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

SMART-RACE was performed after isolating the total RNA of Armillariella tabescens to amplify the full-length cDNA of arabinosidase (GenBank Accession No. AJ620046). Bioinformatics analysis was used to analyze the code frame of arabinosidase, to predict its structure and function. Recombinant plasmid pPIC9-AF was constructed and then electroporated into methylotrophic yeast Pichia pastoris GS115. The secreted 6 ? His fusion protein was purified to analyze its enzymology property. This arabinosidase had high activity at 30-35℃ under acid condition, and was stable within wide range of pH and temperature. It maintained about 80% activity at the range of pH4. 0-8.0 and 20-40℃,wider than many other cloned arabinosidase. So it was worthy to go step further to study this enzyme, and recombinant expression provided a chance of highly expressing arabinosidase.

AIM: To analyze clinical data of hospitalized patients with age - related cataract treated through clinical pathway in our hospital, and to investigate the effectiveness of clinical pathway in standardizing medical behavior, advancing work efficiency, and improving quality of hospital management.
METHODS:Data of patients with age-related cataract, who were treated with phacoemulsification combined with intraocular lens implantation in Xi'an North Hospital, were retrospectively analyzed. In clinical pathway group, 220 patients underwent cataract surgery from October 2012 to May 2013 in accordance with clinical pathway. For control group, 213 patients with cataract receiving surgery from October 2010 to June 2011, comply with conventional process. Effect of cataract surgery, average length of hospitalization, average cost of hospitalization, and degree of satisfaction of patients were comprehensive investigated.
RESULTS: In 220 patients of clinical pathway group, 209 ( 95. 0%) got improved vision, the days of hospitalization was 4. 5 ± 1. 4d, and the average hospitalized consumption was 4 522. 3 ± 285. 1 Yuan. Whereas, in control group, visual acuity in 202 (94. 8%) out of 213 patients were improved, and the length and cost of hospitalization were 5. 1±1. 7d and 4 647. 7±271. 2 Yuan. The difference of the length and cost of hospitalization between these two groups were significant (P<0. 05). However, there was no statistically significance in curative effect and patients satisfaction compare (P>0. 05).
CONCLUSION: Practicing of age - related clinical pathway is helpful in standardizing medical heavier, optimizing clinical process, reducing health cost, improving quality of medical care, and providing evidence for hospital management innovation.

Objective To study the influence of fluoride on the expression of Runx2 in suckling rat osteoblasts. Methods Osteoblasts obtained from calvarium of suckling Wistar rats were cultured in the media supplemented with NaF at different doses(0, 1,2 and 4 rag/L), and Runx2 Mrna expression and protein expression were evaluated by RT-PCR and ELISA, respectively. Results Runx2 Mrna expression in suckling rat osteoblasts cultured in vitro significantly increased after exposure to NaF for 48 h at different doses (0.613±0.055, 0.773±0.070 and 0.775±0.070 for 1,2 and 4 mg/L,respecfively) compared to the control (0.482±0.043 ,P< 0.05). Runx2 Mrna expression further increased after 72 h exposure to NaF(0.969±0.048,1.229±0.061,1.255± 0.063 for 1,2 and 4 mg/L, respectively) ,which is significantly higher than the control(0.724±0.036,P<0.05) and corresponding groups at 48 h. NaF doses and exposure time exhibited a significant synergistic effect on Runx2 Mrna expression (P<0.05). Similarly, NaF also enhanced Bunx2 protein expression in suckling rat osteoblasts cultured in vitro. Significant differences were observed between groups exposed to NaF (1,2 and 4 rag/L) and control at48 h post-exposure (0.141±0.007, 0.143±0.008, 0.143±0.011 vs 0.129±0.012, P<0.05) as well as 72 h post-expesure(0.156±0.014, 0.168±0.018, 0.162±0.0100 vs 0.137±0.016, P<0.05). In addition, Runx2 protein expression at 72 h post-exposure was significantly higher than that at 48 h. Conclusions The results suggested that NaF could increase Runx2 expression in suckling rat osteoblasts with a synergistic effect between the doses and exposure time.

Objective To study the different expressions of receptor activator of nuclear factor-kappa B ligang(RANKL) mRNA in spleens of rats fed with diet of low calcium and high fluoride. Methods A 2× 2×2 factorial design was used and the factors were calcium, fluoride and action time. In the design, 40 Wistar rats [average body mass(118.9±13.5)g] were divided into four groups randomly by weight: control with normal diet (0.790%, calcium), low calcium group with low calcium intake(0.063%, calcium), high fluoride group with normal diet and high fluoride intake(100 mg/L, fluoride) and low calcium and high fluoride group with low calcium and high fluoride intake. After 4 and 8 months, 5 rats of each group were sacrificed and total RNA was extracted from spleen. And the expression levels of RANKL mRNA were determined by reverse transcription polymerase chain reaction (RT-PCR). Results At time of 4 months, the expression level of RANKL mRNA was 0.13± 0.05,0.13± 0.03,0.17±0.02,0.27± 0.05 and at time of 8 months, it was 0.11 ± 0.01,0.16 ± 0.02,0.16± 0.03,0.36 ± 0.07 in control group, low calcium group, high fluoride group, low calcium with high fluoride group, repectively. The factorial design AVONA showed that low calcium and high fluoride had significant effects on RANKL mRNA expression(F = 40.224,56.679, all P < 0.05) while action time had not(F = 2.850, P > 0.05 ). The interactions of low calcium with high fluoride or high fluoride with action time were signifieant(F = 7.247, 18.789, all P < 0.05) while the interaction of high fluoride with action time was not(F = 1.751, P > 0.05). Conclusions Low calcium alone or high fluoride alone or low calcium with high fluoride or low calcium with action time can increase the the RANKL mRNA expression level. High fluoride does not affect the RANKL mRNA level as the action time is prolonged.

Objective To explorethe effect of Bailingcapsule on epithelial-mesenchymal transition(EMT) inrats withadenine-in-duced tubulointerstitial fibrosis .Methods Tubulointerstitial fibrosis ani mal models were established and SDrats were dividedinto mo-del group (n=30) ,treatment group (n=30) andcontrol group(n=30) ,randomly .Experi mental rats were harvested at 7 w,12 w,17 wafter onset of experi ment and functional evaluations were performed. Histology ,i mmunohistology were examined to investigateboth histolopathology changes and the expression of bone morphogenic protein-7 (BMP-7) ,transforming growth factor-?1(TGF-?1)and a-smooth muscle actin (?-SMA) in kidneys at three ti me points mentioned above ,respectively .Results Compared with controlgroup ,24 h urinary proteinin model grouplost increasingly and significantly difference appeared at three ti me points relative to controlgroup(P0 .05) rel-ative to control group.There was significant difference at 12 wand 17 w(P

Objective To explore the effect of Bailing capsule on epithelial-mesenchymal transition( EMT) in rats with adenine-in-duced tubulointerstitial fibrosis. Methods Tubulointerstitial fibrosis animal models were established and SD rats were divided into mo-del group ( n = 30), treatment group ( n = 30) and control group( n = 30), randomly. Experimental rats were harvested at 7 w, 12 w,17 w after onset of experiment and functional evaluations were performed. Histology, immunohistology were examined to investigateboth histolopathology changes and the expression of bone morphogenic protein-7 (BMP-7), transforming growth factor-β1 (TGF-β1 )and a-smooth muscle actin (α-SMA) in kidneys at three time points mentioned above, respectively. Results Compared with controlgroup, 24 h urinary protein in model group lost increasingly and significantly difference appeared at three time points relative to controlgroup ( P ＜ 0.01 ). Urinary NAG in model group was markedly higher than that in control group from 7 w after onset (P ＜ 0.01 ) andwas increasingly raised at 12 w and 17 w (P＜0.01). The value of blood BUN and Cr in model group increased at 7 w (P＞0.05) rel-ative to control group. There was significant difference at 12 w and 17.w (P ＜ 0.01 ). Histologically, kidneys in model group, at 7 w,exhibited tubular casts and gently tubular dilation, granuloma in cortex, mononuclear cells infiltration in tubulointerstitial areas, andmild interstitial fibrosis. At 12 w, the degree of tubular injury and tubulointerstitial fibrosis gradually aggravated. Up to 17 w, diffusetubular dilation or atrophy was observed and focal tubules disappear. Diffuse interstitial fibrosis was exhibited. In normal kidneys, im-munohistochemistry suggested that the light expression of BMP-7 was detected in proximal renal tubular epithelial cells and marked ex-pression was identified in distal tubule, collecting duct, and renal tubular epithelial in junction area between cortex and medulla. How-ever, the expression of BMP-7 in kidneys of model group significantly decreased with increasing tubulointerstitial fibrosis and was nega-tive correlation with the expression of TGF-β1(r = -0. 981 P＜0.01) and α-SMA (r= -0.975 P＜0.01). Bailing capsule ad-ministration protected the expression of BMP-7 and reduced TGF-β1 and α-SMA expression before 12 w(P＜ 0.01 ). Conclusions Ourstudy shows an anti-fibrotic reno-protective function of Bailing capsule in rats with tubulointerstitial fibrosis via prevention of epithelial-mesenchymal transition at early stage. However, the beneficial effect lost with increasing tubulointerstitial fibrosis.

Hearing Loss, Sensorineural ; genetics ; Humans

Adult ; Colonic Neoplasms ; genetics ; DNA, Neoplasm ; genetics ; Female ; Fluorescence ; Humans ; Male ; Microsatellite Instability ; Middle Aged ; Polymerase Chain Reaction ; methods ; Rectal Neoplasms ; genetics