Relevant literatures on the relationship betw een fluorosis and oxidative stress were reviewed.Based on most of the original papers published in recent years,we can see the increased free radicals and oxidative stress may occur in certain stage of fluoride intoxication,but confirmation of the causality between oxidative stress and fluoride-induced damages still remains much work to do.

Objective To observe endoplagmic reticulum stress in bone tissue of fluomsis rats and further explore the pathogenesis of skeletal fluorosis.Methods 48 Wistar rats were divided into 4 groups according to their body mass.The control and low.calcium group were fed with normal diet(0.79%calcium)and low-calcium diet(0.79%calcium)respectively,and both drank tap water(sodium fluoride concentrations＜1 mg/L).High fluoride and low.calcium plus high-fluoride groups were fed with normal diet(0.79%calcium)and low-calcium diet (0.79%calcium)respectively,and both drank tap water containing sodium fluoride(sodium fluoride concentrations 221 mg/L).During experimental period,rats were measured body mass once a week with a stand diet and water available ad libiturn.The experimental period was 3 months.The biochemical techniques were used to test the indicators of oxidative stress and ALP in seFum of fluorosis rats.The total RNA was extracted from the one side of the femur,and the transcription level of Bip,Xbp1,CHOP and PDI were investigated by reverse transcription polymerase chain reaction(RT-PCR).Results The level of MDA in serum of low-calcium plus high-fluoride group wag higher than that of the control[(14.74±3.11)μmol/L vs(10.15±1.96)μmol/L,P＜0.05];the activity of GPx was ma~edly higher in hish-fluoride group compared with the control[(3.87±0.41)×103 U/L vs(2.85± 0.55)×103 U/L,P＜0.05];the level of uric acid in sel'um was significantly lower both in high-fluoride group and low-calcium plus high-fluoride group compared with the respective control and the low-calcium group[(73.95± 9.52)μmol/L vs(110.43±25.48)μmol/L,(54.32±22.09)μmol/L vs(101.71±17.01)μmol/L,P＜0.05]. The activity of ALP wag obviously higher in low-calcium plus high-fluoride group compared with the control [(24.77±4.57)×103U/L vs (12.91±3.97)×103U/L,P＜0.01)].The mRNA expression of Bip/GAPDH in bone tissue was markedly higher in bone of high-fluoride group and low-calcium plus high-fluoride group compared with the control(1.38±0.24,1.35±0.12 vs 1.14±0.06,P ＜ 0.05). The expression of Xbp1/GAPDH in bone tissue significantly increased in low-calcium plus high-fluoride groups compared with the control and the low-calcium group (1.48±0.20 vs 1.02±0.25,1.07±0.25,P ＜ 0.05 or ＜ 0.01);and CHOP/GAPDH in bone tissue significantly increased in low-calcium plus high-fluoride groups compared with the control(0.84±0.18 vs 0.52±0.07,P ＜ 0.05 ). Conclusions Accelerated osteogenetic action is seen in fluorosis rats,accompanied by oxidative stress and bone endoplasmic reticulum stress,which is likely involved in the pathogenesis of skeletal fluorosis.

Bone Density ; physiology ; Bone Diseases ; epidemiology ; etiology ; metabolism ; pathology ; Calcium ; metabolism ; China ; epidemiology ; Endemic Diseases ; Fluoride Poisoning ; epidemiology ; etiology ; metabolism ; pathology ; Humans ; Osteoblasts ; physiology ; Oxidative Stress ; Parathyroid Hormone ; metabolism ; Thioredoxins ; metabolism ; Transcription Factor AP-1 ; metabolism

Objective To investigate the clinical features,treatment response and prognosis in children with Takayasu′s arteritis(TA) in order to improve the understanding of TA.Methods A retrospective study of 10 children with TA was performed.All of them were admitted and diagnosed in Peking University First Hospital from Jan.1998 to Oct.2008.The clinical features,laboratory tests,imaging modalities,treatment response and prognosis were all collected and evaluated.Results There were 3 boys and 7 girls in the 10 patients with TA,and the ratio of male to female was 12.3.The onset was from 4 months to 9 years old,with average age at 5.5 years old.The average duration of diagnosis was 7.6 months.The incidences of hypertension,vascular bruits,albuminuria,convulsion were present in 100%,100%,70% and 40%,respectively.The clinical types included typeⅡ(60%),type Ⅲ(10%) and type Ⅳ(30%).The acute phase inflammatory indices of activity such as erythrocyte sedimentation rate(ESR),C-reactive protein(CRP) and white blood cell(WBC) were not evidently increased.Tuberculosis infection was found in 6 out of 10 patients and anti-tuberculosis treatment was performed.Six patients were treated with steroids and 3 cases of them were also given immunosuppressives cyclophosphamide or methotrexate.Three of the 10 patients received anti-hypertensive and vasodilator.Two patients received percutaneous translurminal angioplasty and 1 patient received nephrectomy.One patient died of renal failure,heart failure and shock.Conclusions The patients with TA had high prevalence of tuberculosis infection,diagnosis as often late because of lack of specific clinical features at the acute inflammatory period.When organic ischaemia occurred,treatment response was usually unsatisfactory.Patients with multi-systemic and multi-viscera lesions should have comprehensive examination,especially for those with hypertension,pulseless and vascular bruits,in order to rule out TA.Early ultrasonography,computed tomography and magnetic resonnance image methods are valued in eariler diagnosis and they are the key factors to improve prognosis.

Background Corneal blindness caused by ocular surface disease is one of the main reasons for the global blinding corneal diseases.With the development and progress of tissue engineering technology,tissueengineered cornea offers a new approach to the treatment of ocular surface disease.Objective This study was to obscrve the growth and differentiation of human umbilical cord mesenchymal stem cclls (UC-MSCs) on thc corneal stroma of receipts and investigate the feasibility of human UC-MSCs differentiated into corneal epithelium-like cells and the reparation of injury cornea.Methods Human UC-MSCs were isolated from human umbilical cord using collagenase Ⅳ digestion and passaged in DMEM/F12 containing fetal bovine serum in vitro.The immunophenotype of cultured human UC-MSCs was evaluated by flow cytometry.The differentiated osteoblasts from the human UC-MSCs by directional induce was identified.Twenty-four New Zealand albino rabbits were randomly divided into 2 groups.The human UC-MSCs were cultured on porcine corneal matrix without corneal epithelium for 4 days and then transplanted onto the 12 left eyes of 12 New Zealand albino rabbits,and porcine corneal matrix without corneal epithelium was transplanted onto the left eyes of other 12 New Zealand albino rabbits as control group.The rabbits received keratoplasty were examined using in vivo confocal microscope through focusing(CMTF).The eyeballs were taken off after 2,4 and 8 weeks,the growth and differentiation,expression of cytokeratin 3 (CK3),CK12 and ATP-binding cassette superfamily G memben 2 (ABCG2)of human UC-MSCs were observed by histopathology and immunofluorescence staining.This use of the experimental animals complied with ARVO Statement.Results Digestive human UCMSCs formed round in shape and was large in size.The attached cells displayed long-fusiform shape like fibroblasts.The cultured human UC-MSCs phenotype was CD105+/CD29+/CD44+/CD34-/CD45-and could be induced toward osteoblast differentiation under the appropriate experimental conditions.Human UC-MSCs grew well on the porcine corneal matrix.The corneal grafts survived wcll without rejection till the experiment end in experimental eyes,but the rejection of corneal graft occurred in control eyes.Confocal microscope could observe corneal epithelium-like cells.The corneal epithelium cells showed the positive response for CK3 and CK12 and absent response for ABCG2.Conclusions Human UC-MSCs with porcine corneal matrix can survive,proliferate and differentiate into corneal epithelium-like cells after transplanting onto the corneal stroma of rabbits.This result suggests that human UC-MSCs is able to repair and reconstruct the injured corneal surfaces.

Objective To establish mild cognitive dysfunction (MCI) models in elderly rats,and to investigate the pathophysiological features.Methods Totally 40 SD rats (14 to 18-month-old) were randomly divided into 2 groups:the model group (n=20) and the sham operation group (n=20).Bilateral carotid artery stenosis was prepared in the model group while bilateral carotid artery was seperated with no bilateral narrowing in the sham operation group.30 days after the operation,Morris water maze test was performed,pathomorphological and electron microscopic observations of the cerebral tissue were examined and the expression of G protein-coupled receptor kinase 2(GRK2) in hippocampus tissue w detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blottin.Results The mortality in model group was only 10％.Pathological morphology and ultrastructure showed that hippocampal tissue structure was almost normal in sham operated group,but in model group group,hippocampal CA1 pyramidal cells were in ischemic demyelination,arranged loose,and part of the cells showed nucleus pyknosis,deeply stained; there was no obvious infarct in white matter,part of the white matter fiher hecame thinner and disorder,nucleolus became smaller and steped aside,cytoplasmic electron density increased,lipofuscin appeared occasionally.Rough endoplasmic reticulum and Golgi were expanded,cytosolic free ribosomes increased,part of mitochondria became swelled,vacuolated.Morris water maze test results showed that the average escape latency in model group was longer than in sham group (P＜0.05).In spatial probe test,the average time of crossing the first original platform in model rats was significantly longer than the sham operated group [(36.80±7.68) s vs.(20.87±6.16)s,P＜0.05].The average number of crossing the original platform in 60 seconds in model group was significantly less than in sham group(1.43±0.51 vs.3.10±1.45,P＜0.05).The expressiones of GRK2 mRNA and protein in the hippocampus were significantly increased in model group rats than in sham group (P＜0.05).Conclusions The model of severe CCA stenosis in elderly rats can be applied for MCI animal models with good stability and repeatability.Compared with sham group,the cells morphology and ultrastructure in model group appeare more obvious pathological changes and mild impairments in cognitive function.GRK2 may play an important role in the development of MCI.

Objective To study the impacts of lipopolysaccharide on expressions of Cx43 and TLR4 proteins in bladder cancer cell lines and on cell proliferation and apoptosis. Methods 5637 cells were cultured in vitro. After stimulation with LPS,expressions of Cx43 and TLR4 proteins were detected by Western blot in bladder cancer 5637 cell. The proliferation and apoptosis in the 5637?control group,5637?LPS group,and 5637?LPS +cisplatin group were detected by CCK?8(cell counting kit)and flow cytometry. Results The gene expression of Cx43 in the 5637?LPS group was significantly higher than that in the 5637?control group(t=3.892,P=0.012). The expressions of TLR4 and Cx43 in 5637?LPS group were significantly higher than those in the 5637?control group(t=7.029,P=0.019;and t=18.17,P=0.003). The proliferation was significantly decreased in the 5637?control group as compared with the 5637?LPS group (t = 8.756,P = 0.018). The apoptotic rate was (8.3 ± 1.58)% in the 5637?control group and (7.8 ± 2.03)% in the 5637?LPS group,with no significant statistical difference(t = 2.935,P = 0.099). However,the rate of the 5637?cisplatin group(60 ± 4.35)%was higher than that in 5637?cisplatin + LPS group(52 ± 6.25)%,the difference was statistically significant(t = 6.992,P =0.019). Conclusions Under the stimulation of LPS,bladder tumor cells may induce tumor cells to escape immune surveillance by increasing the expressions of TLR4 and Cx43.

The effects of Cinnamon granules on pharmacokinetics of berberine in Rhizoma Coptidis granules in healthy male volunteers, and the compatibility mechanism of Jiao-Tai-Wan (JTW) composed of Rhizoma Coptidis granules and Cinnamon granules were investigated. The concentration of berberine in plasma of healthy male volunteers was determined directly by high performance liquid chromatography (HPLC) after an oral administration of Rhizoma Coptidis granules alone or combined with Cinnamon granules (JTW). The plasma concentration-time curves of berberine were plotted. The data were analyzed with Drug and Statistics (DAS) 2.0 pharmacokinetic program (Chinese Pharmacology Society) to obtain the main pharmacokinetic parameters. The results showed that the plasma concentration-time curve of berberine was described by a two-compartment model. The C(max), T(max), t(1/2) and CLz/F of berberine in Rhizoma Coptidis granules were 360.883 μg/L, 2.0 h, 3.882 h, 119.320 L·h(-1)·kg(-1) respectively, and those of berberine in JTW were 396.124 μg/L, 1.5 h, 4.727 h, 57.709 L·h(-1)·kg(-1) respectively. It was suggested that Rhizoma Coptidis granules combined with Cinnamon granules could increase the plasma concentration of berberine, promote berberine absorption and lengthen the detention time of berberine in healthy male volunteers.

· AIM :To investigate the role of pericytes in growth of retinal microvascular endothelial cells with a co-culture system in order to understand some mechanism of angiogenesis in hypoxia induced retinal neovascular disorders.(RMECs) were isolated by a modified protocol using CD31 coated Dynabeads, and identified by immunocytochemical staining with anti-Factor Ⅷ and CD31 antibodies. Rat retinal pericytes were isolated and characterized by immunofluorescent staining with PDGFR-β; and desmin antibodies. Pericytes and RMECs were cultured in a contact co-culture system both under normoxia and hypoxia by Millicell chamber. RMECs proliferation was evaluated by MTT and cell cycle assay with flow cytometry. RT-PCR was used to detect the alteration of KDR/Flk-1 mRNA level in RMECs under normoxia or hypoxia in the co-culture system.harvested with the modified isolating method. The two cell types were identified by positive Factor Ⅷ, CD31 and PDGFR-β, desmin cytochemical staining respectively.RMECs proliferated significantly under hypoxia from 3 to 9d with a maximal rate on day 6 (24.9%, P ＜ 0.01) by MTT. In the co-culture system, the proliferation of RMECs was inhibited by pericytes. After 6d exposure to hypoxia,the fraction of S-phase RMECs number was greatly increased by 43.9% (P ＜ 0.01). In the co-culture system,RMECs proliferation was inhibited by pericytes through decreasing the fraction of S-phase cell number both under normoxia (3.6%, P＜0.05) and under hypoxia (15.1%,P＜0.01). KDR/Flk-1 mRNA level in single cultured RMECs was shown to increase approximately 1.3-fold when exposed to hypoxia. Compared with single cultured RMECs, co-culture with pericytes could decrease KDR/Flk-1 mRNA by 45.1% (P＜0.05) and 27.7% (P ＜ 0.05) under normoxia and hypoxia condition respectively.pericytes could inhibit proliferation of RMECs under both normoxia and hypoxia. The inhibition effects of pericytes maybe, at least in part, due to downregulation of KDR/Flk-1 of RMECs. These findings confirm that pericytes could be a potential inhibitor in the pathogenesis of retinal neovascularization.

Objective To observe the status of oxidative stress and activity of alkaline phosphatase(ALP) in rats exposed to high fluoride for the different periods and to analyze the effect of fluoride on the activity of ALP and oxidative stress in fluorosis rats. Methods Twenty-four Wistar rats were divided into control and high-fluoride groups according to their body mass, 12 rats in each group. The control group drank tap water(sodium fluoride concentrations < 1 mg/L), and high-fluoride group drank tap water containing sodium fluoride(sodium fluoride concentrations 221 mg/L). On a standard diet and water available ad hbitum, each rat was measured body weight once a week in 1,4,8,12 week. The biochemical techniques were used to test the indicators of oxidative stress including malonaldehyde(MDA), superoxidedismutase(SOD), glutathione peroxidase(GPx), uric acid and activity of ALP in serum of fluorosis rats. Results There was a interaction between fluoride and time in the activity of ALP (F = 4.690,P < 0.05). The activity of ALP was obviously higher in rats exposed to fluoride for 1,12 week [ (19.29± 3.69), (15.72 ± 0.79)kU/L] compared to the control[ (14.08 ± 1.99),(12.91 ± 3.97)kU/L, all P< 0.05] ; the level of MDA was obviously higher in rats exposed to fluoride for 1,4 week [ ( 13.37 ± 4.38 ), ( 11.82 ± 2.08) μmol/L ] compared to the respective control[ (8.75 ± 3.24), (7.42 ± 2.62)μmol/L, all P < 0.05]; difference of SOD and GPx between control and high-fluoride groups was not statistically significant(all P > 0.05); the level of uric acid in serum was significantly higher in high-fluoride group for 1,4 week[ (89.53 ± 13.21 ), (88.47 ± 19.78 )μmol/L] compared to the control [ (77.79 ± 11.43 ), (65.42 ± i 3.42) μ mol/L, all P < 0.05 ], but the level of uric acid showed lower in high-fluoride group for 8,12 week [(67.21 ± 9.44), (73.95 ± 9.52)μmol/L] compared to the control [(77.79 ± 11.43), (65.42 ± 13.42)μmol/L]. Conclusions Effect of overdose fluoride on ALP is time-dependant. On the other hand,overdose fluoride stimulates the status of oxidative stress in a way unrelated to the exposure period.