1.Relieving efefct of Dexmedetomidine on myocardial injury induced by renal ischemia-reperfusion at differ-ent time points in rats
Xinling LI ; Bing ZHANG ; Zhong LEI ; Jing DU
Journal of Medical Postgraduates 2015;28(10):1028-1033
Objectiv e Renal ischemia-reperfusion ( I/R) may cause myocardial injury and dexmedetomidine ( DEX) is a new alpha-2 adrenergic agonist with the effects of antisympathia , seda-tion, and analgesia.This study was to investigate the effect of DEX on the myocardial tissue of rats at different time points after renal I/R. Methods Forty male Wistar rats were randomized into 5 groups of equal number,sham operation, 60 min renal ischemia and 3 h reperfusion (I/R1), 120 min ischemia and 3 h reperfusion (I/R2 ), 60 min ischemia and DEX+3 h reperfusion (D1), 120 min ischemia and DEX+3 h reperfusion ( D2) .Renal I/R was induced by removal of the right kidney and ligation of the left re-nal artery and vein followed by 3 hours of reperfusion.Meanwhile, intraperitoneal injection of DEX at 50μg/kg was given to the ani-mals in groups D1 and D2 at 60 at 120 min respectively after ischemia.After 3 hours of reperfusion, blood samples were collected for measurement of the concentrations of serum creatinine (Cr) and blood urea nitrogen (BUN), and renal and myocardial tissues harvested for observation of pathological changes under the light microscope and determination of the expressions of TNF-αand IL-10 by ELISA.Results Significant increases were observed in the concentrations of serum Cr and BUN , the expressions of TNF-αand IL-10 in the renal tissues and those in the myocardial tissues in groups I /R1([84.67 ±9.62] μmol/L, [8.55 ±1.08] mmol /L), I/R2 ([167.11 ±18.81] μmol/L, [13.42 ±1.25] mmol/L), D1 ([69.67 ±9.52] μmol/L, [7.56 ±0.70] mmol/L), and D2 ([114.29 ±12.50] μmol/L, [10.27 ±0.78] mmol/L), as compared with the sham operation group ([53.20 ±9.21] μmol/L, [3.75 ±0.78] mmol/L), (all P <0.05).Significant decreases was observed in the sham operation group as compared with other groups in the expressions of TNF-αand IL-10 (P<0.05).Significant decreases was observed in the D1 and D2 groups compared with other groups in the expressions of TNF-α, but increasing in IL-10.②Injury was reduced in the D1 and D2 groups compared with other groups.③The horizontal stripes of myocardial tissue disappeared in I/R1 and I/R2 decreases of inflammatory cells was observed in D1 and D2 groups compared with others. Conclusion Dexmedetomidine can attenuate myocar-dial injury induced by renal ischemia-reperfusion in rats and its inhibitory effect on inflammatory factors may be involved in the mechanism.
2.The Construction and Expression of Phage Display scFv Library from the Spleen Cells of Mice Immunized With B3HM Cells
Jing XU ; Lei ZHANG ; Shi-Yong DIAO ; Bin LIU ; Lei MENG ; Xue-Ying JING ; Zhong-Chao HAN ;
China Biotechnology 2006;0(07):-
To construct a scFv library by phage display technique from the spleen cells of mice immunized with B3HM cells. Three mice were immunized with B3HM cells, and their spleen cells were harvested. The genes of VH and Vk were amplified by RT-PCR from the cDNA of the immunized spleen cells and a scFv-phage display antibody library was constructed. The capacity of library was measured,and the variety of the library was analyzed by digesting with restriction endonuclease BstNI.ScFv phage clones were randomly picked and identified phage-scFv clone by binding B3HM cells using immunofluorescein.A scFv library containing 5?106 individual clones which showed different patterns after digested with restriction endonuclease BstNI was produced. Individnal phage-scFv clone showed B3HM cells positive using immunofluorescein. A scFv library of anti-B3HM cell surface molecules has been constructed. It will be useful for finding out some novel genes of causing leukemia, and establishs the infarctate foundation of clarifying the pathogenesis of leukemiagenesis.
3.A case-control study on the risk factors of leukemia in mining areas of rare-earth in South Jiangxi.
Lei WU ; Yue-Ping ZHOU ; Hong-Jing ZHONG
Chinese Journal of Epidemiology 2003;24(10):879-882
OBJECTIVEIn order to explore the correlation on radioactive contamination of lanthanon to leukemia, and provide clues for the causes and prevention of leukemia in mining areas of rare-earth elements.
METHODS1:1 matched case-control study was used. A total of 51 clinically confirmed leukemia cases, individually matched with controls from general population, were interviewed in mining areas of rare-earth in South Jiangxi from November to December, 2001. Data were analyzed, using conditional logistic regression.
RESULTSThe main risk factors would include frequently drinking water from river (OR = 5.543), distance from residence to rare-earth mine and years for living in the area (OR = 3.308), exposure to organophosphorus pesticide (OR = 3.014). Tea drinking habit appeared to be a protective factor.
CONCLUSIONSLeukemia seemed to be related to environmental pollution with rare-earth elements around the residential areas and organophosphorus pesticide exposure. The protective factor of tea drinking habit seemed to be unique in this study, which called for further studies.
Adolescent ; Adult ; Aged ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Humans ; Insecticides ; toxicity ; Leukemia ; etiology ; Logistic Models ; Male ; Metals, Rare Earth ; toxicity ; Middle Aged ; Mining ; Organophosphorus Compounds ; Risk Factors ; Tea ; Water Pollutants, Chemical ; toxicity
4.Expression changes of cardiac sinus node Cx45,Cx31.9 on sinus bradycardia model caused by dexmedetomidine in rabbit
Yu ZHU ; Lei TIAN ; Yanruo HUANG ; Jing SHI ; Yi ZHONG ; Yongqiang YIN
The Journal of Clinical Anesthesiology 2017;33(8):801-804
Objective To investigate the changes of cardiac sinus node connexin 45 (Cx45), connexin 31.9(Cx31.9) of gap junction in rabbits'' sinus bradycardia model caused by dexmedetomidine, and to discuss whether the negative frequency and negative conduction caused by Dexmedetomidine are related to the expression changes of connexin.Methods Forty-eight healthy adult New Zealand rabbits were divided into three groups (n=16).Sinus bradycardia rabbits were prepared by intravenous injecting Dexmedetomidine through ear vein.After rabbits were anesthetized by sodium pentobarbital, basic procedures were quickly completed in order to monitor MAP and ECG.Rabbits in group C were injected with normal saline.Rabbits in group D1 were injected a loading dose of dexmedetomidine 10 μg/kg for 10 min, and then continuously pumped 5 μg·kg-1·h-1 for 50 min.Rabbits in group D2 were pumped a loading dose of dexmedetomidine 60 μg/kg for 10 min, and then continuously pumped 30 μg·kg-1·h-1 for 50 min.After observation hearts were quickly removed and sinus node tissue was dissected.The average optical density of sinus node Cx45, Cx31.9 were detected by immunohistochemistry, and the genes expression were detected by real-time quantitative.Results Cx45 gene expression of group D2 showed remarkable increase than groups C and D1(P<0.05), there were no significant differences between groups D1 and C, Cx45 average optical density change were consistent with the gene expression.Cx31.9 gene expression of group D1 showed a more remarkable increase than group C(P<0.05), there were no significant differences between groups D2 and C,D1 and D2, Cx31.9 average optical density changes were consistent with the gene expression.Conclusion Dexmedetomidine increases the expression of low electrical conductivity Cx45, Cx31.9 of gap junction on rabbits'' sinus node, which is one of the possibly reasons that slow down cardiac conduction velocity in sinus node.
5.Effects of different doses of dexmedetomidine on myocardial injury induced by renal ischemia-reperfusion in rats
Xinling LI ; Zhong LEI ; Jing DU ; Xiaoyan JIA ; Wei TIAN ; Jian ZHANG ; Bing ZHANG
Journal of Medical Postgraduates 2015;(8):809-814
Objective Renal ischemia-reperfusion may cause myocardial injury and dexmedetomidine ( DEX)is a new alpha-2 adrenergic agonist which has the effects of antisympathia , sedation and analgesia.The article was to observe the effects of different doses of dexmedetomidine on myocardial injury induced by renal ischemia -reperfusion(I/R) in Rats. Methods 40 male Wistar rats were randomized into 5 groups(n=8 each)using a random number table: sham operation group(isolation of bilateral renal pedicles without ligation), I/R group (3 hours′reperfusion 120 minutes after the right nephrectomy and the ligation of left renal artery ), DEX low dose group, DEX middle dose group and DEX high dose group (DEX 25, 50, 100 μg/kg were respectively injected intraperitone-ally in rats of the three groups plus 3 hours′reperfusion after 120 minutes′ischemia ) .Blood samples were collected at 3 hours′reper-fusion to determine serum creatinine(Cr) and blood urea nitrogen(BUN) concentrations.Kidney and myocardial specimens were ex-tracted for microscopic examination and IL-10,TNF-αcontent were detected by ELISA. Results In sham operation group, renal structure was normal.In I/R group, a great amount of erythrocyte infiltration and interstitial infiltrating inflammatory cells were found in glomerulus and a lot of exfoliative cells were found in renal tubules .In DEX low dose group , erythrocyte infiltration and interstitial infiltrating inflammatory cells were found in glomerulus and a few exfoliative cells were found in renal tubules .In DEX middle dose group, erythrocyte infiltration and interstitial infiltrating inflammatory cells were found in partial glomerulus and a few exfoliative cells were found in renal tubules .In DEX high dose group , erythrocyte infiltration and interstitial infiltrating inflammatory cells in partial glomerulus were found and rare exfoliative cells were found in renal tubules .In sham operation group , cardiomyocytes were arranged in perfect order and normal structure , and chromatins and cytoplasms were in uniform distribution .In I/R group, edema and spongiform were obvious in cardiomyocytes , and focal coagulative necrosis was observed along with a great amount of infiltrating inflammatory cells and rare flaky bleeding .In DEX low dose group , edema and spongiform were found in cardiomyocytes , and focal coagulative necrosis was observed along with a great amount of infiltrating inflammatory cells and rare flaky bleeding .In DEX middle dose group , edema was found in cardiomyocytes , and mini focal coagulative necrosis was observed along with a small amount of infiltrating inflammatory cells.In DEX high dose group , edema was found in cardiomyocytes along with a small amount of infiltrating inflammatory cells .Com-pared with sham operation group , Cr, BUN concentrations in serum and IL-10,TNF-αcontents in kidney tissue and myocardium signif-icantly increased in I/R group, low dose group, middle dose group and high dose group(P<0.05).Compared with I/R group, IL-10 contents kidney tissue and myocardium significantly increased in small dose group (P<0.05).Cr ([167.11 ±18.81, 135.46 ± 9.80, 114.29 ±12.50, 100.15 ±8.81]μmol/L), BUN ([13.42 ±1.25, 11.73 ±1.15, 10.27 ±0.78, 9.28 ±0.52] mmol/L) concentrations in serum and TNF-αcontents in kidney tissue ([578.45 ±30.59, 530.76 ±20.59, 482.23 ±27.12, 423.14 ± 21.16]ng/L) and myocardium ([565.00 ±37.66, 517.82 ±36.89, 469.99 ±32.43, 407.41 ±23.77] ng/L) significantly de-creased in a dose-dependent manner in low , middle and high groups (P<0.05).Microscopic examination showed that the pathological changes of both kidney tissue and myocardium were significantly attenuated in low , middle and high dose group . Conclusion Dexmedetomidine can allenuate myocardial tissue injury induced by renal ischemia -reperfusion in a dose-dependent manner in rats and its mechanism may be involved with the inhibition of inflammatory factors .
6.Inhibition of FK506 on the expression of vascular endothelial growth factor in retinal Müller cells cultured by high concentration glucose
Wei, XIA ; Jing, XIA ; Xiaofeng, ZHANG ; Lei, ZHONG ; Zhengtai, SUN ; Yingming, WANG
Chinese Journal of Experimental Ophthalmology 2013;32(11):998-1003
Background Retinal Muller cells participate in the pathological process of diabetic retinopathy (DR) through expressing vascular endothelial growth factor (VEGF).It is reported that FK506 inhibits the expression of VEGF in solid tumors and experimental corneal neovascularization,but whether FK506 exerts its role on retinal Müller cells or not is still unclear.Objective This study aimed to investigate how FK506 affects the expression vascular endothelial growth factor (VEGF) in rat retinal Müller cells under the condition of high glucose.Methods Immortalized rat retinal Müller cell line was regularly cultivated and logarithmic phase of cells were incubated in 96-well plate with the cell density of 1 × 104/ml.Different concentrations of FK506 (800.00,400.00,200.00,100.00,75.00,50.00,25.00,12.50 and 6.25 pg/ml) (100 μl/well) were added into the culture medium to determine the half maximal inhibitory concentration (IC50) of FK506 by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT).The cell lines were cultured with DMEM medium (containing D-glucose of 5.5 mmol/L) or high glucose DMEM (containing D-glucose of 50 mmol/L),and 75 pg/ml FK506 were added into DMEM,respectively,and the cells were divided into the normal control group,FK506 group,high glucose culture group and high glucose + FK506 group.ELISA was employed to assay the content of VEGF protein in the cell supernatant.The expressions of VEGF mRNA and protein in the cells were detected by reverse transcription PCR (RT-PCR) and Western blot,respectively.Results The cells grew well in the normal control group,FK506 group,high glucose culture group and high glucose+FK506 group in 12,24 and 48 hours after culture with the polygon-like shape.The IC50 of FK506 was 75 pg/ml.The contents of FK506 in the cell supernatant were (966.46± 13.59) pg/ml,(1 059.42±67.43) pg/ml,(16 243.11 ±3 926.38) pg/ml and (9 467.25± 1 525.56) pg/ml in the normal control group,FK506 group,high glucose culture group and high glucose+FK506 group,respectively,showing a significant difference among the four groups (F =20.51,P =0.00).The VEGF levels in cell supernatant were significantly higher in the high glucose group than those of the normal group and the high FK506 group (P =0.00,P =0.02),but no significant difference was found in the VEGF level in cell supernatant between the control group and FK506 group (P =0.08).The expressions of VEGF mRNA and protein in the cells were significantly different among the four groups (F=126.06,P=0.00;F=5.44,P=0.01),and the relative expressing values of VEGF mRNA and protein in the cells of the high glucose group were significantly higher than those of the control group and the high+ FK506 group (all at P<0.01).The relative expressing values of VEGF mRNA and protein were 0.64±0.09 and 0.68±0.18 in the FK506 group,which were lower than those of the normal control group (0.84±0.07 and 0.75± 0.03).However,no significant differences were seen between the two groups (P =0.05,0.07).Conclusions The expression of VEGF in rat retinal Müller cells up-regulates under the high glucose condition.FK506 plays inhibitory effects on VEGF expression to certain extent in vitro.
7.Experimental study on the eyelid reconstruction in situ with the silk fibroin membrane
Lei, ZHONG ; Xiao-feng, ZHANG ; Zheng-tai, SUN ; Jing, XIA ; Ying-ming, WANG
Chinese Journal of Experimental Ophthalmology 2011;29(11):968-972
Background Autologous and allograft renal transplantation exist some disadvantages of less donor source and rejection.As a scaffold of cell in tissue engineering,fibroin was determined to have a good biocompatibility.But whether the fibroin membrane can become a substitution for tissue defect is seldom reported.Objective This experiment aimed to investigate the feasibility of silk fibroin membrane in the rabbit eyelid reconstruction in situ.Methods A 4 mmx3 mm tarsi defect model was created on the upper eyelids of 18 healthy New Zealand white rabbits.The eyelid reconstruction in situ was performed with regenerated silk fibroin membrane material in the right upper eyelids (silk fibroin group ) and allogenic sclera material (sclera group ) on the upper eyelids of fellow eyes.The grafts were clinically examined for the evaluation of inflammation and implant exposure at the first,second and forth week after operation.The inflammation response and collagen distribution were examined by hematoxylin & eosin staining and Masson staining.Expression of basic fibroblast growth factor(bFGF) in the grafts was detected by immunohistochemistry,and ImagePro Plus software was used for statistical analysis.Results All eyelid defects showed a primary healing.The surface of palpebral conjunctival was smooth and the inflammation of ocular surface was mild.The eyelid margin in the sclera group was more notch than that in the silk fibroin group.Results of pathological examination revealed that the arrangement of collagen fibers in the sclera group was more disordered,but that in the silk fibroin group was regular.The expression level(A value) of b-FGF in the operative area in silk fibroin group were 0.027 67±0.004 69,0.051 73±0.008 72,0.058 72±0.006 88,and those in the sclera group were 0.056 48±0.009 14,0.072 83 ± 0.009 17 and 0.078 73 ±0.010 84 in 1,2,4 weeks after operation,showing statistically significant differences between two groups in various time points ( t =- 6.38,t =- 4.99,t =- 2.87,P <0.05 ).Conclusions Silk fibroin membrane can reconstruct the eyelid shape in situ with the less inflammation response and good biocompatibility.Silk fibroin membrane could be used to support the eyelid as a new tarsal repairing materials.
8.Influence of monochromatic light exposure on the growth of Müller cell and the expression of myopia-related cell factors in vitro
Jing, XIA ; Xiao-feng, ZHANG ; Wei, XIA ; Lei, ZHONG ; Zheng-tai, SUN ; Ying-ming, WANG
Chinese Journal of Experimental Ophthalmology 2013;(5):446-451
Background Research showed that exposure of 530 nm monochromatic light can induce myopia in animal,and retinal Müller cells participate in the formation of myopia.However,the effect and mechanism of retinal Müller cells during the formation of monochromatic light induced-myopia is below understood.Objective This study was to investigate biologic characteristics of rat retina Müller cells and the expression of cell factors in Müller cells after being illuminated by the 530 nm monochromatic light,and discuss the role of the retina Müller cells in myopia induced by monochromatic light.Methods Immortalized rat retinal Müller cells were cultured with DMEM containing 10% fetal bovine serum in a self-made cell incubator with monochromatic light by adjusting luminance of 530 nm LED source.The cells were exposed to 125,250 and 500 lx luminance respectively for 6,12 and 24 hours,and the cells without light-irradiation were used as control.The growth of the cells under the different light time and different illuminations was described by MTT as the absorbance at the wavelength 570 nm (A570),and cell cycle analysis of Müller cells was performed by flow cytometry 48 hours after cultured,and the expression of transforming growth factor-beta 1 (TGF-β1),tyrosine hydroxylase(TH),inducible nitric oxide synthase(iNOS) and basic fibroblast growth factor(bFGF)in the cells were detected by reverse transcription PCR(RT-PCR),respectively.Results The Müller cells were uniform in size with polygonal shape and defined edges.No statistically significant difference was found in the A570 value in the cells of the 125 lx and 250 lx illuminated groups compared with the control group in various time points(P>0.05).However,significant lowing was seen in the A570 value in the cells of the 500 lx illuminating for 12 hours and 24 hours in comparison with the control group (P =0.013,0.001).Compared with the control group,the ratio of the number between G2 and G1 phase was not significantly declined in 125 lx,250 lx illuminating for 48 hours (P =0.073,0.330),and the ratio in the 500 lx illuminating group was significantly lower than those in the 250 lx illuminated group and the control group (P =0.028,0.038).RT-PCR revealed that the expression of TGF-β1 mRNA in the cells was higher in the 250 lx illuminated group than that of the 500 lx illuminated group (P=0.006).The expression of iNOS mRNA was gradually upregulated in the 250 lx illuminated group compared with the control group (P =0.001),but that in the 500 lx illuminated group was downregulated (P =0.000).The expression of bFGF mRNA was raised in the 125 lx and 250 lx groups but reduced in the 500 lx group when compared with the control group(P=0.002,0.000,0.005).Also,the expression of TH mRNA was significantly increased in the 250 lx group(P=0.000),but decreased in the 500 lx group(P=0.000,P=0.001).Conclusions The monochromatic light of 530 nm can inhibit the growth of rat Müller cells and downregulate the expression of myopia-related cell factors and therefore exert effect in the formation of myopia.
9.Expression of inflammatory factors in conjunctiva and assessment of dry-eye associated examination in ovariectomized rat
Lei, ZHONG ; Xiaofeng, ZHANG ; Jing, XIA ; Zhengtai, SUN ; Yingming, WANG ; Wei, XIA
Chinese Journal of Experimental Ophthalmology 2014;32(9):807-812
Background Dry eye is increasing gradually recently,but its etiology and manifestation are very diverse.Studies showed that menopause of adult females was one of the risk factors of dry eye.In addition,some inflammatory factors also participate in the pathogenesis and development.But the study on the relationship of sex hormone with inflammation and ocular surface damage is still below understanding.Objective This study was to investigate the expressing changes of interleukin (IL) and tumor necrosis factor-α (TNF-α) in conjunctiva and the manifestation of ocular surface in ovariectomized rat model.Methods Twenty clear female SD rats were randomized into the ovariectomized group and the sham operative group according to randomized number table.Ovariectomy was performed in the ovariectomized group,and abdominal myotomy without ovariectomy was performed in the sham operative group.Serum estrogen and androgen levels were detected by radiation immunoassay 3 months after operation.Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescence staining were carried out in the rats before operation and 1 month,2 and 3 months after operation.The morphology of conjunctival epithelial cells was examined by hematoxylin & eosin staining at the 3rd month after operation.The expressions of IL-17A,IL-1 β,IL-6 and TNF-α in conjunctiva were semi-quantitative analyzed by immunohistochemistry and Western blot.The use and care of the animals complied with State Science and Technology Commission Regulations for the Administration of Affair Concerning Experimental Animals.Results Serum estrogen levels were (23.53 ± 1.65) pg/L and (47.89 ± 1.05) pg/L 3 months after surgery in the ovariectomized group and the sham operative group,respectively; the serum androgen levels were (1.84±0.09) ng/L and (2.47±0.12)ng/L in the ovariectomized group and the sham operative group,respectively,showing a significant decline of serum estrogen and androgen levels in the ovariectomized group compared with the sham operative group (t=-35.37,-12.13,both at P<0.01).No significant differences were seen in S Ⅰ t between the two groups among various time points (Fgroup =0.38,P =0.55 ; Ftime =0.13,P =0.72 ; Finteraction =0.39,P =0.76).No obvious fluorescence staining was seen in the cornea of both the ovariectomized group and the sham operative group.The histopathological examination showed that the layers of rat conjunctival epithelial cells increased with the disordered arrangement in the ovariectomized group.Immunochemistry showed that the expressions of IL-17A,IL-1β,IL-6 and TNF-α (A values) were significantly higher in the ovariectomized group than those in the sham operative group (IL-17A:t=8.22,P<0.01 ;IL-1β:t=16.43,P<0.01 ;IL-6:t=13.44,P<0.01 ;TNF-α:t=16.26,P<0.01).Western blot assay showed the similar results (IL-17A:t=19.41,P<0.01 ;IL-1β:t=12.63,P<0.01 ;IL-6:t=17.17,P<0.01 ;TNF-α:t=15.19,P<0.01).Conclusions Serum estrogen and androgen levels drop obviously,and there is an up-regulation of IL and TNF-α expression in conjunctiva tissue in the ovariectromized SD rats.However,no obvious dry eye-related sign occurs.
10.The curative effect and safety of collagen sponge with different pH and content of protein implanted into orthopedic bone defect.
Gang TONG ; Jing LEI ; Cuihong ZHONG ; Weixin CAI ; Xuewei LIN
Journal of Biomedical Engineering 2012;29(6):1125-1130
This paper presents a 3mm fracture model of radial bone of rabbits using operation method. The bone defect was treated with collagen sponge with different pH and different content. After the operation, the body temperature, blood routine measurement, serum antibody, X-ray examination and histological observation in each group were examined to evaluate and study the curative effect and safety of collagen sponge. Collagen sponge had a good result of safety, but there was slightly change in blood routine, serum antibody, and histological observation, etc, with the pH changing and different content of collagen. The results showed that there was no obvious influence of safety to tissues after treatment of the collagen sponge at different pH implanted into bone defect. Collagen sponge at lower pH could promote the healing of bone defect partly, while the safety of collagen sponge with lower content was better.
Animals
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Collagen Type I
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adverse effects
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chemistry
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therapeutic use
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Hydrogen-Ion Concentration
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Male
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Prostheses and Implants
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Proteins
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analysis
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Rabbits
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Radius Fractures
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surgery