Blood Research Laboratory

Chengdu, China

Blood Research Laboratory

Chengdu, China

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PubMed | Blood Research Laboratory, Chengdu Blood Center and Purdue University
Type: Journal Article | Journal: Archives of virology | Year: 2016

Hepatitis C virus (HCV) is a significant pathogen of global concern. The virus is usually spread through blood contact, such as transfusion, hemodialysis and injection of illegal drugs. HCV genotypes have a geographic distribution in different areas. In this paper, we focus on the distribution of HCV genotypes from volunteer blood donors in Chengdu. The prevalence of genotypes was analyzed using phylogenetic analysis. Phylogenetic trees were constructed based on the HCV core and NS5B regions from 313 sequences. HCV sequences were classified into six subtypes, and HCV genotypes were determined with the following results: 1b in 283, 2a in 14, 3b in seven, 3a in three, 6a in five and 6u in one. Subtype 1b was the most common and accounted for approximately 90.41% (283/313), and a virus of subtype 6u was isolated for the first time from the Chengdu area. Genotypes 4 and 5 were not detected.


Xin X.,Red Cross | Gong T.,Blood Research Laboratory | Hong Y.,Blood Research Laboratory | Dang H.,Red Cross
International Journal of Clinical and Experimental Medicine | Year: 2016

This study was designed to explore the influence of long-term corneal contact lens (CL) wearing on the stability and quality of tear film in Qinghai. Thirty subjects with CL over five years and thirty myopia subjects without CL as control were recruited in our study. Basic tear secretion test, tear film break-up time (TBUT), and corneal fluorescein staining were determined. For further evaluating the quality of tear film of two groups, the mucin levels including mucin 1 (MUC1) and mucin 5AC (MUC5AC) were measured by enzyme-linked immunosorbent assay (ELISA), meanwhile MUC1 and MUC5AC mRNA expression were processed by real-time polymerase chain reaction (RT-PCR) after samples were collected. Compared with control group, basic tear secretion and TBUT decreased, obvious corneal fluorescein staining presented in CL group. The mean protein quantity of MUC1 and MUC5AC in CL group (1.1612±0.2119 ng/ml, 1.6731±0.2457 ng/ml respectively) was less than that of control group (1.2369±0.1825 ng/ml and 1.7892±0.2461 ng/ml respectively), there were significant differences between the two groups (t=2.098, P=0.038; t=2.586, P=0.011 respectively). Both the mRNA expression of MUC1 and MUC5AC in the CL group (1.3944±0.1212 and 1.622±0.1988 respectively) decreased in comparison with control group (1.4630±0.1028 and 1.7056±0.1638 respectively), significant differences was observed between the two groups (t=3.343, P=0.001; t=2.514, P=0.013 respectively). These results suggest that long-term CL wearing in Qinghai might pose an impact on the stability and quality of tear film. © 2016, E-Century Publishing Corporation. All rights reserved.


Zhou C.,Xavier University of Louisiana | Zhou C.,Blood Research Laboratory | Zhong Q.,Xavier University of Louisiana | Rhodes L.V.,Tulane University | And 8 more authors.
Breast Cancer Research | Year: 2012

Introduction: Acquired tamoxifen resistance involves complex signaling events that are not yet fully understood. Successful therapeutic intervention to delay the onset of hormone resistance depends critically on mechanistic elucidation of viable molecular targets associated with hormone resistance. This study was undertaken to investigate the global proteomic alterations in a tamoxifen resistant MCF-7 breast cancer cell line obtained by long term treatment of the wild type MCF-7 cell line with 4-hydroxytamoxifen (4-OH Tam).Methods: We cultured MCF-7 cells with 4-OH Tam over a period of 12 months to obtain the resistant cell line. A gel-free, quantitative proteomic method was used to identify and quantify the proteome of the resistant cell line. Nano-flow high-performance liquid chromatography coupled to high resolution Fourier transform mass spectrometry was used to analyze fractionated peptide mixtures that were isobarically labeled from the resistant and control cell lysates. Real time quantitative PCR and Western blots were used to verify selected proteomic changes. Lentiviral vector transduction was used to generate MCF-7 cells stably expressing S100P. Online pathway analysis was performed to assess proteomic signatures in tamoxifen resistance. Survival analysis was done to evaluate clinical relevance of altered proteomic expressions.Results: Quantitative proteomic analysis revealed a wide breadth of signaling events during transition to acquired tamoxifen resistance. A total of 629 proteins were found significantly changed with 364 up-regulated and 265 down-regulated. Collectively, these changes demonstrated the suppressed state of estrogen receptor (ER) and ER-regulated genes, activated survival signaling and increased migratory capacity of the resistant cell line. The protein S100P was found to play a critical role in conferring tamoxifen resistance and enhanced cell motility.Conclusions: Our data demonstrate that the adaptive changes in the proteome of tamoxifen resistant breast cancer cells are characterized by down-regulated ER signaling, activation of alternative survival pathways, and enhanced cell motility through regulation of the actin cytoskeleton dynamics. Evidence also emerged that S100P mediates acquired tamoxifen resistance and migration capacity. © 2012 Zhou et al.; licensee BioMed Central Ltd.


News Article | January 25, 2016
Site: www.biosciencetechnology.com

A single injection. That’s all someone with a factor VII deficiency would need for a life-long cure, thanks to a new gene therapy treatment developed in a collaboration of researchers at the University of North Carolina (UNC) and The Children’s Hospital of Philadelphia (CHOP). Factor VII deficiency is a rare blood disorder that can lead to profuse bleeding after injuries, including minor cuts. Currently, patients must be injected regularly with protein factors. But the new gene therapy approach, details of which were published in the journal Blood, would require only one injection and occasional follow up to ensure that the body is still producing proper amounts of factor VII. “For many people living in the developing world, constant injections aren’t practical or at least not feasible right now; but a single injection could be,” said Tim Nichols, M.D., who led the UNC team and is professor of medicine and pathology at the UNC School of Medicine. “We’ve now shown that this treatment is safe and effective for dogs with inherited factor VII deficiency. This is exciting because our other gene therapy work on bleeding disorders, such as hemophilia, has demonstrated that safety and efficacy in dogs has translated well in human clinical trials.” Certain populations of dogs have factor VII deficiency, and Nichols said he wouldn’t hesitate to treat them with this new gene therapy. In people, factor VII deficiency is rare, occurring in an estimated 1 in 300,000 to 500,000 births. Both parents must carry the defective gene for their children to have the disorder. Without enough factor VII, people cannot form clots properly. Babies are typically diagnosed within six months of life, usually after an episode of profuse bleeding in the brain or the gastrointestinal tract. After early childhood, people with factor VII deficiency typically do not experience spontaneous bleeding, but they are susceptible to serious bleeding after surgery, and they bruise easily. A cut or injury on the skin or in the mouth or nose can lead to severe bleeding. Women with factor VII deficiency can suffer from heavy bleeding during periods. For the study published in Blood, Paris Margaritis, DPhil, the lead researcher at The Children’s Hospital of Philadelphia and Penn’s Perelman School of Medicine and the senior author of the study, and his colleagues cloned the canine factor VII gene and developed a method to package a substantial number of factor VII genes inside an adeno-associated virus (AVV) vector – a sort of transport system, a virus that’s been rendered harmless. Meanwhile Nichols, the director of the Francis Owen Blood Research Laboratory at the UNC School of Medicine, characterized the factor VII deficiency in each of four dogs. Then he treated each of them with a different amount of the AAV therapy. The amount of factor VII produced was directly proportional to the amount of vector given to the individual dogs. Over the next three years, Nichols’s team characterized the health of the dogs. Nichols also worked with The CHOP researchers to analyze the factor VII gene expression data. One dog received the amount of AAV associated with the maximum amount that a human could likely tolerate before increasing the risk of an immune system response against the therapy itself. This dog produced 30 percent of the amount of factor VII that a normal dog would produce. This amount of factor VII would be more than enough to cure the dog. “This is the only large animal model that represents the majority of factor VII mutation types,” Margaritis said. “Our data are the first to demonstrate feasibility, safety, and long-term duration of AAV gene therapy for factor VII deficiency. The table is now set to propose clinical trials that would treat people who suffer from this disease.”


Gong T.X.,Blood Research Laboratory
Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology | Year: 2012

The purpose of this study was to find the rare individual JK(a-b-) phenotype of proband family and explore its molecular mechanism and the genetic background, in order to provide base for searching compatible donor to blood transfusion of the individuals with rare JK(a-b-) phenotype. Urea lysis test was used to screen the JK(a-b-) phenotype and results were confirmed with serological method. The genotypes were detected with PCR-SSP. The 4-11 exons and their flanking intron regions of JK gene were amplified and sequenced. The results showed that her elder brother has a same phenotype JK(a-b-) and genotypes JK(a)/JK(b) with proband. The phenotype and genotypes of their parent is JK (a+b-) and JK(a)/JK(b), respectively; and the younger sister's is JK (a+b-) and JK(a)/JK(a). Acceptor site of intron 5 3' g > a mutation was detected in proband and her elder brother, which may cause the JK(a-b-) phenotype of proband and her elder brother. There is g/a and a at this site in their parent and younger sister, respectively. Additionally, the SNP (ncbi:rs8090908) a > g at nt-99 in intron 3 was found in proband and her elder brother, it needs to be explored whether the SNP is related to JK(a-b-) phenotype. This SNP was not found in their parent and younger sister. This JK(a-b-) phenotype abides by the rule of dominant inheritance in the family, suggesting that there is higher probability to find homology phenotype and genotype by investigating in their family, especially in their siblings.


Chen X.,Blood Research Laboratory
Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology | Year: 2012

This study was aimed to establish the real-time multiple-PCR with melting curve analysis for Duffy blood group Fy-a/b genotyping. According to the sequence of mRNA coding for β-actin and Fy-a/b, the primers of β-actin and Fy-a/b were synthesized. The real-time multiple-PCR with melting curve analysis for Fy-a/b genotyping was established. The Fy-a/b genotyping of 198 blood donors in Chinese Chengdu area has been investigated by melting curve analysis and PCR-SSP. The results showed that the results of Fy-a/b genotype by melting curve analysis were consistent with PCR-SSP. In all of 198 donors in Chinese Chengdu, 178 were Fy(a) (+) (89.9%), 19 were Fy(a) (+) Fy(b) (+) (9.6%), and 1 was Fy(b) (+) (0.5%). The gene frequency of Fy(a) was 0.947, while that of Fy(b) was 0.053. It is concluded that the genotyping method of Duffy blood group with melting curve analysis is established, which can be used as a high-throughput screening tool for Duffy blood group genotyping; and the Fy(a) genotype is the major of Duffy blood group of donors in Chinese Chengdu area.


Hong Y.,Blood Research Laboratory | Gong T.-X.,Blood Research Laboratory | Zhou C.-H.,Blood Research Laboratory
Chinese Journal of Medical Genetics | Year: 2012

Objective: To study the molecular genetics characteristics of Jk(a-b-) phenotype of blood donors from Chengdu. Methods: Exons 4-11 of the JK genes and their flanking intronic regions for 8 Jk(a-b-) samples were analyzed with PCR-sequence specific primers (PCR-SSP) and DNA sequencing. Results: All samples had AA genotype at position 838 of exon 9 predicting a null Jkb-like alleles. Sequence analysis has revealed 4 mutant alleles, which included: (1) IVS5 - 1G>A, A to G at position 588 (Pro196Pro) of exon 7; (2) G to A at position 896 (Gly299Glu) of exon 9, A to G at position 588 (Pro196Pro) of exon 7; (3) IVS5-1G>A, C>A at position 222 (Asn74Lys) of exon 5, A to G at position 499 (Met167Val) of exon 7, A to G at position 588 (Pro196Pro) of exon 7; and (4) IVS5-1G> A, G to A at position 896 (Gly299Glu) of exon 9, A to G at position 588 (Pro196Pro) of exon 7. Conclusion: IVS5-1G>A, C to A at position 222 (Asn74Lys) of exon 5 and G to A at position 896 (Gly299Glu) of exon 9 might have been the molecular genetic mechanisms underlying Jk(a-b-) phenotype of the selected blood donors.


PubMed | Blood Research Laboratory
Type: Journal Article | Journal: Zhongguo shi yan xue ye xue za zhi | Year: 2012

This study was aimed to establish the real-time multiple-PCR with melting curve analysis for Duffy blood group Fy-a/b genotyping. According to the sequence of mRNA coding for -actin and Fy-a/b, the primers of -actin and Fy-a/b were synthesized. The real-time multiple-PCR with melting curve analysis for Fy-a/b genotyping was established. The Fy-a/b genotyping of 198 blood donors in Chinese Chengdu area has been investigated by melting curve analysis and PCR-SSP. The results showed that the results of Fy-a/b genotype by melting curve analysis were consistent with PCR-SSP. In all of 198 donors in Chinese Chengdu, 178 were Fy(a) (+) (89.9%), 19 were Fy(a) (+) Fy(b) (+) (9.6%), and 1 was Fy(b) (+) (0.5%). The gene frequency of Fy(a) was 0.947, while that of Fy(b) was 0.053. It is concluded that the genotyping method of Duffy blood group with melting curve analysis is established, which can be used as a high-throughput screening tool for Duffy blood group genotyping; and the Fy(a) genotype is the major of Duffy blood group of donors in Chinese Chengdu area.


PubMed | Blood Research Laboratory
Type: Journal Article | Journal: Zhongguo shi yan xue ye xue za zhi | Year: 2012

The purpose of this study was to find the rare individual JK(a-b-) phenotype of proband family and explore its molecular mechanism and the genetic background, in order to provide base for searching compatible donor to blood transfusion of the individuals with rare JK(a-b-) phenotype. Urea lysis test was used to screen the JK(a-b-) phenotype and results were confirmed with serological method. The genotypes were detected with PCR-SSP. The 4-11 exons and their flanking intron regions of JK gene were amplified and sequenced. The results showed that her elder brother has a same phenotype JK(a-b-) and genotypes JK(a)/JK(b) with proband. The phenotype and genotypes of their parent is JK (a+b-) and JK(a)/JK(b), respectively; and the younger sisters is JK (a+b-) and JK(a)/JK(a). Acceptor site of intron 5 3 g > a mutation was detected in proband and her elder brother, which may cause the JK(a-b-) phenotype of proband and her elder brother. There is g/a and a at this site in their parent and younger sister, respectively. Additionally, the SNP (ncbi:rs8090908) a > g at nt-99 in intron 3 was found in proband and her elder brother, it needs to be explored whether the SNP is related to JK(a-b-) phenotype. This SNP was not found in their parent and younger sister. This JK(a-b-) phenotype abides by the rule of dominant inheritance in the family, suggesting that there is higher probability to find homology phenotype and genotype by investigating in their family, especially in their siblings.


PubMed | Blood Research Laboratory, PhD corresponding author . and MA.
Type: Journal Article | Journal: Immunohematology | Year: 2015

The detection of polymorphism is the basis of blood group genotyping and phenotype prediction. Genotyping may be useful to determine blood groups when serologic results are unclear. The development and application of different methods for blood group genotyping may be needed as a substitute for blood group typing. The purpose of this study is to establish an approach for blood group genotyping based on a melting curve analysis of real-time polymerase chain reaction (PCR). Using DNA extracted from whole blood, we developed and validated a DNA typing method for detecting DO*01/DO*02, DO*01/DI*02, LU*01/LU*02, and GYPB*03/GYBP*04 alleles using a melting curve analysis. All assays were confirmed with a commercial reagent containing sequence-specific primers (PCR-SSP), and a cohort of the samples was confirmed with sequencing. Results for all blood groups were within the range of specificity and assay variability. Genotypes of 300 blood donors were fully consistent with PCR-SSP data. The obtained genotype distribution is in complete concordance with existing data for the Chinese population. There are several advantages for this approach of blood group genotyping: lower contamination rates with PCR products in this laboratory, ease of performance, automation potential, and rapid cycling time.

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