Objective To investigate the relation between a set of single nucleotide polymorphisms (SNP) in core gene of HBV in chronic hepatitis B patients and HBV DNA levels. Methods PCR restriction fragment length polymorphism(PCR-RFLP) assay and restriction enzyme Tsp509I were adopted to determine HBV SNP in HBV core gene. Nucleotide sequences of core gene were determined using the dideoxy chain termination method. HBV DNA levels were quantitated with real-time PCR. Results Five typical RFLP patterns, RFLP-C, RFLP-D, RFLP-E, RFLP-G and RFLP-C/G mixture were found and the distribution of HBV RFLP patterns was as follows: C, 61. 5% ; D, 2. 6% ; E, 9.6%; G, 16.7%; C/G mixture, 9.6%. Five SNPs, A165T, A336C, A336T, T337C and T385C, were found to be associated with RFLP patterns change and only SNP A336C or A336T caused the substitution of Glu-83 with Asp in HBcAg. The serum HBV DNA levels in RFLP-C group were higher than that in RFLP-G (P =0. 02) and RFLP-C/G group(P = 0. 006) , respectively, furthermore, the positive rate of serum ALT in RFLP-C/G group was lower than that in RFLP-C, RFLP-E and RFLP-G group, respectively. Under the condition of HBeAg-positive, the serum HBV DNA levels in RFLP-C group were higher than that in RFLP-G (P = 0. 015) and RFLP-C/G group(P =0.008) , respectively. Conclusion PCR-RFLP used in this study can be adopted to determine HBV SNPs, not genotypes in Chinese patients with chronic hepatitis B. The serum HBV DNA level in RFLP-C group higher than that in RFLP-G or RFLP-C/G group maybe associated with amino acid mutation, Glu83 Asp.

ObjectiveTo explore the expression of MRP1 and CD34 in colorectal carcinoma tissue and the relationship with clinicopathological factors.MethodsImmunohistochemical streptavidin-perosidase method was used to examine the expression of MRP1 and CD34 in 53 cases with colorectal carcinoma and normal colorectal tissue.The correlation between the levels of MRP1and CD34 expression and clinicopathological factors were analyzed.ResultsThe positive expression rates of MRP1 in the carcinoma group and normal colorectal tissue group were 49.1％ and 15.1％ respectively,and there was a significant difference of the positive expression between the two groups( x2 =14.029,P ＜ 0.01 ).The expression of MRP1 had no correlation with the degree of differentiation,the depth of invasion,the metastasis of lymph node and all the other clinicopathological factors ( P ＞ 0.05 ).CD34 value in the carcinoma group and normal colorectal tissue group were ( 35.63 ± 12.23 ) MVD/HP and ( 6.12 ± 0.97) MVD/HP,respectively,and there was a significant difference between the two groups (t =17.565,P ＜ 0.01 ).CD34 was not correlated with age,sex,tumor size,localization of the primary tumor ( P ＞ 0.05 ),but correlated with Dukes staging,lymph node metastasis,differentiation of the tumor,depth of invasion( all P ＜ 0.05).ConclusionThe overexpression of MRP1 and CD34 protein may involve in colorectal carcinogenesis;MRP1 may involve in the primary multidrug resistance in colorectal carcinoma.; CD34 may involve in the colorectal carcinoma invasion and metastasis.Investigating the expression of MRP1 and CD34 in colorectal carcinoma simultaneously can provide new referential indexes for the treatment and prognosis of colorectal carcinoma.