Relationship between catecholamine level and gene polymorphism of β1 adrenergic receptor G1165C in children with EV71 infection in hand foot and mouth disease

OBJECTIVE@#To investigate the relationship between the levels of plasma adrenaline and norepinephrine and gene polymorphism of β1 adrenergic receptor G1165C in children with enterovirus 71 (EV71) infection in hand foot and mouth disease (HFMD).@*METHODS@#The polymerase chain reaction (PCR) was used to detect the expression of gene polymorphism of β1 adrenergic receptor G1165C in vitro. The levels of plasma adrenaline and norepinephrine were measured by enzyme-linked immunosorbent assay (ELISA).@*RESULTS@#The plasma norepinephrine level of severe group was significantly higher than the mild group in children with EV71 infection in HFMD (P  0.05); There was no significant difference in the distribution of β1 adrenergic receptor G1165C genotype and allele between EV71 infection group and healthy control group (P > 0.05). Further analysis of EV71 infection group by dividing it into mild and severe groups showed that there was no significant difference in the distribution of genotype and allele between these two groups as well (P > 0.05). There was no significant difference in the levels of epinephrine and norepinephrine in different genotypes of EV71 infection group (P > 0.05), and in the levels of plasma epinephrine and norepinephrine in the mild and severe groups (P > 0.05).@*CONCLUSIONS@#As the disease gets worse, the plasma norepinephrine level has a rising trend in children with EV71 infection in HFMD, which is an important indicator to evaluate the progress of the disease. However, the gene polymorphism of β1 adrenergic receptor G1165C have no significant correlation, not only with the susceptibility and severity of EV71 infection in hand, foot and mouth disease, but also with the levels of catecholamine.

Relationship between catecholamine level and gene polymorphism of β1 adrenergic receptor G1165C in children with EV71 infection in hand foot and mouth disease

Objective To investigate the relationship between the levels of plasma adrenaline and norepinephrine and gene polymorphism of β1 adrenergic receptor G1165C in children with enterovirus 71 (EV71) infection in hand foot and mouth disease (HFMD). Methods The polymerase chain reaction (PCR) was used to detect the expression of gene polymorphism of β1 adrenergic receptor G1165C in vitro. The levels of plasma adrenaline and norepinephrine were measured by enzyme-linked immunosorbent assay (ELISA). Results The plasma norepinephrine level of severe group was significantly higher than the mild group in children with EV71 infection in HFMD (P 0.05); There was no significant difference in the distribution of β1 adrenergic receptor G1165C genotype and allele between EV71 infection group and healthy control group (P > 0.05). Further analysis of EV71 infection group by dividing it into mild and severe groups showed that there was no significant difference in the distribution of genotype and allele between these two groups as well (P > 0.05). There was no significant difference in the levels of epinephrine and norepinephrine in different genotypes of EV71 infection group (P > 0.05), and in the levels of plasma epinephrine and norepinephrine in the mild and severe groups (P > 0.05). Conclusions As the disease gets worse, the plasma norepinephrine level has a rising trend in children with EV71 infection in HFMD, which is an important indicator to evaluate the progress of the disease. However, the gene polymorphism of β1 adrenergic receptor G1165C have no significant correlation, not only with the susceptibility and severity of EV71 infection in hand, foot and mouth disease, but also with the levels of catecholamine.

[Effect of UBE2C overexpression on the proliferation of 293T cell line].

AIM: To establish a 293T cell line with stable expression of UBE2C and to investigate the effect of UBE2C overexpression on the proliferation of 293T cells. METHODS: Recombinant plasmid pcDNA3.1(-)/UBE2C successfully constructed in previous time was transfected into 293T cells by lipofectamine 2000. After screened with G418 for 4 weeks, Western blot was used to detect the protein level of UBE2C and MTT assay to detect the proliferation of 293T cells. RESULTS: Compared with pcDNA3.1(-) transfected group, Western blot analysis demonstrated that the level of UBE2C was significantly increased in pcDNA3.1(-)/UBE2C transfected group (P < 0.05). MTT assay showed that 293T cells overexpressing UBE2C proliferated faster than control 293T cells. CONCLUSION: A 293T cell line with UBE2C stable expression is successfully established and UBE2C promoted the proliferation of 293T cells.

[Construction of recombinant eukaryotic expression vector pDsRed1-C3/LOC51255 and subcellular localization of LOC51255 protein in HePG2 cell line].

AIM: To construct the recombinant eukaryotic expression vector pDsRed1-C3/LOC51255 and investigate the subcellular localization of LOC51255 protein in HePG2 cell line. METHODS: The cDNA of LOC51255 was amplified from the recombinant plasmid pET28b/LOC51255 by PCR. The aim gene fragment LOC51255 from pMD18-T/LOC51255 was subcloned into eukaryotic expression vector pDsRed1-C3. The recombinant plasmid pDsRed1-C3/LOC51255 was identificated by BamH I/Xho I double digestion and sequence analysis, and then transfected into HePG2 cell line by lipofectamine 2000. The expression of LOC51255 protein and its subcellular localization were detected by fluorescence microscope. RESULTS: The construction of the recombinant plasmid pDsRed1-C3/LOC51255 was proved by restriction enzyme digestion analysis and DNA sequencing. Its red fluorescent protein was mainly detected in the cytoplasm of HePG2 cell line. CONCLUSION: The eukaryotic expression plasmid pDsRed1-C3/LOC51255 has been successfully constructed and expressed in the HepG2 cell line. LOC51255 protein has been proved to be located in the cytoplasm of HePG2 cell line. Our research will help further studies on the function of human gene LOC51255.