Objective To study the significance of the sample S/Co ratio when using a domestic reagent for anti-hepatitis C virus(HCV)antibody detection and to explore the procedure and standard of anti-HCV antibody diagnosis by using this domestic reagent.Methods Anti-HCV antibody was detected in 295 000 blood donors by a domestic anti-HCV reagent with enzyme-linked immunosorbent assay(ELISA)method and the reactive samples were tested again by ortho anti-HCV antibody reagent.The samples which anti-HCV antibodies were determined as positive by ortho anti-HCV rea- gent were examined by recombinant immunoblot assay(RIBA)reagent and 106 samples of them were also tested for HCV RNA.Results Six hundred and eighty-one samples were reactive in 295 000 samples screened by the domestic ELISA reagent,the reactive ratio was 0.23 %.Among the reactive samples screened by the domestic ELISA reagent,367 samples were determined as positive by ortho anti-HCV reagent while 66.2% of them showed a S/Co ratio≥3.8.The consistency rate between positive results determined by the domestic reagent and RIBA reagent respectively was 53.8%.For the samples showing S/Co ratio≥3.8 by ortho anti-HCV reagent,94.2% had a S/Co ratio≥8.0 when using the domestic ELISA reagent,while the percentage of samples showing S/Co ratio

Carcinoma, Hepatocellular ; genetics ; pathology ; secondary ; Diagnosis, Differential ; Humans ; Liver Neoplasms ; genetics ; pathology ; secondary ; Loss of Heterozygosity ; Male ; Microsatellite Repeats ; Middle Aged ; Neoplasms, Multiple Primary ; genetics

OBJECTIVETo observe blood uric acid levels and Goldstein grading, as well as their correlation in Wilson's disease (WD) patients with different Chinese medical syndrome types.

METHODSTotally 906 WD patients in line with inclusive criteria were assigned to 6 groups, i.e., the heart spirit confused by phlegm group (HSCP, 26 cases), the phlegm-fire disturbing heart group (PFDH, 90 cases), the retention of damp-heat group (RDH, 113 cases), deficiency of qi and blood group (DQB, 168 cases), the deficiency of Gan-yin and Shen-yin group (DGYSY, 327 cases), the deficiency of Gan and Shen group (DGS, 182 cases) due to different Chinese medical syndrome types. Recruited were another 160 healthy subjects having similar ages and diet structures, who came for medical examinations, as the healthy control group. Venous blood was collected from the medial cubital vein of each-patient on an empty stomach in early mornings to detect blood uric acid levels. Results Blood uric acid levels were lower in each syndrome type group than in the healthy control group (146.08 +/- 67.24 micromol/L in the HSCP group; 157.08 +/- 69.77 micromol/L in the PFDH group; 162.58 +/- 97.72 micromol/L in the RDH group; 156.20 +/- 62.63 micromol/L in the DQB group; 161.83 +/- 111.23 micromol/L in the DGYSY group; 194.41 +/- 90.01 micromol/L in the DGS group; 242.39 +/- 87.55 micromol/L in the healthy control group, P < 0.01). Blood uric acid levels were higher in the DGYSY group than in the other 5 syndrome groups (P < 0.01). Correlation analyses between Goldstein grading and blood uric acid showed that, along with increased Goldstein grade (that was aggravating disease conditions), WD patients' blood uric acid levels decreased (P < 0.01).

CONCLUSIONSWD patient's blood uric acid levels decreased more. Blood uric acid levels and Goldstein grading were different in various Chinese medical syndrome types. Blood uric acid levels had certain value in assessing the severity of WD.

Asian Continental Ancestry Group ; Heart ; Hepatolenticular Degeneration ; blood ; classification ; diagnosis ; Humans ; Medicine, Chinese Traditional ; Syndrome ; Uric Acid ; blood

This study was purposed to construct the recombinant retroviral vector containing human homeobox gene hoxA10 and to establish the packaging cell lines which stably produce viruses. The whole coding region of hoxA10 gene was amplified by PCR and inserted into the retroviral vector MSCVneo. The recombinant vector was identified by DNA sequencing. The recombinant and control retroviral vectors were transfected into the packaging cell line PT67 by liposome Lipofectamine(TM) 2000. These cell lines stably producing retrovirus were isolated following G418 selection. The viral suspension was harvested and the viral titer was determined by NIH3T3 cells. The results showed that the recombinant retroviral vector was proved to encode hoxA10 genes by sequencing. The cell lines efficiently producing virus were screened by G418 and designated as PT67/MSCVneo and PT67/MSCVneo-hoxA10. The titers of them were 5 x 10(5) CFU/ml and 4 x 10(4) CFU/ml respectively. It is concluded that the recombinant retroviral vector containing homeobox gene hoxA10 and the stably packaging cell lines which efficiently and correctly produce viruses are successfully constructed, which provides a basis for further exploration of the hoxA10 gene function in the regulation of hematopoiesis.
Cell Line ; Genes, Homeobox ; Genetic Vectors ; Homeodomain Proteins ; genetics ; Humans ; Retroviridae ; genetics ; Transfection

The study was aimed to establish a standard procedure for human umbilical cord blood bank. The hematopoietic nucleated cells in cord blood were processed by using sedimentation and centrifugation method. After finishing CD34(+) cell counting, hematopoietic progenitor cell assay, microbial culture, infectious disease test and HLA typing, cord blood units were stored in the liquid nitrogen for further application. The results showed that nucleated cells of cord blood were (10.94 +/- 2.74) x 10(8) per unit; recovery rate of nucleated cells was (79.82 +/- 17.76)%. CD34(+) cells in cord blood were counted as (51.62 +/- 30.53) x 10(5) per unit. Eight units of cord blood were thawed after two years of cryopreservation, the recovery rate of nucleated cells, CD34(+) cells and CFU-GM were (91.4 +/- 6.0)%, (84.6 +/- 20.0)% and (85.8 +/- 14.9)% respectively. It is suggested that the methods and procedure reported for processing and cryopreservation of hematopoietic stem/progenitor cells in the human umbilical cord blood is effective.
Antigens, CD34 ; blood ; Blood Preservation ; methods ; Cell Separation ; methods ; Colony-Forming Units Assay ; Cryopreservation ; methods ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; immunology ; Humans

The aim of this study was to clone human RHD gene and to investigate its expression in transduced K562 cells. Total RNA was extracted from reticulocyte of cord blood. RHD and RHCE genes were amplified using RT-PCR method. The amplified products were cloned into pGEM-T plasmid by TA ligation and several clones were screened by direct sequencing method in order to obtain the RHD gene. RHD gene was subcloned into pcDNA3.1(-) expression vector, then the recombined plasmids were transduced into K562 cells with superfect transfection reagent kit. Finally transcription and expression of RHD gene in K562 cells were detected. The result showed that RHD gene has been cloned sucessfully, the inserted sequence and direction of RHD cDNA in its recombined pcDNA3.1(-) vector were identified using enzyme cutting and sequencing method. After transduced with recombined pcDNA3.1(-) vector, K562 cells could transcribe RHD mRNA in its cytoplasm and express RhD antigen on its membrane surface. In conclusion, RhD antigen can expressed in K562 cells with RHD cDNA transduction, and the expression system in vitro may be helpful to further investigate the molecular basis of RhD variants.
Base Sequence ; Cell Membrane ; metabolism ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Gene Expression ; Genetic Vectors ; genetics ; Humans ; K562 Cells ; Molecular Sequence Data ; RNA, Messenger ; biosynthesis ; genetics ; Rh-Hr Blood-Group System ; biosynthesis ; genetics ; Sequence Analysis, DNA ; Transfection

To study the effects of short-term cryopreservation of cord blood hematopoietic cells in liquid nitrogen, the viability and function of cord blood hematopoietic cells were investigated by using each of 8 samples cryopreserved for six months, one and two years after thawing respectively. Nucleated cells (NC) were detected by blood cell analyzer. CD34+ cells were analyzed by flow cytometry, CFU-GM were cultured and detected in vitro, the survival rate was determined by trypan blue staining. The results showed that the differences of recovery rate of NC, CD34+, CFU-GM were nonsignificant at three different cryopreserved times. In conclusion, the short-term storage in liquid nitrogen showed a good effect on cord blood hematopoietic cell without any significant change of activities and number of the cryopreserved hematopoietic cells.
Cell Survival ; Cryopreservation ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; physiology ; Humans

We were introducing a log-linear model for case-parent triad study.Data from a previous study of the association between MTHFR C677T aud cleft lip with or without cleft palate (CL/P) was analyzed,in order to investigate the maternal effect,fetal effect and their interaction,using a series of log-linear models.Our results showed that mothers who were carrying two copies of MTHFR C677T variant alleles appeared to have reduced the risk of CL/P in offspring,comparing to those with homozygous of wild-type allele.With S2=0.43 (95％ CI:0.19-0.95).No significant association was found for fetal genotype and maternal-fetal iuteraction with CL/P.Log-linear model method seemed to be useful in the estimation of maternal effect,fetal effect and maternal-fetal interaction,in the case-parent triad study design.This approach showed specific benefit in studies that related to genetic effects on complex diseases such as pregnancy complications and diseases originated from fetus.

This study was purposed to investigate the molecular genetics basis for para-Bombay phenotype. The para-Bombay phenotype of two probands was identified by routine serological techniques. The full coding region of alpha (1, 2) fucosyltransferase gene (FUT1 and FUT2) in the probands was amplified by polymerase chain reaction and the amplified fragments were directly sequenced, meanwhile the mutations of FUT1 were also identified by TOPO TA cloning sequence method. The results indicated that two heterozygous mutations were detected by directly sequencing in two probands: AG deletion at position 547 - 552 and C to T mutation at position 658. Two different mutations were confirmed to be true compound heterozygotes with each mutation on a separate homologous chromosome by TOPO TA cloning sequence method. AG deletion at position 547 - 552 caused a reading frame shift and a premature stop codon. C658T mutation resulted in Arg-->Cys at amino acid position 220. It is suggested that the FUT1 mutation of two probands are compound heterozygous mutation with different chromosomes, which are named h1h3 and may be the genetics basis of para-Bombay phenotype.
ABO Blood-Group System ; genetics ; Alleles ; Frameshift Mutation ; Fucosyltransferases ; genetics ; Gene Deletion ; Heterozygote ; Humans ; Male ; Mutation, Missense

To investigate the molecular genetics basis for one para-Bombay phenotype, the red blood cell phenotype of the proband was characterized by standard serological techniques. Exon 6 and 7 of ABO gene, the entire coding region of FUT1 gene and FUT2 gene were amplified by polymerase chain reaction from genomic DNA of the proband respectively. The PCR products were purified by agarose gels and directly sequenced. The PCR-SSP and genescan were performed to confirm the mutations detected by sequencing. The results showed that the proband ABO genotype was A(102)A(102). Two heterozygous mutations of FUT1 gene, an A to G transition at position 682 and AG deletion at position 547-552 were detected in the proband. A682G could cause transition of Met-->Val at amino acid position 228, AG deletion at position 547-552 caused a reading frame shift and a premature stop codon. The FUT2 genotype was heterozygous for a functional allele Se(357) and a weakly functional allele Se(357), 385 (T/T homozygous at position 357 and A/T heterozygous at 385 position). It is concluded that the compound heterozygous mutation--a novel A682G missense mutation and a 547-552 del AG is the molecular mechanism of this para-Bombay phenotype.
ABO Blood-Group System ; genetics ; China ; DNA Mutational Analysis ; Female ; Fucosyltransferases ; genetics ; Genotype ; Humans ; Male ; Mutation ; Mutation, Missense ; Pedigree ; Phenotype ; Sequence Deletion