1.Research advances on microchimerism.
Xue-Dong SUN ; Jian-Li SANG ; Hui-Sheng AI
Journal of Experimental Hematology 2014;22(4):1153-1156
The microchimerism is a status of the microcell or DNA of an individual in another one with genetic differences. Taking an overall view about the discovery and research of the microchimerism, it was found that although the study of the microchimerism emphasizes the formation, origin, distribution, type, relationship to disease and several other aspects, the objects of the study are always the microchimerism that obtained naturally. As it is known to all, the microchimerism can also be produced in some clinical treatment, such as in the transplant and transfusion, but compared with the microchimerism gained naturally, obviously, the study for the iatrogenic microchimerism formed in the treatment is not elaborate enough. The curative effect of micro transplantation, a new technique for leukemia treatment, is obvious, but its mechanism is unclear, whether that is related to microchimerism still needs further research. This review summarizes the study history and perspective of the microchimerism so as to provide some ideas for studying the action mechanism of microchimerism in micro transplantation.
Chimerism
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DNA
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genetics
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Humans
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Transplantation Chimera
2.Characterization of a pair of twins as blood group chimeras.
Yuli ZHU ; Wenhui MA ; Haiyan WANG ; Bin HU ; Zhihui FENG
Chinese Journal of Medical Genetics 2020;37(12):1399-1402
OBJECTIVE:
To delineate the blood group for a pair of twins with inconclusive ABO blood typing result.
METHODS:
Serological test for blood group was carried out by using ABO and Rh Blood Grouping Cards (Microcolumn Gel Immunoassay). Sequence specific primer-PCR (PCR-SSP), direct sequencing and TA clone sequencing were used to analyze the ABO gene. Genetic status was analyzed by using 16 short tandem repeat (STR) markers.
RESULTS:
Red blood cells of the twins displayed 2+ mixed agglutination phenomenon with anti-A, anti-A1 and anti-E. PCR-SSP and DNA sequencing of exons 6 to 7 revealed that they have an ABO*O.01.01/ABO*O.01.02 genotype. DNA sequencing of microsatellite enhancer region revealed presence of A gene. STR analysis revealed more than two haplotypes for 9 loci between the twins. After clustered by anti-A, the red blood cells were divided into two groups: A, CcDEe and O, CcDee, respectively.
CONCLUSION
Serological and molecular techniques have characterized the twins as blood group chimeras.
ABO Blood-Group System/genetics*
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Alleles
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Chimera/genetics*
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Genotype
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Humans
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Twins/genetics*
3.Short tandem repeat panel settled for quantitative chimerism analysis following allogeneic stem cell transplantation and its application.
Ying YANG ; Wen WU ; Ling-Ling WANG ; He-Ping CHEN ; Zi-Yan ZHU
Journal of Experimental Hematology 2009;17(3):706-712
To evaluate the roles of 8 short tandem repeats (STR) loci as STR panel in quantitative analysis of chimerism following transplantation, the primers were synthesized and marked with different dyes for D3S3045, D4S2366, D4S2639, D5S818, D13S317, D18S1002, D20S481 and D22S689. The blood samples of 15 cases received allogeneic stem cell transplantation were collected before and after transplantation, then DNA was extracted and amplified with these primers, and was further analysed under ABI Genetic Analyser 3100 to select suitable informative STR locus. Donor/recipient dilution series were prepared to get standard curves in selected loci, the DNAs extracted at different days after transplantation were used to quantitatively analyze the chimerism in patients according to the values of peak area or peak height of fluorescent signals. The standard curves can be used to calculate the chimerism by plotting the respective R/D quotient value against the percentage of recipient DNA. The results indicated that the calculated chimerism was in concordance with the donor/recipient dilution. The STR panel succeeded in identifying at least one informative marker and quantitative monitoring the chimerism after HSCT in 15 donor-recipient pairs and a relapsed case was diagnosed. It is concluded that the STR panel and its detection method can accurately and quantitatively monitor the chimerism after allogeneic HSCT, which is more economical and flexible than using commercial kits.
DNA
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genetics
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DNA Primers
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Hematopoietic Stem Cell Transplantation
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Humans
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Microsatellite Repeats
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Transplantation Chimera
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genetics
4.A rare case of dicentric ring chromosome and derivative ring chromosome Chimera.
Junzhen ZHU ; Xiaoping YU ; Xiaofeng QI ; Qinying CAO ; Wenshuang ZHU ; Dan YANG ; Haoyu ZHANG ; Zhanyun SONG ; Shibo WANG ; Cuixia WANG
Chinese Journal of Medical Genetics 2022;39(5):534-536
OBJECTIVE:
Utilize high-resolution chromosome analysis and microarray detection to determine the genetic etiology of infertility of a 32-year old female patient.
METHODS:
The peripheral blood of the patient was cultured for high-resolution chromosome G and C banding karyotype analysis, and then 750K SNP-Array chip detection was performed.
RESULTS:
Karyotype analysis results showed that the patient's karyotype was 45,XX,-13 [7]/46,XX,r(13) (p13q34) [185]/46,XX,dic r(13;13)(p13q34;p13q34) [14]/ 47,XX,+der(13;13;13;13) (p13q34;p13q34;p13q34; p13q34), dic r(13;13) [1]/ 46,XX [3]. The microarray results showed that the patient had a 3.3 Mb deletion in the 13q34 segment of chromosome 13, which may be related to infertility.
CONCLUSION
Infertility of the patient reported in this article may be related to the deletion of chromosome segment (13q34-qter).
Adult
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Chimera
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Chromosome Banding
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Chromosome Deletion
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Chromosome Disorders/genetics*
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Dacarbazine
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Female
;
Humans
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Infertility/genetics*
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Ring Chromosomes
5.Clinical correlations of peripheral blood microchimerism after liver transplantation.
Kyung Mo KIM ; Eun Jung KIM ; Han Wook YOO ; Jong Jin SEO ; Sung Gyu LEE
Journal of Korean Medical Science 2000;15(3):260-264
The aim of this study was to evaluate microchimerism after human liver transplantation (LT). This study included 13 female recipients who received hepatic allograft from male donors at Asan Medical Center. A nested PCR specific for Y-chromosome gene (DYZ3) was used to analyze the small number of male cells in the peripheral blood mononuclear cells of the female recipients. Microchimerism was observed in 6 of 13 recipients and 16 out of 35 samples. Only 3 patients showed microchimerism 3 months after LT. There was no statistical difference between the presence of microchimerism and clinical findings such as type of donor, type of immunosuppression, episode of rejection and age of recipient. This study did not show any clinical relevance of microchimerism and further larger study are needed to confirm the results.
Adolescence
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Adult
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Animal
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Child
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Child, Preschool
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Female
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Human
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Infant
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Liver Transplantation*
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Lymphocytes/immunology*
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Male
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Transplantation Chimera/immunology*
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Transplantation Chimera/genetics
;
Y Chromosome
6.Clinical correlations of peripheral blood microchimerism after liver transplantation.
Kyung Mo KIM ; Eun Jung KIM ; Han Wook YOO ; Jong Jin SEO ; Sung Gyu LEE
Journal of Korean Medical Science 2000;15(3):260-264
The aim of this study was to evaluate microchimerism after human liver transplantation (LT). This study included 13 female recipients who received hepatic allograft from male donors at Asan Medical Center. A nested PCR specific for Y-chromosome gene (DYZ3) was used to analyze the small number of male cells in the peripheral blood mononuclear cells of the female recipients. Microchimerism was observed in 6 of 13 recipients and 16 out of 35 samples. Only 3 patients showed microchimerism 3 months after LT. There was no statistical difference between the presence of microchimerism and clinical findings such as type of donor, type of immunosuppression, episode of rejection and age of recipient. This study did not show any clinical relevance of microchimerism and further larger study are needed to confirm the results.
Adolescence
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Adult
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Animal
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Child
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Child, Preschool
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Female
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Human
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Infant
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Liver Transplantation*
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Lymphocytes/immunology*
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Male
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Transplantation Chimera/immunology*
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Transplantation Chimera/genetics
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Y Chromosome
7.Detection of hematopoietic chimera by real-time fluorescent quantitative PCR with erythrocyte Kidd blood group gene.
Shu CHEN ; Xian-Guo XU ; Ying LIU ; Xiao-Zhen HONG ; Fa-Ming ZHU ; Hang-Jun LÜ ; Li-Xing YAN
Journal of Experimental Hematology 2012;20(3):676-678
This study was aimed to establish the real-time fluorescent quantitative PCR (RT-qPCR) with erythrocyte Kidd blood group gene for detecting the hematopoietic chimera and to investigate the feasibility of this method. The TaqMan MGB probes and special primers were designed on basis of difference of erythrocyte Kidd blood group alleles, the hematopoietic chimerism was detected by RT-qPCR, the DNA chimerism was simulated by means of dilution of multiple proportions, and the sensitivity analysis was performed. The results showed that the RT-qPCR with erythrocyte Kidd blood group gene could effectively distinguish JK*A and JK*B alleles. There was no significant difference between the theoretic value and the practical measured value by this method (P > 0.05). As 156 donor's cells could be discriminated from 10(4) chimeric cells, this method may effectively detect donor's cells with correlation coefficient 0.998. It is concluded that the established RT-qPCR with erythrocyte Kidd blood group gene shows the feasibility for quantitative detection of hematopoietic chimera, and may be used to quantitatively detect chimera in a certain range.
Chimera
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Erythrocytes
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Humans
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Kidd Blood-Group System
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genetics
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Real-Time Polymerase Chain Reaction
8.Strategy Optimization and Clinical Analyze of Multiple Nucleotide Polymorphism Analysis in the Chimerism Detection after Allogeneic Hematopoietic Stem Cell Transplantation.
Zhe ZOU ; Rui SUN ; Yang-Wei LI ; Zhen GUO ; Rui-Hua FAN ; Xiao-Dong LYU
Journal of Experimental Hematology 2022;30(3):913-918
:
AbstractObjective: To investigate the sample selection, result correction and clinical application value of multi nucleotide polymorphism chimerism detection method based on Next-generation sequencing.
METHODS:
The chimerism samples from November 2018 to June 2020 were collected, and Pearson correlation coefficient (r) was used to analyze the consistency of bone marrow and peripheral blood results detected by MNPseq; according to the different information integrity before transplantation, the calibration model was constructed to analyze the correction value of the micro chimerism results in each model; the clinical results were retrospectively analyzed to verify the reliability and practicability of chimerism results correction and the clinical value of MNPseq method.
RESULTS:
The results of bone marrow and peripheral blood chimerism detected by MNPseq method were consistent with each other and showed significant correlation (r=0.985, P<0.01). The three groups of calibration models were constructed according to different pre-transplant information. For the no donor and pre-transplant patients information group, the correction value was 1%; while for the group with pre-transplant patients and without donor information, 0.61% of the chimerism rate and 13 heterotopic points were used as the correction value; 0.26% of the chimerism rate and 21.57% of the heterotopic points were used as the correction value for the group with pre-transplantation patients and donor information. After correction, the number of the patients with incomplete chimerism decreased from 276 (74.19%) to 141 (37.91%) (P<0.01). Among 18 (18/141, 12.77%) patients with incomplete chimerism, the results of MNPseq in the patients were 25-39 days earlier than those in STR and flow MRD, and the result showed statistical significance.
CONCLUSION
MNPseq method can be used to monitor chimerism with peripheral blood instead of bone marrow samples, and the results can be corrected to detect the changes of graft status in vivo in a more timely manner.
Chimerism
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Hematopoietic Stem Cell Transplantation
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Humans
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Nucleotides
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Reproducibility of Results
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Retrospective Studies
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Transplantation Chimera/genetics*
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Transplantation, Homologous
9.Morphological Characteristics of Normal and Gynandromorphic Hyalomma asiaticum Schulze and Schlottke, 1930.
Ze CHEN ; You Quan LI ; Qiao Yun REN ; Jin LUO ; Yonghong HU ; Kai LI ; Guang Yuan LIU ; Jian Xun LUO ; Jingze LIU ; Hong YIN
The Korean Journal of Parasitology 2015;53(3):361-364
Gynandromorphic ticks are extremely rare, and often attract parasitologists' attention. During our examination of tick specimens, an engorged gynandromorph of Hyalomma asiaticum was noticed. This is the first record of gynandromorphic ticks from China. In this study, several important morphological structures of normal and gynandromorphic H. asiaticum were analyzed. Comparing to the normal H. asiaticum, the gynandromorphic specimen was a typical bipartite protogynander. Its right side showed normal female characteristics, whereas the left side had normal male traits. Different from other gynandromorphic ticks containing 1 anus, this tick reported here had 2 complete anuses, and the anus of the male part had a single adanal plate.
Animals
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Chimera/*anatomy & histology/genetics
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China
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Female
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Ixodidae/*anatomy & histology/genetics
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Male
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Sheep
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Sheep Diseases/*parasitology
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Tick Infestations/parasitology/*veterinary
10.A Pichia pastoris with alpha-1, 6-mannosyltransferases deletion and its use in expression of HSA/GM-CSF chimera.
Yue WANG ; Xin GONG ; Shao-Hong CHANG ; Bo LIU ; Miao SONG ; Hai-Hua HUANG ; Jun WU
Chinese Journal of Biotechnology 2007;23(5):907-914
Yeast is a widely used host for recombinant protein expression. However, glycoproteins derived from yeast contain N-glycan of high mannose type and are usually hyperglycosylated. alpha-1,6-mannosyltransferases gene (och1) encodes the enzyme that initiates the first step of out-chain elongation of high mannose type N-glycan in yeast, which is different from that in human. So, a high efficient method to knockout target gene by two-step recombination was established and was used to delete och1. In the first recombinant, a plasmid with och1::ADE1 and ura3 gene was linearized in the downstream of och1 and inserted to the och1 site of P. pastoris genome, where the upstream and downstream of och1 were duplicated. In the second recombinant, the duplicated fragments of och1 were exchanged and the och1 deletion strains were selected on the plates containing 5-FOA, but no adenine. Then the och1 deletion strain was applied to express an human serum albumin (HSA) granulocyte-macrophage colony-stimulating factor (GM-CSF) chimera. Different with the hyperglycosylated HSA/GM-CSF chimera expressed in wild type P. pastoris, the chimera expressed in the och1 deletion strain, contained smaller N-glycan. The results suggested that the och1 mutant yeast may be more suitable for production of recombinant glycoproteins. And the och 1 deletion strain could be used for further re-engineering to produce complex human glycoproteins.
Chimera
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Gene Deletion
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Gene Knockout Techniques
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Granulocyte-Macrophage Colony-Stimulating Factor
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biosynthesis
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genetics
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Mannosyltransferases
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genetics
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Pichia
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enzymology
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genetics
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metabolism
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Recombinant Fusion Proteins
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biosynthesis
;
genetics
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Serum Albumin
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biosynthesis
;
genetics