| BackgroundWith the improvement of people's living standards and lifestyle changes,the prevalence of hyperuricemia(HUA)is increasing year by year.The prevalence of HUA in coastal and economically developed areas in China is between 10% and 20%.A cross-sectional study of 24 hospitals in Jiangsu,Zhejiang and Anhui provinces between 2010 and 2015 found that the rate of HUA in Jiangsu was 14.84%.The cause of elevated uric acid(UA)may be related to renal dysfunction caused by treatment during radiotherapy and chemotherapy,malignant tumor treatment is often accompanied by HUA.Bone metastasis is the metastasis of extramedullary malignant tumors to bone tissue,which occurs mostly in the middle and late stages of cancer.As treatments continue to evolve,patient survival is significantly prolonged,and opportunities for bone metastases are significantly increased.Early detection of bone metastasis is important to prevent and avoid inappropriate treatment and improve the quality of life of patients.Current tests for bone metastases include radionuclide bone scans,X-rays,and CT,but the problem is poor specificity.Serological testing has the advantages of simple operation and suitable for early and continuous monitoring,and is widely favored by clinicians and researchers.Serological markers related to bone metabolism mainly include bone alkaline phosphatase(BALP),tartrate-resistant phosphatase 5b(TRACP 5b)and the like.In the 2014 expert consensus on bone metastasis of lung cancer,the serum marker BALP,which reflects the level of osteogenesis metabolism,was specifically mentioned and its application value in the diagnosis and detection of bone metastasis progress.However,it is worth noting that the detection concentration of BALP is often underestimated in the routine testing of BALP clinical specimens.A similar problem has occurred in studies where the serum marker TRACP 5b is interfered by HUA.TRACP 5b is a serum marker that monitors bone resorption metabolism.Under the HUA conditions,TRACP 5b results are falsely reduced.However,the correlation between HUA and BALP results has not been reported.This study attempts to explore the practical problems and related mechanisms of clinical findings.Methods The sample was collected from patients who were admitted to the hospital from August 2014 to February 2016.The exclusion criteria were cancer,hepatitis,renal insufficiency and inflammatory diseases.The uric acid in vitro interference experiment was carried out: BALP standard and bone transfer patient serum were used to detect the measured concentrations of BALP under different dilutions of uric acid.The uric acid concentration in vitro was observed to have any effect on the BALP test results.The human osteosarcoma cell line(Saos-2)and the human embryonic kidney 293 cell line(HEK293)were cultured using cell culture techniques,all of which were obtained from a US standard strain library,harvested by trypsinization,and subcultured twice a week.Construction of BALP Recombinant Plasmid Technology: Design of forward and reverse primers using Olie 7.0 software,PCR technique using Prime STAR? HSDNA Polymerase to amplify BALP fragments(relative to transcription start site-473 to +154bp),Kpn I and Bgl II The digestive enzyme was digested and recombined into the promoterless vector p GL3-Basic,and the recombinant plasmid was sequenced to verify that the positive clone was successfully obtained.HEK293 cells were then transiently transfected with the recombinant plasmid.Finally,the luciferase reporter assay was used to determine the inhibitory effect of HUA on the activity of the BALP promoter region.Results 1)The levels of BALP in the HUA group were significantly lower than those in the healthy control group,and were not related to gender.2)There was no statistically significant difference in the actual detection results of BALP under different UA concentrations in vitro.3)Comparing after hemodialysis in patients caused by hyperuremic academia with before hemodialysis: the concentration of UA decreased,but concentration of BALP increased at the same time.4)UA inhibited the expression of m RNA and protein levels of BALP in Saos-2 cells.5)BALP promoter activity was inhibited by high UA.ConclusionsThere was an inverse correlation between UA levels and BALP levels in HUA patients.High uric acid interferes with BALP expression levels,possibly by inhibiting BALP promoter activity and affecting its expression level. |
PDF Full Text Request