Effect of enhanced green fluorescent protein fusion on Ano1 physiological feature / 生理学报

The aim of the present study was to investigate whether the physiological features of Ano1 were affected by enhanced green fluorescent protein (EGFP) fusing at Ano1 C-terminal. The eukaryotic expression vectors of Ano1 and EGFP-Ano1 were constructed, and these plasmids were transfected into Fischer rat thyroid follicular epithelial (FRT) cells using liposome. The expression and location of Ano1 were examined by using inverted fluorescence microscope. The ability of Ano1 to transport iodide was detected by kinetics experiment of fluorescence quenching. The results showed that both Ano1 and EGFP-Ano1 were expressed on FRT cell membrane and could be activated by Ca(2+). There was no significant difference of the ability to transport iodide between Ano1 and EGFP-Ano1. These results suggest Ano1 and EGFP-Ano1 have similar physiological feature.

Inhibition of rapamycin on proliferation of human acute promyelocytic leukemia HL60 cells / 中国药学杂志

OBJECTIVE: To investigate the effect of rapamycin on proliferation, autophagy and apoptosis of human acute promyelocytic leukemia HL60 cells. METHODS: The inhibitory effect of rapamycin on HL60 cell was detected by MTT assay. The expressions of LC3-II autophagy-related protein were determined by Western blot. The cell cycles were analyzed by flow cytometry (FCM). Apoptosis were detected by DNA agarose gel electrophoresis. RESULTS: After treatment with rapamycin of 0, 1, 5, 10, 20, 40 nmol · L-1 for 24 h, proliferation of HL60 cells were inhibited in a dose-dependent manner (r=0.97, P < 0.05). Western-blot shows that the expression of LC3-II protein of each concentration group was higher than that of the control group (P < 0.01). Rapamycin has a role in promoting cell autophagy. Compared with control group, more cells were arrested at G0/G1 phases (P < 0.05) and fewer cells were at S phases (P < 0.05). Agarose gel electrophoresis shows no apoptosis DNA fragments. CONCLUSION: In given concentration range, rapamycin can inhibit the proliferation of human acute promyelocytic leukemia HL60 cells by promoting autophagy rather than inducing apoptosis.

Analysis of vWF gene A1381T polymorphism in patients with coronary heart disease / 中国实验血液学杂志

This study was purposed to investigate the vWF gene A1381T polymorphism in patients with coronary heart disease (CHD). A case-control study was designed, including 104 continuously hospitalized patients with CHD, aging from 40 to 75 years (average 59) and 96 persons underwent physical examination in outpatient department as controls, aging from 39 to 70 years (average 56). The plasma vWF: Ag level of CHD patients and control persons was detected by ILISA. vWF gene A1381T polymorphism was analyzed by the polymerase chain reaction-restriction fragment length polymorphism and sequencing when it is necessary. The data were grouped by gender, blood group and/or genotype in CHD group and control groups. The difference of plasma vWF level between male and female was analyzed by independent sample t test; one way ANOVA was used to analyze the difference of vWF level between different blood group genotypes, while the factorial design ANOVA was used to test the difference of vWF level in plasma between A1381T genotype and/or ABO blood groups. χ(2) Crosstabs were used to test the CHD susceptibility. The results showed that the frequencies of GG genotype (wild type) of vWF gene A1381T polymorphism were 62.5% in CHD group and 67.7% in control group, and the frequencies of AG genotype (heterozygous variant) were 37.5% in CHD group and 32.3% in control group. χ(2) Crosstabs showed no significant correlation between vWF gene A1381T polymorphism (AG) and CHD (OR = 1.258, 95% CI = 0.702 - 2.255, χ(2) = 0.595, p = 0.440). The plasma vWF level in CHD group was statistically very higher than that in control group (p < 0.001), even though the relationship of vWF A1381T polymorphism (rs216311) and susceptibility of CHD in CHD group was not found. The plasma vWF level of AG or GG genotype was higher in CHD group than in control group (p < 0.001). The plasma vWF level of AG genotype was higher than that of GG in CHD group (p < 0.05), but not in control group. The plasma vWF of O blood group was lower than that of A, B and AB blood groups (p < 0.05), while among A, B, AB blood groups, the vWF level was not different (p > 0.05). Among O, A, B, AB blood groups in CHD group, vWF level was not different (p > 0.05). Although the two-way analysis of variance ANOVA showed no interaction of A1381T genotype and ABO blood groups on plasma vWF level, the plasma vWF level in AG mutant of vWF A1381T gene polymorphism with O blood group was higher than that of GG mutant (p = 0.023) in CHD group, not different in other blood groups. It is concluded that there is no association between vWF gene A1381T polymorphism and CHD susceptibility. The plasma vWF level in CHD group interrelated with ABO blood group and A1381T polymorphism, in which the plasma vWF level in AG genotype increase mostly. Plasma vWF level in vWF gene A1381T polymorphism with AG mutant was significantly much higher than GG mutant in CHD. This change may be beneficial to further study the effect of A1381T polymorphism on vWF gene expression and activity.