The Investigation Association Between Protein S100A8 And The Urinary Calcium Oxalate Stone

Objective: To explore the role of protein S100A8 in calcium oxalate stone formation by observing the different concentrations of calcium oxalate stones in the urine between patients and the normal.And contrast the different concentrations of protein S100A8 in the urine between patients and the normal.Constructing calcium oxalate crystal model in vitro,and then detect inhibition of growth index of calcium oxalate monohydrate crystal with different concentrations of protein S100A8 in vitro on a kind of crystal technology.Detect the crystallization induction time,nucleation and coalescence rate.Observing the effect of different concentrations of protein in S100A8 to calcium oxalate stone formation types?COM and COD?in vitro.Methods:?1?Gathering specimens of 50 cases with ureteroscopy and percutaneous nephroscope lithotomy and postoperative surgical calculi ingredient analysis from September 2014 to September 2015.collecting Morning urine specimen phase of 50 cases from calcium oxalate stones patients before operation.And urine samples of 50 cases from normal person?not calculi disease?.Determination protein S100A8 concentration of specimens between patients with urinary stones and normal in urine sample separately using the way of ELISA.?2?Building several groups of stone crystal model in vitro with the kind of crystal technology,and setting up several groups of different concentration gradient of S100A8 protein,and add them to the above model,then protein S100A8 will react with calculus model fully mixed,to detect the absorbance and the concentration of Ca2+ in the model with atomic absorption spectrophotometer.?3?Atomic absorption spectrophotometer were used to detect the calcium oxalate solution in different concentration S100A8 protein absorbance value,and nucleation aggregation curve drawing,calculation of calcium oxalate crystal growth induction time,nucleation rate and accumulation rate.?4?Adding different concentrations of protein S100A8 to the calcium oxalatemodel built in vitro,Using SEM to observe formation and the proportion of different crystal of calcium oxalate crystal type?COM and COD?.?5?To show the statistical data with x?±s,and process statistical with the software SPSS19.0,then do statistical analysis.Using two independent samples test statistical analysis to compare two groups of data,using statistical analysis and comparison of the single factor analysis of variance of three sets of data.Results:?1?Protein S100A8 is contained in calcium oxalate stone specimens,and its density range is: 6.231 ±1.251 ng/m L.?2?Protein S100A8 exist calcium oxalate stones patients in the urine?not calculi disease?and normal urine specimens,but protein S100A8 concentration range in the patients with calcium oxalate stones urine specimens as follows: 56.258±0.229 ng/m L,and S100A8 and protein in normal person?not calculi disease?in urine sample concentration range is: 13.589± 0.236 ng/m L.The experimental group compared with normal group,the difference has statistically significant?P < 0.05?.We can see that calcium oxalate stones protein S100A8 concentration in urine sample of patients was obviously higher than that in normal urine specimen.?3?vitro experiments show that inhibiting effect is increasing from different concentrations of protein S100A8 on calcium oxalate crystals with the increase of protein concentration,in vitro.Its inhibition for the growth of calcium oxalate crystal decreased,moreover,differences between groups have statistical significance?P < 0.05?.?4?By spectrophotometer will be between groups with different concentrations of S100A8 protein calcium oxalate solution for testing.To calculate crystal maximum induction time Tmax,nucleation rate SN and aggregation rate SA parameter value,we find that when we add different concentration S100A8 protein,T_max,S_N and S_A are increased,and with the increase of protein concentration,T_max decreases,and S_N and S_A increased gradually.?5?The cross section for the hexagonal crystal and double rectangular pyramid of crystal are present in calcium oxalate stone in S100A8 protein intervention model,But the proportion in the stone of crystallization have difference.With the increase ofS100A8 protein concentration,the proportion of the cross section for the hexagonal crystal gradually increase,and the proportion of double rectangular pyramid of crystal decrease.Conclusions:?1?Urinary calcium oxalate stone contains a certain amount of Protein S100A8.?2?Calcium oxalate stones in patients and healthy people in urine?people without calculi disease?are contain S100A8 protein in the urine specimen,but its concentration is different,and calcium oxalate stones patients urine S100A8 protein concentration is much higher than healthy people.?3?Protein S100A8 can promote effectively urinary calcium oxalate stone formation in vitro.and with higher protein concentration of protein S100A8,its inhibition index decreased.?4?Crystal type is COD formed by the low concentration protein S100A8 intervention,and easy to deposition,are difficult to drain out.However,calcium oxalate crystal types for COM are formed by high concentrations of protein S100A8 intervention,not easy to drain out.?5?The protein S100A8 expression overly in calculi specimens of patients with calcium oxalate stones,and protein S100A8 concentration increases in urine,they both may be important factors of calcium oxalate stone formation.?6?Crystal technology in vitro can effectively build calcium oxalate calculus model,and it can test protein molecules for the function in the process of stone formation.