| OBJECTIVEThe part one was to investigate the distributions and differences of Nm23 genotype and phenotype by analyzing the erythrocytic NDPK activity, the Nm23 m RNA expression, as well as the genotypes and alleles of one SNPs(rs2302254C/T) in the promoter region and three SNPs(rs34214448G/T, rs11868380C/G, rs2318785G/A) in intron area in healthy Chinese Hans populations and patients after solid organ transplantation, which including renal transplantation and other types, in Hubei province. Finally, to evaluate the correlations among Nm23 gene polymorphism, m RNA expression level and human erythrocytic NDPK activity. The part two was to investigate the associations among Nm23 genotype, phenotype and some biochemical indexes in renal transplantation population in Hubei province.METHODS(1) Correlations of Nm23 Genotype-Phenotype in patients after solid organ transplantation in Hubei Province:In this case-control study, 1000 healthy subjects and 604 solid organ recipients(314 renal transplantations and 290 other types) with immunosuppressant treatment in Tongji and Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, P.R. China were enrolled as DNA sample donors, from February to March 2014. The 4 SNPs of Nm23 gene were genotyped by real time Taq Man-MGB genotyping assay and validated by direct gene sequencing. According to certain principles, part of the samples whose genotypes had been texted were chosen for determination of m RNA expression level and NDPK activity level. The fluorescence quantitative PCR assay was used for determination of Nm23 m RNA expression. The HPLC method was used for determination of NDPK activity level. The statistical package SPSS version 19.0 was used for the analysis of genotypic and allelic distributions and differences as well as the correlation between genotype and penotype.(2) Correlations of Nm23 tag SNPs genotype-phenotype with biochemical indexes in renal transplant patients in Hubei Province: 205 renal transplantation recipients with immunosuppressant treatment in Wuhan General Hospital of Guangzhou Military, Wuhan, P.R. China were enrolled as DNA sample donors, from December 2012 to November 2013. All of the subjects had complete clinical data, including name, gender, age, smoking history, drinking history, history of hypertension and history of diabetes, blood routine data and blood chemistry data. Finally, to evaluate the correlations between Nm23 gene polymorphism, human erythrocytic NDPK activity and biochemical Indexes.RESULTS(1) The frequencies of rs34214448 TT and GT genotypes were higher in control group than that in kidney transplantation group when compared to GG genotypes(Padj= 0.039, adjusted OR=0.760, 95% CI=0.585-0.987). Nm23 polymorphisms in rs2302254, rs2318785 and rs11868380 exhibited no significant differences between kidney transplant group and control group. There were no significant differences of these four SNPs in the allele frequencies and genotype distributions between non-renal transplantation group and control group.(2) There were no statistically significant differences of Nm23 m RNA expression among the healthy group, renal transplantation group and non-renal transplantation group. rs2302254 CC genotype was associated with lower m RNA level when compared to CT and TT genotypes(P < 0.001). rs2318785 AA genotype was associated with lower m RNA level when compared to AG and GG genotypes(P<0.001). rs34214448 GG genotype was associated with lower m RNA level when compared to GT and TT genotypes(P<0.001). The analysis did not reveal any significant association between the expression level of Nm23 and rs11868380 SNP(P=0.319).(3) There were significant differences in NDPK activity levels of the healthy group, renal transplantation group and non-renal transplantation group. NDPK activity levels in healthy subjects were significantly higher than those in non-renal recipients(P=0.005). And NDPK activity levels in non-renal recipients were significantly higher than those in renal recipients(P < 0.001). There were no significant correlation between the genotypes of these four genotypes and NDPK activity level. No significant correlation between Nm23 m RNA expression and NDPK activity level was observed as well.(4) The associations among the four selected SNPs, NDPK activity and some biochemical indexes(TB(total bilirubin), DB(direct bilirubin), AST(aspartate aminotransferase), ALT(alanine aminotransferase), TP(total protein), A(albumin), G(globulin), A/G(albumin/globulin), BUN(blood urea nitrogen), UA(uric acid), Cr(creatinine), Glu(FPG), CO2CP(carbon dioxide combining power), e GFR(estimated glomerular filtration rate), WBC(white blood cells), RBC(red blood cells), PLT(platelets), Hb(Hemoglobin), HCT(hematocrit)) were not significant.CONCLUSIONS(1) Rs34214448 G allele may increase the risk of renal parenchymal disease or uremia resulted from chronic renal failure(the major indications for kidney transplantation). Nm23 genetic polymorphisms can potentially affect Nm23 m RNA expression level.(2) Statistically significant differences were found in Nm23 m RNA expression levels among different genotypes of rs2302254, rs2318785 and rs34214448 in the analysis of genotype-phenotype correlations. Rs2302254-CC, rs2318785-AA and rs34214448-GG may associated with lower Nm23 m RNA expression levels when compared to rs2302254-CT/TT, rs2318785-GG/GA and rs34214448-GT/TT.(3) There were significant differences in NDPK activity levels of the healthy group, renal transplantation group and non-renal transplantation group. NDPK activity levels in healthy subjects were significantly higher than those in non-renal recipients(P=0.005). And NDPK activity levels in non-renal recipients were significantly higher than those in renal recipients(P<0.001). NDPK activity may be correlated with physical condition.(4) There were no significant correlations between Nm23 tag SNPs genotype-phenotype and biochemical indicators, while further research is needed for ensurement. |
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