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Qin F.,Zhejiang Provincial Blood Center
Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology | Year: 2010

The aim of this study was to investigate the feasibility of using fetal short tandem repeat (STR) loci in maternal plasma as gender-independent fetal DNA marker. DNA from maternal plasma sample was extracted using QIAamp DNA Kit. AmpF1 STR profiler box was used to amplify 9 different polymorphic short tandem repeat (STR) loci (D3S1358, VWA, FGA, D5S818, D13S317, D7S820, D8S1179, D21S11, D18S51), the multiplex fluorescent PCR was used to amplify the STR alleles of fetal DNA in 36 pregnant plasma samples of pregnant women at different pregnancy. Their husbands' DNA isolated from whole blood samples were amplified at the same time. The PCR products were electrophoresis by ABI Prism 377 sequencer, the results of electrophoresis were analysed by Genscan. The presence of fetal DNA in maternal plasma by Paternally inherited fetal alleles were detected. The results showed that paternally inherited fetal alleles were detected in 4 cases in early pregnancy (4/6), 19 cases in middle pregnancy (19/20) and 9 cases in late pregnancy (9/10) respectively, the paternally inherited fetal alleles in 4 of 36 cases could not be detected. It is concluded that fluorescent multiplex PCR can be used for amplification of male and female fetal STRs in maternal plasma to obtain genetic information, which may have implication for non-invasive prenatal diagnosis of certain hereditary diseases independent of the fetal sex. Source

Xu X.G.,Zhejiang Provincial Blood Center
Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology | Year: 2010

The aim of study was to explore the feasibility of quantitative chimerism analysis of regulatory T (Treg) cells using immune sorting coupling short tandem repeat (STR) method. 14 sets of artificial chimera samples were prepared by mixed lymphocytes according to different proportion. The CD4(+)CD25(+) Treg cells were harvested by negative and positive selection of immunomagnetic beads, then the STR polymorphisms of 16 loci in sorted Treg cells was analyzed. The results showed that the DNA amount extracted from sorted Treg cells was fit for STR detection. All STR alleles specific for recipient or donor could be detected and the quantitative results were consistent with theoretic values in over 10% recipient chimeras. But only partial recipient alleles could be detected and the quantitative results were different from theoretic values in less then 1% recipient chimeras. It is concluded that a quantitative chimerism analysis of Treg cell based on immune sorting is established. The sensitivity and accuracy for chimera detection are 1% to 10%, and this method can be used to monitoring hematopoietic chimerism following allogeneic hematopoietic stem cell transplantation. Source

Xia X.-H.,Zhejiang Provincial Blood Center | Huang D.,Zhejiang University of Science and Technology
World Chinese Journal of Digestology | Year: 2014

AIM: To explore the significance of joint detection of blood follistain-like protein 1 (FSTL1), C-reactive protein (CRP) and D-dimer in the early diagnosis of ulcerative colitis. METHODS: Venous blood samples were collected from 67 patients with ulcerative colitis and 60 healthy people. Serum levels of FSTL1, CRP and D-dimer were detected by ELISA, and the significance of combined detection of the three indexes in the early diagnosis of ulcerative colitis was analyzed. RESULTS: Serum levels of FSTL1, CRP and D-two dimer were significantly higher in UC patients than in normal controls, and in patients with active UC than in those in remission (t = 10.671, 10.398, 31.873, P < 0.05 for all). With the aggravation of the disease, serum levels of FSTL1, CRP and D-dimer significantly increased in patients with UC, and the levels were significantly higher in the severe group than in the moderate and mild groups (t = 5. 766, 5.821, 2.307, 2.213, 2.789, 2.363, P < 0.05 for all). Serum levels of FSTL1, CRP, and D-dimer were significantly correlated with UC disease activity index (DAI) (r = 0.511, 0.312, 0.317, P < 0.05 for all). The sensitivity and negative predictive value of combined detection of FSTL1, CRP and D-dimer were significantly higher than those of detection of any one of the indexes (χ2 = 0.013, 4. 823, P < 0.05). CONCLUSION: The joint detection of serum FSTL1, CRP and D-dimer can be used for early diagnosis of ulcerative colitis. © 2014 Baishideng Publishing Group Co., Limited. All rights reserved. Source

Ying Y.,Zhejiang Provincial Blood Center | He Y.,Zhejiang Provincial Blood Center | Tao S.,Zhejiang Provincial Blood Center | Han Z.,Zhejiang Provincial Blood Center | And 6 more authors.
Immunogenetics | Year: 2013

The polymorphism of major histocompatibility complex class I chain-related gene B (MICB) and variations in MICB alleles in a variety of populations have been characterized using several genotyping approaches. In the present study, a novel polymerase chain reaction sequence-based typing (PCR-SBT) method was established for the genotyping of MICB exons 2-6, and the allelic frequency of MICB in the Zhejiang Han population was investigated. Among 400 unrelated healthy Han individuals from Zhejiang Province, China, a total of 20 MICB alleles were identified, of which MICB005:02:01, MICB002:01:01, and MICB004:01:01 were the most predominant alleles, with frequencies of 0.57375, 0.1225, and 0.08375, respectively. Nine MICB alleles were detected on only one occasion, giving a frequency of 0.00125. Of the 118 distinct MICB ∼ HLA-B haplotypes identified, 42 showed significant linkage disequilibrium (P < 0.05). Haplotypes MICB005:02:01 ∼ B46:01, MICB005:02:01 ∼ B40:01, and MICB008 ∼ B58:01 were the most common haplotypes, with frequencies of 0.0978, 0.0761, and 0.0616, respectively. Five novel alleles, MICB005:07, MICB005:08, MICB027, MICB028, and MICB029 were identified. Compared with the MICB005:02:01 sequence, a G > A substitution was observed at nucleotide position 210 in MICB005:07, and a 1,134 T > C substitution in MICB005:08 and an 862 G > A substitution in MICB027 were detected. In addition, it appears that MICB028 probably arose from MICB004:01:01 with an A to G substitution at position 1,147 in exon 6. MICB029 had a G > T transversion at nucleotide position 730 in exon 4, compared with that of MICB002:01:01. On the basis of the new PCR-SBT assay, these observed results demonstrated MICB allelic variations in the Zhejiang Han population. © 2013 Springer-Verlag Berlin Heidelberg. Source

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