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1.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 29(4): 1318-1324, 2021 Aug.
Article in Chinese | MEDLINE | ID: mdl-34362523

ABSTRACT

OBJECTIVE: To study the serological characteristics and molecular biological basis of 8 individuals with Para-Bombay phenotypes in Guangxi area. METHODS: Serological tests were used to identify the blood groups of red cells. Molecular biological methods, including PCR-SSP for ABO genotyping and DNA sequencing for FUT1, were used to detect the genotypes of ABO and FUT1 which determined the expression of H antigen. RESULTS: Eight individuals in the study were all the Para-Bombay phenotypes, including 4 cases of Bmh and 4 cases of Amh. The DNA sequencing for FUT1 showed that 6 cases were h3h3 [c.658C>T (p.Arg220Cys) homozygous mutation], 1 was h832h832 [c.832G>A (p.Asp278Asn) homozygous mutation], and 1 was h328h3 [compound heterozygous mutations of c.328G>A (p.Ala110Thr) and c.658C>T (p.Arg220Cys)]. CONCLUSION: There are varieties of molecular genetic mechanisms for Para-Bombay phenotypes. In this study, the FUT1 mutations that cause Para-Bombay phenotypes in Guangxi area are mainly h3, h328, and h832, among which h3 is the most common mutant.


Subject(s)
ABO Blood-Group System , Fucosyltransferases , ABO Blood-Group System/genetics , Alleles , China , Fucosyltransferases/genetics , Genotype , Humans , Mutation , Phenotype , Galactoside 2-alpha-L-fucosyltransferase
2.
Theor Appl Genet ; 134(10): 3225-3236, 2021 Oct.
Article in English | MEDLINE | ID: mdl-34132847

ABSTRACT

KEY MESSAGE: Decisive role of reduced vrs1 transcript abundance in six-rowed spike of barley carrying vrs1.a4 was genetically proved and its potential causes were preliminarily analyzed. Six-rowed spike 1 (vrs1) is the major determinant of the six-rowed spike phenotype of barley (Hordeum vulgare L.). Alleles of Vrs1 have been extensively investigated. Allele vrs1.a4 in six-rowed barley is unique in that it has the same coding sequence as Vrs1.b4 in two-rowed barley. The determinant of row-type in vrs1.a4 carriers has not been experimentally identified. Here, we identified Vrs1.b4 in two-rowed accessions and vrs1.a4 in six-rowed accessions from the Qinghai-Tibet Plateau at high frequency. Genetic analyses revealed a single nuclear gene accounting for row-type alteration in these accessions. Physical mapping identified a 0.08-cM (~ 554-kb) target interval on chromosome 2H, wherein Vrs1 was the most likely candidate gene. Further analysis of Vrs1 expression in offspring of the mapping populations or different Vrs1.b4 and vrs1.a4 lines confirmed that downregulated expression of vrs1.a4 causes six-rowed spike. Regulatory sequence analysis found a single 'TA' dinucleotide deletion in vrs1.a4 carriers within a 'TA' tandem-repeat-enriched region ~ 1 kb upstream of the coding region. DNA methylation levels did not correspond to the expression difference and therefore did not affect Vrs1 expression. More evidence is needed to verify the causal link between the 'TA' deletion and the downregulated Vrs1 expression and hence the six-rowed spike phenotype.


Subject(s)
Chromosome Mapping/methods , Chromosomes, Plant/genetics , Gene Expression Regulation, Plant , Hordeum/growth & development , Hordeum/genetics , Phenotype , Plant Proteins/metabolism , DNA Methylation , Phylogeny , Plant Proteins/genetics
3.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 28(6): 2056-2065, 2020 Dec.
Article in Chinese | MEDLINE | ID: mdl-33283742

ABSTRACT

OBJECTIVE: To study two novel CD36 gene mutations at the CD36 splicing sites found in Guangxi population, as well as the molecular basis and population incidence of them. METHODS: DNA sequencing and cDNA clonal sequencing were used to detect CD36 exon sequence and the protein coding region sequence of CD36 mRNA for 2 CD36 deficient individuals (HHC and WGM) found in Guangxi population. Eukaryotic expression cell lines were established for the discovery of CD36 mRNA abnormal transcripts and Western blot assay was used to verify the effect of abnormal CD36 mRNA transcripts on CD36 expression. A DNA PCR-SSP genotyping method was established for the two CD36 novel mutations, and the population distribution was investigated among 110 CD36 deficient individuals in Guangxi region and 296 random individuals in Guangxi population. RESULTS: Novel mutation of c.430 -1G>C was found at the CD36 splicing site in HHC and WGM individuals, and novel mutation of c.1006 +2T>G at the CD36 splicing site was also found in the WGM individual. CD36 cDNA clonal sequencing showed that CD36 c.430 -1G>C could lead to the production of the two CD36 mRNA transcript variants: c.429_430insï¼»430-17_430-2;Cï¼½(p.Ala144fsTer1) and c.430_609del(p.Ala144_Pro203del)(GenBank:HM 217023.1); and CD36 c.1006 +2T>G could lead to the production of CD36 mRNA transcript variant of c.819_1006 del (p.Ser274GlufsTer16) (GenBank: HM217025.1). It was verified that all the three transcript variants could lead to CD36 deficiency by establishment of eukaryotic expression cell lines and Western blot assay. A study of the population incidence of two novel CD36 splicing site mutations found showed that in 110 CD36 deficient individuals and in 296 random individuals in Guangxi region, the mutation rate of CD36 c.430 -1G>C was 10.91% (12/110) and 1.35% (4/296), respectively, while CD36 c.1006 +2T>G was 2.73% (3/110) and 0 (0/296), respectively. CONCLUSION: This study identifies two novel CD36 mutations at CD36 splicing site, and preliminary clarified their molecular basis for the CD36 deficiency and the distribution characteristics in Guangxi population as well. It provides an experimental and theoretical basis for studying the molecular mechanism and characteristics of CD36 deficiency in Chinese population.


Subject(s)
Blood Platelet Disorders , CD36 Antigens , CD36 Antigens/genetics , China , Genetic Diseases, Inborn , Humans , Mutation , RNA Splicing
4.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 26(2): 541-546, 2018 Apr.
Article in Chinese | MEDLINE | ID: mdl-29665929

ABSTRACT

OBJECTIVE: To analyse the cases of platelet transfusion refractoriness after received HLA-matched unrelated donor hematopoietic stem cell transplantation, to analyze and identify the phenotype and genotype of CD36 in both the patient and stem cell donor, as well as the characteristic of antibody induced platelet transfusion refractoriness, and to analyse the efficacy of matched CD36-deficiency platelets transfusions. METHODS: The CD36 expression on platelet and monocyte was analyzed by flow cytometry (FCM) in both patient and donor. Polymerase chain reaction sequence-based typing (PCR-SBT) was used to analyze the exons sequence of CD36 and HPA. Fast monoclonal antibody-specific immobilization of platelet antigen (F-MAIPA) and FCM were used to identify platelet antibodies in the patient. Short tandem repeat polymerase chain reaction (STR-PCR) was applied to monitor engraftment evidence. The platelet level was monitored. CD36- deficiency donor's platelets were selected from CD36- deficiency donor blood bank. RESULTS: The donor was CD36 positive and the patient was typed I CD36 deficiency. The anti-CD36 antibody was identified in patient's serum (after transplantation), while the HLA and HPA-related antibodies were excluded. Sequence analysis of CD36 exon in the patient showed Exon 6 -1G>C(Change in splicing site) homozygote, which was a novel CD36 mutation. STR, HPA and CD36 of the patient (complete chimerism) were conversed to that of donor gene types on day 18 after allo-HSCT. The positive CD36 expression on platelet and monocyte in the patient was observed on day 96 after allo-HSCT. The patient showed the platelet transfusion refractoriness which was significantly improved after platelets transfusions from CD36 deficiency donors. CONCLUSION: Stem cell transplants resulted in anti-CD36 and caused platelet transfusion refractoriness, that was first reported in China. To ensure the efficacy of platelet transfusion, the CD36-deficiency patient should receive CD36 deficiency platelets for transfusion.


Subject(s)
Platelet Transfusion , Antigens, Human Platelet , Blood Platelet Disorders , Blood Platelets , CD36 Antigens , China , Humans , Thrombocytopenia
5.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 22(2): 399-402, 2014 Apr.
Article in Chinese | MEDLINE | ID: mdl-24763012

ABSTRACT

This study was aimed to investigate the detection and diagnosis of the neonatal alloimmune thrombocytopenia purpura (NAITP) caused by anti-HPA-5b antibody. The platelet count and clinical manifestation in the newborn were examined. The HPA-1-21bw genotypes of the newborn and her parents were detected by multiple-PCR and DNA sequencing. The HPA-specific antibody in the sera of newborn and her mother were detected and identified by flow cytometry (FCM) and monoclonal antibody-specific immobilization of platelet antigens (MAIPA). The results indicated that the clinical manifestations of the newborn were lighter. The HPA genotyping showed that the genotype of the newborn was HPA-5ab, while that of her mother and father were HPA-5aa and HPA-5ab, respectively. The antibody against the platelet of newborn's father existed in the newborn's mother sera. The HPA antibody of the mother was identified as anti-HPA-5b. It is concluded that the newborn with neonatal alloimmune thrombocytopenia purpura was caused by the antibody against HPA-5b.


Subject(s)
Antigens, Human Platelet/genetics , Purpura, Thrombocytopenic, Idiopathic/diagnosis , Thrombocytopenia, Neonatal Alloimmune/diagnosis , China , Female , Genotype , Humans , Infant, Newborn , Purpura, Thrombocytopenic, Idiopathic/genetics , Thrombocytopenia, Neonatal Alloimmune/genetics
6.
Zhonghua Xue Ye Xue Za Zhi ; 34(1): 45-8, 2013 Jan.
Article in Chinese | MEDLINE | ID: mdl-23597464

ABSTRACT

OBJECTIVE: To explore the diagnosis and treatment of a case of neonatal alloimmune thrombocytopenia purpura (NAITP) caused by anti HPA-3a antibody. METHODS: The platelet counts and purpuric symptom in the newborn were clinical examined. The HPA-1-21bw genotypes of the newborn and his parents were detected by multiple DNA-PCR, gene sequencing and genotyping. The HPA specificity antibody in the sera of newborn and his mother were detected by flow cytometry (FCM), and the HPA specificity antibody was identified by monoclonal antibody-specific immobilization of platelet antigens (MAIPA). RESULTS: The newborn had the typical symptom of NAITP, multiple subcutaneous petechia, hematuria and coffee-like vomitus. The HPA genotype of the newborn was HPA-3ab, while that of his mother and his father were HPA-3bb and HPA-3aa, respectively. The sera of newborn and his mother existed antibody against the platelet of newborn's father. The HPA antibody of the newborn and his mother were identified as anti HPA-3a. The newborn was approved a patient of NAITP caused by anti HPA-3a antibody. CONCLUSION: The diagnosis and treatment for NAITP newborn caused by anti HPA-3a antibody in this study was the first domestic report. It could provide successful experiences and references for the similar cases.


Subject(s)
Antigens, Human Platelet/immunology , Isoantibodies/adverse effects , Thrombocytopenia, Neonatal Alloimmune/etiology , Antibody Specificity/immunology , Genotype , Humans , Infant, Newborn , Isoantibodies/immunology , Male , Thrombocytopenia, Neonatal Alloimmune/immunology
7.
Article in English | WPRIM (Western Pacific) | ID: wpr-251386

ABSTRACT

Cigarette smoke is associated with the development of several diseases, such as chronic obstructive pulmonary disease (COPD). The purpose of this study was to investigate genotoxicity and heat shock protein 70 (Hsp70) in human airway smooth muscle cells (HASMCs) exposed to cigarette smoke extract (CSE). HASMCs was exposed to CSE with different doses for 24 h. The level of 8-hydroxydeoxyguanosine (8-OHdG) was determined by using HPLC-ECD, the DNA damage was analyzed by using comet assay, and apoptosis was examined by using Annexin-FITC/PI staining. The production of Hsp70 after CSE stimulation was tested. Results indicated that CSE significantly increased the level of 8-OHdG, DNA damage and cell apoptosis, and reduced the production of Hsp70. In particular, levels of Hsp70 were inversely correlated with 8-OHdG, DNA damage and cell apoptosis. It was concluded that cigarette smoke induced genotoxicity and decreased the production of cell protective protein Hsp70, which may contribute to the development of some airway diseases.


Subject(s)
Humans , Apoptosis , DNA Damage , Deoxyguanosine , Metabolism , HSP70 Heat-Shock Proteins , Genetics , Metabolism , Lung , Cell Biology , Myocytes, Smooth Muscle , Metabolism , Smoke , Nicotiana , Toxicity , Tumor Cells, Cultured
8.
Article in English | WPRIM (Western Pacific) | ID: wpr-636470

ABSTRACT

Cigarette smoke is associated with the development of several diseases, such as chronic obstructive pulmonary disease (COPD). The purpose of this study was to investigate genotoxicity and heat shock protein 70 (Hsp70) in human airway smooth muscle cells (HASMCs) exposed to cigarette smoke extract (CSE). HASMCs was exposed to CSE with different doses for 24 h. The level of 8-hydroxydeoxyguanosine (8-OHdG) was determined by using HPLC-ECD, the DNA damage was analyzed by using comet assay, and apoptosis was examined by using Annexin-FITC/PI staining. The production of Hsp70 after CSE stimulation was tested. Results indicated that CSE significantly increased the level of 8-OHdG, DNA damage and cell apoptosis, and reduced the production of Hsp70. In particular, levels of Hsp70 were inversely correlated with 8-OHdG, DNA damage and cell apoptosis. It was concluded that cigarette smoke induced genotoxicity and decreased the production of cell protective protein Hsp70, which may contribute to the development of some airway diseases.

9.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 42(5): 589-93, 2011 Sep.
Article in Chinese | MEDLINE | ID: mdl-22007478

ABSTRACT

OBJECTIVE: To study the effect of RhoE on the transcriptional regulation of cd44 and in vivo tumorigenicity of nude mice. METHODS: cd44 promoter was amplified from human embryonic kidney HEK293 cells with PCR and insert into Dual-Luciferase Reporter plasmid pGL3-Basic. After confirmed with sequence analysis, the generated recombinant was transfected into SW480 and LoVo cells to monitor their activity. Colon cancer SW480 and LoVo cells were cotransfected with pGL3-CD44 promoter along with pcDNA3. 1-RhoE and pcDNA3. 1 respectively. SW480 and LoVo cells were stably transfected with pcDNA3. 1-RhoE and the control group and were inoculated into nude mice to observe tumor growth. Immunohistochemistry assay was applied to observe the morphology of tumor cells and the expression of CD44 molecules. RESULTS: The cd44 promoter sequence was amplified correctly, Dual-Luciferase Reporter Assay showed that the constructed reporter gene has promoter activity. The expression of cd44 promoter sequence containing reporter gene in pcDNA3. 1-RhoE expression positive LoVo cells was inhibited; HE staining demonstrated that the pcDNA3. 1-RhoE transfected tumor cells was significantly smaller than that in the control group, and consistent size and shape tumor cells were observed but no tumor giant cells, the corresponding volume of the tumor nuclei were also small. CONCLUSION: RhoE could partially reverse the malignant biological behavior of tumors by inhibiting the transcriptional regulation of cd44 promoter.


Subject(s)
Colorectal Neoplasms/genetics , Genes, Tumor Suppressor/physiology , Hyaluronan Receptors/genetics , Promoter Regions, Genetic/genetics , rho GTP-Binding Proteins/genetics , Animals , Cell Line, Tumor , Colorectal Neoplasms/pathology , Humans , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Transplantation , Transfection , rho GTP-Binding Proteins/biosynthesis , rho GTP-Binding Proteins/physiology
10.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 42(4): 445-50, 2011 Jul.
Article in Chinese | MEDLINE | ID: mdl-21866623

ABSTRACT

OBJECTIVE: To investigate the effect of RhoA to VEGF, HIF-1alpha and MVD (microvascular density) and the effect of MG132 to RhoA. METHODS: The constitutively-active mutant vectors of RhoA (pCEFL-GST-V14RhoA) were transfected into gastric cancer cell line MKN-45 by Lipofectamine 2000, single clones were selected by G418 and identified with western blot. The content of VEGF in the conditioned media was detected by ELISA. Constitutively-active RhoA nude mice models were established and treated with MG132. The effect of RhoA and MG132 on expression of HIF-1alpha, VEGF and CD31 were detected by immunohistochemistry. RESULTS: Cell line of stable-transfected constitutively-active RhoA was established and constitutively-active RhoA could stimulate secretion of VEGF but MG132 inhibited that. Constitutively-active RhoA could obviously induce growth of tumor (P < 0.05), but MG132 inhibited it (P < 0.05). Constitutively-active RhoA could promote protein of HIF-1alpha, VEGF and CD31 but MG132 inhibited the function of RhoA (P < 0.05). CONCLUSION: Our studies indicates that MG132 could affect angiogenesis of tumors through inhibition the regulating function of RhoA on HIF-1alpha, VEGF and CD31.


Subject(s)
Angiogenesis Inhibitors/pharmacology , Leupeptins/pharmacology , Proteasome Inhibitors , Stomach Neoplasms/blood supply , rhoA GTP-Binding Protein/antagonists & inhibitors , Animals , Cell Line, Tumor , Humans , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Mice , Mice, Nude , Neoplasm Transplantation , Neovascularization, Pathologic , Stomach Neoplasms/pathology , Transfection , Vascular Endothelial Growth Factor A/metabolism , rhoA GTP-Binding Protein/pharmacology
11.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 42(4): 451-4, 460, 2011 Jul.
Article in Chinese | MEDLINE | ID: mdl-21866624

ABSTRACT

OBJECTIVE: To observe the affect of Small interfering RNA Rac1b (Si-Rac1b) on the malignant biological behaviors of colorectal cancer cell including the proliferation, migration, invasion and apoptosis of the cells. METHODS: Mediated by lipofectamine 2000, Si-Rac1b was transfected into colorectal cancer cell line SW1116 (with overexpression of Rac1b). The expression of Rac1b was detected by Western blotting and RT-PCR. The CCK-8 assay was used to analyze the cell proliferation, the Wound-healing assay and invasion assay were respectively applied to analyze the cell migration and invasion, and the Hoechst 33258 was used to evaluate the apoptotic index. RESULTS: Si-Rac1b can knock down the Rac1b but not Rac1 both in the level of mRNA and protein. In addition, Si-Rac1b could singnificantly facilitate the cell proliferation, migration, invasion and control the cell apoptosis. CONCLUSION: Si-Rac1b could partically reverse the malignant phenotypes of colorectal cancer cell.


Subject(s)
Apoptosis/genetics , Colorectal Neoplasms/genetics , RNA, Small Interfering/genetics , rac1 GTP-Binding Protein/genetics , Cell Line, Tumor , Cell Movement , Cell Proliferation , Colorectal Neoplasms/pathology , Humans , Transfection
12.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 42(4): 466-70, 2011 Jul.
Article in Chinese | MEDLINE | ID: mdl-21866627

ABSTRACT

OBJECTIVE: To determine the effect of cell division cycle42 (Cdc42) on the multidrug resistance of oxaliplatin-resistant human colon cancer cells. METHODS: The protein expression levels of Cdc42 in oxaliplatin-resistant colon cancer cells and parental cells were examined with Western blot. pDEST26-His-Cdc42 was transfected by lipofectamine 2000 into SW480 and Colo320 cells with low expression of Cdc42. Cdc42 siRNA was transfected by lipofectamine 2000 into SW480/L-OHP and Colo320/L-OHP cells with high expression of Cdc42. The expression of Cdc42 in these cell lines were examined by Western blot and RT-PCR 48 hours after transfection. The sensitivity of colon cancer cells to antitumor drugs was evaluated using CCK-8 assay. The concentration of each drug that caused a 50% reduction in the numbers of cells (IC50) was calculated. The expression of P-gp, MRP1 in SW480/Cdc42 and Colo320/ Cdc42 cells were examined with Western blot. RESULTS: Cdc42 was over-expressed in the SW480/L-OHP and Colo320/L-OHP cell lines. Over-expression of Cdc42 significantly enhanced the resistance of colon cancer cells to multiple antitumor drugs and up-regulated the expression of P-gp and MRP1. CONCLUSION: Cdc42 enhances the resistance of colon cancer cells to several antitumor drugs. It might become a potential target for reversing multidrug resistance of colon cancer.


Subject(s)
Colonic Neoplasms/pathology , Drug Resistance, Multiple , Drug Resistance, Neoplasm , Organoplatinum Compounds/pharmacology , cdc42 GTP-Binding Protein/metabolism , Antineoplastic Agents/pharmacology , Cell Line, Tumor , Colonic Neoplasms/metabolism , Humans , Oxaliplatin , cdc42 GTP-Binding Protein/genetics
13.
Bioinformatics ; 25(8): 981-8, 2009 Apr 15.
Article in English | MEDLINE | ID: mdl-19237446

ABSTRACT

MOTIVATION: Identification of disease-related genes using high-throughput microarray data is more difficult for complex diseases as compared with monogenic ones. We hypothesized that an endophenotype derived from transcriptional data is associated with a set of genes corresponding to a pathway cluster. We assumed that a complex disease is associated with multiple endophenotypes and can be induced by their up/downregulated gene expression patterns. Thus, a neural network model was adopted to simulate the gene-endophenotype-disease relationship in which endophenotypes were represented by hidden nodes. RESULTS: We successfully constructed a three-endophenotype model for Taiwanese hypertensive males with high identification accuracy. Of the three endophenotypes, one is strongly protective, another is weakly protective and the third is highly correlated with developing young-onset male hypertension. Sixteen of the involved 101 genes were highly and consistently influential to the endophenotypes. Identification of SLC4A5, SLC5A10 and LDOC1 indicated that sodium/bicarbonate transport, sodium/glucose transport and cell-proliferation regulation may play important upstream roles and identification of BNIP1, APOBEC3F and LDOC1 suggested that apoptosis, innate immune response and cell-proliferation regulation may play important downstream roles in hypertension. The involved genes not only provide insights into the mechanism of hypertension but should also be considered in future gene mapping endeavors.


Subject(s)
Hypertension/epidemiology , Hypertension/genetics , Neural Networks, Computer , Oligonucleotide Array Sequence Analysis , Age of Onset , Computer Simulation , Humans , Male , Phenotype
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