Objective: To clone the 5＇-flanking region of the human CD34 gene containing transcriptional regulatory sequence (TRS). Methods: According to the registered 5＇-flanking region of CD34 gene, two pairs of primers were designed and net-PCR was used to amplify 661 bp long TRS of CD34 gene. The CD34 TRS fragment was cloned into reported plasmid pEGFP-1. The role of the regulating the specific expression of recombinant plasmid pCD34 EGFP in hematopoietic and non-hematopoietic cells was observed. Results: Restrictive endonuclease identification and DNA sequencing provedthat the CD34 promoter cloned was consistent with the sequence reported to a large extent. It could induce the EGFP gene to express in hematopoietic cell line K562 specifically, while has no effect on hepatocellular carcinoma cell HepG-2. Conclusion: The cloned CD34 gene TRS has the effect of regulating gene expression specifically. The study established the fundament for the construction of specific gene expression vector used in hematopoietic system cells.

Objective: To analyze the expression of HLA class Ⅰ molecules and MHC classⅠ related chain A and B (MICA/MICB) in human nasopharyngeal carcinoma cell line (CNE2) and multi-drug resistant nasopharyngeal carcinoma cell line (CNE2/ DDP), and to assess their influence on NK cell-mediated lysis.Methods: Expression of HLA classⅠ molecules and MICA/MICB on the surface of CNE2 and CNE2/DDP cell lines was analyzed by flow cytometry. Cytotoxicity of NK cells (isolated from 3 healthy persons) against CNE2 and CNE2/DDP cells were detected by LDH releasing assay at different effect-to-target cell ratios (E∶T). In blocking experiments, anti-MHC class Ⅰ monoclonal antibody (mAb) (W6/32, a pan anti-HLA class Ⅰ antibody) and anti-MHC class I chain related molecules mAb (BAMO-1, specificly against MICA and MICB) were added to the target cells at a E∶T ratio of 10∶1. Results:It was found that the expression of HLA class Ⅰ molecules and MICA/MICB on CNE2 was higher than that on CNE2/DDP(P

Objective:To study the inhibitory effect of allogeneic natural killer(NK) cells on subcutaneously transplanted human multi-drug resistant nasopharyngeal carcinoma cells(CNE2/DDP) in BALB/c nude mice.Methods:Human leucocyte antigen(HLA) genotypes of CNE2/DDP cells and the genotypes of inhibitory killer cell immunoglobulin-like receptor(KIR) in NK cells(isolated from 3 healthy persons by immuno-magnetic microbead technique) were analyzed by PCR-SSP.Twelve BALB/c nude mice were evenly divided into 2 groups:the control group and the treatment group.Mice in the treatment group were injected subcutaneously with 1?106 CNE2/DDP cells together with 3?107 NK cells via the tail veins;mice in the control group were injected with 1?106 CNE2/DDP cells subcutaneously.The tumor formation time,tumor formation rate and changes of tumor size were observed.Three weeks after tumor formation,all the mice were killed and human NK cells in peripheral blood were analyzed by flow cytometry;the tumors were weighed and the tumor inhibitory rates were calculated.Results:The HLA genotypes of CNE2/DDPcells were A2,24,B18,35,Cw4,and 7;the KIR genotypes of the 3 healthy persons were KIR2DL1,KIR2DL3,KIR3DL1,and KIR3DL2.There were mismatches between the KIRs expressed in NK cells and HLA class Ⅰ molecules expressed in the CNE2/DDP cells.NK cells obviously inhibited the growth of CNE2/DDP xenograft in nude mice.The tumor formation periods of control group and NK cell group were(17.17?1.17) d and(24.83?1.47) d,respectively(P

OBJECTIVEThis study aimed to investigate the feasibility of gene therapy for severe G6PD deficiency.

METHODSThe recombinant retroviral vector bearing normal human G6PD cDNA was constructed and transferred into the erythroleukemia cell line K562. Author identified the integration of NeoR gene in the targeted cellular DNA by means of specific PCR. Quantitative method was used to measure the expression of G6PD in the targeted cells.

RESULTSConstruction of the recombinant retroviral vector was successfully established. PCR indicated the integration of NeoR gene in the targeted genomic DNA of the cells. The vector was also shown to be capable of expressing the foreign gene compared to the control (P<0.01).

CONCLUSIONSThe recombinant retroviral vector is competent for transferring and expressing the G6PD gene.

Gene Expression ; Genetic Therapy ; Genetic Vectors ; Glucosephosphate Dehydrogenase ; genetics ; Glucosephosphate Dehydrogenase Deficiency ; therapy ; Humans ; K562 Cells ; Retroviridae ; genetics ; Transfection

BackgroundSchizophrenia and the use of antipsychotic medications are identified to be the likely contributors to the development of metabolic syndrome （MS） and cardiovascular disease， and jeopardize the prognosis of schizophrenia. Therefore， effectively preventing or reducing the risk of developing MS in patients with schizophrenia is critical. ObjectiveTo explore the efficacy of aripiprazole combined with olanzapine for male schizophrenia patients and its effect on MS， so as to provide a certain reference for the selection of antipsychotic drugs for schizophrenia patients. MethodsMale patients （n=80） who were hospitalized in The Third People's Hospital of Meizhou from February to June 2023 and fulfilling the International Classification of Diseases， tenth edition （ICD-10） diagnostic criteria for the schizophrenia were enrolled， and grouped using random number table method， each with 40 cases. Study group was treated with aripiprazole combined with olanzapine， while control group was given aripiprazole monotherapy. The treatment lasted for 6 continuous weeks in both groups. At the baseline， Positive and Negative Symptom Scale （PANSS） score， MS-related indices ［fasting plasma glucose （FPG）， hemoglobin A1c （HbA1c）， body mass index （BMI）， waist-to-hip ratio （WHR）， lipid profile］， S100 calcium-binding protein B （S100B） and high sensitivity C-reactive protein （hs-CRP） were recorded. Then the PANSS scores at the end of the 2^nd， 4^th and 6^th week of treatment， the Clinical Global Impression （CGI） scores at the end of the 2^nd and 6^th week of treatment， as well as the MS-related indices， S100B， hs-CRP， Treatment Emergent Symptom Scale （TESS） score and Rating Scale for Extrapyramidal Side Effects （RSESE） score at the end of the 6^th week of treatment were recorded in all participants. ResultsAnalysis on PANSS score revealed a significant group effect， time effect and group×time interaction effect （F=18.092， 634.780， 2.917， P<0.05 or 0.01）. Analysis on CGI score revealed a significant group effect and time effect （F=20.492， 99.190， P<0.01）. At the end of the 6^th week of treatment， study group detected lower serum concentrations of HbA1c and triglyceride （TG） compared with control group （t=-3.495， -3.293， P<0.05）. The post-treatment hs-CRP level was lower in study group than that in control group （t=-3.916， P<0.05）. Study group scored lower on TESS compared with control group （t=-4.684， P<0.01）. ConclusionAripiprazole combined with olanzapine can effectively alleviate psychotic states in male schizophrenia patients， and the combination therapy yields less impact on MS-related indices than olanzapine monotherapy. ［Funded by Meizhou Science and Technology Plan Project （number， 2022B116）］