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1.
Chinese Journal of Otorhinolaryngology Head and Neck Surgery ; (12): 479-484, 2022.
Article in Chinese | WPRIM | ID: wpr-936237

ABSTRACT

Objective: To investigate the sensitization characteristics of Juniperus chinensis pollen in patients with allergic rhinitis and/or allergic asthma in Beijing area, and to explore the characteristics of Juniper chinensis pollen sensitized population. Methods: Patients with suspected allergic rhinitis and/or asthma from January 2017 to December 2019 in the outpatient department of Allergy Department of Beijing Shijitan Hospital were selected in this study. Skin prick test (SPT) was performed with Juniper chinensis pollen allergen reagent to compare different age and disease allergen distribution, and to observe the sensitization characteristics of its population. All of the analyses were performed using SAS software version 9.4. Results: A total of 8 380 patients were enrolled in the end. The total positive rate of Juniper chinensis pollen SPT reached 49.92% (4 183/8 380). The positive rate of Juniper chinensis pollen SPT was highest in the 10-14 age group, reaching 60.99% (283/464). Compared with other age groups, there was a statistical difference (χ²=266.77, P<0.01). The SPT positive rate of patients aged less than 10 years increased with the increase of age, while the SPT positive rate of patients aged over 40 years decreased with the increase of age. Single Juniper chinensis pollen was less allergenic, accounting for about 25.05% (1 048/4 183), and the patients' age was (35.21±12.39) years. Regardless of single Juniper chinensis pollen or other pollen allergies, allergic rhinitis was the main disease. Among the patients with SPT positive Juniper chinensis pollen combined with other inhaled pollen allergens, willow pollen accounted for the first (74.99%). The positive rate of Juniper chinensis pollen was the highest in patients with single allergic rhinitis, accounting for 52.05% (3 797/7 295), and the rate in patients with single allergic asthma was the lowest, accounting for 17.49% (53/303), with statistically difference (χ²=138.99, P<0.01). Conclusions: Juniper chinensis pollen is highly sensitized in patients with allergic rhinitis and/or allergic asthma in Beijing . The positive rate of SPT is highest among 10-14 age group, most of which showed strong positive reaction, and allergic rhinitis is more common in Juniper chinensis pollen sensitization diseases.


Subject(s)
Adolescent , Adult , Child , Humans , Allergens , Asthma , Juniperus , Pollen , Rhinitis, Allergic , Skin Tests
2.
Chinese Journal of Biotechnology ; (12): 331-341, 2021.
Article in Chinese | WPRIM | ID: wpr-878566

ABSTRACT

Genetic and epigenetic alterations accumulate in the process of hepatocellular carcinogenesis, but the role of genomic spatial organization in HCC is still unknown. Here, we performed in situ Hi-C in HCC cell line PLC/PRF/5 compared with normal liver cell line L02, together with RNA-seq and ChIP-seq of SMC3/CTCF/H3K27ac. The results indicate that there were significant compartment switching, TAD shifting and loop pattern altering in PLC/PRF/5. These spatial changes are correlated with abnormal gene expression and more opening promoter regions of the HCC cell line. Thus, the 3D genome organization alterations in PLC/PRF/5 are important in epigenetic mechanisms of HCC tumorigenesis.


Subject(s)
Humans , Carcinoma, Hepatocellular/genetics , Cell Line , Cell Line, Tumor , Genomics , Liver Neoplasms/genetics
3.
World Journal of Emergency Medicine ; (4): 244-247, 2019.
Article in English | WPRIM | ID: wpr-783832

ABSTRACT

@#Though notorious for its tendency to induce recurrent neck abscess, pyriform sinus fistula is of importance despite its rarity. It usually presents diagnostic and therapeutic challenges. Because of post-infectious fibrosis, the embryologic origin of pyriform sinus fistula is difficult to appreciate in certain cases. Here we present a case with empyema and mediastinal abscess caused by pyriform sinus fistula and share our experience in the treatment of this patient.

4.
Journal of Southern Medical University ; (12): 395-401, 2018.
Article in Chinese | WPRIM | ID: wpr-690456

ABSTRACT

<p><b>OBJECTIVE</b>To determine whether erythropoietin (EPO) promotes rapid proliferation of glioma through Akt pathway.</p><p><b>METHODS</b>We detected the expression of EPO in human glioma tissues using immunohistochemistry. A nude mouse model bearing human glioma U87 cell xenograft was established and given intraperitoneal injection of EPO or saline every other day, and the tumor growth was observed. In the in vitro experiment, U87 cells were treated with PBS (control), EPO, or EPO with Akt inhibitor, and the expression of p-Akt and cyclin D1 was detected using Western blotting; the cell proliferation rate was determined using cell counting kit-8 and clone formation assay, and the cell cycle changes were analyzed with flow cytometry.</p><p><b>RESULTS</b>Compared with low-grade glioma tissues, high-grade glioma tissues exhibited a significantly increased EPO expression (P=0.0002). In the tumor-bearing mice, EPO treatment significantly increased the expression of EPO (P=0.0006) and p-Akt (P=0.0003) in the tumor and obviously increased the tumor volume (P<0.0001) and weight (P=0.0003). In U87 cells cultured in vitro, EPO treatment obviously accelerated the cell proliferation (P=0.020 on day 3 and 0.028 on day 5), promoted clone formation (P=0.0010), and increased proliferation index (P=0.0028); EPO significantly enhanced the protein expression of p-Akt (P=0.0020) and cyclin D1 (P=0.0022). The application of Akt inhibitor significantly suppressed the effect of EPO in enhancing cyclin D1 and p-Akt expression (both P<0.0001) and promoting cell proliferation.</p><p><b>CONCLUSION</b>EPO can significantly accelerate the proliferation of glioma through Akt pathway.</p>

5.
Chinese Journal of Analytical Chemistry ; (12): 288-292, 2018.
Article in Chinese | WPRIM | ID: wpr-692248

ABSTRACT

The macroporous microspheres were prepared through suspension polymerization and based on a copolymer of glycidyl methacrylate and ethylene glycol dimethacrylate.The effect of porogen on the microspheres structure was evaluated in terms of pore size and surface area.Porogen contained dichloromethane (δ=9.7 (cal/cm3)1/2) and N-octanol (δ=10.3 (cal/cm3)1/2) which corresponded to a good and poor solvent,respectively.The solubility parameter of porogen was controlled in the range of 9.89-10.09 (cal/cm3)1/2.The pore size of microspheres increased with the difference value of solubility parameter between the polymer and the porogen.On the contrary,the surface area of microspheres decreased in this study.The anion exchange media was prepared through coupling poly(ethylene imine) in the microspheres,and the proteins transport was determined by frontal analysis method.The macroporous microspheres with 257 nm pore size could still afford a high proteins capacity (45.1 mg/mL).These macroporous supports showed a large potential in a rapid separation of proteins.

6.
Fudan University Journal of Medical Sciences ; (6): 733-737, 2017.
Article in Chinese | WPRIM | ID: wpr-668601

ABSTRACT

Epigenetics is the study of heritable traits that are not involved in the changes in DNA sequences.It has been widely recognized that epigenetics plays multiple regulatory roles in diverse cellular activities and is associated with the occurrence and development of a variety of diseases.Following the human genome project (HGP),discoveries in the field of epigenetics reveal the complexity and accuracy of genetic regulation.Institute of Biomedical Sciences (IBS),which is affiliated with Fudan University Shanghai Medical School,has been attaching great importance to "Medical Epigenetics" since the establishment of IBS.In the past 12 years,a number of influential discoveries in medical epigenetics have been made,raising China's status in the field of epigenetics and contributing greatly to the disciplinary construction and talent cultivation of biomedical-related disciplines in Fudan University.In this review,a list of accomplishments in epigenetics in IBS were summarized.

7.
Experimental & Molecular Medicine ; : e325-2017.
Article in English | WPRIM | ID: wpr-212097

ABSTRACT

Post-translational modifications, such as methylation, acetylation and phosphorylation, of histone proteins play important roles in regulating dynamic chromatin structure. Histone demethylation has become one of the most active research areas of epigenetics in the past decade. To date, with the exception of histone H3 lysine 79 methylation, the demethylases for all major lysine methylation sites have been discovered. These enzymes have been shown to be involved in various biological processes, with embryonic development being an exciting emerging area. This review will primarily discuss the involvement of these demethylases in the regulation of mammalian embryonic development, including their roles in embryonic stem cell pluripotency, primordial germ cell (PGC) formation and maternal-to-zygotic transition.


Subject(s)
Female , Pregnancy , Acetylation , Biological Phenomena , Chromatin , Embryonic Development , Embryonic Stem Cells , Epigenomics , Germ Cells , Histone Demethylases , Histones , Lysine , Methylation , Phosphorylation , Protein Processing, Post-Translational
8.
Chinese Medical Journal ; (24): 804-810, 2015.
Article in English | WPRIM | ID: wpr-350399

ABSTRACT

<p><b>BACKGROUND</b>Retroperitoneal fibrosis (RPF) is an uncommon disease that is characterized by development of fibrosclerotic tissues involving retroperitoneal structures. This study aimed to investigate the clinical features of 30 patients with RPF in a single center in Beijing in a 10-year period.</p><p><b>METHODS</b>We retrospectively analyzed clinical data on demographic characteristics, clinical manifestations, laboratory findings, radiological findings, modalities of treatments, outcomes and prognosis of 30 patients with RPF. Patients were treated in Beijing Chao-Yang Hospital between January 2003 and December 2013.</p><p><b>RESULTS</b>The mean age of patients with RPF was 56.7 ± 14.4 years. Twenty-three patients were men and seven patients were women. Acute phase reactants were elevated in most patients. Rheumatic factor was positive in 4/25 (16.0%) patients, and antinuclear antibody was positive in 6/22 (27.3%) patients. Elevation of IgG4 was observed in 9/22 (40.9%) patients. The most common type was I + III (n = 13), followed by I + II + III (n = 12). Five patients undertook an 18 F-fluoro-deoxy-D-glucose positron emission tomography examination and increased uptake was detected in four patients. Eight patients received combination therapy with glucocorticoids and tamoxifen. Surgical intervention treatments included intraureteral double-J stent implantation (n = 26), percutaneous nephrostomy (n = 2), open ureterolysis and intraperitonealization of the ureters (n = 5) and laparoscopic ureterolysis and intraperitonealization of the ureters (n = 5). Three patients underwent hemodialysis because of renal failure.</p><p><b>CONCLUSIONS</b>Clinical characteristics of RPF patients in our study are similar to those previously reported. Steroids and immunosuppressive therapy combined with ureterolysis could be a viable choice of treatment for RPF. More prospective, multi-center studies with a longer follow-up are warranted.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Glucocorticoids , Therapeutic Uses , Immunoglobulin G , Blood , Retroperitoneal Fibrosis , Blood , Diagnosis , Drug Therapy , General Surgery , Retrospective Studies , Tamoxifen , Therapeutic Uses , Treatment Outcome
9.
Journal of Experimental Hematology ; (6): 1386-1390, 2015.
Article in Chinese | WPRIM | ID: wpr-274030

ABSTRACT

<p><b>OBJECTIVE</b>To detect platelet anti-HPA-1a and -1b antibodies using recombinant GPIIIa fragments coupled to Luminex beads.</p><p><b>METHODS</b>The sensitivity of 2 techniques, monoclonal antibody specific immobilization of platelet antigen (MAIPA) and Luminex bead assay, was compared using 12 twofold-serial dilutions (from neat to 1 in 2048) of an anti-HPA-1a WHO international standard. The specificity of Luminex assay to identify anti-HPA-1a and -1b antibodies was assessed using 8 negative or positive controls and 36 blinded samples provided by WHO Platelet Workshop.</p><p><b>RESULTS</b>The sensitivity of MAIPA and Luminex bead assay to detect anti-HPA-1a was dilution 1/64 (i.e. 1.56 IU/ml) and far more than dilution 1/2048 (i.e. 0.049 IU/mL), respectively. The Luminex bead assay could specifically identify negative and positive controls of anti-HPA-1a and -1b. All results of 36 blinded samples by Luminex assay were accordant to reference results except one sample which contained high concentration antithetical antibody and resulted in false positive of anti-HPA-1b. Cross-reactivity was also not observed with the samples containing HLA, ABO or other platelet antibodies.</p><p><b>CONCLUSION</b>The Luminex beads coupled with recombinant GPIIIa fragments can be used to detect HPA-1 system antibodies with sufficient sensitivity and specificity, that is suitable for the detection of platelet alloantibodies in clinical alloimmune thrombocytopenia.</p>


Subject(s)
Humans , Antibodies, Monoclonal , Antigens, Human Platelet , Allergy and Immunology , Blood Platelets , Integrin beta3 , Chemistry , Isoantibodies , Blood , Purpura, Thrombocytopenic, Idiopathic , Diagnosis , Recombinant Proteins , Chemistry , Sensitivity and Specificity
10.
Journal of Experimental Hematology ; (6): 57-61, 2013.
Article in Chinese | WPRIM | ID: wpr-325212

ABSTRACT

The purpose of this study was to detect the minimal residual disease (MRD) in peripheral blood of newly diagnosed patients with acute myeloid leukemia (AML) on day 8 of induction chemotherapy and analyze the correlation between day 8 MRD (D8RD) and therapeutic effectiveness. 29 adult patients (13 males and 16 females, aged 16 - 75 years, median 41 years) with AML diagnosed and treated in West China Hospital from September 2009 to June 2010 were analyzed and followed up in the study. The leukemia-associated aberrant immunophenotype (LAIP) of all the patients were detected by multiparameter flow cytometry (FCM) before therapy. The level of MRD in the peripheral blood at day 8 of induction chemotherapy was detected by FCM based on the LAIP. The overall survival curve was drawn by calculation using Kaplan-Meier method using, and the comparison between different groups was carried out by Log-rank test. The results indicated that after first course therapy, the levels of peripheral D8RD in 7 out of 29 AML cases were lower than 0.01% (negative group), and that in another 22 cases were higher than 0.01% (0.08% - 55%, positive group). The sex, age, WBC, LDH, percentage of bone marrow blasts at diagnosis in these groups were not statistically different. 6 cases achieved CR (86%) in D8RD negative group, and also 6 cases achieved CR (27%) in D8RD positive group, CR rate in D8RD negative group was higher than in D8RD positive group (86% vs 27%, P < 0.05). The median follow-up of 29 cases lasted for 15 months; the 1-year overall survival rate of D8RD negative and D8RD positive groups was 100% and 39.4%, respectively (P < 0.01). It is concluded that MRD level in peripheral blood at day 8 of induction chemotherapy is an early index to predict clinical efficacy of induction therapy in AML.


Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Early Diagnosis , Flow Cytometry , Leukemia, Myeloid, Acute , Blood , Drug Therapy , Mortality , Neoplasm, Residual , Diagnosis , Drug Therapy , Mortality , Prognosis , Survival Rate , Treatment Outcome
11.
Journal of Experimental Hematology ; (6): 1042-1045, 2013.
Article in Chinese | WPRIM | ID: wpr-283985

ABSTRACT

The individual with the deficiency of CD36 antigen on platelet displayed the risk of anti-CD36 immune reaction induced by transfusion, which is one of the reasons for platelet transfusion refractoriness (PTR). This study was purposed to detect the expression level of CD36 antigen on platelet by flow cytometry among apheresis platelet donors of Hangzhou area, and the frequency of CD36 deficiency was analyzed. Platelet-rich plasma (PRP) was separated from fresh anticoagulant whole blood by centrifugation, then the platelets were washed and adjusted to 1×10(6). The platelets were incubated with FITC-labeled CD36 and PE-labeled CD41 monoclonal antibodies, then the expression level of CD36 was detected by flow cytometry. The CD36 expression on monocytes for the samples of CD36-deficiency on the platelets was further analyzed. The results showed that 7 samples with CD36 antigen deficiency were found in 192 apheresis platelet donors. The frequency of CD36 deficiency was 3.6% and all of them were typeII deficiency. The significant difference of CD36 antigen expression was observed in the platelet donors of Hangzhou population, among them 59 individuals with low expressed CD36 antigen and 126 individuals with highly expressed CD36 antigen were found according to the geometric mean fluorescence intensity. It is concluded that the CD36 antigen deficient phenotype existed in the population, these data will provide the information for research of the CD36 antigen distribution and help to solve the platelet transfusion refractoriness.


Subject(s)
Humans , Blood Platelet Disorders , Diagnosis , Blood Platelets , Metabolism , CD36 Antigens , Metabolism , Flow Cytometry , Methods , Genetic Diseases, Inborn , Diagnosis
12.
Journal of Experimental Hematology ; (6): 749-752, 2012.
Article in Chinese | WPRIM | ID: wpr-263310

ABSTRACT

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.


Subject(s)
Humans , Asian People , Genetics , Blood Group Antigens , Genetics , Blood Grouping and Crossmatching , Methods , Erythrocytes , Allergy and Immunology , Molecular Biology , Phenotype
13.
Chinese Journal of Medical Genetics ; (6): 548-551, 2011.
Article in Chinese | WPRIM | ID: wpr-326894

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the molecular basis for an individual with ABx09 phenotype of ABO subtype.</p><p><b>METHODS</b>The 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.</p><p><b>RESULTS</b>Both 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.</p><p><b>CONCLUSION</b>G to A at nt889 of alpha-1,3 galactosyltransferasegene can result in Bx09 phenotype, with the presence of anti-B antibody in serum.</p>


Subject(s)
Adolescent , Female , Humans , ABO Blood-Group System , Genetics , Metabolism , Alleles , Base Sequence , Blood Grouping and Crossmatching , Gene Frequency , Genotype , Molecular Sequence Data , Pedigree , Phenotype
14.
Journal of Experimental Hematology ; (6): 702-705, 2011.
Article in Chinese | WPRIM | ID: wpr-313912

ABSTRACT

The objective of this study was to analyze the molecular genetic basis for 2 individuals with A2B phenotype of ABO subtype. The ABO group antigens on red blood cells were identified by monoclonal antibodies and the ABO antibodies in serum were detected by the standard A, B, O cells. The exon 5 to exon 7 coding region of ABO gene was amplified by polymerase chain reaction (PCR) and the PCR product was sequenced directly after the enzymes digested. The amplified product was also cloned by TOPO TA cloning sequencing kit to split the 2 alleles apart and chosen colonies were sequencing bidirectionally for exon 6 to 7 of ABO gene. The results showed that both A and B antigen were identified on red blood cells of the individuals and there was anti-A1 antibody in their serum. There was no 261G deletion and showed 297A/G, 467C/T, 526C/G, 657C/T, 703G/A, 742C/T, 796C/A, 803G/C, 930G/A heterozygotes by direct DNA sequencing. After cloning and sequencing, it was obtained the B101 and one novel A allele. The novel allele has one nucleotide change at 742 position C to T compared with A102, which results in an amino acid Arg to Cys at 248 position and was nominated as A213. It is concluded that C742T mutation of the α1, 3 N-acetyl-D-galactosaminyl-transferase gene can lead to A2 phenotype and with anti-A1 antibody in serum.


Subject(s)
Humans , ABO Blood-Group System , Genetics , Allergy and Immunology , Alleles , Antibodies, Anti-Idiotypic , Allergy and Immunology , Blood Donors , Exons , Genotype , Heterozygote , Molecular Sequence Data , Mutation , N-Acetylgalactosaminyltransferases , Genetics , Phenotype , Sequence Analysis, DNA
15.
Chinese Journal of Medical Genetics ; (6): 37-41, 2011.
Article in Chinese | WPRIM | ID: wpr-234322

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the polymorphisms of platelet membrane glycoprotein genes related to human platelet alloantigen (HPA)-1 to 17w.</p><p><b>METHODS</b>The DNA segments of platelet membrane glycoprotein genes related to HPA-1 to 17w were amplified using author's designed primers. The amplification products were purified and directly sequenced to identify the HPA genotype and glycoprotein gene polymorphisms.</p><p><b>RESULTS</b>Thirteen new single nucleotide polymorphisms (SNPs) and a micro-satellite sequence were found in the glycoprotein genes from the 112 random samples, in which two SNPs (1333G/A and 1960G/A) in ITGB3 gene result in two amino acid change (V419M and E628K) on glycoprotein GPIIIa.</p><p><b>CONCLUSION</b>New variants in platelet membrane glycoprotein genes were identified, which may lead to structure change of platelet membrane glycoprotein and affect the accuracy of partial HPA genotyping method.</p>


Subject(s)
Humans , Antigens, Human Platelet , Genetics , Isoantigens , Genetics , Platelet Membrane Glycoproteins , Genetics , Polymorphism, Single Nucleotide
16.
Chinese Journal of Medical Genetics ; (6): 83-87, 2011.
Article in Chinese | WPRIM | ID: wpr-234312

ABSTRACT

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>ABO 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.</p><p><b>RESULTS</b>Twenty 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.</p><p><b>CONCLUSION</b>There were polymorphisms in the 5'-UTR of ABO gene and the nucleotide sequences near the proximal promoter were conservative.</p>


Subject(s)
Humans , 5' Untranslated Regions , Genetics , ABO Blood-Group System , Genetics , Cloning, Molecular , Genotype , Haplotypes , Polymorphism, Genetic , Sequence Analysis, DNA
17.
Journal of Experimental Hematology ; (6): 223-226, 2011.
Article in Chinese | WPRIM | ID: wpr-244950

ABSTRACT

This study was purposed to investigate the molecular basis of a para-Bombay phenotype for screening and identification of rare blood group. ABO and H phenotypes of the proband were identified by serological techniques. The exon 6 to exon 7 of ABO gene and full coding region of α-1,2-fucosyltransferase (fut1) gene of the proband were analyzed by polymerase chain reaction and direct sequencing of the amplified fragments. The haplotype of compound heterozygote of fut1 was also identified by cloning sequencing. The results indicated that a rare para-Bombay phenotype was confirmed by serological techniques. Two deletion or insertion variant sites near nucleotide 547 and 880 were detected in fut1 gene. The results of cloning sequence showed that one haplotype of fut1 gene was two bases deletion at 547-552 (AGAGAG→AGAG), and another one was two bases deletion at position 880-882 (TTT→T). Both two variants caused a reading frame shift and a premature stop codon. It is concluded that a rare para-Bombay phenotype is found and confirmed in blood donor population. The molecular basis of this individual is compound heterozygote of two bases deletion on fut1 gene which weaken the activity of α-1, 2-fucosyltransferase.


Subject(s)
Female , Humans , ABO Blood-Group System , Genetics , Alleles , Base Pairing , Fucosyltransferases , Genetics , Genotype , Heterozygote , Mutation , Phenotype , Sequence Deletion
18.
Journal of Experimental Hematology ; (6): 1613-1616, 2010.
Article in Chinese | WPRIM | ID: wpr-332308

ABSTRACT

In order to explore the effects of 35C > T and 682A > G mutations on the activity of alpha-(1,2) fucosyltransferase, the coding region of fut1 gene was amplified by polymerase chain reaction (PCR) from genomic DNA. PCR product was ligated into expression vector using TOPO TA cloning kit to obtain the recombinant plasmids. The recombinant plasmids were transfected into COS-7 cells by liposome method. After screening by using G418, H antigen expression on the COS-7 was tested by flow cytometry and fut1 mRNA was detected by real-time PCR. The results indicated that three kinds of recombinant plasmids pcDNA3.1/V5-His-wild (35C + 682A), pcDNA3.1/V5-His-35T and pcDNA3.1/V5-His-35T-682G were successfully constructed. After transfection, the H antigen expressed on membrane of COS-7 cells at the second day, with the maximum level of expression at the fourth day. When compared with pcDNA3.1/V5-His-wild transfected cells, the H antigen expression level of the 35T and 682G + 35T recombinant plasmids in the transfected cells was 52.7% and 13.3% on the fourth day, respectively. Although the level of fut1 mRNA decreased with prolonging of time, the mRNA expressed on the pcDNA3.1/V5-His-35T-682G transfected cells reached to 14% of the wild plasmids on the first day. It is concluded that 682A > G mutation obviously reduces the activity of alpha-(1,2) fucosyltransferase, while 35C > T mutation leads to partial reduction of H antigen in vitro expression.


Subject(s)
Animals , Antigens, Bacterial , Genetics , COS Cells , Chlorocebus aethiops , Fucosyltransferases , Genetics , Genetic Vectors , Mutation , Plasmids , RNA, Messenger , Genetics , Transfection
19.
Journal of Experimental Hematology ; (6): 898-902, 2010.
Article in Chinese | WPRIM | ID: wpr-237628

ABSTRACT

The aim of this study was to investigate the effect of Celastrol on induction of HL-60 cell apoptosis and its possible mechanism. The proliferative activity of HL-60 cells treated with 0.25 - 8.0 μmol/L of Celastrol for 24 - 72 hours was assayed by MTT method, the effects of Celastrol on apoptosis and cell cycle of HL-60 were detected by TUNEL staining and flow cytometry with Annexin V-FITC/PI double labeling, the expression of pAkt and cyclin D1 at protein and gene level in HL-60 cells treated with Celastrol were measured by Western blot and RT-PCR. The results showed that the Celastrol could obviously inhibit the proliferation of HL-60 cells in concentration-and time-dependent manners, the IC₅₀ value of Celastrol for 24 hours was 6.21 ± 0.242 μmol/L. The Celastrol concentration-dependently induced the apoptosis of HL-60 cells, accompanying with morphological changes of apoptotic cells, which may be related with arrest of cells in G₀/G₁ phase. The Celastrol suppressed the expression of pAkt and Cyclin D1 in HL-60 cells to a varying degree which showed obvious concentration-and time-dependent manners. It is concluded that the Celastrol inhibits the proliferation and induced the apoptosis of HL-60 cells. Its mechanism may be related with down-regulation of p-Act and cyclin D1 expressions.


Subject(s)
Humans , Apoptosis , Cyclin D1 , Metabolism , Down-Regulation , Gene Expression Regulation, Leukemic , HL-60 Cells , Proto-Oncogene Proteins c-akt , Metabolism , Triterpenes , Pharmacology
20.
Chinese Journal of Pathology ; (12): 45-50, 2008.
Article in Chinese | WPRIM | ID: wpr-349979

ABSTRACT

<p><b>OBJECTIVE</b>To investigate vasculogenic potential of endothelial progenitor cells (EPCs) derived from human umbilical cord blood and their contribution to the neovascularization of malignant glioma in vivo.</p><p><b>METHODS</b>EPCs were isolated from human umbilical cord blood by density gradient centrifugation. After 7-10 days of culture, EPCs were investigated for CD34 and VEGFR-2 expression by direct immunofluoresent staining. The proliferative activity, migratory capability and forming capillary-like tubules were also monitored after stimulation with VEGF(50 mg/L) in vitro. Moreover, EPCs were administered into tumor-bearing mice, and the tumor and mouse organs were examined under confocal laser scanning microscope to visualize the distribution and localization of transplanted EPCs. In order to quantity the incorporation of EPCs into tumor vessels, cryosections of the tumor tissue were double-labelled with antihuman CD31 and anti-mouse CD31.</p><p><b>RESULTS</b>After 7 to 10 days of culture, EPCs assumed cobblestone-like monolayer growth pattern with nearly complete confluence, and expressed CD34 and VEGFR-2. Significant proliferative activity, increased migratory capability and forming capillary-like tubules were observed when stimulated with VEGF. The transplanted EPCs in vivo specifically homed to solid tumor tissue and incorporated into the tumor's endothelium. Quantitative analysis revealed that human EPCs contributed significantly to tumor neovascularization by incorporation into tumor vasculature (18.68 +/- 1.32)% of the total vessels.</p><p><b>CONCLUSION</b>EPCs possess the potential to form neovascular network in tumor and play a role in the phenotypical heterogeneity of tumor microvascular architecture.</p>


Subject(s)
Animals , Humans , Mice , Antigens, CD34 , Allergy and Immunology , Endothelial Cells , Pathology , Physiology , Endothelium, Vascular , Pathology , Fetal Blood , Cell Biology , Glioma , Pathology , Neovascularization, Pathologic , Pathology , Platelet Endothelial Cell Adhesion Molecule-1 , Allergy and Immunology , Stem Cells , Pathology , Physiology , Vascular Endothelial Growth Factor Receptor-2 , Allergy and Immunology
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