1.Promoter methylation status of hPer3 gene in AML patients and the in vitro effect of decitabine on the status.
Ye-kai WANG ; Ji-hang ZHOU ; Shi-quan ZHOU ; Guo-an FANG ; Yi-wei LI ; Lei QIU ; Xu-chun YANG ; Xiao-guang LIU
Chinese Journal of Hematology 2011;32(5):317-321
OBJECTIVETo investigate the clinical significance of promoter methylation status of hPer3 gene in acute myeloid leukemia (AML) patients and the in vitro effect of decitabine (DCA) on AML cell lines HL-60 and U937.
METHODSThe promoter methylation status of hPer3 gene and mRNA expression levels in bone marrow of 206 AML and 40 iron deficiency anemia (IDA) patients (as control) were detected by methylation specific PCR (MS-PCR) and real-time PCR (RT-PCR). The HL-60 and U937 cell lines were treated with different concentrations of DCA for 48 and 72 h. The inhibition rates of cell proliferation were detected by methyl thiazolyl tetrazolium (MTT); the early apoptosis rates by staining with Annexin V and PI; the CD14 and CD11b expressions by flow cytometry (FCM); the promoter methylation status of hPer3 gene by MS-PCR; and the hPer3 mRNA expressions levels by RT-PCR.
RESULTSThe promoter methylation rates of hPer3 in newly diagnosed (ND) group, partial remission(PR) group, complete remission (CR) group, relapse (R) group and control group were 93.65% (59/63), 54.39% (31/57), 24.66% (18/73), 61.54% (8/13) and 0% (0/40), and the hPer3 mRNA expression levels were 0.19 ± 0.08, 6.28 ± 2.11, 52.76 ± 14.17, 8.18 ± 4.36, 75.03 ± 18.16, respectively. There was a significant statistic difference between any two group (P < 0.01) excepting for between PR and R group (P > 0.05). After DCA treatment, the promoter hypermethylation status of hPer3 was reduced and the mRNA and CD14, CD11b expression levels were up regulated in a dose dependent manner with an induction of cell apoptosis.
CONCLUSIONSPromotor methylation status and mRNA expression of hPer3 gene may be indicators for evaluating AML. DCA can induce the expression of hPer3 gene and cells apoptosis in AML.
Adolescent ; Adult ; Aged ; Azacitidine ; analogs & derivatives ; pharmacology ; Cell Proliferation ; DNA Methylation ; Female ; HL-60 Cells ; Humans ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Period Circadian Proteins ; genetics ; Promoter Regions, Genetic ; U937 Cells ; Young Adult
2.Rare blood group screening by serological and molecular methods in Zhejiang Han population.
Hong ZHU ; Ying LIU ; Xiao-Zhen HONG ; Xiao-Guo XU ; Xiao-Fei LAN ; Kai-Rong MA ; Ji HE ; Fa-Ming ZHU ; Hang-Jun LU
Journal of Experimental Hematology 2012;20(3):749-752
This study was aimed to investigate the distribution of rare blood group in Zhejiang Han population. The H(-) (H system), GPA(-) and s(-) (MNS), Rhnull, Rhmod, D--, CCDEE, CCdEE (variations of Rh), GPC(-) (Gerbich), i(+) (I), Lu(b-) (Lutheran), Js(b-) and k(-) (Kell), Fy(a-) (Duffy), Ok(a-) (Ok), Di(b-) (Diego) phenotypes were screened by serological or molecular methods. Jk (a-b-) phenotype was detected by urea hemolytic test. The results showed that one Di (a+b-) individual was found in 1618 blood donors, three Fy (a-b+) individuals in 1007 donors and one CCdEE individual in 633 Rh negative donors. No Jk (a-b-), H(-), GPA(-), s(-), GPC(-), i(+) (adult), Lu(b-), k(-), Js(b-), Lu(b-) and Ok(a-) phenotypes were found in this large scale survey. It is concluded that Di (a+b-), Fy (a-b+), CCdEE phenotypes are confirmed in the blood donors and this study provides the distribution data of erythrocyte rare blood group in Zhejiang Han population.
Asian Continental Ancestry Group
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genetics
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Blood Group Antigens
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genetics
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Blood Grouping and Crossmatching
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methods
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Erythrocytes
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immunology
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Humans
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Molecular Biology
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Phenotype
3.Clinical observation of 100 patients with malignant lymphoma treating with different preconditioning regimens followed by autologous hematopoietic stem cell transplantation.
Lan-Lan SHAO ; Xiu-Bin XIAO ; Kai-Li ZHONG ; Yun LU ; Xi-Lin CHEN ; Yong DA ; Jing LIU ; Shi-Hua ZHAO ; Yi MA ; Qiu-Shi YANG ; Hang SU ; Wei-Jing ZHANG
Journal of Experimental Hematology 2012;20(3):598-602
This study was designed to compare the curative effect, prognosis and safety of different preconditioning regimens for patients who received autologous hematopoietic stem cell transplantation (AHSCT) for malignant lymphoma (ML). The clinical data of 100 ML patients (Sep 1992 to Aug 2010 in 307 Hospital) were retrospectively analyzed, and were divided into two groups by different preconditioning regimens: the high-dose chemotherapy preconditioning group and high-dose chemotherapy/radiotherapy preconditioning group. The overall survival (OS) rate, progress free survival (PFS) rate and adverse effect were analyzed. The results showed that until Feb 2011, the median follow-up was 33.5 months. All patients were engrafted and their hematopoiesis was reconstituted. The median time of WBC recovery up to > 1.0×1.0(9)/L in high-dose chemotherapy preconditioning group and high-dose chemotherapy/radiotherapy preconditioning group were (6.0 ± 0.4) d and (8.2 ± 0.4) d, platelet up to > 20.0×1.0(9)/L in two groups were (7.1 ± 0.8) d and (11.4 ± 2.5) d (P < 0.05). The 3-year OS rate of the two groups were 67.3% and 68.9%. 5-year OS rates of two groups were 62.8% and 60.6%, 10-year OS rates of two groups were 57.6% and 56.2% respectively; 3-year PFS of two group were 63.6% and 63.2%, 5-year of two group were 59.4% and 58.3%, 10-year of two group were 50.8% and 55.3% respectively (P > 0.05). Meanwhile, the incidence of fever, infection, bleeding, secondary cancer between two groups was not significant different (P > 0.05). It is concluded that the hematopoietic reconstitution of high-dose chemotherapy/radiotherapy preconditioning group is later than that of high-dose chemotherapy preconditioning group. However, there is no significant difference in curative effect and prognosis between the two groups.
Adolescent
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Adult
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Aged
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Child
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Combined Modality Therapy
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Female
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Hematopoietic Stem Cell Transplantation
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Humans
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Lymphoma
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surgery
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therapy
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Male
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Middle Aged
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Prognosis
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Retrospective Studies
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Transplantation Conditioning
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methods
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Transplantation, Autologous
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Young Adult
4.Identification of an ABx09 phenotype of ABO subtype.
Xiao-zhen HONG ; Yan-ling YING ; Xian-guo XU ; Kai-rong MA ; Xiao-fei LAN ; Ying LIU ; Fa-ming ZHU ; Hang-jun LV ; Li-xing YAN
Chinese Journal of Medical Genetics 2011;28(5):548-551
OBJECTIVETo analyze the molecular basis for an individual with ABx09 phenotype of ABO subtype.
METHODSThe ABO group antigens on red blood cells of the proband were identified by monoclonal antibodies, and the ABO antibody in serum was detected by standard A, B, O cells. The exons 1 to 7 of ABO gene were amplified by polymerase chain reaction (PCR) respectively and the PCR products were sequenced directly. The amplified products for exons 5 to 7 were also cloned by TOPO TA cloning sequencing kit to split the two alleles apart, selected colonies were sequenced bidirectionally for exons 5 to 7 of the ABO gene. The samples of the proband's parents were collected, then serological test of the blood group and sequence analysis for exons 6 and 7 of ABO gene were preformed.
RESULTSBoth A and B antigens were detected on red blood cells of the proband and there was anti-B antibody in the serum. There was no G deletion at position 261, while 297AG in exon 6, 467CT, 526CG, 657CT, 703GA, 796CA, 803GC, 889GA and 930GA heterozygote in exon 7 were detected by direct DNA sequencing, which can be assigned for A102Bx09 genotype. After cloning and sequencing, two alleles A102 and Bx09 were obtained. The sequence of Bx09 had one nucleotide changes (G to A) at position 889 compared with that of B101, which resulted in an amino acid change of Glu to Lys at 297 position. The Bx09 in the proband was inherited from her mother by family investigation.
CONCLUSIONG to A at nt889 of alpha-1,3 galactosyltransferasegene can result in Bx09 phenotype, with the presence of anti-B antibody in serum.
ABO Blood-Group System ; genetics ; metabolism ; Adolescent ; Alleles ; Base Sequence ; Blood Grouping and Crossmatching ; Female ; Gene Frequency ; Genotype ; Humans ; Molecular Sequence Data ; Pedigree ; Phenotype
5.Survey of schistosomiasis KAP and influencing factors of behaviors among residents in Jiangsu Province
rong De HANG ; xin Yi HUANG ; bao Yao LIU ; Xu ZHANG ; qing YI XIE ; gang Wei YIN ; Kai TANG ; heng Lian ZHANG ; kuan Dao SUN
Chinese Journal of Schistosomiasis Control 2017;29(5):574-578
Objective To understand the schistosomiasis control knowledge,attitude,and practice(KAP),and influenc-ing factors of behaviors among residents in Jiangsu Province,so as to provide the evidence for making effective health education and health promotion models. Methods The probability proportionate to size sampling(PPS)and multi-stage sampling meth-ods were adopted to sample the research objects. A questionnaire survey of schistosomiasis control KAP was conducted in the res-idents of 16 to 69 years old in schistosomiasis endemic areas of Jiangsu Province,and the results were statistically analyzed. Re-sults The total awareness rate of the participants was 95.98%for schistosomiasis control knowledge. The correct rates of atti-tude and practice were 89.06%and 77.43%,respectively. The awareness/correct rates of knowledge,attitude and practice re-duced in turns significantly(χ2=1282.96,P<0.01). The knowledge awareness rate of fishermen and boatmen was 90.98%, but their attitude correct rate was only 53.81%(χ2=120.52,P<0.01). The unconditional logistic regression analysis showed that with the education level increasing,their practice correct rate rose,and the participants with the college degree or above had a higher correct rate compared to illeterate ones(OR=6.411,95%CI:4.896-8.395). The practice correct rate of the fisher-men and boatmen was only 5.1%of the rate of the farmers(OR=0.051,95%CI:0.029-0.091). Conclusions The total aware-ness rate of basic knowledge of schistosomiasis prevention and control in the residents of Jiangsu Province has reached the re-quirements in the"National Schistosomiasis Control Long-term Planning Outline(2004-2015)",but the correct rate of behav-iors is low. The education level,occupation and residential areas affect the health behaviors of schistosomiasis prevention and control. Therefore,it is necessary to carry out targeted health promotion activities to promote the formation of healthy lifestyle and behaviors.
6.Preparation, characterization and antitumor of cyclodextrin inclusion of an anti-cancer drug regorafenib.
Liu KAI-HANG ; Sun MENGYING ; Tang GUPING ; H U XIURONG
Journal of Zhejiang University. Medical sciences 2017;46(2):151-159
In order to improve the drug's solubility, dissolution and bioavailability, RG-β-CD, RG-γ-CD and RG-Hp-β-CD were prepared by co-crystallization between Regorafenib (RG) and β-cyclodextrin (β-CD), γ-cyclodextrin (γ-CD) and Hydroxypropyl-β-cyclodextrin (Hp-β-CD).Three inclusion complexes were prepared by recrystallization and solvent evaporation methods and characterized by fourier transform infrared spectroscopy (FT-IR), thermal analysis (TG), differential scanning calorimetry (DSC), powder X-ray diffraction (PXRD),H nuclear magnetic resonance (H-NMR), nuclear overhauser effect spectroscopy (NOESY).experiments, tumor suppression assay were made with SW620 colon cancer cell.The ability of solubility and dissolution were improved after inclusion with three kinds of cyclodextrins. The regorafenib-β-cyclodextrin inclusionis proved to have the best stability. The less enhanced was regorafenib-γ-cycl-odextrin inclusion. The best dissolution of regorafenib-β-cyclodextrin inclusion complex was to bring as the tumor suppression assay, the result shows that regorafenib inclusion with β-cyclodextrin is better than regorafenib itself.The bioavailability of regorafenib by inclusion with cyclodextrin can enhance due to the solubility enhancement of RG, which can provide an effective method for improving solubility and dissolution of insoluble drug in clinical medication.
7.Autologous peripheral blood stem cell transplantation combined with autologous bone marrow transplantation for treating refractory lymphoma.
Xi-Lin CHEN ; Hang SU ; Kai-Li ZHONG ; Yong DA ; Xiu-Bin XIAO ; Jing LIU ; Jing-De YU ; Wei-Jing ZHANG
Journal of Experimental Hematology 2009;17(1):155-159
This study was aimed to investigate the differences of therapeutic efficiencies, side effects and recovery rates of immune function in refractory lymphoma patients treated with autologous peripheral blood hematopoietic stem cell transplantation (APBHSCT), autologous bone marrow transplantation (ABMT) and combination of APBHSCT with ABMT (APBHSCT + ABMT) by retrospective analysis, and to evaluate the merits and demerits of 3 kinds of transplantation for treatment of refractory lymphoma. 68 patients with malignant lymphoma were treated with autologous hematopoietic stem cells transplantation. Out of 68 patients 10 cases were treated with autologous bone marrow transplantation (ABMT), 46 cases were treated with autologous peripheral blood hematopoietic stem cell transplantation (APBHSCT), and 12 cases were treated with autologous peripheral blood hematopoietic stem cells transplantation combined with autologous bone marrow transplantation (APBHSCT + ABMT). The results indicated that the therapeutic response rates and survival rates at 1, 3, 5 years for each transplant regimen were 90% and 75%, 57.1%, 33.3%; 86.4% and 74.4%, 54.2%, 38.1%; 83.3% and 72.7%, 55.6%, 40%. The times of ANC > or = 0.5 x 10(9)/L were 13, 11 and 9 days, times of platelet >/= 20 x 10(9)/L were 17, 14 and 10 days. The recovery rates of T cell subtypes in patients received ABMT, APBHSCT and APBHSCT + ABMT on 3 months, 6 months, 1 year were (0%, 33.3%, 60%), (10.8%, 32%, 73.9%), (27.3%, 55.6%, 85.7%) respectively. In conclusion, the efficacy and side effects of APBHSCT + ABMT as compared with ABMT and APBHSCT are roughly the same, but ABMT + APBHSCT can result in more rapid hematopoietic reconstitution and less restrictions with contributes to widen choice of transplant regimen for patients with alder age and impaired hematopoietic functions.
Adolescent
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Adult
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Bone Marrow Transplantation
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Female
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Humans
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Lymphoma
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surgery
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Male
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Middle Aged
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Peripheral Blood Stem Cell Transplantation
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Retrospective Studies
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Transplantation, Autologous
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Young Adult
8.A Sensor for Detection of Breast Tumor with Three-dimensional Electrical Impedance Tomography
Kai LIU ; An-Qi LI ; Fang LI ; Cheng-Jun ZHU ; Hang TIAN ; Jia-Feng YAO
Chinese Journal of Analytical Chemistry 2024;52(2):248-255,中插16-中插18
An intensive breast array sensor was designed based on three-dimensional electrical impedance tomography in this work.Firstly,an electrical impedance sensor for detection of breast cancer was developed.The sensor adopted the integrated design of excitation electrode array and ground electrode to achieve structural simplification.It realized electric field densification through conical matrix and double-layer circumferentially arranged electrode array and improved the detection accuracy of target object through taper optimization.Secondly,the imaging system was designed,and the sensor was optimized by numerical simulation.The simulation results showed that halving the number of electrodes did not affect imaging accuracy of the sensor,but could improve the imaging speed.Finally,the performance of the sensor was verified by experiment.The signal-to-noise ratio and channel consistency of the system were at a good level.The sensor was used to reconstruct three-dimensional image of the experimental model with relative volume of the detection field of 0.4%.The image correlation coefficient of the single target imaging was above 0.6 and the position of the double target object could be clearly identified,and thus the visual detection of breast cancer was realized.
9.Experimental study of T lymphocyte reactivity inhibited by allogeneic bone marrow mesenchymal stem cells.
Jian-lin CHEN ; Kai FENG ; Zi-Kuan GUO ; Ren-na U ; Chen XU ; Yu-hang LI ; Xiao-dan LIU ; Ning MAO ; Hu CHEN
Chinese Journal of Hematology 2005;26(12):740-742
OBJECTIVETo address the question whether bone marrow mesenchymal stem cells (MSCs) could lower responsiveness of allogeneic T lymphocytes against alloantigens, and explore a feasible strategy for prevention of graft versus host disease (GVHD) occurred in allogeneic bone marrow transplantation.
METHODST cells were co-cultured with (60)Co-irradiated bone marrow MSCs from different individuals. The proliferative activity of T cells and their reactivity to allogeneic cells and ConA were evaluated with (3)H-TdR incorporation assay.
RESULTST cells could not be activated upon primary or even secondary exposure to allogeneic MSCs (compared the CPM value of 27,529 +/- 969 of T cell alone with that of primary and secondary exposures to allogeneic MSCs were 9,126 +/- 654 and 13,260 +/- 874, respectively). When MSCs were induced to express HLA-DR, they still could not elicit T cell activation. The proliferation rate of allogenous T cells exposed to MSCs was dramatically declined when T cells from the same donor's MSCs were used as stimulator (CPM value decreased from 45,876 +/- 5285 before coculture to 9850 +/- 1618 after coculture). Furthermore, the results remained unchanged even ConA was added into the culture system.
CONCLUSIONSHeterogenetic MSCs could suppress T cell activation. MSCs pretreatment might be useful in the prevention of GVHD in HLA-mismatched bone marrow transplantation.
Bone Marrow Cells ; immunology ; Cell Communication ; immunology ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Graft vs Host Disease ; immunology ; prevention & control ; Humans ; Lymphocyte Activation ; Mesenchymal Stromal Cells ; immunology ; T-Lymphocytes ; immunology
10.Cloning and sequence analysis of 5'untranslated region of human ABO gene.
Ying LIU ; Xian-guo XU ; Kai-rong MA ; Xiao-fei LAN ; Xiao-zhen HONG ; Fa-ming ZHU ; Hang-jun LV ; Li-xing YAN
Chinese Journal of Medical Genetics 2011;28(1):83-87
OBJECTIVETo clone and investigate the polymorphism of the 5'-untranslated region (5'-UTR) of the human ABO gene, in order to provide the basis for exploring the transcriptional regulation of the human ABO histo-blood group genes.
METHODSABO phenotypes of 30 unrelated healthy blood donors were determined by serological technique, their genotypes were analyzed by sequencing the exons 6 and 7 of the ABO gene. Nearly 5 kb of the 5'-UTR of ABO gene was obtained by PCR amplification and sequencing was performed bidirectionally. Haplotypes of samples with heterozygous sites in the 5'-UTR of ABO gene were separated and analyzed after cloning.
RESULTSTwenty polymorphic sites were identified in these samples where ABO genotypes were consistent with serological phenotypes. It included sixteen nucleotide sequence variations, one 8 bp deletion, one 6 bp deletion/insertion, one 36 bp insertion and one 43 bp repeats. Among them, 11 were novel polymorphic sites. Seven different haplotypes of 5'-UTR of ABO gene were defined to the cis/trans linkage of those mutations.
CONCLUSIONThere were polymorphisms in the 5'-UTR of ABO gene and the nucleotide sequences near the proximal promoter were conservative.
5' Untranslated Regions ; genetics ; ABO Blood-Group System ; genetics ; Cloning, Molecular ; Genotype ; Haplotypes ; Humans ; Polymorphism, Genetic ; Sequence Analysis, DNA