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Objective:To analyze the gene mutation types and haematological characteristics of αβ compound thalassemia, non-delectable α-thalassemia and Hemoglobin H Disease (HbH disease) in Foshan.Methods:Using the method of retrospective analysis, we selected the population who had been tested for thalassemia gene in Foshan Second People's Hospital Affiliated to Southern Medical University from January 2011 to November 2019. Sysmex XT-5000 automatic hematology analyzer was used for routine blood analysis. α-, β- thalassemia genes were detected by PCR + diversion hybridization method.Results:A total of 4 563 people were tested, of which 1 829 were diagnosed as thalassaemia through genetic diagnosis. αβ compound thalassaemia was detected in 81 cases with a positive rate of 1.8%; non-delectable α-thalassemia was detected in 18 cases with a positive rate of 0.4%; HbH disease was detected in 23 cases with a positive rate of 0.5%. The most common genotypes of αβ compound thalassemia were -- SEA/αα\β41-42/βN (17.3%, 14/81), -α 3.7/αα\β41-42/βN (14.8%, 12/81), -- SEA/αα\β654/βN (11.1%, 9/81). The main manifestations of hematology were normal to mild anemia (93.8%, 76/81). Only β-thalassemia with double heterozygotes and α-thalassemia showed severe anemia. αα CS/αα\βN/βN genotypes were common in the local non delectable α-thalassemia (50.0%, 9/18), and the non delectable α-thalassemia was characterized by non-positive phenotype or typical small-cell hypochromatosis in hematology. The genotypes of local HbH patients were -α 3.7/-- SEA\βN/βN (65.2%, 15/23), and simple HbH manifested as moderate anemia (87.0%, 20/23). Patients with HbH disease and β-thalassemia had normal or mild anemia (13.0%, 3/23). Conclusions:The genotypes of αβ compound thalassemia in Foshan area are diverse and complex, and hematology mainly manifests as mild anemia or normal. Non-delectable α-thalassaemia is common in the genotype of αα CS/αα\βN/βN, and clinical manifestations are asymptomatic gene carriers. The genotype of local HbH patients is mainly -α 3.7/-- SEA\βN/βN, and the hematology mainly shows moderate anemia.
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Objective To investigate the clinical value in changes of serum glutamic acid decarboxylase antibody (GAD-Ab) ,islet cell antibodies(ICA) ,insulin autoantibodies (IAA) and high-sensitivity C-reactive protein (hs-CRP) and renal function in elderly type 2 diabetic patients .Methods 122 cases of endocrine inpatient in our hospital had been diagnosed with type 2 diabetes were chosen from January 2012 to December 2012 .They were divided into islet autoimmunity antibody positive group (n=21) and islet autoimmunity antibody negative group (n=101) according to the antibody test results ,Fasting C-peptide(FCP) ,2-hour postprandial C-peptide(2 h CP) ,glycosylated hemoglobin(HbA1c) ,high-sensitivity-CRP(hs-CRP) and renal function[urea (UREA) ,creatinine (Cr) ,microalbuminuria(urinary mALB) ,urinary β2-microglobulin (urinary β2-MG)]were detected .Test results were statistically analyzed and compared .Results At least one Islet autoimmune antibodies were found in 21 cases of 122 elderly patients with type 2 diabetes .The positive rate was 17 .21% .GAD-Ab was detected positive in 14 cases(11 .47% ) ,ICA was detected positive in 10 ca-ses(8 .19% ) ,IAA was detected positive in 1 case(0 .82% ) ,Two antibodies were detected positive together in 4 patients(3 .27% ) , Three antibodies were not detected positive together .The levels of hs-CRP ,UREA and Cr in Islet autoantibodies positive group were higher then in islet autoimmunity antibody negative group ,the difference was statistically significant (P0 .05) . Conclusion Chronic inflammation and the appearance of islet autoantibodies are closely related to the damage of islet cell function . It has a higher value in monitoring complications and efficacy through understanding islet autoantibodies ,inflammation and changes in renal function in elderly type 2 diabetes .
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BACKGROUND:There is a linker between Hsp65 and Esat6 coding a segment of water repellent polypeptide.It is very gentle and easy to be folded,which is profitable to translate the keno-folding correctly between the two proteins and to make the space structure of fusional proteins consistent with those two native ones.Then,a correct structure is formed and the immunogenicity of the fusional proteins is improved.OBJECTIVE:This study was to clone the fusional genes Hsp65-Esat6 from mycobacterium tuberculosis(MTB)H37Rv, then to construct into a eukaryotic expressing vector which contains an enhanced green fluorescent protein(EGFP) reporter gene,and finally to identify the expression of Hsp65 protein and Esat6 protein by IHC methods.DESIGN:Single sample experiment.SETTING:Department of Microbiology ＆ Immunology,Medical College of Jinan University.MATERIALS:Plasmid pVAE was donated by Professor Cao from South China University of Technology.MTB H37Rv,E.coil DH5 α,expressing plasmid pEGFP-C1 and Hela cells were donated by Doctor Hu Ping.METHODS:The experiment was conducted at the Department of Microbiology ＆ Immunology and the Medical Experimental Center of Jinan University between September 2005 and June 2006.The whole-genome was extracted from MTB H37Rv by molecular cloning technique,and used it as template to amplify Hsp65 (no terminator) gene by polyrnerase chain reaction (PCR),to recombine it with pEGFP-C1 vector after purification to construct pEGHsp65(no terminator)recombination vector.pVAE vector was used as template to amplify Linker-Esat6 gene(with terminator)by PCR, and then to recombine it with pEGHSP65(no terminator)to construct an eukaryotic expressing vector pEGHsp65-Esat6 with the fusional genes Hsp65-Esat6 inside.Finally,genes Hsp65-Esat was checked by molecule biology methods such as PCR, restriction endonuclease and DNA sequencing.Hela cells were transfected with pEGHsp65-Esat6 and the expression of EGFP and the efficiency of transfection were observed.The expressions of Hsp65 protein and Esat6 protein were detected by IHC methods.MAIN OUTCOME MEASURES:①The size of pEGHsp65-Esat6.②The DNA sequencing result of the pEGFP-C1.③The expression of EGFP in the transfected Hela cells.④The IHC results of Hsp65 protein and Esat6 protein in Hela cells.RESULTS:①The size of pEGFP-C1 was 4.7 kb,that of pEGHsp65 was 6.4 kb,and that of pEGHsp65-Esat6 was 6.7 kb.There were differences between their speeds In agarose electrophoresis.②The results showed that it was the same as reported Hsp65 sequence and Esat6 sequence of MTB H37Rv.③After transfecting pEGHsp65-Esat6 for 24 hours,EGFP was found in 30% of Hela through Laser scanning confocal microscope. But there was no EGFP in non-transfected Hela.④Hsp65 protein and Esat6 protein with biological activities were detected in transfected Hela cells by IHC methods.CONCLUSION: Using Hsp65 and Esat6 as immunogens,we have successfully cloned and constructed a eukaryotic expressing vector which contain fusionaI genes Hsp65-Esat6 of MTB H37Rv and EGFP.