| Objective1. To study a most suitsble pH value for preventing the form of urate calculi and promoting the drainage of urate in urine by observing the changing in shape of monosodium urate.2. The urate crystal appeared in urine is a marker of uric acid in super-saturation. It is very important to detect urate crystal in urine of Gout patient for diagnosis and assessment of Gout.3. To observe the effect of fructose in different concentrations on the expression of hURAT1 gene in HK-2 cells.Method1. Crystal was collected from gout tophi. It had confirmed that are monosodium urate crystals by polarizing microscope. This crystal 200 ul was put in tub. The crystal was diluted by using pH 5.5 PBS buffer to be 8mg/L in concentration and put it in 37℃for 30 minutes. And then the crystal in shape was observed by polarizing microscope. The same method were used by using pH 5.7,6.1,6.5,7.2,7.6 and 8.0 to dilute crystal. The crystal in shape were observed in six different pH value. All images of crystal in shape were saved by Leica Application Suite software and analyzed.2.56 cases with Gout without nephrolithiasis and 23 health control were selected. Every one was collected 50ml morning urine from testing. All urine were done Centrafugation. The sediment was observed urate crystal by polarize-ng microscope.3. HK-2 cells were divided into control(cultured with DMEM/F12),F1 (cultured with DMEM/F12,100μmol fructose),F2 (cultured with DMEM/F12, 500μmol fructose)groups. In each group,8 bottles of the cells were cultured in vitro for 48 hours. Then the relative quantity of hURAT1mRNA in the HK-2 cells was detected by real-time fluorescent quantitative PCR assay.Results1. The urate crystal in pH 5.5-6.1 was needle-like and appeared a congregating phenomenon. Along with the pH value enhanced of buffer, the urate crystal in shape was going to small and seperated in pH value 6.5-7.6 liquids. It was in PH 8.0, the crystal was become granule or dessolved.2.54 of 56 cases were detected urate crystal (96.43%). This crystal is needle-like or stars-like. And two of 23 health control were also detected (8.7%). This crystal is stars-like. It is significative difference in statistics that positive rate of urate crystal in two group (P<0.05)3. hURAT1 mRNA could be detected in all the samples. And the level of hURAT1mRNA in the control group was significantly lower than that in the other groups. The quantity of hURAT1mRNA in the F1 group was 146.6% of that in the.control group, and that in the F2 group was 178.5% of that in the control group.Conclusion1. The urate crystal is not to be conglomeration in a trending alkalescent environment. It may benefit to prevent the uric acid nephrolithiasis forming in urine and to promote the uric acid draining from urine for gout patient same time to maintain the renal condition in physiology, the pH value 7.0 of urine is regulated.2. Although patient with Gout has no nephrolithiasis, the urate crystal detected in urine has a important clinical significance for Gout diagnosis. The urate crystal in urine appears uric acid is in supersaturation and it is risk factor to form urate calculi and make lesion in kidney. It also guide how to reasonable choose a low uric acid agent. In order to prevent nephrolithiasis formed in kidney, the uricosuric aengt is unreasonable for positive patient of urate crystal in urme.3. Fructose can up-regulate the expression of hURAT1 gene.
|
PDF Full Text Request