The Diversity Research Of Urinary Protein Of Calcium Oxalate And Calcium Oxalate Mixed Carbonate Apatite Stones

The first part:The diversity research of urinary protein of the acetone methodand freeze-drying method extracting urinary calculi Objective:Discussion acetone precipitation and freeze-dried urine protein extraction of urinary stone patients gel strips difference.Methods:(1)Collect urinary stones male patients and female patients each two cases first morning urine 50 ml, after removal of cell debris repackaging,-80℃ cryopreservation;(2)Urine protein concentration using lyophilization and acetone precipitation method;(3)Urinary protein concentration was measured Bradford;(4)SDS-PAGE comparison between groups of Protein;(5)Lane 1D gel image analysis software to analyze the optical density of both protein gel,SPSS17.0 for data analysis.Result:(1)Acetone method to extract protein concentration in urine of patients urinary tract stones0.92±0.13mg/ml),Lyophilization extract concentration(2.28±0.55mg/ml),The difference was statistically significant(P<0.05);(2)Acetone precipitation method to extract urine of patients with urinary calculi protein gel, about 4 to 7 protein bands, mainly in the 55~95KD, 20~34 KD;extracting protein gel, about 3 to 5 protein bars lyophilization belt, mainly nearby72 KD and 26 KD;(3)Acetone precipitation method to extract urine of patients with urinary calculi protein gels average optical density values(4.52×107±1.31×107),Lyophilized protein gel extraction mean optical density value(3.03×107±0.79×107),The difference was not statistically significant.(1)Lyophilization urine protein concentration is higher than the extracted acetoneprecipitation method, and the difference was statistically significant(P<0.05);(2)Acetone precipitation method to extract the number of urine protein species higher than lyophilization, but the difference was not statistically significant;Conclusion:The second part:The diversity research of urinary protein of calcium oxalateand calcium oxalate mixed carbonate apatite stones Objective:To investigate the urinary system calcium oxalate stones, calcium oxalate stones were mixed carbonate apatite urine protein and urine protein healthy population differences.Methods:(1)Collect healthy adult male and female in 2 cases, calcium oxalate stones male patients and female patients all 6 cases, the mixed carbonate apatite calcium oxalate stones male patients and female patients all 6 cases, the first early morning fasting urine 50 ml, removal after packing frozen cell debris;(2)Each set of experiments to extract a sample of urine protein Acetone precipitation;(3)Comparison with the use of SDS-PAGE protein gel strip difference between the groups;(4)Data Analysis.Result:(1)Men protein concentration in each group: Ca Ox protein concentration 1.91 ±0.46ug/ul, Ca Ox + CH protein concentration 2.04 ± 0.87 ug / ul, protein concentration in normal control group 1.25 ± 0.11 ug / ul; protein concentration difference between the groups was not statistically significance(P <0.05);(2)The protein concentration of women in each group: Ca Ox protein concentration of 1.61 ± 0.65 ug / ul, Ca Ox + CH protein concentration 1.72 ± 0.81 ug /ul, protein concentration in the control group 1.07 ± 0.45 ug / ul; protein concentrationdifference between the groups was not statistically significance(P <0.05);(3)Male urine protein gel photo optical density of calcium oxalate stones were3.20×107±1.25×107, the healthy population of urine protein gel photo optical density0.63 × 107, the difference was statistically significant(P <0.05);(4)female patients with urinary calcium oxalate stones protein gel photo optical density value 2.42×107±1.42×107, the healthy population of urine protein gel photo optical density of 0.41×107, the difference was not statistically significant(P <0.05);(5)male patients with calcium oxalate stones mixed carbonated apatite urine protein gel photo optical density value of 1.96×107±0.71×107, the healthy population of urine protein gel photo optical density of 0.71×107, the difference was not statistically significant(P <0.05);(6)female mixed calcium oxalate stones in the urine of patients carbonated apatite protein gel photo optical density value 4.45×107±0.89×107, the healthy population of urine protein gel photo optical density of 0.84×107, the difference between the two statistically significant(P <0.05).Conclusion:(1) acetone precipitation method to extract calcium oxalate stones, calcium oxalate mixed carbonate apatite and healthy population difference was not statistically significant urinary protein concentration(P <0.05);(2) male urine protein gel optical density of calcium oxalate stones in patients than in healthy populations of urine protein gel optical density values, both statistically significant(P <0.05);(3) female mix carbonated apatite calcium oxalate in urine protein gel photo optical density value is higher than the healthy population of patients with urinary calculi protein gel photo optical density values, the difference was statistically significant(P <0.05).The third part:The quantitative analysis of urinary calcium oxalate and calciumoxalate mixed carbonate apatite stones related to five kinds of urinary protein Objective:Quantitative Comparison of healthy people and calcium oxalate stones, calcium oxalate stones were mixed carbonate apatite urine OPN, THP, NGAL, Albumin andα-1-microglobulin in content.Methods:(1) collect healthy adult men and women of all 10 cases, calcium oxalate stones in male patients(n = 16) and female patients(n = 13), a mixed carbonate apatite calcium oxalate stones in male patients(n = 13) and female patients(n = 16), the first early morning fasting urine 50 ml, after removal of cell debris repackaging,-80 ℃cryopreservation;(2) the concentration of each protein within the Luminex Luminex detection system analysis of samples in each group;(3) Data analysis.Result:1, THP:(1) male group comparison of data between the groups were not significantly different(P <0.05);(2) female groups: normal group(8.46±1.64 ug / ml) contrast Ca Ox group(1.91±0.43 ug / ml) was significantly(P <0.05); normal contrast Ca Ox + CH group(1.98±0.37 ug / ml) was significantly(P <0.05);2, NGAL:(1)Male groups: normal group(63.32 ± 12.23 ng / ml) contrast Ca Ox group(427.79 ± 90.17 ng / ml) was significantly(P <0.05); normal contrast Ca Ox + CH(196.93±38.50 ng / ml) group the difference was statistically significant(P <0.05);(2)Female groups: normal group(30.32 ± 3.92 ng / ml) contrast Ca Ox group(503.82±174.35 ng / ml) was significantly(P <0.05); normal contrast Ca Ox + CH(552.85±156.78 ng / ml) group the difference was statistically significant(P <0.05);3, α-1-microglobulin: comparison of data between men and women in each group had no significant difference(P <0.05).4, Albumin:(1) male groups: normal group(48.17 ± 6.21 ug / ml) contrast Ca Ox group(309.38 ± 33.44 ug / ml) was significantly(P <0.05); normal contrast Ca Ox + CH group(194.20±33.85 ug / ml) was significantly(P <0.05);(2) female groups: normal group(33.55 ± 3.51 ug / ml) contrast Ca Ox group(80.69±8.78 ug / ml) was significantly(P <0.05); normal contrast Ca Ox + CH group(91.72±9.60 ug / ml) was significantly(P <0.05);5, OPN:(1) male groups: normal group(4.65±0.99 ug / ml) contrast Ca Ox group(1.15±0.40 ug / ml) was significantly(P <0.05); normal contrast Ca Ox + CH group(0.90±0.12 ug / ml) was significantly(P <0.05);(2) female groups: normal group(1.82±0.28 ug / ml) contrast Ca Ox group(0.28±0.09 ug / ml) was significantly(P <0.05); normal contrast Ca Ox + CH group(0.15±0.09 ug / ml) was significantly(P <0.05);Conclusion:Compared with the normal people, urinary calcium oxalate and calcium oxalate mixed carbonate apatite the urine of patients with low concentration of THP there was no statistically significant difference(male), NAGL high concentration and high concentration of Albumin, OPN concentration is low, the AMBP concentration difference.