Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assist...Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assisted methods are important approaches for RNA direct detection,but its specificity will be limited when the fidelity of ligases is not ideal.The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection.Methods In this study,a dualcompetitive-padlock-probe(DCPLP)assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation.To verify the method,we employed dual competitive padlock probe-mediated rolling circle amplification(DCPLP-RCA)to genotype the CYP2C9 gene.Results The specificity was well improved through the competition and strand displacement of dual padlock probe,with an 83.26%reduction in nonspecific signal.By detecting synthetic RNA samples,the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L.Furthermore,clinical samples were applied to the method to evaluate its performance,and the genotyping results were consistent with those obtained using the qPCR method.Conclusion This study has successfully established a highly specific direct RNA SNP detection method,and provided a novel avenue for accurate identification of various types of RNAs.展开更多
The relationship of three Brachionus calyciflorus morphotypes (two short-spined, single-spined and spineless ones) was studied in the laboratory. Newly hatched females of each morphotype were cultured individually i...The relationship of three Brachionus calyciflorus morphotypes (two short-spined, single-spined and spineless ones) was studied in the laboratory. Newly hatched females of each morphotype were cultured individually in an inorganic or Asplanchna-conditioned mediums, and the morphotype of their offspring were checked at 200 x magnification. Females of each morphotype, cultured in the inorganic medium, could produce offspring of these three morphotypes in a single generation. Females of all three morphotyes could respond to specific Asplanchna-substances when they were cultured in Asplanchna-conditioned medium. Moreover, we observed that these three morphotype females could coexist in the medium and had three types of egg-bearing females. Based on the above observations we suggest that B. calyciflorus was a morphotype complex and has three basic morphotypes.展开更多
Objective To investigate the relationship of four single nucleotide polymorphism (SNP) haplotypes in the angiotensinogen (AGT) gene to the primary hypertension with or without cerebral infarction in the Li nationa...Objective To investigate the relationship of four single nucleotide polymorphism (SNP) haplotypes in the angiotensinogen (AGT) gene to the primary hypertension with or without cerebral infarction in the Li nationality of Hainan, China. Methods Total 300 subjects were allocated into three different groups: Groupl, 100 patients who have primary hypertension; Group 2, 100 patients who have primary hypertension with cerebral infarction; and control group, 100 healthy individuals. The genotypes of all subjects were determined by PCR-sequencing to analyze the four poly- morphisms at position - 152 (G-A), -20 (A-C), - 18 (C-T), and -6 (A-G) in the promoter region of AGT. Results The frequen- cies ofCT genotype of AGT-18 and T allele in Group 1 (P = 0.003, P = 0.004) and Group 2 (P = 0.002, P = 0.002) were both significantly higher than in healthy controls. The frequency of G allele of AGT-6 was significantly higher in Group 2 than in the control group (P = 0.016), while there is no significant difference between Group 1 and the control. Haplotype analysis revealed that H6 haplotype frequency which included -20C and -6G was significantly increased in Group 2 (P = 0.003) compared with the control group, while H5 haplotype frequency which included -20C and -18T was signifi- cantly increased in Group 1 (P = 0.006) versus the control. Conclusion The -20 (A-C) and - 18 (C-T) of the AGT may play an important role in pathogenesis of primary hypertension; and -20 (A-C), -18 (C-T), and -6 (A-G) may be the genetic risk factors for the onset of primary hypertension with cerebral infarction in the Li nationality of Halnan, China.展开更多
In the present study with Tan sheep, small-tailed Han sheep, Hu sheep, Tong sheep, and Wadi sheep, we detected the distribution of gene frequency of several microsatellite sites in different chromosomes, the result sh...In the present study with Tan sheep, small-tailed Han sheep, Hu sheep, Tong sheep, and Wadi sheep, we detected the distribution of gene frequency of several microsatellite sites in different chromosomes, the result showed that: 1) Hu sheep was in the status of Hardy-Weinberg extreme disequilibrium (P 〈 0.01), while populations including Tong sheep, small-tailed Han sheep, Tan sheep, and Wadi sheep were in Hardy-Weinberg equilibrium (P 〉 0.05). 2) Variance analysis of the heterozygosity and poly- morphic information content at rnicrosatellite makers showed that there were not significant differences among populations as to heterozygosity and PIC (P 〉 0.05), as to effective number of alleles there were not significant differences both among Tan sheep, Hu sheep, Tong sheep, and Wadi sheep, and between Wadi sheep and small-tailed sheep (P 〉 0.05), but between the former three populations and the latter two populations, there were significant differences (0.01〈 P 〈0.05). The variation levels of small-tailed Han sheep was the highest in the five populations based on microsatellite maker data, subsequently followed by Wadi sheep, Tong sheep, Tan sheep, and then Hu sheep. 3) The phylogenetic relationships of the five sheep populations in this study did not meet the mechanism of isolation by distance, and the genetic differentiation relationships among five sheep populations were not closely linearly correlative with their geography distribution. Our findings supported related records in literature: The five populations originated on different time stage from the primogenitor population and communicated genetically with each other thereafter in the process of natural and artificial selection and on different ecological environment.展开更多
The authors investigated the possible association of -4522C/T variation of adiponectin gene with coronary heart disease (CHD) and type 2 diabetes mellitus (T2DM). Genotyping of SNP --4522C/T in 304 patients with C...The authors investigated the possible association of -4522C/T variation of adiponectin gene with coronary heart disease (CHD) and type 2 diabetes mellitus (T2DM). Genotyping of SNP --4522C/T in 304 patients with CHD, 389 patients with T2DM, and 405 age and sex-matched healthy control subjects was carried out by means of PCR-RFLP approach. No significant difference in the genotype or allele frequencies was found, either between patients with CHD and control subjects, or between patients with T2DM and control subjects. However, in the subgroup analysis, an association of the TAr genotype and T allele with type 2 diabetes combined with obesity (BMI ≥ 25 kg/m2) was found (P = 0.014 and P = 0.034, respectively). Also the homeostasis model assessment of insulin resistance (HOMA-IR) in T2DM patients with T/T genotype was significantly higher than that in T2DM patients carrying C allele (P = 0.0069). The authors' findings for the first time demonstrated that SNP --4522 in the adiponectin gene was associated with T2DM that combined with obesity and higher insulin resistance index in patients with T2DM. This indicated that the variation might associate with an increased susceptibility to type 2 diabetic obesity and insulin resistance. But -4522C/T polymorphism did not contribute to the susceptibility of CHD.展开更多
The polymorphism of MHC class II B gene in 14 Chinese alligators was analyzed, which came from three different areas: a wild population from Xuancheng, Anhui, a captive population from Changxing, Zhejiang, and a capt...The polymorphism of MHC class II B gene in 14 Chinese alligators was analyzed, which came from three different areas: a wild population from Xuancheng, Anhui, a captive population from Changxing, Zhejiang, and a captive population from Anhui Research Center for Reproduction of Chinese Alligators. The gene fragment was amplified using a pair of specific primers designed from the MHC gene sequence of the spectacled caiman. A total of 34 sequence haplotypes of exon 3 were detected in the sampled Chinese alligators. The numbers of haplotypes of the 3 Chinese alligator populations were 15, 10, and 9, respectively. The overall estimation of the MHC polymorphism in the Chinese alligator population was higher than those in mammals and in cypdnid fish, The rates of nonsynonymous substitutions (dN) occurred at a significantly lower frequency than that of synonymous substitutions (ds), which were not consistent with the common rule. This result might suggest that the polymorphism of exon 3 seemed not to be maintained by the balancing selection. The neutrality test of Tajima excluded the null hypothesis that the polymorphism of exon 3 was generated by a random drift, and the fact that D = -0.401 indicated an excess of rare mutations in the Chinese alligator. The nucleotide diversity of the sequences and the phylogenetic relations were also analyzed, and the results suggested that there was no significant difference in genetic diversity among the 3 populations of Chinese alligator.展开更多
In the present paper, RAPD was used to study the genetic polymorphism of fisheswith different genome combinations. Our results indicated that four of the 26 random primersproduced distinct and reproducible electrophor...In the present paper, RAPD was used to study the genetic polymorphism of fisheswith different genome combinations. Our results indicated that four of the 26 random primersproduced distinct and reproducible electrophoretic patterns which were genome-specific andcould distinguish different biotypes. This enabled us to derive a diagnostic profile, from whichwe constructed a molecular marker key for different biotypes. By the analysis of the data ofRAPD patterns, the genetic relationship was constructed with UPGMA (unweighted pair-groupmethod with arithmetical averages). Our experiments also concluded that RAPD was moresuccessful in variety identification than protein polymorphism analysis and serohematology for itstechnological simplicity and sensitivity.展开更多
In order to find the completeness threshold which offers a practical method of making bounded model checking complete, the over-approximation for the complete threshold is presented. First, a linear logic of knowledge...In order to find the completeness threshold which offers a practical method of making bounded model checking complete, the over-approximation for the complete threshold is presented. First, a linear logic of knowledge is introduced into the past tense operator, and then a new temporal epistemic logic LTLKP is obtained, so that LTLKP can naturally and precisely describe the system's reliability. Secondly, a set of prior algorithms are designed to calculate the maximal reachable depth and the length of the longest of loop free paths in the structure based on the graph structure theory. Finally, some theorems are proposed to show how to approximate the complete threshold with the diameter and recurrence diameter. The proposed work resolves the completeness threshold problem so that the completeness of bounded model checking can be guaranteed.展开更多
A method for the multi target locating and tracking with the multi sensor in a field artillery system is studied. A general modeling structure of the system is established. Based on concepts of cluster and closed ba...A method for the multi target locating and tracking with the multi sensor in a field artillery system is studied. A general modeling structure of the system is established. Based on concepts of cluster and closed ball, an algorithm is put forward for multi sensor multi target data fusion and an optimal solution for state estimation is presented. The simulation results prove the algorithm works well for the multi stationary target locating and the multi moving target tracking under the condition of the sparse target environment. Therefore, this method can be directly applied to the field artillery C 3I system.展开更多
AIM: To investigate the prevalence and genotype distribution of Torque teno virus (TTV) in patients with different liver diseases and chronic renal failure treated at a referral hospital in North India. METHODS: W...AIM: To investigate the prevalence and genotype distribution of Torque teno virus (TTV) in patients with different liver diseases and chronic renal failure treated at a referral hospital in North India. METHODS: Whereas prevalence of TFV was based on amplification of conserved region of ORF2 of TTV genome, the genotyping of TFV was carried out using restriction fragment length polymorphism (RFLP) procedure on the N22 region of ORFI. RESULTS: TTV-DNA was detected in 137 of 513 (26.7%) patients with liver diseases and 38 of 65 (58.5%) patients with chronic renal failure. Trv was also detected in 2/7% of healthy controls. The sequence analysis of the PCR product from 10 randomly selected cases failed to show a significant sequence divergence when compared with that of the TRM1 isolate of TTV genotype 1. The results of genotyping in 55 randomly selected patients showed the presence of genotype 1 (G1) in 53 (96.4%) and genotype 2 (G2) in 2 cases (3.6%), respectively. Other genotypes were not identified in this patient subgroup, suggesting that G1 is predominant in this area. The results of genotyping by RFLP were also supported by phylogenetic tree analysis, where G1 was found to be the major genotype. CONCLUSION: These results indicate that TTV is moderately present in Indian patients, with G1 to be the major genotype in North India. The pathogenicity and etiological role of TTV in different diseases is still a question mark and warrant further studies.展开更多
AIM: To investigate the influence of IL-1B-511 gene polymorphism on IL-1B mRNA expression and gastric acid output in individual with or without Helicobacter pylori (H pylori) infection.METHODS: IL-1B mRNA expression a...AIM: To investigate the influence of IL-1B-511 gene polymorphism on IL-1B mRNA expression and gastric acid output in individual with or without Helicobacter pylori (H pylori) infection.METHODS: IL-1B mRNA expression and gastric acid secretion in 117 health volunteers were assayed using semi-quantitative RT-PCR and gastric juice assay, respectively. Pepsinogen (PG) Ⅰ and Ⅱ of 255 subjects (including 117 health volunteers) were also examined.RESULTS: T/T genotype individuals with H pylori infection had a more decreased PG Ⅰ/Ⅱ ratio. In gastric antrum mucosa, the individuals with H pylori infection had higher IL-1B expression than those without H pylori infection, but there was no obvious difference among each genotype. In gastric corpus, the individuals with H pylori infection had a significantly higher IL-1B expression than those without H pylori infection. IL-1B-511T/T genotype was markedly higher as compared with the other two genotypes. Both maximal acid output and basic acid output were similar among each genotype in IL-1B-511 gene locus, regardless of H pylori infection.CONCLUSION: IL-1B-511 T allele does not decrease gastric acid output, although it has a stimulated influence on IL-1B expression. Consequently, the pathway,through which IL-1B plays a central role in gastric cancer development, might not depend on low acid, but on the other regulation mechanisms.展开更多
AIM: To characterize the biochemical and immunological properties of an experimental ISCOMS vaccine prepared from a novel therapeutic polypeptide based on T cell epitopes of HBsAg, and a heptatis B-ISCOMS was prepared...AIM: To characterize the biochemical and immunological properties of an experimental ISCOMS vaccine prepared from a novel therapeutic polypeptide based on T cell epitopes of HBsAg, and a heptatis B-ISCOMS was prepared and investigated. METHODS: An immunostimulating complexes(ISCOMS)-based vaccine containing a novel therapeutic hepatitis B polypeptide was prepared by dialysis method, and its formation was visualized by electron microscopy and biochemically verified by SDS-polyacrylamide gel electrophoresis. Amount of the peptide within ISCOMS was determined by Bradford assay, and specific CTL response was detected by ELISPOT assay. RESULTS: Typical cage-like structures of submicroparticle with a diameter of about 40nm were observed by electron microscopy. Results from Bradford assay showed that the level of peptide incorporation was about 0.33g.L(-1). At the paralleled position close to the sixth band of the molecular weight marker(3480kDa) a clear band was shown in SDS-PAGE analysis, indicating successful incorporation of polypeptide into ISCOMS. It is suggested that ISCOMS delivery system could efficiently improve the immunogenicity of polypeptide and elicit specific immune responses in vivo by the results of ELISPOT assay, which showed that IFN-gamma producing cells(specific CTL responses) were increased(spots of ISCOMS-treated group: 47+/-5, n =3; control group: 5+/-2, n =3). CONCLUSION: ISCOMS-based hepatitis B polypeptide vaccine is successfully constructed and it induces a higher CTL response compared with short polypeptides vaccine in vivo.展开更多
AIM: To determine common NOD2/CARD15 mutations and TLR4 D299G polymorphism in Hungarian patients with CD. METHODS: A total of 527 unrelated patients with CD (male/female: 265/262, age: 37.1 (SD 7.6) years) and 200 hea...AIM: To determine common NOD2/CARD15 mutations and TLR4 D299G polymorphism in Hungarian patients with CD. METHODS: A total of 527 unrelated patients with CD (male/female: 265/262, age: 37.1 (SD 7.6) years) and 200 healthy subjects were included. DNA was screened for possible NOD2/CARD15 mutations by denaturing high-performance liquid chromatography (confirmed by direct sequencing). TLR4 D299G was tested by PCR-RFLP. RESULTS: NOD2/CARD15 mutations were found in 185 patients (35.1%) and in 33 controls (16.5%,P<0.0001). SNP8/R702W (10.8% vs 6%, P= 0.02), SNP13/3020insC (19.4% vs 5%, P<0.0001) and exon4 R703C (2.1% vs 0%, P= 0.02) mutations were more frequent in CD, while the frequency of SNP12/G908R was not increased. The frequency of TLR4 D299G was not different (CD: 9.9% vs controls: 12.0%). Variant NOD2/CARD15 allele was associated with an increased risk for CD (ORhet=1.71, 95%CI=1.12-2.6, P= 0.0001, ORtwo-risk alleles = 25.2, 95%CI =4.37- ,P<0.0001), early disease onset (carrier: 26.4 years vs non-carrier: 29.8 years, P=0.0006), ileal disease (81.9% (?) 69.5%, OR = 1.99, 95%CI = 1.29-3.08, P= 0.02, presence of NOD2/CARD15 and TLR4: 86.7% vs 64.8%), stricturing behavior (OR = 1.69,95%CI = 1.13-2.55, P= 0.026) and increased need for resection (OR=1.71, 95%CI: 1.13-2.62, P= 0.01), but not with duration, extra-intestinal manifestations, familial disease or smoking. TLR4 exhibited a modifier effect: age of onset in wt/TLR4 D299G carriers: 27.4 years vs NOD2mut/TLR D299G: 23 years (P = 0.06), in NOD2mut/wt: 26.7 years. CONCLUSION: These results confirm that variant NOD2/ CARD15 (R702W, R703C and 3020insC) alleles are associated with earlier disease onset, ileal disease, stricturing disease behavior in Hungarian CD patients. In contrast, although the frequency of TLR4 D299G polymorphism was not different from controls, NOD2/TLR4 mutation carriers tended to present at earlier age.展开更多
AIM: To explore the susceptibility of children to intrauterine HBV infection by studying the relationship between IFN-γ gene polymorphism, including IFN-γ+874A/T single nucleotide polymorphism(SNP) and CA repeat...AIM: To explore the susceptibility of children to intrauterine HBV infection by studying the relationship between IFN-γ gene polymorphism, including IFN-γ+874A/T single nucleotide polymorphism(SNP) and CA repeat microsatellite polymorphism and intrauterine HBV infection. METHODS: A TaqMan fluorescence polymerase chain reaction in the IFN-γ+874A/T single nucleotide polymorphism was tested in the intrauterine HBV infection group(group Ⅰ) and the normal immune children group(group Ⅱ). Capillary electrophoresis was performed in the above two groups to assay the IFN-γ, CA repeat microsatellite polymorphism. RESULTS: Frequencies of AA, AT and TT genotypes were 67.4%, 19.6% and 13.0% in the intrauterine HBV infection group, and 45.2%, 30.1% and 24.7% in the normal immune children group, respectively. A significant difference was found in the frequency distribution of IFN-γ+874 genotype between the two groups (x^2 = 5.102, P = 0.02389). In the intrauterine HBV infection group the AA genotype was more common than in the normal immune group. Frequency of IFN-γ+874A allele was 77.17% in the intrauterine HBV infection group, and 60.27% in the normal immune children group. In the intrauterine HBV infection group the IFN-γ+874A allele was more common than in normal immune group. A significant difference was found in the frequency distribution between the two groups (x^2= 7.238, P= 0.02389, OR = 2.228, 95% CI = 1.244-3.992). (CA12)^+/(CA12)^+ of IFN-γ CA microsatellite polymorphism was 11.90% in the intrauterine HBV infection group and 26.47% in the normal immune children group. A significant difference was found in the frequency distribution between the two groups (x^2 = 5.64, P = 0.0176). Frequency of IFN-γ CA repeat was 25% in the intrauterine HBV infection group and 43.38% in the normal immune children group. The frequency of IFN-γ CA repeat was less in the intrauterine HBV infection group than in normal immune group. A significant difference was found in the frequency distribution between the two groups (x^2 = 7.548, P= 0.0060). CONCLUSION: There is a relationship between IFN-γ+874A/T SNP and intrauterine HBV infection as well as between IFN-γ CA microsatellite polymorphism and intrauterine HBV infection. IFN-γ gene polymorphism might be important in determining individual's susceptibility to intrauterine HBV infection.展开更多
AIM: To investigate the allelic and haplotypic association of reg1α gene with tropical calcific pancreatitis (TCP). Since TCP is known to have a variable genetic basis, we investigated the interaction between mutatio...AIM: To investigate the allelic and haplotypic association of reg1α gene with tropical calcific pancreatitis (TCP). Since TCP is known to have a variable genetic basis, we investigated the interaction between mutations in the susceptibility genes, SPINK1 and CTSB with reg1α polymorphisms. METHODS: We analyzed the polymorphisms in the reg1α gene by sequencing the gene including its promoter region in 195 TCP patients and 150 ethnically matched controls, compared their allele and haplotype frequencies, and their association with the pathogenesis and pancreaticolithiasis in TCP and fibro-calculous pancreatic diabetes. RESULTS: We found 8 reported and 2 novel polymo-rphisms including an insertion-deletion polymorphism in the promoter region of reg1α. None of the 5' UTR variants altered any known transcription factor binding sites, neither did any show a statistically significant association with TCP. No association with any reg1α variants was observed on dichotomization of patients based on their N34S SPINK1 or L26V CTSB status. CONCLUSION: Polymorphisms in reg1α gene, including the regulatory variants singly or in combination with the known mutations in SPINK1 and/or CTSB genes, are not associated with tropical calcific pancreatitis.展开更多
An algebraic diagonalization method is proposed. As two examples, the Hamiltonians of BCS ground stateunder mean-field approximation and XXZ antiferromagnetic model in linear spin-wave frame have been diagonalized byu...An algebraic diagonalization method is proposed. As two examples, the Hamiltonians of BCS ground stateunder mean-field approximation and XXZ antiferromagnetic model in linear spin-wave frame have been diagonalized byusing SU(2), SU(1,1) Lie algebraic method, respectively. Meanwhile, the eigenstates of the above two models are revealedto be SU(2), SU(1,1) coherent states, respectively. The relation between the usual Bogoliubov Valatin transformationand the algebraic method in a special case is also discussed.展开更多
AIM: To explore the association between single nucleotide polymorphisms (SNPs) at 8q24 and gastric cancer risk. METHODS: A case-control investigation including 212 gastric cancer patients and 377 healthy controls was ...AIM: To explore the association between single nucleotide polymorphisms (SNPs) at 8q24 and gastric cancer risk. METHODS: A case-control investigation including 212 gastric cancer patients and 377 healthy controls was conducted. The genotypes of SNPs (rs6983267, rs7008482 and rs10808555) were examined and established through polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Multivariate logistic regression models were used to evaluate the association between SNPs and gastric cancer. RESULTS: The genotype frequencies of rs6983267 in gastric cancer patients were obviously different from those in the control (P = 0.005). GT genotype of rs6983267 was associated with an increased risk of gastric cancer compared with GG genotype (adjusted odds ratio = 2.01, 95% confidence interval: 1.28-3.14). Further stratified analysis indicated that rs6983267 GT genotype facilitated the risk of gastric cancer of non-cardiac and intestinal type (OR: 2.638, 95% CI: 1.464-4.753; OR: 1.916, 95% CI: 1.166-3.150, respectively). CONCLUSION: This study demonstrates for the first time that rs6983267 is involved in susceptibility to gastric cancer, although further large-sample investigations are still needed.展开更多
AIM: To evaluate the effect of single nucleotide poly- morphisms of interleukin (IL)-28B, rs12979860 on progression and treatment response in chronic hepatitis C. METHODS: Patients (n = 64; 37 men, 27 women; mean...AIM: To evaluate the effect of single nucleotide poly- morphisms of interleukin (IL)-28B, rs12979860 on progression and treatment response in chronic hepatitis C. METHODS: Patients (n = 64; 37 men, 27 women; mean age, 44 + 12 years) with chronic hepatitis C, genotype 1, received treatment with peg-interferon plus ribavirin. Genotyping of rs12979860 was per- formed on peripheral blood DNA. Histopathological assessment of necroinflammatory grade and fibrosis stage were scored using the METAVIR system on a liver biopsy sample before treatment. Serum viral load, ami- notransferase activity, and insulin level were measured. Insulin resistance index, body mass index, waist/hip ratio, percentage of body fat and fibrosis progression rate were calculated. Applied dose of interferon and ribavirin, platelet and neutrophil count and hemoglobin level were measured. RESULTS: A sustained virological response (SVR) was significantly associated with IL28B polymorphism (CC vs -l-r allele: odds ratio (OR), 25; CC vs CT allele: OR, 5.4), inflammation activity (G 〈 1 vs G 〉 1: OR, 3.9), fibrosis (F 〈 1 vs F 〉 1: OR, 5.9), platelet count (〉 200 × 109/L vs 〈 200 ×109/L: OR, 4.7; OR in patients with genotype CT: 12.8), fatty liver (absence vs presence of steatosis: OR, 4.8), insulin resistance index (〈 2.5 vs 〉 2.5: OR, 3.9), and baseline HCV viral load (〈 106 IU/mL vs 〉 106 IU/mL: OR, 3.0). There was no association with age, sex, aminotransferases activity, body mass index, waist/hip ratio, or percentage body fat. There was borderline significance (P = 0.064) of increased fibrosis in patients with the I-I allele, and no differences in the insulin resistance index between groups of patients with CC, CT and -IF alleles (P = 0.12). Spearman's rank correlation coefficient between insulin resistance and stage of fibrosis and body mass index was r = 0.618 and r = 0.605, respectively (P 〈 0.001). Significant dif- ferences were found in the insulin resistance index (P = 0.01) between patients with and without steatosis. Patients with the C-I- allele and absence of a SVR had a higher incidence of requiring threshold dose reduction of interferon (P = 0.07). CONCLUSION: IL28B variation is the strongest host factor not related to insulin resistance that determines outcome of antiviral therapy. Baseline platelet count predicts the outcome of antiviral therapy in CT allele patients.展开更多
文摘Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assisted methods are important approaches for RNA direct detection,but its specificity will be limited when the fidelity of ligases is not ideal.The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection.Methods In this study,a dualcompetitive-padlock-probe(DCPLP)assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation.To verify the method,we employed dual competitive padlock probe-mediated rolling circle amplification(DCPLP-RCA)to genotype the CYP2C9 gene.Results The specificity was well improved through the competition and strand displacement of dual padlock probe,with an 83.26%reduction in nonspecific signal.By detecting synthetic RNA samples,the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L.Furthermore,clinical samples were applied to the method to evaluate its performance,and the genotyping results were consistent with those obtained using the qPCR method.Conclusion This study has successfully established a highly specific direct RNA SNP detection method,and provided a novel avenue for accurate identification of various types of RNAs.
文摘The relationship of three Brachionus calyciflorus morphotypes (two short-spined, single-spined and spineless ones) was studied in the laboratory. Newly hatched females of each morphotype were cultured individually in an inorganic or Asplanchna-conditioned mediums, and the morphotype of their offspring were checked at 200 x magnification. Females of each morphotype, cultured in the inorganic medium, could produce offspring of these three morphotypes in a single generation. Females of all three morphotyes could respond to specific Asplanchna-substances when they were cultured in Asplanchna-conditioned medium. Moreover, we observed that these three morphotype females could coexist in the medium and had three types of egg-bearing females. Based on the above observations we suggest that B. calyciflorus was a morphotype complex and has three basic morphotypes.
基金the Science Foundation of the Health Department of Hainan Province, China (No. 2005-65).
文摘Objective To investigate the relationship of four single nucleotide polymorphism (SNP) haplotypes in the angiotensinogen (AGT) gene to the primary hypertension with or without cerebral infarction in the Li nationality of Hainan, China. Methods Total 300 subjects were allocated into three different groups: Groupl, 100 patients who have primary hypertension; Group 2, 100 patients who have primary hypertension with cerebral infarction; and control group, 100 healthy individuals. The genotypes of all subjects were determined by PCR-sequencing to analyze the four poly- morphisms at position - 152 (G-A), -20 (A-C), - 18 (C-T), and -6 (A-G) in the promoter region of AGT. Results The frequen- cies ofCT genotype of AGT-18 and T allele in Group 1 (P = 0.003, P = 0.004) and Group 2 (P = 0.002, P = 0.002) were both significantly higher than in healthy controls. The frequency of G allele of AGT-6 was significantly higher in Group 2 than in the control group (P = 0.016), while there is no significant difference between Group 1 and the control. Haplotype analysis revealed that H6 haplotype frequency which included -20C and -6G was significantly increased in Group 2 (P = 0.003) compared with the control group, while H5 haplotype frequency which included -20C and -18T was signifi- cantly increased in Group 1 (P = 0.006) versus the control. Conclusion The -20 (A-C) and - 18 (C-T) of the AGT may play an important role in pathogenesis of primary hypertension; and -20 (A-C), -18 (C-T), and -6 (A-G) may be the genetic risk factors for the onset of primary hypertension with cerebral infarction in the Li nationality of Halnan, China.
基金This work was supported by the International Cooperation Item of the National Natural Science Foundation of China (No. 30213009, 30310103007, 30410103150)Natural Science Foundation of Jiangsu Province of China (No. BK2007556)+1 种基金Basic Natura Science Foundation for Colleges and Universities Jiangsu Province (No. NK051039, 06KJD230203)the New Century Talent Project of Yangzhou University in China.
文摘In the present study with Tan sheep, small-tailed Han sheep, Hu sheep, Tong sheep, and Wadi sheep, we detected the distribution of gene frequency of several microsatellite sites in different chromosomes, the result showed that: 1) Hu sheep was in the status of Hardy-Weinberg extreme disequilibrium (P 〈 0.01), while populations including Tong sheep, small-tailed Han sheep, Tan sheep, and Wadi sheep were in Hardy-Weinberg equilibrium (P 〉 0.05). 2) Variance analysis of the heterozygosity and poly- morphic information content at rnicrosatellite makers showed that there were not significant differences among populations as to heterozygosity and PIC (P 〉 0.05), as to effective number of alleles there were not significant differences both among Tan sheep, Hu sheep, Tong sheep, and Wadi sheep, and between Wadi sheep and small-tailed sheep (P 〉 0.05), but between the former three populations and the latter two populations, there were significant differences (0.01〈 P 〈0.05). The variation levels of small-tailed Han sheep was the highest in the five populations based on microsatellite maker data, subsequently followed by Wadi sheep, Tong sheep, Tan sheep, and then Hu sheep. 3) The phylogenetic relationships of the five sheep populations in this study did not meet the mechanism of isolation by distance, and the genetic differentiation relationships among five sheep populations were not closely linearly correlative with their geography distribution. Our findings supported related records in literature: The five populations originated on different time stage from the primogenitor population and communicated genetically with each other thereafter in the process of natural and artificial selection and on different ecological environment.
基金the Chinese High Tech Programs (863) from the Ministry of Science and Technology (No. 2002BA- 711A08)the National Natural Science Foundation of China (No. 30671155, and 39993420)+1 种基金Grant FMU-RT002 of Program for Innovative Research Team in Science and Technology in Fujian Province Universitythe Science Foundation from the Depart-ment of Education of Fujian Province (No. JA05251, and JB06215).
文摘The authors investigated the possible association of -4522C/T variation of adiponectin gene with coronary heart disease (CHD) and type 2 diabetes mellitus (T2DM). Genotyping of SNP --4522C/T in 304 patients with CHD, 389 patients with T2DM, and 405 age and sex-matched healthy control subjects was carried out by means of PCR-RFLP approach. No significant difference in the genotype or allele frequencies was found, either between patients with CHD and control subjects, or between patients with T2DM and control subjects. However, in the subgroup analysis, an association of the TAr genotype and T allele with type 2 diabetes combined with obesity (BMI ≥ 25 kg/m2) was found (P = 0.014 and P = 0.034, respectively). Also the homeostasis model assessment of insulin resistance (HOMA-IR) in T2DM patients with T/T genotype was significantly higher than that in T2DM patients carrying C allele (P = 0.0069). The authors' findings for the first time demonstrated that SNP --4522 in the adiponectin gene was associated with T2DM that combined with obesity and higher insulin resistance index in patients with T2DM. This indicated that the variation might associate with an increased susceptibility to type 2 diabetic obesity and insulin resistance. But -4522C/T polymorphism did not contribute to the susceptibility of CHD.
基金the National Natural Science Foundation of China (No. 30270213 and 30470244)a grant of Special Scholar Foundation from Anhui Province (No. 04043049)the Key Laboratory of Biotic Environment and Ecological Safety in Anhui Province.
文摘The polymorphism of MHC class II B gene in 14 Chinese alligators was analyzed, which came from three different areas: a wild population from Xuancheng, Anhui, a captive population from Changxing, Zhejiang, and a captive population from Anhui Research Center for Reproduction of Chinese Alligators. The gene fragment was amplified using a pair of specific primers designed from the MHC gene sequence of the spectacled caiman. A total of 34 sequence haplotypes of exon 3 were detected in the sampled Chinese alligators. The numbers of haplotypes of the 3 Chinese alligator populations were 15, 10, and 9, respectively. The overall estimation of the MHC polymorphism in the Chinese alligator population was higher than those in mammals and in cypdnid fish, The rates of nonsynonymous substitutions (dN) occurred at a significantly lower frequency than that of synonymous substitutions (ds), which were not consistent with the common rule. This result might suggest that the polymorphism of exon 3 seemed not to be maintained by the balancing selection. The neutrality test of Tajima excluded the null hypothesis that the polymorphism of exon 3 was generated by a random drift, and the fact that D = -0.401 indicated an excess of rare mutations in the Chinese alligator. The nucleotide diversity of the sequences and the phylogenetic relations were also analyzed, and the results suggested that there was no significant difference in genetic diversity among the 3 populations of Chinese alligator.
文摘In the present paper, RAPD was used to study the genetic polymorphism of fisheswith different genome combinations. Our results indicated that four of the 26 random primersproduced distinct and reproducible electrophoretic patterns which were genome-specific andcould distinguish different biotypes. This enabled us to derive a diagnostic profile, from whichwe constructed a molecular marker key for different biotypes. By the analysis of the data ofRAPD patterns, the genetic relationship was constructed with UPGMA (unweighted pair-groupmethod with arithmetical averages). Our experiments also concluded that RAPD was moresuccessful in variety identification than protein polymorphism analysis and serohematology for itstechnological simplicity and sensitivity.
基金The National Natural Science Foundation of China (No.10974093)the Scientific Research Foundation for Senior Personnel of Jiangsu University (No.07JDG014)the Natural Science Foundation of Higher Education Institutions of Jiangsu Province (No.08KJD520015)
文摘In order to find the completeness threshold which offers a practical method of making bounded model checking complete, the over-approximation for the complete threshold is presented. First, a linear logic of knowledge is introduced into the past tense operator, and then a new temporal epistemic logic LTLKP is obtained, so that LTLKP can naturally and precisely describe the system's reliability. Secondly, a set of prior algorithms are designed to calculate the maximal reachable depth and the length of the longest of loop free paths in the structure based on the graph structure theory. Finally, some theorems are proposed to show how to approximate the complete threshold with the diameter and recurrence diameter. The proposed work resolves the completeness threshold problem so that the completeness of bounded model checking can be guaranteed.
文摘A method for the multi target locating and tracking with the multi sensor in a field artillery system is studied. A general modeling structure of the system is established. Based on concepts of cluster and closed ball, an algorithm is put forward for multi sensor multi target data fusion and an optimal solution for state estimation is presented. The simulation results prove the algorithm works well for the multi stationary target locating and the multi moving target tracking under the condition of the sparse target environment. Therefore, this method can be directly applied to the field artillery C 3I system.
基金Indian Council of Medical Research (ICMR),New Delhi-110049 for financial grant
文摘AIM: To investigate the prevalence and genotype distribution of Torque teno virus (TTV) in patients with different liver diseases and chronic renal failure treated at a referral hospital in North India. METHODS: Whereas prevalence of TFV was based on amplification of conserved region of ORF2 of TTV genome, the genotyping of TFV was carried out using restriction fragment length polymorphism (RFLP) procedure on the N22 region of ORFI. RESULTS: TTV-DNA was detected in 137 of 513 (26.7%) patients with liver diseases and 38 of 65 (58.5%) patients with chronic renal failure. Trv was also detected in 2/7% of healthy controls. The sequence analysis of the PCR product from 10 randomly selected cases failed to show a significant sequence divergence when compared with that of the TRM1 isolate of TTV genotype 1. The results of genotyping in 55 randomly selected patients showed the presence of genotype 1 (G1) in 53 (96.4%) and genotype 2 (G2) in 2 cases (3.6%), respectively. Other genotypes were not identified in this patient subgroup, suggesting that G1 is predominant in this area. The results of genotyping by RFLP were also supported by phylogenetic tree analysis, where G1 was found to be the major genotype. CONCLUSION: These results indicate that TTV is moderately present in Indian patients, with G1 to be the major genotype in North India. The pathogenicity and etiological role of TTV in different diseases is still a question mark and warrant further studies.
文摘AIM: To investigate the influence of IL-1B-511 gene polymorphism on IL-1B mRNA expression and gastric acid output in individual with or without Helicobacter pylori (H pylori) infection.METHODS: IL-1B mRNA expression and gastric acid secretion in 117 health volunteers were assayed using semi-quantitative RT-PCR and gastric juice assay, respectively. Pepsinogen (PG) Ⅰ and Ⅱ of 255 subjects (including 117 health volunteers) were also examined.RESULTS: T/T genotype individuals with H pylori infection had a more decreased PG Ⅰ/Ⅱ ratio. In gastric antrum mucosa, the individuals with H pylori infection had higher IL-1B expression than those without H pylori infection, but there was no obvious difference among each genotype. In gastric corpus, the individuals with H pylori infection had a significantly higher IL-1B expression than those without H pylori infection. IL-1B-511T/T genotype was markedly higher as compared with the other two genotypes. Both maximal acid output and basic acid output were similar among each genotype in IL-1B-511 gene locus, regardless of H pylori infection.CONCLUSION: IL-1B-511 T allele does not decrease gastric acid output, although it has a stimulated influence on IL-1B expression. Consequently, the pathway,through which IL-1B plays a central role in gastric cancer development, might not depend on low acid, but on the other regulation mechanisms.
基金the National Natural Science Foundation of China,No.39789010
文摘AIM: To characterize the biochemical and immunological properties of an experimental ISCOMS vaccine prepared from a novel therapeutic polypeptide based on T cell epitopes of HBsAg, and a heptatis B-ISCOMS was prepared and investigated. METHODS: An immunostimulating complexes(ISCOMS)-based vaccine containing a novel therapeutic hepatitis B polypeptide was prepared by dialysis method, and its formation was visualized by electron microscopy and biochemically verified by SDS-polyacrylamide gel electrophoresis. Amount of the peptide within ISCOMS was determined by Bradford assay, and specific CTL response was detected by ELISPOT assay. RESULTS: Typical cage-like structures of submicroparticle with a diameter of about 40nm were observed by electron microscopy. Results from Bradford assay showed that the level of peptide incorporation was about 0.33g.L(-1). At the paralleled position close to the sixth band of the molecular weight marker(3480kDa) a clear band was shown in SDS-PAGE analysis, indicating successful incorporation of polypeptide into ISCOMS. It is suggested that ISCOMS delivery system could efficiently improve the immunogenicity of polypeptide and elicit specific immune responses in vivo by the results of ELISPOT assay, which showed that IFN-gamma producing cells(specific CTL responses) were increased(spots of ISCOMS-treated group: 47+/-5, n =3; control group: 5+/-2, n =3). CONCLUSION: ISCOMS-based hepatitis B polypeptide vaccine is successfully constructed and it induces a higher CTL response compared with short polypeptides vaccine in vivo.
文摘AIM: To determine common NOD2/CARD15 mutations and TLR4 D299G polymorphism in Hungarian patients with CD. METHODS: A total of 527 unrelated patients with CD (male/female: 265/262, age: 37.1 (SD 7.6) years) and 200 healthy subjects were included. DNA was screened for possible NOD2/CARD15 mutations by denaturing high-performance liquid chromatography (confirmed by direct sequencing). TLR4 D299G was tested by PCR-RFLP. RESULTS: NOD2/CARD15 mutations were found in 185 patients (35.1%) and in 33 controls (16.5%,P<0.0001). SNP8/R702W (10.8% vs 6%, P= 0.02), SNP13/3020insC (19.4% vs 5%, P<0.0001) and exon4 R703C (2.1% vs 0%, P= 0.02) mutations were more frequent in CD, while the frequency of SNP12/G908R was not increased. The frequency of TLR4 D299G was not different (CD: 9.9% vs controls: 12.0%). Variant NOD2/CARD15 allele was associated with an increased risk for CD (ORhet=1.71, 95%CI=1.12-2.6, P= 0.0001, ORtwo-risk alleles = 25.2, 95%CI =4.37- ,P<0.0001), early disease onset (carrier: 26.4 years vs non-carrier: 29.8 years, P=0.0006), ileal disease (81.9% (?) 69.5%, OR = 1.99, 95%CI = 1.29-3.08, P= 0.02, presence of NOD2/CARD15 and TLR4: 86.7% vs 64.8%), stricturing behavior (OR = 1.69,95%CI = 1.13-2.55, P= 0.026) and increased need for resection (OR=1.71, 95%CI: 1.13-2.62, P= 0.01), but not with duration, extra-intestinal manifestations, familial disease or smoking. TLR4 exhibited a modifier effect: age of onset in wt/TLR4 D299G carriers: 27.4 years vs NOD2mut/TLR D299G: 23 years (P = 0.06), in NOD2mut/wt: 26.7 years. CONCLUSION: These results confirm that variant NOD2/ CARD15 (R702W, R703C and 3020insC) alleles are associated with earlier disease onset, ileal disease, stricturing disease behavior in Hungarian CD patients. In contrast, although the frequency of TLR4 D299G polymorphism was not different from controls, NOD2/TLR4 mutation carriers tended to present at earlier age.
基金Supported by the National Natural Science Foundation of China, No.30271365
文摘AIM: To explore the susceptibility of children to intrauterine HBV infection by studying the relationship between IFN-γ gene polymorphism, including IFN-γ+874A/T single nucleotide polymorphism(SNP) and CA repeat microsatellite polymorphism and intrauterine HBV infection. METHODS: A TaqMan fluorescence polymerase chain reaction in the IFN-γ+874A/T single nucleotide polymorphism was tested in the intrauterine HBV infection group(group Ⅰ) and the normal immune children group(group Ⅱ). Capillary electrophoresis was performed in the above two groups to assay the IFN-γ, CA repeat microsatellite polymorphism. RESULTS: Frequencies of AA, AT and TT genotypes were 67.4%, 19.6% and 13.0% in the intrauterine HBV infection group, and 45.2%, 30.1% and 24.7% in the normal immune children group, respectively. A significant difference was found in the frequency distribution of IFN-γ+874 genotype between the two groups (x^2 = 5.102, P = 0.02389). In the intrauterine HBV infection group the AA genotype was more common than in the normal immune group. Frequency of IFN-γ+874A allele was 77.17% in the intrauterine HBV infection group, and 60.27% in the normal immune children group. In the intrauterine HBV infection group the IFN-γ+874A allele was more common than in normal immune group. A significant difference was found in the frequency distribution between the two groups (x^2= 7.238, P= 0.02389, OR = 2.228, 95% CI = 1.244-3.992). (CA12)^+/(CA12)^+ of IFN-γ CA microsatellite polymorphism was 11.90% in the intrauterine HBV infection group and 26.47% in the normal immune children group. A significant difference was found in the frequency distribution between the two groups (x^2 = 5.64, P = 0.0176). Frequency of IFN-γ CA repeat was 25% in the intrauterine HBV infection group and 43.38% in the normal immune children group. The frequency of IFN-γ CA repeat was less in the intrauterine HBV infection group than in normal immune group. A significant difference was found in the frequency distribution between the two groups (x^2 = 7.548, P= 0.0060). CONCLUSION: There is a relationship between IFN-γ+874A/T SNP and intrauterine HBV infection as well as between IFN-γ CA microsatellite polymorphism and intrauterine HBV infection. IFN-γ gene polymorphism might be important in determining individual's susceptibility to intrauterine HBV infection.
基金Supported by the Council of Scientific and Industrial Research, Ministry of Science and Technology, Government of India
文摘AIM: To investigate the allelic and haplotypic association of reg1α gene with tropical calcific pancreatitis (TCP). Since TCP is known to have a variable genetic basis, we investigated the interaction between mutations in the susceptibility genes, SPINK1 and CTSB with reg1α polymorphisms. METHODS: We analyzed the polymorphisms in the reg1α gene by sequencing the gene including its promoter region in 195 TCP patients and 150 ethnically matched controls, compared their allele and haplotype frequencies, and their association with the pathogenesis and pancreaticolithiasis in TCP and fibro-calculous pancreatic diabetes. RESULTS: We found 8 reported and 2 novel polymo-rphisms including an insertion-deletion polymorphism in the promoter region of reg1α. None of the 5' UTR variants altered any known transcription factor binding sites, neither did any show a statistically significant association with TCP. No association with any reg1α variants was observed on dichotomization of patients based on their N34S SPINK1 or L26V CTSB status. CONCLUSION: Polymorphisms in reg1α gene, including the regulatory variants singly or in combination with the known mutations in SPINK1 and/or CTSB genes, are not associated with tropical calcific pancreatitis.
文摘An algebraic diagonalization method is proposed. As two examples, the Hamiltonians of BCS ground stateunder mean-field approximation and XXZ antiferromagnetic model in linear spin-wave frame have been diagonalized byusing SU(2), SU(1,1) Lie algebraic method, respectively. Meanwhile, the eigenstates of the above two models are revealedto be SU(2), SU(1,1) coherent states, respectively. The relation between the usual Bogoliubov Valatin transformationand the algebraic method in a special case is also discussed.
基金Supported by Shanghai "Phosphor" Science Foundation, China, No. 09QB1403100the National High Technology Researchand Development Program of China, No. 2006AA020704 and 2006AA02A407the Funds for Key Programs of Ministry ofHealth of China, No.2008ZX10002-017
文摘AIM: To explore the association between single nucleotide polymorphisms (SNPs) at 8q24 and gastric cancer risk. METHODS: A case-control investigation including 212 gastric cancer patients and 377 healthy controls was conducted. The genotypes of SNPs (rs6983267, rs7008482 and rs10808555) were examined and established through polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Multivariate logistic regression models were used to evaluate the association between SNPs and gastric cancer. RESULTS: The genotype frequencies of rs6983267 in gastric cancer patients were obviously different from those in the control (P = 0.005). GT genotype of rs6983267 was associated with an increased risk of gastric cancer compared with GG genotype (adjusted odds ratio = 2.01, 95% confidence interval: 1.28-3.14). Further stratified analysis indicated that rs6983267 GT genotype facilitated the risk of gastric cancer of non-cardiac and intestinal type (OR: 2.638, 95% CI: 1.464-4.753; OR: 1.916, 95% CI: 1.166-3.150, respectively). CONCLUSION: This study demonstrates for the first time that rs6983267 is involved in susceptibility to gastric cancer, although further large-sample investigations are still needed.
文摘AIM: To evaluate the effect of single nucleotide poly- morphisms of interleukin (IL)-28B, rs12979860 on progression and treatment response in chronic hepatitis C. METHODS: Patients (n = 64; 37 men, 27 women; mean age, 44 + 12 years) with chronic hepatitis C, genotype 1, received treatment with peg-interferon plus ribavirin. Genotyping of rs12979860 was per- formed on peripheral blood DNA. Histopathological assessment of necroinflammatory grade and fibrosis stage were scored using the METAVIR system on a liver biopsy sample before treatment. Serum viral load, ami- notransferase activity, and insulin level were measured. Insulin resistance index, body mass index, waist/hip ratio, percentage of body fat and fibrosis progression rate were calculated. Applied dose of interferon and ribavirin, platelet and neutrophil count and hemoglobin level were measured. RESULTS: A sustained virological response (SVR) was significantly associated with IL28B polymorphism (CC vs -l-r allele: odds ratio (OR), 25; CC vs CT allele: OR, 5.4), inflammation activity (G 〈 1 vs G 〉 1: OR, 3.9), fibrosis (F 〈 1 vs F 〉 1: OR, 5.9), platelet count (〉 200 × 109/L vs 〈 200 ×109/L: OR, 4.7; OR in patients with genotype CT: 12.8), fatty liver (absence vs presence of steatosis: OR, 4.8), insulin resistance index (〈 2.5 vs 〉 2.5: OR, 3.9), and baseline HCV viral load (〈 106 IU/mL vs 〉 106 IU/mL: OR, 3.0). There was no association with age, sex, aminotransferases activity, body mass index, waist/hip ratio, or percentage body fat. There was borderline significance (P = 0.064) of increased fibrosis in patients with the I-I allele, and no differences in the insulin resistance index between groups of patients with CC, CT and -IF alleles (P = 0.12). Spearman's rank correlation coefficient between insulin resistance and stage of fibrosis and body mass index was r = 0.618 and r = 0.605, respectively (P 〈 0.001). Significant dif- ferences were found in the insulin resistance index (P = 0.01) between patients with and without steatosis. Patients with the C-I- allele and absence of a SVR had a higher incidence of requiring threshold dose reduction of interferon (P = 0.07). CONCLUSION: IL28B variation is the strongest host factor not related to insulin resistance that determines outcome of antiviral therapy. Baseline platelet count predicts the outcome of antiviral therapy in CT allele patients.