[Combined transgenic inhibition of CaMKII and Ik1 on cardiac remodeling].

This study was aimed to establish an experimental mouse model of combined transgenic inhibition of both multifunctional Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) and inward rectifier potassium current (Ik1), and to observe whether the specific inhibition of both CaMKII and Ik1 can bring about any effects on cardiac remodeling. Mice were divided into 4 groups: wild type (WT), CaMKII inhibited (AC3-I), Ik1 inhibited (Kir2.1-AAA) and combined inhibition of both CaMKII and Ik1 (AC3-I+Kir2.1-AAA). Mice in each group received electrocardiogram (ECG) and echocardiography examination. ECG in the condition of isoproterenol (ISO) injection was also checked. The whole cell patch clamp technique was used to measure Ik1 and the transient outward potassium current (Ito) from enzymatically isolated myocytes of left ventricle. In the condition of basal status, no significant changes of heart rate, PR interval and QRS interval were observed. No mouse showed ventricular arrhythmias in all of the 4 groups. After ISO injection, each group presented no significant ventricular arrhythmias either. The indexes measured by M-mode (motion-mode) and two-dimensional echocardiography had no significant differences among the four groups. Ik1 in AC3-I group was significantly higher than those in other three groups (P < 0.01) because of the results brought about by CaMKII inhibition. Among the latter three groups, both Kir2.1-AAA group and AC3-I+Kir2.1-AAA group had a significant reduced Ik1 compared with that of WT group, which was due to the Ik1 inhibition (P < 0.01). Ito in AC3-I group was higher than that of the other three groups (P < 0.01), but there were no significant differences in Ito among WT, Kir2.1-AAA and AC3-I+Kir2.1-AAA groups. Thus, combined transgenic myocardial CaMKII and Ik1 inhibition eliminated the up-regulation of Ik1 in CaMKII inhibited mice, and had no effects on cardiac remodeling including heart structure and function as well as arrhythmias at the basic and ISO conditions. The results of this study may provide a basis for the further investigation of combined inhibition of CaMKII and Ik1 in pathogenic cardiac remodeling.

[Linkage disequilibrium analysis of -1516, -574 and 4259 single nucleotide polymorphisms in TIM-3 gene].

OBJECTIVE: To investigate the frequencies of -1516,-574 and 4259 single nucleotide polymorphisms (SNPs) of T cells immunoglobulin mucin -3(TIM-3) gene in Hubei population and address the question whether they are in linkage disequilibrium(LD) . METHODS: Genotypes and allele frequencies of TIM-3 gene were examined by allele-specific polymerase chain reaction (AS-PCR) methods in 147 healthy Hubei Han individuals. Hardy-Weinberg equilibrium and Two-point LD analyses and haplotype frequencies were evaluated with Arlequin v3.1 software. RESULTS: The allele frequencies of the 3 SNPs were in agreement with Hardy-Weinberg equilibrium. Minor allelic frequencies of TIM-3 -1516G/T,-574T/G and 4259G/T were 8.5%,1.0% and 2.0%,respectively. The dominant haplotypes comprising the three loci were G-G-G(2.0%),G-G-T(88.4%), T-G-T(8.5%) and G-T-T(1.0%). LD analyses revealed that all of the coefficient of linkage disequilibrium (D') were 1. CONCLUSION: The -1516,-574 and 4259 loci of TIM-3 gene are in complete linkage disequilibrium. Our study has provided population genetic data on TIM-3 gene in Chinese Hubei Han population and a basis for searching immune-mediated disease-related TIM-3 haplotype.

[The relationship between the haplotype of T cells immunoglobulin mucin-3 gene and allergic asthma in the Han population from Hubei province of China].

OBJECTIVE: To investigate the single nucleotide polymorphisms (SNPs) of -1516G/T in the promoter region and 4259G/T in the exon-3 region of the T cells immunoglobulin mucin-3 (TIM-3) and their linkage disequilibrium, and therefore to detect their haplotype relationship with allergic asthma of the Han population from Hubei province of China. METHODS: The two polymorphisms were detected with allelic specific polymerase chain reaction (ASPCR). In the 175 asthmatic subjects and in the 202 healthy controls collected from June, 2004 to October 2007 in the Han population from Hubei province. The genotype and allele frequencies, the D' value between the two SNPs sites, the haplotype and their frequencies were calculated and analyzed, respectively. RESULTS: The genotype frequencies of GG, GT and TT in -1516G/T polymorphism of TIM-3 gene were 82.7% (167/202), 17.3% (35/202), 0 (0/202) respectively in the 202 controls, and 82.9% (145/175), 17.1% (30/175), 0 (0/175) respectively in the 175 asthmatic subjects. The genotype frequencies of GG, GT and TT in 4259G/T polymorphism of TIM-3 gene were 0.5% (1/202), 2.5% (5/202), 97.0% (196/202) respectively in the 202 controls and 0.6% (1/175), 5.7% (10/175), 93.7% (164/175) respectively in the 175 asthmatic subjects. The control group: D' = 1.0, the asthma group D' = 0. 9. The 3 haplotypes were G-G, G-T, T-T in the Han population from Hubei province of China, and their haplotype frequencies were distributed similarly in asthma 3.4% (12/350), 88.0% (308/ 350), 8.6% (30/350) and in the controls 1.7% (7/404), 89.6% (362/404), 8.7% (35/404). None of these differences were statistically significant (chi2 = 2.15, 0.47, 0.003 respectively, all P > 0.05). CONCLUSION: There are strong linkage disequilibrium between the two SNPs sites in TIM-3 gene in Han population from Hubei province, but the haplotypes G-G, G-T and T-T are not associated with asthma susceptibility of this population. We cannot exclude the possibility that the haplotypes of TIM-3 may be associated with asthma susceptibility in other ethnic populations or the susceptibility to other atopic diseases and autoimmunity diseases.

[Study on the association of single nucleotide polymorphisms and haplotypes of GPR154 gene with allergic asthma in Han nationality in Hubei Chinese population].

OBJECTIVE: To investigate whether two polymorphism sites of the G protein-coupled receptor 154 gene (GPR154) are associated with asthma in Han nationality of Hubei province in China. METHODS: The polymorphisms were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 145 cases of allergic asthma and 120 healthy controls. RESULTS: (1)The genotype frequencies of SNP563704 were 0.324 for CC, 0.524 for CT, 0.152 for TT in allergic asthma patients in a Chinese population. The genotype frequencies of SNP522363 were 0.289 for CC, 0.521 for CG, 0.190 for GG in allergic asthma patients in a Chinese population. (2)No significant differences in the distributions of SNP563704 and SNP522363 polymorphisms were found between allergic asthma and healthy control subjects (chi square is 1.880, P> 0.05; chi square is 0.700, P> 0.05, respectively). (3)No significant difference in serum total IgE was found between patients with allergic asthma with different genotypes of SNP563704 and SNP522363 (F is 0.714, P> 0.05; F is 0.083, P> 0.05, respectively). (4) The distribution of frequencies of 4 haplotypes showed significant difference(chi square is 16.50,P< 0.01). The haplotype frequencies of CT and GT were remarkably higher in asthma subjects than those in controls. CONCLUSION: The polymorphisms of SNP563704 and SNP522363 are not associated with allergic asthma in Han nationality in Hubei Chinese population. However, the haplotypes are associated with allergic asthma.

[Study on relationship between polymorphism sites of TIM4 and allergic asthma in the population of Han nationality from Hubei province of China].

OBJECTIVE: To investigate two single nucleotide polymorphism sites of T cell immunoglobulin domain and mucin domain protein-4 (TIM4) and to detect their relationship with allergic asthma in a population of Hans from Hubei province of China. METHODS: The polymorphisms (8570G > A and 11515C > A) were detected with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 145 cases of allergic asthma and 130 healthy controls. The genotype and allele frequencies were calculated and analyzed. RESULTS: The genotype frequencies of GG, GA and AA in 8570G > A polymorphism were 0.985, 0.015 and 0 respectively in the healthy population and 0.931, 0.069 and 0 respectively in the allergic asthma population. There was significant difference in genotype and allele frequencies between the allergic asthma patients and control subjects (P=0. 030, P=0.032). The polymorphism of 11515C>o A was not detected. CONCLUSION: The polymorphism of 8570G > A in TIM4 may be associated with allergic asthma in the population of Han nationality from Hubei province of China.

[The research on the correlation between eotaxin-3 gene polymorphisms and allergic asthma].

OBJECTIVE: To study in the linkage between eotaxin-3 gene polymorphisms and allergic asthma susceptivity, blood plasma IgE or peripheral blood eosinophil in adult population of Han nationality from Hubei province of China. METHODS: Polymerase chain reaction (PCR), single strand conformation polymorphism (SSCP), tetra-primer PCR technique and restriction analysis were applied to identify the single nucleotide polymorphism. RESULTS: The allele frequency of eotaxin-3 +2497 G, total levels of plasma IgE and peripheral blood eosinophil counts revealed the significant difference between control and allergic asthma group, that the P value was 0.011, 0.021 or 0.029 respectively. The allele frequency of eotaxin-3 +77 T and total levels of plasma IgE showed to have no significant difference between control and allergic asthma group, that the P value was 0.824 and 0.473 respectively. However, the peripheral blood eosinophil counts was significantly different between control and allergic asthma group, and the P value was 0.044. CONCLUSION: Single nucleotide polymorphism of eotaxin-3 +2497 is associated with the asthma susceptibility, peripheral eosinophil counts and total levels of plasma IgE in adult population from Hubei province, and polymorphism of +77 is associated with peripheral eosinophil counts.

[Study on relationship between polymorphism sites of TIM-3 and allergic asthma in a population of adult Hans from Hubei province of China].

OBJECTIVE: To investigate two single nucleotide polymorphism sites of the promoter region in T cells immunoglobulin domain and mucin domain protein-3 (TIM-3) and detect their relationship with allergic asthma in a population of adult Hans from Hubei province of China. METHODS: The polymorphisms were detected with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and allelic specific polymerase chain reaction (ASPCR). The genotype and allele frequencies were calculated and analyzed. RESULTS: The genotype frequencies of CC, CT and TT in -1541 C/T polymorphism were 0.961, 0.039 and 0 respectively in the healthy population and were 0.935, 0.065 and 0 respectively in the allergic asthma population. No significant difference in genotype and alleles frequencies was found between the allergic asthma patients and control subjects (P=0.314, P=0.321). The genotype frequencies of GG, GT and TT in -574 T/G polymorphism were 0.992, 0.008 and 0 respectively in the healthy population and were 0.941, 0.059 and 0 respectively in the allergic asthma population. There was significant difference in genotype and allele frequencies between the allergic asthma patients and control subjects (P=0.046, P=0.048). CONCLUSION: There are polymorphism sites of the promoter region in TIM-3 , and one of these sites, the -574 G/T polymorphism site, may be associated with allergic asthma in the population of adult Hans from Hubei province of China.

[Study on the high-affinity IgE receptor beta gene polymorphism and its association with the susceptibility to allergic asthma in Han nationality of Hubei province].

OBJECTIVE: To determine whether 2 polymorphism sites of the high-affinity IgE receptor beta (Fc epsilon RI beta) gene were associated with allergic asthma in Han nationality of Hubei province in China. METHODS: DNA and clinical data were obtained from allergic asthma patients and were compared with those from a group of healthy control subjects. The subjects were genotyped for the -109C/T and a coding variant Glu237Gly in Fc epsilon RI beta gene by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: (1) The genotype frequencies were 0.403 for -109T/T, 0.491 for -109T/C and 0.106 for -109C/C in allergic asthma patients in a Chinese population. No significant difference in the distribution of -109C/T polymorphism was found between allergic asthma patients and healthy control subjects; however, a homozygosity for the -109T allele was associated with increased total plasma IgE levels in patients with allergic asthma (F=7.523, P<0.01). (2) The frequency of Gly237Gly genotype was 0.051 in patient group, and 0.020 in control group. The allele frequencies of Gly237 in the patients and control were 0.236 and 0.136 respectively. There was a significant association between Gly/Gly genotype and allergic asthma. Among allergic asthma patients Gly237Gly was significantly associated with high IgE. CONCLUSION: These data suggested that the Gly237Gly genotype of the Fc epsilon RI beta gene conferred genetic susceptibility to allergic asthma in Chinese, which affected the total plasma IgE levels in the allergic asthma patients. And a homozygosity for the -109T allele was associated with increased total plasma IgE.

[Exploring the possibility of a relationship between polymorphism in TIM-1 and allergic asthma in children of the Hans from Hubei province of China].

OBJECTIVE: To investigate two polymorphism sites of exon 4 in T cells, immunoglobulin domain and mucin domain protein-1 (TIM-1, also human hepatitis A virus cellular receptor-1) and to detect whether they are associated with allergic asthma in children of the Hans in Hubei province of China. METHODS: The ins/del and IVS 8+9 G/A polymorphisms in TIM-1 were detected with polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). The genotypes and alleles frequencies were calculated and analyzed. RESULTS: (1) Two alleles, a wide type del and a variant allele ins were identified in the TIM-1 exon 4. The genotype frequencies of ins/ins, ins/del, and del/del were 0.065,0.326, and 0.608 respectively in the healthy population of the Hans. Another IVS 8/9 G/A polymorphism was also found. The genotype frequencies of A/A, G/A, G/G were 0.022, 0.196 and 0.783, respectively. (2) The genotype frequencies of ins/ins, ins/del, and del/del were 0.045, 0.318, and 0.636 respectively in the allergic asthma population in children of the Hans. No significant difference in ins/del polymorphism was found between allergic asthma patients and control subjects. Another 8/9 IVS G/A polymorphism was also found. The genotype frequencies of A/A, G/A and G/G were 0.009, 0.209 and 0.782 respectively in allergic asthma. No significant difference in IVS G/A polymorphism was found between allergic asthma patients and control subjects. CONCLUSION: The genotype and allele frequencies in the two polymorphism sites in TIM-1 in healthy population of the Hans from Hubei province of China were similar to those in Japanese. The two polymorphism sites of TIM-1 are not associated with allergic asthma in Chinese children.

[Polymorphism of regulated upon activation, normal T cell expressed and secreted promoter region -28 position in Chinese allergic asthmatic children].

OBJECTIVE: To study the effect of regulated upon activation, normal T cell expressed and secreted (RANTES)-28 polymorphism on allergic asthma in Chinese children. METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to identify the polymorphism at position -28 of the RANTES promoter in 100 asthmatic children (group A). Levels of plasma IgE and RANTES were detected by chemiluminescence and ELISA respectively. The number of peripheral eosinophils was counted. Data were compared with those of the control group of 90 healthy children (group B). RESULTS: (1) Both C allele and G allele were detected at -28 of the RANTES promoter; the prevalence of the G allele in group A was 19.5%, as compared to 10.6% in group B; the prevalence of RANTES-28G allele was significantly different between the two groups (P < 0.05). (2) For genotypes C/C, C/G and G/G in asthmatics, levels of plasma RANTES were (289 +/- 199) ng/L, (515 +/- 119) ng/L and (1 071 +/- 138) ng/L, respectively, the difference being significant among the three genotypes (P < 0.01). (3) For genotypes C/C, C/G and G/G in asthmatics, total levels of plasma IgE (lg IgE) were 2.45 +/- 0.12, 2.77 +/- 0.07 and 3.16 +/- 0.09, respectively, the difference being not significant among the three genotypes (P > 0.05). (4) The numbers of peripheral eosinophils were (2.9 +/- 1.4) x 10(8)/L, (6.4 +/- 0.8) x 10(8)/L and (9.9 +/- 2.3) x 10(8)/L in genotypes C/C, C/G and G/G in asthmatics respectively, and the difference being significant among the three genotypes (P < 0.01). CONCLUSION: Polymorphism of RANTES C-28G was associated with susceptibility of asthma in children, and may aggravate the disease through increasing the level of RANTES and eosinophils.

[Correlation between activation-induced cytidine deaminase gene polymorphism and atopic asthma and plasma IgE in adult].

AIM: To explore whether the activation-induced cytidine deaminase(AICDA) gene 8408 C/T polymorphism is related to adult atopic asthma and the level of plasma IgE. METHODS: The polymorphism of AICDA gene was detected by PCR and restriction fragment length polymorphism (PCR-RFLP). RESULTS: The frequencies of 8408T/T genotype had significant difference (P<0.05) between adult asthma patients and control group, while the frequencies of T allele between two groups were not significantly different. The total plasma IgE level in adult atopic asthma patients with 8408T/T genotype was higher than that in the patients with C/C and C/T genotypes(P<0.05). CONCLUSION: The 8408 T/T genotype of AICDA is correlated with atopic asthma and total plasma IgE level in adult.