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ObjectiveTo investigate the relationship between HLA-DRB1 alleles and chronic urticaria with positive autologous serum skin test (ASST) in Guangxi Zhuang Autonomous Region.MethodsASST was conducted in 144 patients with chronic urticaria,who were subsequently divided into two groups according to the test result:positive group (n =62) and negative group (n =82).PCR amplification with sequence-specific primers was used to determine the genotypes of HLA-DRB1 alleles in the patients and 199 normal human controls.Chi-square test was performed to analyse the difference in the frequency of HLA-DRB1 alleles between the 3 groups by using the SPSS 13.0 statistical software package.ResultsThere were significant differences in the frequency of HLA-DRB1*01,*1401 and *16 alleles among the patients with positive and negative ASST and the controls (x2 =10.92,Pc =0.032;x2 =35.34,Pc ＜ 0.01 ;x2 =12.69,Pc =0.032).Paired comparison revealed significant differences in the frequency of HLA-DRB1*1401 allele between the patients with positive ASST and controls(RR =17.09,Pc ＜ 0.01 ) as well as between the patients with positive and negative ASST (RR =7.20,Pc ＜ 0.01).ConclusionHLA-DRB1*1401 allele may be,or be linked to,the predisposing gene of chronic urticaria with positive ASST in Guangxi Zhuang Autonomous Region.
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Objective To construct a recombinant adenovirus vector containing the gene of major histocompatibility complex(MHC)class Ⅱ transactivator(C Ⅱ TA).Methods The restriction fragment of CIITA was inserted into pUC57 vector with EeoR Ⅰ and Xho Ⅰ.Then,recombinant plasmid pShutde-GFP-CMV CⅡTA was constructed with EcoR Ⅰ and Sal Ⅰ,and was confirmed by restriction enzyme digestion and sequeneing.After the treatment with Ⅰ-Ceu Ⅰ and Ⅰ-See Ⅰ,the fragment C Ⅱ TA from recombinant plasmid DShuttle-GFP-CMV.CⅡTA Was inserted into vector pAdxsi.And the pAdxsi-GFP-C Ⅱ TA wag packed into liposome,and was transfected to 293 cens.Results Recombinant plasmid pShuttle-GFP-CMV-C Ⅱ TA Was constructed successfully. After packed into vector pAdxsi, and transfected to 293 cells, significant virus Dlaques were observed,which showed the successful homologous recombination.The titer of the purified AdC Ⅱ TA was 2.0×10~(11) PFU/ml.Conclusions Recombinant adenovirus AdC Ⅱ TA containing gene of MHC class Ⅱ transactivator was established successfully.
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Objective To observe the regulatory effect of recombinant human DCN on the expression of HLA class Ⅰ, Ⅱ molecule on hepatoma carcinoma cell HepG2 and investigate the relatively immune mechanism of human DCN on enhancing anti-tumor effect. Methods After transfected HepG2 cells with pcDNA3.1 (+)-DCN by liposome transfection, the expression of HLA class Ⅰ , Ⅱ molecule and the apoptotic indexes were analyzed by the flow cytometer. Results The expression of HLA class Ⅰ , Ⅱ molecule on the HepG2 cells transfected with pcDNA3.1 (+)-DCN was up-regulated with the apoptosis indexes being significantly higher than that of other control groups. Conclusion The human DCN can induce HepG2 apoptosis and up-regulate the expressions of HLA class Ⅰ , Ⅱ molecule which may contribute to its antitumor effect.
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Objective To investigate the association of HLA-DRB31*03,*04 and *11 alleles with alopecia areata(AA)in Han Nationality in East China.Methods Polymerase chain reaction-sequence specific primer(PCR-SSP)method was conducted in 158 Chinese Han patients with AA as well as in 172 healthy human controls in East China.The relationships of HLA-DRB1 polymorphism to age of onset,episode frequency,clinical course,family history,and severity of AA were evaluated.Results No significant differences were observed for the frequency of HLA DRB1*03,*11 alleles between the patients and human controls,while increased frequency of HLA-DRB1*04 was observed in patients(OR=1.99,Pc=0.01).Multiple logistic regression analysis revealed that HLA-DRB1*04 was more prevalent in patients with an onset after 16 years of age(OR=1.94,Pc=0.02),those without family history(OR=1.97,Pc=0.02),those with recurrent AA(OR=2.49,Pc=0.02),those with a clinical course of more than 1 year(OR=2.94,Pc=0.01),those with severe AA(OR=3.53,Pc=0.00)and tbose with single episode of AA(OR=1.83,Pc=0.04)in comparison with the normal human controls.Conclusion This study demonstrates that HLA-DRB1*04 allele is associated with the occurrence and clinical types of AA in Han Nationality in East China.
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Objective To investigate the genetic association of HLA class II DRB1,DQB1,DQA2 and DQB2 locus with schizophrenia in the Han people of Northern China.Methods The polymerase chain reaction(PCR)and restrictive fragment length polymorphism(RELP) were applied to detect the gene types in the 195 Han family trios including patients with schizophrenia and their healthy parents in the north of China.The chi-square(?2) test,transmission disequilibrium test(TDT) and the haplotype relative risk(HRR) analysis were used to process the genotyping data statistically.Results The genotypic frequency of four genes did not deviate from Hardy-Weinberg equilibrium in both case and control groups;The SNP rs2239800,a G to A base change,presented in the DQA2 locus.Its allelic frequencies showed significant difference between case and control(?2=5.021,P=0.025).And the AA genotype of rs2239800 showed associations with schizophrenia.Conclusion The polymorphism of rs2239800 in the DQA2 gene may be associated with schizophrenia.
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Summary: In order to investigate the association of genotypes of HLA-DRB1 and HLA-DQB1 alleles with the genetic susceptibility of chronic urticaria (CU), genotypes of HLA-DRB1 and HLA-DQB1 genes were detected by polymerase chain reactions with sequence-specific primers (PCR-SSP) in 42 patients with CU (19 men and 23 women, mean age 30.67±12.45 y old as well as 193 racially matched healthy persons in ethnic Han from Hubei provinece. Gene frequencies of HLA-DRB1*12, *0901 (RR=3.11, χ2=7.579, P=0.006; RR=2.47, χ2=5.684, P=0.017) were significantly increased in CU patients as compared with that in healthy people. Gene frequencies of HLA-DQB1*05 (RR＝0.26, χ2=6.683, P=0.01) were significantly decreased in CU patients. It was suggested that CU was found strongly associated with HLA-DRB1*12, *0901 and HLA-DQB1*05, the former might be the genetic markers for susceptibility to CU, but the latter might play a resistive role.
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Objective To investigate the association of HLA class Ⅱ genes and vitiligo in the eastern China Han nationality. Methods Ninety-eight patients with vitiligo and 150 healthy controls were studied for HLA-DRB1, DQA1 and DQB1 locus alleles by PCR-SSOP typing. Results The frequency of HLA-DQA1*03 increased significantly (Pc = 0.008) and DQA1*05 decreased significantly (Pc = 0.016) in the patients with vitiligo. Conclusions The results suggest that there exists a correlation between HLA class Ⅱ genes and vitiligo, and DQA1*03 allele may be a susceptible gene or have a close linkage with susceptible genes, while DQA1*05 allele may be a protective gene in the eastern China Han nationality.
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Objective To develop an oligoneucleotide array for HLA-DR52 group rapid genotyping.Methods According to the special allele sequences of HLA-DRB loci in Chinese Han's population, HLA-DR52 group typing probes which were immobilized on a glass supports were synthesized. A pair of group-special primers labeled by the Cy5-dCTP were designed and were used in the PCR. The labeled PCR products with Cy5 were hybridized with array. The signals were scanned by a scanner and analyzed by Image software. 83 samples were typed by this array and the results were compared with PCR-SSP typing.Results Among 57 HLA-DR52 group loci typed by PCR-SSP,2 samples had no HLA-DR52 loci typed by array,3 DR52 group homozygotes typed by PCR-SSP were actually heterozygotes by array. The other 1 non-DR52 group homozygote identified by PCR-SSP was a heterozygote with one DR52 group locus. Conclusion The oligoneucleotide array technique is a precise, rapid molecular method for HLA-DR52 genotyping. Compared with PCR-SSP method, the genotyping chip is more sensitive and intuitionistic and suitable for clinic practice.
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Objective To study the HLA-DRB1 genotype and their r elation with HBV infection among Han patients in shanxi area. Methods HLA-DRB1 genotyping was conducted in 54 patients HBV infection and 108 health controls, as well as 32 asymtomatic HBsAg carriers by using polymerase ch ain reaction/sequence specific primer (PCR-SSP) method. All the patients, asy mtomatic HBsAg carriers, and healthy subjects were inhabitants of Shan'xi area o f Han nationality. The association between HLA-DRB1 genotype and different repl ication of HBV were also studied. Results DRB1*04,DRB1*09,DRB 1*12, DRB1*15 were the most common genotypes in the Shan'xi Han inbabitants wit h the frequency of 16.2%,12.5%,11.6% and 13.4% respectively. Compared to 108 healthy controls, the allele frequency of HLA-DRB1*03 was 11.1% in HBV patien ts versus 3.7% in healthy controls, with odds ratio=3.57 and Pc=0.014 ( P
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Objectives To investigate the association between HLA-DQB1 alleles and Han Chinese with generalized pustular psoriasis (GPP) from Shandong. Methods The distributing frequencies of HLA-DQB1 alleles were detected with polymerase chain reaction-sequence specific primers (PCR-SSP) in 38 GPP patients and 94 healthy subjects from Shandong. Results ①The frequencies of HLA-DQB1*0201 and *0603 alleles increased significantly in GPP patients compared with those of the healthy controls(Pc = 0.005 and Pc = 0.013 , respectively), and the frequency of HLA- DQB1*0604 alleles decreased (OR = 0.08, Pc = 0.039). ②There were significant association between the GPP patients with a previous history of psoriasis vulgaris and HLA-DQB1*0201 and *0603 alleles (OR = 32.31, Pc = 0.005; OR = 12.42, Pc = 0.005, respectively), and between the GPP patients without the history and HLA-DQB1*0602 alleles (OR = 5.60, Pc = 0.039). Conclusions HLA-DQB1*0201 and *0603 alleles are significantly associated with the Chinese patients with GPP in Han population from Shandong. There is a significant heterogeneity in the GPP patients with and without the history of psoriasis vulgaris.
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Objective To identify the association of HLA-DQA1 and-DQ B1 alleles with vitiligo in Chinese Hans in Anhui province.Methods Polymerase chain reaction sequence-specific primer (PCR-SSP) method was used to analyze the distribution of HLA-DQA1 and-DQB1 alleles among 187 patients with vitiligo and 273 healthy controls.Results The frequencies of HLA-DQA1*0302,-DQB1*0303,-DQB1*0503 alleles were significantly increased in the patients with vitiligo compared with those in the controls,and HLA-DQA1*0501 allele frequency was mark edly decreased.HLA-DQA1*0302,-DQA1*0601,-DQB1*0303,-DQB1*0503 alleles were p ositively associated,whereas HLA-DQA1*0501 allele was negatively associated wit h childhood vitiligo,and HLA-DQB1*0303 allele was positively associated with ad ult vitiligo,in comparison with those in the controls.The frequency of HLA-DQB 1*0303 allele was significantly increased in localized vitiligo patients compare d with that in the controls,whereas frequencies of HLA-DQA1*0302,-DQB1*0303,-DQB1*0503 alleles were significantly increased and the frequency of HLA-DQA1*050 1 allele was markedly decreased in generalized vitiligo patients.Conclusions HLA-DQA1*0302,-DQA1*0601,-DQB1*0303 and-DQB1*0503 alleles could be the suscep tible alleles of vitiligo,while HLA-DQA1*0501 allele could be the protective al lele in Chinese Hans in Anhui province.There may be different genetic backgroun ds between childhood and adult patients,and between localized and generalized vitiligo.