This study was aimed to discriminate the alleles in the HLA-C*07:01:01G and HLA-C*07:02:01G groups and analyze their associations with HLA-B locus. Samples previously typed as HLA-C*07:01:01G and HLA-C*07:02:01G were collected. The nucleotide sequences in exons 1 to 7 of the HLA-C locus were sequenced by polymerase chain reaction sequence-based typing (PCR-SBT) and HLA-B genotyping was also preformed by PCR-SBT in these samples. The results showed that 4 samples (30.8%) were confirmed as HLA-C*07:01:01 and 9 samples (69.2%) were HLA-C*07:06 among 13 samples previously typed as HLA-C*07:01:01G. Linkage disequilibrium (LD) analysis showed that HLA-C*07:06 allele was strongly related with HLA-B*44:03. All samples were typed as C*07:02:01 among 102 individuals previously typed as C*07:02:01G. LD analysis showed that C*07:02:01 was strongly related with HLA-B*51:01, B*46:01, B*39:01, B*40:01, B*38:02, B*15:02 alleles. It is concluded that HLA-C*07:01:01 and HLA-C*07:06 alleles are confirmed in the HLA-C*07:01:01G group and HLA-C*07:02:01 is a preferred allele in the HLA-C*07:02:01G.
Alleles ; Base Sequence ; Exons ; HLA-B Antigens ; genetics ; HLA-C Antigens ; genetics ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA

This study was purposed to investigate the nucleotide sequences of a novel HLA-B*15:124 allele and its molecular mechanism. The genomic DNA from whole blood was extracted by using commercial DNA extraction kit. The sequences of exon 2, 3 and 4 of HLA-B locus in the proband were amplified by PCR with group-specific primers, the PCR products were purified by enzymes digestion, then exon 2 to 4 of HLA-B locus for both orientations was sequenced. The results showed that 2 HLA-B alleles of proband were gained after amplification and sequencing of group-specific primers, among them one was a B*40:03, another was a novel allele. After BLAST analysis, the novel allele showed nucleotides different from HLA-B*15:52 in exon 3 at nucleotide position 427 A > T and 440 G > T which resulted in amino acid change from Thr to Ser at codon 143 and Trp to Leu at conon 147. It is concluded that a novel HLA-B allele has two different nucleotides. This HLA-B allele is identified and has been officially named B*15:124 by the WHO Nomenclature Committee.
Alleles ; Base Sequence ; Exons ; Female ; HLA-B Antigens ; classification ; genetics ; Humans ; Sequence Analysis, DNA

OBJECTIVETo investigate the recombination events between human leukocyte antigen (HLA) loci within two families.

METHODSIdentification of HLA-A, -C, -B, -DRB1 and -DQB1 loci was firstly carried out using polymerase chain reaction-sequence specific oligonucleotide. Then HLA high resolution typing was performed using polymerase chain reaction sequencing-based typing. The recombination between HLA loci was identified by family genetic analysis. The parentage possibility was analyzed by short tandom repeat technique.

RESULTSRecombination between the HLA-A and C loci was identified within two families. One individual inherited a paternal haplotype that was the result of a recombination event between the father's HLA-A and -C loci on his chromosomes. The other individual inherited a maternal haplotype that was the result of a recombination event between the mother's HLA-A and -C loci. The high parentage possibilities were obtained in the family members.

CONCLUSIONThe recombination events of HLA-A and -C have been found in two Chinese families, which may help further study on the mechanism of HLA recombination.

Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; Ethnic Groups ; genetics ; Female ; Genetic Loci ; genetics ; HLA-A Antigens ; genetics ; HLA-C Antigens ; genetics ; Haplotypes ; genetics ; Humans ; Male ; Pedigree ; Recombination, Genetic ; genetics

OBJECTIVETo analyze the sequence of the exons 2-4 of human leukocyte antigen (HLA) novel allele HLA-B*15:129.

METHODSDNA of the proband was extracted from whole blood by commercial DNA extraction kit. The amplification for HLA-B exons 2-4 was performed separately by polymerase chain reaction (PCR) with allele group specific primers. The PCR products were digested with enzymes and then directly sequenced for exons 2-4 of HLA-B locus in both directions.

RESULTSSequencing results showed the HLA-B alleles of the proband included B*07:02 and a novel allele. The sequence of the novel allele has been submitted to GenBank (accession no. EF473219) and the allele has been officially named B*15:129 by the WHO Nomenclature Committee. Comparing with the HLA-B*15:01:01:01, the sequence of exons 2-4 of HLA-B*15:129 showed three nucleotide difference in exon 3 at positions 362 and 363 from GG to AT and positions 369 from C to T, which resulted in an amino acid change from Arg to Asn at codon 97.

CONCLUSIONA novel HLA-B allele was identified and has been officially named B15:129 by the WHO Nomenclature Committee.

Alleles ; Base Sequence ; DNA Primers ; Exons ; HLA-B Antigens ; genetics ; Humans ; Male ; Molecular Sequence Data ; Molecular Typing ; Polymerase Chain Reaction

This study was aimed to investigate the molecular genetics basis of a novel allele HLA-A * 2459 in Chinese population. DNA was extracted from whole blood by PEL-FREEZ DNA extraction kit. The amplification of HLA-A exons 2 - 4 of the proband was preformed by allele specific primer PCR and the amplified product was sequenced bidirectionally with primers. The sequencing results showed HLA-A alleles of the proband as A * 1101 and the novel allele. The sequences of the novel allele have been submitted to GenBank (DQ313255, DQ313256, DQ313257). After Blast HLA analysis, the novel allele showed only one nucleotide differences with HLA-A * 24020101 at nucleotide position 527 T to C in exon 3. This results in an amino acid changes from Val to Ala at codon 152. In conclusion, this allele is a novel one and has been officially named HLA-A * 2459 by the WHO Nomenclature Committee.
Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Blood Donors ; China ; HLA-A Antigens ; genetics ; Humans ; Molecular Sequence Data ; Sequence Analysis, DNA

Objective To investigate the relationship and the influence between pre-diabetes mellitus (PDM) and hyperuricemia (HUA).Methods 157 PDM patients,aged 20 to 75 years old were selected from the Second Clinical Medical College of Harbin Medical University,from 2009 February to 2010 February and were divided into HUA group (76 cases) and NUA group (81 cases).All the patients had not been on thiazide drugs.T-test and Pearson correlation analysis were used to calculate the differences and correlation between uric acid and biochemical indicators.Results In the HUA group,BMI was (27.74 ± 2.88) kg/m2,waist to height ratio (WSR) was (0.55 ± 0.41),TC was (6.61 ± 0.73) mmol/L,TG was (3.94 ± 1.97) mmol/L,LDL-C was (3.60 ± 0.45) mmol/L and homeostasis model assessment-insulin resistance index (HOMA-IR) was (3.09± 1.20).There were significant differences noticed in BMI,TG,TC,LDL-C,HOMA-IR at higher level in the HUA group than those in the NUA group.Pre-diabetes uric acid levels were positively correlated with TG,TC,LDL-C while HOMA-1R (TG:r=0.29,TC:r=0.33,LDL-C:r=0.49,HOMA-IR:r=0.51,P＜0.05)was negatively correlated (r=－0.30,P＜0.05) with the HbAlc.Conclusion The levels of PDM uric acid might both be related with TC,TG,LDL-C and HOMA-IR.The High level of uric acid status in vivo appeared closely related to HOMA-IR,which could further promote the progress of pre-diabetic patients to diabetes and causing dyslipidemia.Our findings suggested that the levels of pre-diabetes uric acid levels should be under concern.

BACKGROUND: Although more and more accurate and personalized detection and assistive tools are used in the field of orthopedics with the development of technology, the preoperative CT is still the first-line examination choice of many orthopedic surgeons. Therefore, to master the lumbar spine CT imaging performance is not only the basis of spine surgeons, but also the cornerstone of spinal nail surgery. OBJECTIVE: Through CT imaging technique, we measured the parameters of lumbar pedicle imaging, and understood its internal structure as the basis for the choice of screws, in order to improve the safety and accuracy of nail surgery. METHODS: Totally 36 cases without lumbar vertebrae-related disease undergoing lumbar vertebra and abdominal cavity CT scan were randomly selected, including 20 males and 16 females at the age of (43.3±12.3) years. PACS was used to reconstruct lumbar vertebrae, and the related imaging parameters were measured. RESULTS AND CONCLUSION: (1) There was no significant difference between the left and right sides of the pedicle for paired t-test (P >0.05). (2) Pedicle sagittal angle had downward trend, the level of change was small; L4increased relative to L3; L5level was minimum at the average of -1.3°. Pedicle transverse angle had gradually increased trend, and maximum at L5; the average was 25.4°. (3) The length of the pedicle channel increased first and then decreased in the lumbar segment; the longest average was 52.2 mm in the L2segment, the shortest at L5, with an average of 47.4 mm. (4) The pedicle height was slightly narrowed at L1-L3levels; the smallest was 15.4 mm (L3mean), and reached the maximum in the L5at 22.7 mm (L5mean). Cancellous bone height also showed the same trend, the minimum in the L3at 10.2 mm, the maximum in the L5at 17.9 mm. (5) Lumbar pedicle width showed an upward trend, L1minimum of 7.7 mm, L5maximum of 12.5 mm. Cancellous bone width also became a similar upward trend; the narrowest L1, averagely 4.9 mm; the maximum L5, averagely 8.5 mm. (6) Overall, the upper lumbar pedicle channel was slender, while the lower lumbar spine was slightly short. (7) The parameters of the pedicle can be used as the basis for selecting the appropriate screw. At the same time, lumbar pedicle morphology has a certain difference, and there is a certain law. Accurately grasping its structure and image characteristics of pedicle screw fixation is the basis of surgery. The preoperative imaging data collection and assessment, combined with the general situation of patients, and the development of personalized programming, will help in improving the safety and accuracy of screw placement.

OBJECTIVETo discriminate and analyze the relative frequencies of alleles in HLA-DRB1*12:01:01G(HLA-DRB1*12:01:01/12:06/12:10/12:17) and HLA-DRB1*14:01:01G (DRB1*14:01:01/14:54) groups and assess their associations with HLA-DRB3 and HLA-DQB1 loci.

METHODSA total of 115 DNA samples previously typed as HLA-DRB1*12:01:01G and 108 samples from HLA-DRB1*14:01:01G were selected. DNA sequences for exons 1 to 3 of the HLA-DRB1 locus were analyzed for HLA-DRB1*12:01:01G, and exons 2 to 3 were analyzed for HLA-DRB1*14:01:01G by polymerase chain reaction sequence-based typing (PCR-SBT). Genotyping of HLA-DRB3 and HLA-DQB1 were achieved by PCR-SBT.

RESULTSAmong 115 samples previously typed as HLA-DRB1*12:01:01G, 101 (87.8%) were confirmed as HLA-DRB1*12:01:01 and 14 (12.2%) were HLA-DRB1*12:10, but HLA-DRB1*12:06 and HLA-DRB1*12:17 alleles were not identified. For 108 samples previously typed as HLA-DRB1*14:01:01G, all were typed as HLA-DRB1*14:54. HLA-DRB1*12:01:01 was linked with HLA-DRB3*01:01:02 and HLA-DQB1*03:01, while HLA-DRB1*12:10 was strongly linked with HLA-DRB3*02:02:01 and HLA-DQB1*03:01. HLA-DRB1*14:54 was strongly linked with HLA-DRB3*02:02:01 and two different HLA-DQB1*05:02, *05:03 alleles.

CONCLUSIONHLA-DRB1*12:01:01 was more prevalent than HLA-DRB1*12:10 in the HLA-DRB1*12:01:01G group, and HLA-DRB1*14:54 was the dominant allele for HLA-DRB1*14:01:01G.

Alleles ; Exons ; Gene Frequency ; Genotype ; HLA-DQ beta-Chains ; genetics ; HLA-DRB1 Chains ; genetics ; HLA-DRB3 Chains ; genetics ; Humans

Objective Using the Back Propagation (BP) Neural Network Model to discover the relationship between meteorological factors and mortality of intracerebral hemorrhage,to provide evidence for developing an intracerebral hemorrhage prevention and control program,in Harbin.Methods Based on the characteristics of BP neural network,a neural network Toolbox of MATLAB 7.0 software was used to build Meteorological data of 2007-2009 with intracerebral hemorrhage mortality to predict the effect of BP neural network model,and to compare with the traditional multivariate linear regression model. Results Datas from the multivariate linear regrcssion indicated that the cerebral hemorrhage death mortality had a negative correlation with maximum temperatureand minimum humidity while having a positive correlation with the average relative humidity and the hours of sunshine.The linear correlation coefficient of intracerebral hemorrhage mortality was 0.7854,with mean absolute percentage (MAPE) as 0.21,mean square error (MSE) as 0.22,mean absolute error(MAE) as 0.19.The accuracy of forecasting was 81.31％ with an average error rate as 0.19.The Fitting results of BP neural network model showed that non-linear correlation coefficient of intracerebral hemorrhage mortality was 0.7967,with MAPE as 0.19,MSE as 0.21,MAE as 0.18.The forecasting accuracy was 82.53％ with the average error rate as 0.17.Conclusion The BP neural network model showed a higher forecasting accuracy when compared to the multiple linear regression model on intraccrebral hemorrhage mortality,using the data of 2010' s.

OBJECTIVEThis study was to expand the cytotoxic T lymphocytes (CTL) through inducing the differentiation of umbilical blood monomuclear cells (UBMNC) by using various combination of cytokines, and to investigate the functions of expanded CTL.

METHODSThe MNC were isolated by ficoll density gradient centrifugation. Then, the PHA-P, IFN-γ combined with IL-2, IL-15 and other cytokines were used for induction and expansion of the cord blood-derived CTL. The biological function of CTL was examined by phenotype analysis, cytotoxic tests and real-time fluorescence quantitative PCR.

RESULTSAfter expansion for 15 days, the cell number increased by 1522% ± 137%. The content of CD3(-)CD8(-) cells in uncultured cord blood MNC was 95%, and the CD3(+)CD8(+) CTL cells reached 82.77% in cultured cord blood MNC after expansion for 15 days. The expanded CTL cell showed the cytotoxic activity against K562 and HeLa cell line. The killing rate of MNC was 61.88 ± 1.08%. After expansion, the killing rate could reach to 90% with the average value of 90.33 ± 2.02%. The expanded CTL cells highly expressed some key cytokines, such as granzyme A, granzyme B, GM-CSF, granulysin, IFN-γ, TGF-β, TNF-α and perforin. Compared with the control group, the expression of IFN-γ and TGF-β significantly increased (P < 0.05), and the other factors dramatically increased (P < 0.01).

CONCLUSIONThe cord blood-derived CTL can be expanded by different combinations of cytokines. These protocols may provide alternative choices for CTL cell expansion in tumor adoptive immunotherapy.

Cytokines ; Fetal Blood ; Granulocyte-Macrophage Colony-Stimulating Factor ; Granzymes ; Histocompatibility Antigens Class I ; Histocompatibility Antigens Class II ; Humans ; Immunotherapy, Adoptive ; Perforin ; Phytohemagglutinins ; T-Lymphocytes, Cytotoxic