SST (SST glucose) and NaF (NaF glucose) methods. RESULTS: The mean SST glucose was significantly higher than NaF glucose (99.0 mg/dL vs 96.5 mg/dL, P<0.05). NaF glucose showed a negative mean bias of 2.6 mg/dL vs SST glucose but showed high correlation (R=0.9899). There was no significant correlation between the bias of blood glucose concentrations by two methods and the storage time of NaF glucose. CONCLUSIONS: The negative bias associated with the use of NaF tubes may significantly affect the prevalence of diabetes. Serum separation and refrigeration within 30 min after venous sampling is recommended over NaF method, not only to minimize the preanalytical impact on detecting diabetes but also to reduce sample volume and number of tubes.
Blood Glucose/*analysis ; Blood Specimen Collection/*methods ; Diabetes Mellitus/diagnosis ; Female ; Glycolysis/*drug effects ; Humans ; Male ; Nutrition Surveys ; Republic of Korea ; Sodium Fluoride/*pharmacology ; Specimen Handling

Animals ; Apoptosis ; Biflavonoids ; pharmacology ; Catechin ; pharmacology ; Grape Seed Extract ; pharmacology ; Kidney ; drug effects ; Male ; Oxidative Stress ; Proanthocyanidins ; pharmacology ; Protective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sodium Fluoride ; adverse effects

OBJECTIVEThis study was conducted to study the effects of sodium fluoride (NaF) on rat renal apoptosis and proliferation, the antagonistic effect of selenium-zinc preparation (Se-Zn) to NaF.

METHODSWistar rats were provided with distilled water containing NaF (50 mg/L) and administered by gavage with different dosed of Se-Zn for six months. Kidney cell apoptosis and the cell cycle of proliferation were detected by TUNEL (TdT-mediated dUTP Nick End Labelling) and flow cytometry.

RESULTSNaF caused rat renal apoptosis, reduce the cell number of G(2)/M period in cell cycle and decrease the relative content of DNA significantly. Se-Zn inhibited the effects of NaF on apoptosis and increased the cell number of G(2)/M period in cell cycle, but failed to increase relative content of DNA.

CONCLUSIONIt was suggested that NaF could induce apoptosis and change the cell cycle in rat renal cells and Se-Zn could antagonize apoptosis and the changes of cell cycle induced by NaF.

Animals ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; DNA ; drug effects ; genetics ; metabolism ; Drug Antagonism ; Flow Cytometry ; In Situ Nick-End Labeling ; Kidney ; drug effects ; pathology ; Rats ; Selenium ; pharmacology ; Sodium Fluoride ; pharmacology ; Zinc ; pharmacology

OBJECTIVETo investigate the effects of fluoride on Fas expression, caspase-3 and caspase-8 activity and apoptosis in rat incisor cells.

METHODSForty male SD rats were divided into 4 groups randomly and provided with distilled water containing NaF at the doses of 0, 10, 50 and 100 mg/L respectively. Each group had 10 animals. Five animals were sacrificed at 60 and 90 days respectively. Fas expression was measured with immunohistochemistry, and colorimetric assay was used to examine caspase-3 and caspase-8 activity with enzyme-labelled meter. The apoptosis was detected by flow cytometry in mandibular incisor cells.

RESULTSNaF at the doses of 10, 50 and 100 mg/L for 60 d and 90 d caused Fas overexpression, promoted activity of caspase-3 and caspase-8, increased apoptosis rate in mandibular incisor cells. At 60 days, the value of Fas expression was 0.1819 ± 0.0025 for control, 0.2120 ± 0.0084 for 10 mg/L NaF group, 0.2283 ± 0.0183 for 50 mg/L NaF group, 0.2818 ± 0.0233 for 100 mg/L NaF group. At 90 days, the value of Fas expression was 0.2077 ± 0.0289 for control, 0.2216 ± 0.0105 for 10 mg/L NaF group, 0.2377 ± 0.0059 for 50 mg/L NaF group, 0.2775 ± 0.0088 for 100 mg/L NaF group. Statistics analysis yielded close relationship between the dose of NaF in water and the Fas expression, and also between the dose of NaF in water and caspase-3 activities, and the relative coefficient was 0.9728 (60 d, P < 0.01) and 0.9889 (90 d, P < 0.01) for Fas expression, 0.9533 (60 d, P < 0.01) and 0.9849 (90 d, P < 0.01) for caspase-3 activity respectively. Apoptosis rate and caspase-8 activity also had close relationship with the NaF doses, and the relative coefficient was 0.9733 (90 d, P < 0.01) for apoptosis, 0.9928 (90 d, P < 0.01) for caspase-8. At the doses of 10, 50 and 100 mg/L NaF for 60 d and 90 d, obvious relationship was found between Fas expression and caspase-3 activity, and the relative coefficient was 0.9619 (60 d, P < 0.01) and 0.9912 (90 d, P < 0.01). Obvious relationship between Fas expression and apoptosis, between Fas expression and caspase-8 activity was found in groups for 90 d, and the relative coefficient was 0.9841 (P < 0.01) for apoptosis, 0.9767 (P < 0.01) for caspase-8.

CONCLUSIONSFluoride could induce Fas overexpression and mediate caspase activation and apoptosis at the doses of 10, 50 and 100 mg/L for 60 d and 90 d in rat mandibular incisor cells.

Animals ; Apoptosis ; drug effects ; Cariostatic Agents ; administration & dosage ; pharmacology ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Dose-Response Relationship, Drug ; Incisor ; cytology ; metabolism ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Sodium Fluoride ; administration & dosage ; pharmacology ; fas Receptor ; metabolism

OBJECTIVETo study expression of proto-oncogenes c-fos and its accompanying gene c-jun in osteoblasts activated by action of excessive fluoride in vivo and in vitro.

METHODSExperimental Wistar rats were exposed to sodium fluoride (NaF) added to their drinking water, and NaF was also added in cell culture supernatant for osteoblast-like cells in vitro. Expression of both mRNA and protein of c-fos and c-jun in bone-tissue of rats with chronic fluorosis and cultured osteoblast-like cells were determined by hybridization in situ, Western blot and immunohistochemistry at varied time periods after exposure.

RESULTSSodium fluoride could stimulate the proliferation of osteoblast in rats with chronic fluorosis and induce expression of both c-fos and c-jun in all envelops of the spine bone, as compared with its control group. Value of optical absorption in mRNA expression of c-fos and c-jun was 139.63 and 126.37, respectively, in rats with NaF plus high-calcium, significantly lower than that in control group with high-calcium only (107.74 and 117.48, respectively) (P < 0.001). Immunohistochemical analysis showed that protein level of c-fos and c-jun was significantly higher in rats with NaF plus high-calcium than that in control rats with high-calcium only, with values of optical absorption of 139.16, 131.15, 149.98 and 149.19 (P < 0.05), respectively, and protein level of c-fos and c-jun was significantly higher in rats with NaF plus low-calcium than that in control rats with low-calcium only, with values of optical absorption of 117.24, 111.46, 132.46 and 129.79 (P < 0.05), respectively. Western blotting showed that level of protein expression of c-fos and c-jun in periosteal osteoblasts was significantly higher in all rat groups with NaF than that in all control groups, with values of optical absorption of 123.32, 116.60, 115.97 and 108.30, respectively. mRNA expression of c-fos and c-jun in osteoblast-like cells treated with NaF for 12 h increased obviously, and remained at high level 48 h after exposure, with values of optical absorption of 114.80, 161.14, 118.20, and 150.41, respectively, as compared with that in control group (P < 0.001 and P < 0.05).

CONCLUSIONSExposure to excessive fluoride could stimulate activation and proliferation of both osteoblasts in rats and cultured osteoblast-like cells in vitro, and cause enhanced expression of mRNA and protein of both c-fos and c-jun. Over-expression of c-fos could play an important role in development and proliferation of skeletal lesions in rats with chronic fluorosis.

Animals ; Bone Diseases ; metabolism ; pathology ; Calcium ; pharmacology ; Cell Division ; drug effects ; Cells, Cultured ; Female ; Fluoride Poisoning ; metabolism ; pathology ; Gene Expression ; Male ; Osteoblasts ; cytology ; metabolism ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-jun ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Sodium Fluoride ; pharmacology

OBJECTIVETo explore the DNA damage effects and apoptosis in brain cells of rats induced by sodium fluoride.

METHODSSD rats were divided into two groups, i.e. control group and fluoride treated group, which were injected intraperitoneally with distilled water and sodium fluoride (20 mg.kg(-1).d(-1)) respectively. On the hand, 5 mmol/L NaF were used in in vitro study. Single Cell Gel Electrophosis (SCGE or Comet Assay) was utilized to measured DNA damage and apoptosis was detected by the TUNEL method and Flow Cytometry (FCM).

RESULTSThe DNA damage in pallium neurons in rats of the fluoride group was much more serious compared with those of the control group, with the Ridit value being 0.351 and 0.639 respectively (P < 0.01) in vivo, and 0.384 4 and 0.650 1 respectively (P < 0.01) in vitro. TUNEL positive cells were found in pallium, hippocampus and cerebellar granule cells in rats of fluoride group, whereas those in the control group were rare. It was demonstrated by FCM results that the percentages of apoptotic cells both in pallium and hippocampus were significantly higher (P < 0.01) in rats of fluoride group (27.12 +/- 3.08, 34.97 +/- 5.46) than those in control group (4.63 +/- 0.98, 5.35 +/- 0.79), (P < 0.01).

CONCLUSIONSodium fluoride could induce DNA damage and apoptosis in rats brain.

Animals ; Apoptosis ; genetics ; Brain ; drug effects ; metabolism ; pathology ; Comet Assay ; DNA ; drug effects ; genetics ; metabolism ; Flow Cytometry ; In Situ Nick-End Labeling ; Male ; Rats ; Rats, Sprague-Dawley ; Sodium Fluoride ; pharmacology

OBJECTIVETo study the effects of selenium and zinc on oxidative stress, apoptosis, and cell cycle changes in rat renal cells induced by fluoride.

METHODSWistar rats were given distilled water containing sodium fluoride (50 mg/L NaF) and were gavaged with different doses of selenium-zinc preparation for six months. Four groups were used and each group had eight animals (four males and four females). Group one, sham-handled control; group two, 50 mg/L NaF; group three, 50 mg/L NaF with a low dose of selenium-zinc preparation (0.1 mg/kg Na2 SeO3 and 14.8 mg/kg ZnSO4 x 7H2O); and group four, 50 mg/L NaF with a high dose of selenium-zinc preparation (0.2 mg/kg Na2 SeO3 and 29.6 mg/kg ZnSO4 x 7H2O). The activities of serum glutathione peroxidase (GSH-Px), kidney superoxide dismutase (SOD), and the levels of malondialdehyde (MDA) and glutathione (GSH) in the kidney were measured to assess the oxidative stress. Kidney cell apoptosis and cell cycle were detected by flow cytometry.

RESULTSNaF at the dose of 50 mg/L increased excretion of fluoride in urine, promoted activity of urine gamma-glutamyl transpeptidase (gamma-GT), inhibited activity of serum GSH-PX and kidney SOD, reduce kidney GSH content, and increased kidney MDA. NaF at the dose of 50 mg/L also induced rat renal apoptosis, reduced the cell number of G2/M phase in cell cycle, and decreased DNA relative content significantly. Selenium and zinc inhibited effects of NaF on oxidative stress and apoptosis, promoted the cell number of G2/M phase in cell cycle, but failed to increase relative DNA content significantly.

CONCLUSIONSodium fluoride administered at the dose of 50 mg/L for six months induced oxidative stress and apoptosis, and changes the cell cycle in rat renal cells. Selenium and zinc antagonize oxidative stress, apoptosis, and cell cycle changes induced by excess fluoride.

Animals ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Glutathione ; metabolism ; Glutathione Peroxidase ; blood ; Kidney ; drug effects ; metabolism ; Malondialdehyde ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Rats, Wistar ; Selenium ; pharmacology ; Sodium Fluoride ; antagonists & inhibitors ; toxicity ; urine ; Superoxide Dismutase ; metabolism ; Zinc ; pharmacology ; gamma-Glutamyltransferase ; urine

OBJECTIVETo investigate mRNA and protein expressions of OPGL and M-CSF mRNA in bones of rats with experimental fluorosis induced by intake of fluoride in the drinking water, and to study the antagonizing effects of Danlan Xianpeng Liaofu capsules on the gene expression.

METHODSTotally 72 SD rats were randomly assorted into 6 groups including the control group, the fluoride group, the high-dosage (0.8 g/kg×d), mid-dosage (0.4 g/kg×d) and low dosage (0.2 g/kg×d) medication groups and the borax group (borax, 0.8 g/kg×d). The distribution of female and male rats in each group was divided up on a fifty-fifty basis. Except the control group, a NaF containing water (NaF 50 mg/L in concentration) was supplied as the drinking water for all the experimental rats in order to establish experimental fluorosis. The thickness and density of trabecula and the thickness of bone cortex were measured by light microscopy. The fluoride content in urine and bone were analyzed by using fluoride ion selective electrode method. Expressions of OPGL and M-CSF mRNA and protein were studied using RT-PCR and immuno-histochemistry, respectively.

RESULTS(1) 10/12 of the experimental fluorosis rats developed dental fluorosis, and 2/12 of dental fluorosis rats occurred in the low-dosage medication group. Fluoride content in urine and bone of the fluorosis rats increased (P<0.05). (2) Compared with that of the control rats, the bone trabecular depth, cortical thickness and trabecular density in experimental fluorosis rats were remarkably reduced. (3) Compared with that of the control group, mRNA expression of both OPGL and M-CSF was increased in the fluoride group rats. The difference was statistically significant (P<0.05). (4) Compared with that of the fluoride group animals, the expression intensity of OPGL mRNA decreased in animals of the control group, the high, mid- and low- dosage medication groups and the borax group. Among them, except the low-dosage group, the difference between all the other groups and the fluoride group was statistically significant, respectively (P<0.05). There was also a decrease of M-CSF mRNA in all the 3 medication groups and the borax group animals in comparing with that of the fluoride group and the difference was also statistically significant (P<0.05), respectively. (5) Compared with that of the control group. There were an increase of OPGL and a decrease of M-CSF protein expression; and in addition, there were a decrease of OPGL and an increase of M-CSF protein expression in all 3 medication groups and the borax group in comparing with that of the fluoride group anima (P < 0.05).

CONCLUSIONSExcessive fluoride induces an accelerated bone turnover and may promote the absorption activity of osteoclasts by increasing the expression of OPGL and M-CSF. Danlan Xianpeng Liaofu capsule may be capable of regulating bone remodeling through a down-regulation on OPGL and M-CSF expression.

Animals ; Bone Density ; drug effects ; Bone Remodeling ; drug effects ; Bone and Bones ; metabolism ; pathology ; Borates ; pharmacology ; Capsules ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Female ; Fluorides ; metabolism ; urine ; Fluorosis, Dental ; metabolism ; pathology ; Macrophage Colony-Stimulating Factor ; genetics ; metabolism ; Male ; RANK Ligand ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sodium Fluoride ; poisoning