MICA基因第5外显子与乙肝后肝硬变的相关性研究

目的研究MICA基因多态性、sMICA分子与乙肝后肝硬化（livercirrhosis,LC）的相关性。方法采用MICA—STR微卫星基因分型技术检测101例HBsAg阳性LC患者和141例健康对照者MICA基因第5外显子的基因多态性,并进行统计学分析。结果在101例湖南汉族HBsAg阳性LC患者中检测到5种MICA基因第5外显子等位基因,频率分别为：MICA＊A4（10．9％）,MICA＊A5（37．1％）,MICA＊A5．1（34．2％）,MICA＊A6（7．4％）,MICA＊A9（10．4％）。15种基因型分别为MICA＊A4／A4,MICA＊A4／A5．MICA＊A4／A5．1．MICA＊A4／A6,MICA＊A4／A9,MICA＊A5／A5,MICA＊A5／A5．1,MICA＊A5／A6．MICA＊A5／A9,MICA＊A5．1／A5．1,MICA＊A5．1／A6,MICA＊A5．1／A9,MICA＊A6／A6,MICA＊A6／A9,MICA＊A9／A9。MICA＊A5．1基因与LC发病正相关（P=0．042）,OR值为1．398,与患者性别无关（χ^2=0．08,P=0．778）。结论MICA＊A5．1基因与乙肝后LC发病正相关,提示该基因可能是乙肝后出现LC的重要易感基因。     

Serological characteristic and molecular basis of A2 subgroup in the Chinese population

BackgroundA2 phenotype is a common subgroup of blood group A, but the serological characteristic and genetics basis of A2 phenotype currently was rare reported in the Chinese Han population. Here, a large scale study of the serology and genetics of A2 and A2B phenotypes was performed.Methods/materials11263 Chinese individuals with group A and AB phenotypes were determined for A2 antigen with the standard serological method. The full coding region of the ABO gene was sequenced in the individuals with A2 and A2B phenotypes. Some samples including each ABO genotypes were chosen for determining the activity of glycosyltransferase A (GTA) in plasma.Results134 individuals were assigned as A2 and A2B phenotypes in 11263 individuals. There was imbalance in A2 and A2B phenotypes and the proportion of A2B among AB samples was significantly higher than that of A2 in group A samples. All samples of the A2 and A2B phenotypes were classified into A2-related allele group, A1-related allele group and the other group based on kind of the ABO genotype. Four novel A2-related alleles (A217, A218, A219, A220) were identified. The individuals with same genotype showed different agglutination strength with anti-A1 and anti-H on their RBCs. The plasma from individuals with A2-related allele had almost no GTA activity, while plasma from individuals with A1-related allele had some GTA activity.ConclusionA2 and A2B phenotypes could derive from different genotypes and the serological characteristic may be heterogeneity in the Chinese Han population.     

Atrioventricular nodal reentry tachycardia with multiple AH jumps: electrophysiological characteristics and radiofrequency ablation

This article describes the additional use of incremental atrial burst pacing (A1A1) and double atrial extrastimulation with a predefined fast pathway conducted A2 (A1A2A3), rather than single atrial extrastimulation (A1A2) only, to characterize typical atrioventricular nodal reentrant tachycardia (AVNRT). The authors noted an additional 32% of patients had multiple anterograde AV nodal physiology demonstrated when A1A1 or A1A2A3 protocols were deployed compared to more conventional A1A2 protocols. The A2H2max (449 +/- 147 vs 339 +/- 94 ms) and A3H3max (481 +/- 120 vs 389 +/- 85 ms) were higher in 31 patients where multiple jumps in the AV nodal conduction curve were obtained (group 1) compared to 192 patients where only single jump was obtained (group 2) (both P < 0.01). Postablation, the degree of reduction of A2H2max (49%) and A3H3max (50%) in group 1 was greater than in group 2 (38% and 42%, respectively, P < 0.05). In seven of group 1 patients in whom A1A2A3 stimulation was required to reveal multiple jumps, the A2H2max remained unchanged after ablation (237 +/- 89 vs 214 +/- 59, P > 0.05). A3H3max was the only parameter that shortened significantly after ablation. Generally, successful ablation resulted in loss of multiple discontinuities in A1A1/A1H1 or A2A3/A3H3 curves. In conclusion, a combination of A1A2, A1A1, and A1A2A3 are required to fully elucidate AVNRT. Significant shortening of AHmax or loss of multiple jumps after ablation indicates successful elimination of AVNRT in these patients.     

A37 REACTIVITY OF PLATELET AUTOANTIBODIES WITH PAPAIN TREATED PLATELETS AND COMPARISON WITH GLYCOPROTEIN SPECIFICITY.

A1 INFLUENCE OF POSTURE ON REACTIONS IN NEW BLOOD DONORS. A2 A CONFIDENTIAL UNIT EXCLUSION SYSTEM IDENTIFIES DONORS WITH A POTENTIAL FOR HIV INFECTION. A3 A STABLE BLOOD SUPPLY FOR THE FUTURE: THE RECRUITMENT OF 16 TO 18 YEAR OLD DONORS TODAY AND THEI CONTRIBUTION AS COMMITTED REGULAR DONORS OF TOMORROW. A4 APPROACH TO A SUPPLY CRISIS OF HYPERIMMUNE RHESUS PLASMA FOR THE PRODUCTION OF RhD IMMUNOGLOBULIN A5 THE INFLUENCE OF AGE, SEX AND ABO BLOOD GROUP ON THE INCIDENCE OF CMV ANTIBODIES IN SYDNEY BLOOD DONORS. A6 THE INCIDENCE OF CATEGORY VI AMONGST WEAK Rh(D) POSITIVE SYDNEY BLOOD DONORS. A7 A MODIFIED METHOD FOR DETECTING HIGH TITRE ANTI-A AND ANTI-B IN GROUP O DONORS A8 IMPROVING THE CLINICAL SPECIFICITY OF ALANINE AMINO TRANSFERASE (ALT) TEST RESULTS WITHIN THE NORMAL BLOOD DONOR POPULATION OF QUEENSLAND. A9 EXTRACTION OF HCV RNA USING A GUANIDINE ISOTHIOCYNATE METHOD. A10 HEPATITIS C VIRUS (HCV) ANTIBODY DETECTION IN TASMANIAN BLOOD DONORS. A11 EFFECTIVE INTERNAL QUALITY CONTROL FOR ENZYME IMMUNOASSAYS A12 DETECTION OF ANTIBODY TO NON-PATHOGENIC RETROVIRUSES (SPUMAVIRUSES) IN HUMAN SERUM A13 DETECTION OF ANTIBODY TO NON-PATHOGENIC RETROVIRUSES (SPUMAVIRUSES) IN HUMAN SERUM A14 A NOVEL BLOOD BAG SYSTEM WITH POTENTIAL, FOR THE ASIA-PACIFIC MARKET. A15 DESIGN OF CONTAINERS SUITABLE FOR THE TRANSPORT OF RED CELL, PLATELET AND FROZEN PLASMA PRODUCTS. A16 EVALUATION OF INDICATOR LABELS FOR QUALITY ASSURANCE OF IRRADIATION PROCEDURE OF BLOOD PRODUCTS. A17 MOLECULAR TYPING FOR UNUSUAL ABO TYPES. A18 AN EXAMPLE OF THE RARE ABO SUBGROUP, A19 RFLP ANALYSIS OF A RH NULL BLOOD DONOR. A20 A RELATIONSHIP BETWEEN LEWIS ERYTHROCYTE PHENOTYPES AND COLORECTAL CANCER. A21 PATERNITY TESTING USING SINGLE LOCUS DNA PROBES: OBSERVATIONS ON THE REFERENCE DATA BASE SIZE A22 USE OF FAMILY AND POPULATION STUDIES TO DETERMINE THE SPECIFICITY AND INHERITANCE OF NEUTROPHIL ANTIGENS DEFINED BY PLANT LECTINS. A23 SAMPLING PLANS: IS THERE RELEVANCE FOR BLOOD COMPONENT QC? A24 QUALITY MANAGEMENT: HOW DO WE DO IT IN A STATE THE SIZE OF QUEENSLAND? A26 THE ENERGY METABOLISM OF CIRCULATING CELLS. A27 ACETATE UTILISATION RATES AND THE EFFECT OF GLUCOSE-FREE PLASMA IN PLATELET CONCENTRATE STORED IN A MIMIMAL MEDIUM (MPM). A28 IMPROVED LEVELS OF 2,3 DJPHOSPHOGLYCERATE IN RED CELL SUSPENSIONS PREPARED FROM BLOOD COLLECTED INTO DEXTROSE-FREE ANTICOAGULANT. A29 EVALUATION OF RED CELL FREEZING METHODS AS A PRELUDE TO ADOPTING -80° C FREEZING IN HIGH GLYCEROL IN ROUTINE PRACTICE. A30 CLUMPING IN PLATELET CONCENTRATES - AN UNSOLVED PROBLEM. A31 AUTOLOGOUS BLOOD: SAFE FOR OTHERS OR NOT? A32 ESTABLISHMENT OF AN AUSTRALIAN HAEMOPHILIA TREATMENT CENTRE DATA BANK. A33 EXPERIENCE IN THE USE OF ROBOTICS AND MICROPLATE TECHNOLOGY TO SEMI-AUTOMATE A ROUTINE HOSPITAL BLOOD BANK. A34 AN ANTI-IgAl/IgA2 ELISA ASSAY FOR THE INVESTIGATION OF HYPESENSITIVITY TRANSFUSION REACTIONS. A35 THE INFLUENCE OF IgG AGGREGATES AND FRESH NORMAL SERUM ON THE MONOCYTE MONOLAYER ASSAY A36 DETECTION OF Rh(D) POSITIVE FETAL CELLS IN PREGNANT Rh(D)-NEGATIVE WOMEN BY FLOW CYTOMETRY. A38 HAEMOSTAT-IX: A HIGH PURITY FACTOR CONCENTRATE FOR THE TREATMENT OF PATIENTS WITH HAEMOPHILIA B. A39 GRAVITY FILTRATION OF PLASMA FROM DONOR BLOOD UTILISING A HOLLOW FIBRE FILTER MEMBRANE DEVICE A40 The Therapeutic Device Problem Reporting Scheme, and the Victorian Red Cross Blood Bank A43 HIGH FREQUENCY ANTIBODIES AND THE ADVANTAGES OF MANUAL POLYBRENE. A44 FACTS AND FANTASY IN THE DEVELOPMENT OF PLATELET ADDITIVE SOLUTIONS. A45 LACK OF EFFECT OF STORAGE CONTAINER ON STORAGE OF PLATELETS PREPARED FROM DEXTROSE-FREE BLOOD, A46 PLATELETS PREPARED FROM DEXTROSE-FREE BLOOD MAY BE STORED WITHOUT AGITATION. A47 QUALITY OF BED CELL CONCENTRATE IN HOSPITALS COMPARED TO THE BLOOD BANK A48FLOW CYTOMETRIC CHARACTERISATION OF LEUCOCYTE - DEPLETED RED CELL CONCEHTRATES. A49 PRODUCTION AND CHARACTERISATION OF HUMAN MONOCLONAL ANTI-D ANTIBODIES. A50 CD55 AND CD59 SUSCEPTIBILITY TO PROTEASE TREATMENT AND THE RESULTANT EFFECT ON COMPLEMENT LYSIS OF RBCs. A51 DIRECT COMPARISON BETWEEN PLATELET STORAGE CONTAINERS - IMPROVEMENT IN STORAGE CHARACTERISTICS OF TUTA PLATELET BAGS OVER THE PAST FOUR YEARS. A52 IMPROVED SOLID-PHASE MIXED PASSIVE HAENAGGLUTININ ASSAY (MPHA) WITH FROZEN PANEL PLATELETS FOR THE DETECTION OF HUMAN PLATELET ANTIBODIES. A53 DEVELOPMENT OF A SOLVENT DETERGENT TREATED THROMBIN CONCENTRATE AS A COMPONENT OF A FIBRIN GLUE KIT. A54 Autologous blood transfusion: a promotional programme A55 AVAILABILITY OF BLOOD PRODUCTS FOR ACUTELY BLEEDING PATIENTS. A56 REMINISCENCES OF 50 YEARS A. A TRANSFUSION ST. A57 A NATIONAL SYSTEM FOR REPORTING TRANSFUSION REACTIONS TO FRACTIONATED BLOOD PRODUCTS. A58 EFFECT OF FLUORESCENT LIGHTING ON THE VISUAL APPEARANCE OF PLATELET CONCENTRATES A59 USING A MICROWAVE OVEN TO THAW FRESH FROZEN PLASMA. A60 COAGULATION CAPACITY OF POOLED PLATELET PLASMA. A61 A COMPARISON OF IMMUNOHAEMATOLOGY SURVEY PERFORMANCE BETWEEN NEK ZEALAND AND AUSTRALIA A62 COMPATIBILITY TESTING: ARE ENZYME TESTS REQUIRED? A63 AN EVALUATION OF THE DIAMED MEROTYPING SYSTEM FOR THE PERFORMANCE OF THE DIRKT ANTIGLOBUDIN TEST. A64 ASSESSMENT OF PERFORMANCE IN BLOOD GROUP ANTIBODY DETECTION. A65 CHARACTERISATION OF MABS TO THROMBIN-HIRUDIN COMPLEXES WITH IMMUNOASSAY POTENTIAL. A66 MONITORING ANTT-HPA-la (P1^A1) PLATELET ANTIBODY LEVELS DURING PREGNANCY USING THE MAIPA TEST. A67 COMPARISON OF PIFT AND MAIPA TEST“ IN THE DETECTION OF ANTI-HPA-la (PI^A1) PLATELET ANTIBODIES. A68 USE OF PLATELET-CROSSMATCHING IN SUPPORT OF A CASE OF MYELODYSPLASIA WTTH A PLATELET SPECIFIC AND B LYMPHOCYTE ANTIBODY A69 The Pattern of Leucocyte Antibody formation in Transfused Patients. A70 DETECTION OF HPA-Ia ANTIBODY IN BREAST MILK A71 ANALYSIS OF PRENATAL SCREENING. A72 DETECTION OF MINOR POPULATIONS OF ERYTHROCYTES A73 MODIFICATIONS TO THE MCNOCYTE-MEDIATED ADCC ASSAY. A74 AN AUTO ANTI-JMH; GAMMA-CLONE POLYSPECIFIC AHG AS A USEFUL TOOL. A75 CLINICALLY SIGNIFICANT ANTI-A1 DERIVED FROM B LYMPHOCYTES FOLLOWING SINGLE LUNG TRANSPLANTATION. A76 CONFIRMATION THAT ANTI-ELO CAUSES HAEMOLYTIC DISEASE OFTHE NEWBORN. A77 ANTI-Do^a STIMULATED BY PREGNANCY. A78 DONOR IgM ANTI-A ASSOCIATED WITH HAEMOLYTIC TRANSFUSION REACTION A79 COLLECTION OF GRANULOCYTES AND PLATELETS USING FENWALL CS 3000 AND HAEMONETICS 30 CELL SEPARATORS - A COMPARISON. A80 APPARENT LYMPHOPENIA IN PLASMAPHERESIS DONORS     

中国北方汉族人群A204等位基因的发现及家系调查

目的了解中国北方汉族人群ABO血型系统A2亚型的基因型分布特点及A204的家系遗传规律。方法应用血清学试验筛选出A亚型标本,采用PCR-SSP法对ABO血型A2亚型作基因分型检测,随后直接DNA测序;对A204先证者作家系调查。结果从3 395名献血者中筛选出23例A亚型:13例为A205、8例为A201、1例为A204、1例不能确定;A205、A201、A204基因测序结果均与genebank的参考序列相同,1例基因分型不能确定者,基因测序结果与genebank的参考序列比对,确定其基因型为A102;A204先证者的A204是由其母亲遗传而来。结论 A2等位基因在所调查的人群(中国北方汉族)中以A205为主,其次为A201,存在A204,且A204可以在家系内遗传。     

Identification of Francisella tularensis subsp. tularensis A1 and A2 infections by real-time polymerase chain reaction

Francisella tularensis subsp. tularensis (type A) is subdivided into clades A1 and A2. Human tularemia infections caused by A1 and A2 differ with respect to clinical outcome; A1 infections are associated with a higher case fatality rate. In this study, we develop and evaluate TaqMan polymerase chain reaction (PCR) assays for identification of A1 and A2. Both assays were shown to be specific to either A1 or A2, with sensitivities of 10 genomic equivalents. Real-time PCR results for identification of A1 and A2 were in complete agreement with results obtained by pulsed field gel electrophoresis analysis or conventional PCR when specimens from sporadic tularemia cases and a tularemia outbreak involving both A1 and A2 were tested. In addition, outbreak samples not previously typed to the clade level could be classified as A1 or A2. The assays described here provide new diagnostic tools with a level of sensitivity not previously available for identification of A1 and A2 infections.     

成年男性血红蛋白水平升高与空腹血糖、血尿酸及血脂检测结果分析

目的探讨进行健康体检的成年男性血红蛋白(Hb)水平升高与其空腹血糖(FBG)、血尿酸(UA)及血脂[三酰甘油(TG)与总胆固醇(TC)]检测结果。方法将2018年10-11月1 007例Hb升高的成年男性按Hb水平不同分为A1、A2、A3、A4、A5组,另选取同期85例Hb水平正常体检者作为对照组,对所有研究对象FBG、UA及血脂水平进行检测,并对检测结果进行分析。结果 A1、A2、A3、A4、A5组FBG、UA及血脂水平均值及异常百分数均高于对照组,差异均有统计学意义(P<0.05)。均值比较:A1、A2、A3、A4、A5组血脂及A3组UA水平与对照组比较,差异均有统计学意义(P<0.05);A1组与A3组UA与TG水平比较,差异均有统计学意义(P<0.05),其他各组UA与TG水平比较,差异均无统计学意义(P>0.05);异常百分数比较:除A1、A2、A3、A4、A5组TG,A1、A2、A3、A4组TC与对照组及A1组与A4组FBG与TC差异有统计学意义(P<0.05)外,其余组间比较,差异均无统计学意义(P>0.05)。结论成年男性Hb水平升高与其生化项目FBG、UA及血脂的变化有一定相关性,与FBG不存在统计学上的差异,但整体呈上升趋势,对非传染性流行病有辅助诊断价值。     

CYP17基因单核苷酸多态与子宫内膜异位症和子宫腺肌病易感性的关系

目的探讨CYP17基因T-C单核苷酸多态与子宫内膜异位症(内异症)和子宫腺肌病(腺肌病)易感性的关系。方法蛋白酶K消化-饱和酚氯仿法提取外周血白细胞基因组DNA,聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)方法对61例内异症(内异症组)、59例腺肌病(腺肌病组)和65例健康女性(对照组)进行基因分型。结果①内异症组和腺肌病组A1A1基因型频率及A1等位基因频率均高于对照组,差异均有统计学意义;内异症组与腺肌病组之间差异均无统计学意义。②A1A1和A1A2基因型个体发生内异症的相对危险度(OR)分别是A2A2型个体的4.615倍(1.704-12.504,P=0.002)和3.467倍(1.339-8.977,P=0.009),发生腺肌病的相对危险度分别是A2A2型个体的3.692倍(1.337-10.196,P=0.010)和3.867倍(1.505-9.933,P=0.004),差异均有统计学意义。结论 A1A1、A1A2基因型增加内异症和腺肌病的易感性,A1等位基因可能是内异症和腺肌病的遗传易感基因,内异症和腺肌病发病机理相似。     

Comparison of the antigenic spectrum A1, A2, and Ax erythrocytes: inhibition of mab with glucoconjugates of lipid and protein nature with different isoelectric properties

The inhibition of monoclonal antibodies (MAbs) (anti-A1 2-24, anti-A 2-22, anti- 2-19, and anti- A 2-18; Workshop IV) by glycotopes (glycoconjugates) obtained by the enzymatic treatment of A1, A2, and Ax erythrocytes and by chloroform-methanol method from the membranes of these erythrocytes, followed by ion-exchange gel chromatography was evaluated. According to specificity and isoelectric properties, glycoconjugates (in the sequence of desorption from an anion exchanger) of glycoprotein (A(pr-00), A(pr-0), A(pr-1), A(pr-3)) and glycosphingolipid (A1p,00, A(1p-0), A(1p-1), A(1p-3), A(1p-3)) origin were identified. A1 erythrocytes showed the broad-spectrum inhibiting activity of glycoconjugates, which being at maximum with acid A(1p-00) and A(pr-3) (the ionic points of pH 6.55 and 6.45), as well as with A(lp-00) and A(1p-00) 1 The inhibition of A(pr-3) from A2 was significantly weaker that that from A1. An A(pr-3) fraction from Ax erythrocytes inhibited as strongly as in A1 and A2. The inhibition of A(pr-3p) from Ax either failed to develop at all (MAb 2-19) or turned out be weakened (MAbs 2-22 and 2-18). Thus, the cascade decrease in agglutinogenicity in the order of A1, A2, and Ax correlated with the reduction in the inhibiting capacity of A(pr-3) in all the study groups, including Ax.     

Novel short-acting A2A adenosine receptor agonists for coronary vasodilation: inverse relationship between affinity and duration of action of A2A agonists.

Several potent and selective A2A adenosine receptor agonists are currently available. These compounds have a high affinity for the A2A receptor and a long duration of action. However, in situations where a short duration of action is desired, currently available A2A receptor agonists are less than ideal. From a series of recently synthesized A2A receptor agonists, two agonists (CVT-3146 and CVT-3033) with low affinity were selected for further characterization as selective and short-acting coronary vasodilators. Both compounds were selective for the A2A adenosine receptor (AdoR) versus the A1, A2B, and A3AdoR in binding and functional studies. CVT-3146 and CVT-3033 appeared to be weak partial agonists to cause cAMP accumulation in PC12 cells, but were full and potent agonists to cause coronary vasodilation, a response that has a very large A2A receptor reserve. However, the durations of action of CVT-3146 and CVT-3033 were remarkably shorter than those of the high-affinity agonists CGS21680 or WRC0470, presumably due to the relative lower affinity of CVT-3146 and CVT-3033 for the A2A receptor. Indeed, an inverse relationship was found between the affinity of the various agonists for the A2A receptor and the duration of their actions. These data indicate that low-affinity agonists can produce a response that is of equivalent magnitude but more rapid in termination than that caused by a high-affinity agonist. Hence, the low-affinity A2A agonists CVT-3146 and CVT-3033 may prove to be superior to currently available high-affinity agonists as coronary vasodilators during myocardial imaging with radionuclide agents.     

Identification and phenotype characterization of two CYP3A haplotypes causing different enzymatic capacity in fetal livers

BACKGROUND: The fetal liver cytochrome P450 (CYP) 3A enzymes metabolize potentially toxic and teratogenic substrates and drugs in addition to endogenous hormones and differentiation factors. CYP3A7 is the most abundant CYP in the human liver during fetal stages and the first months of postnatal age and shows a large interindividual variability of unknown molecular basis. METHODS: A new variant gene (CUpsilonP3A7*2), which carries a mutation in exon 11 of CUpsilonP3A7 causing a T409R substitution, was identified by direct sequencing. Genotype analysis was performed by use of polymerase chain reaction followed by restriction enzyme analysis. CYP3A7.2 activity was assessed in heterologous expression systems and human fetal liver microsomes. RESULTS: The frequency of CUpsilonP3A7*2 was 8%, 17%, 28%, and 62% in white, Saudi Arabian, Chinese, and Tanzanian individuals, respectively. By use of human HEK293 cells, no significant differences in expression between CYP3A7.1 and CYP3A7.2 were found and fetal livers homozygous for CUpsilonP3A7*2 had similar or higher CYP3A7 protein contents than CUpsilonP3A7*1 livers. Kinetic studies showed that CYP3A7.2 was a functional enzyme with a significantly higher catalytic constant (kcat) as compared with CYP3A7.1 (P < .05). Interestingly, fetal livers that expressed CYP3A7.2 also expressed CYP3A5 protein, and we found a linkage disequilibrium between the CUpsilonP3A7*2 and CUpsilonP3A5*1 alleles that was subject to interethnic differences. Determination of the alprazolam 1-hydroxylation rate revealed that CYP3A5 plays a significant role in the metabolism of CYP3A substrates in the fetal liver. CONCLUSION: We have identified 2 different CYP3A phenotypes in the fetal liver--one that is the result of a CUpsilonP3A7*1/CUpsilonP3A5*3 haplotype causing CYP3A7.1 but no CYP3A5 expression and another with higher detoxification capacity, inherent in the CUpsilonP3A7*2/CUpsilonP3A5*1 haplotype, where CYP3A5 and a more active form of CYP3A7 are expressed.     

Relationship between interindividual differences in nicotine metabolism and CYP2A6 genetic polymorphism in humans

BACKGROUND: Nicotine is mainly metabolized to cotinine by cytochrome P450 (CYP) 2A6. Previously, we found that the CYP2A6 gene was deleted homozygously in one subject who was deficient in cotinine formation from nicotine. OBJECTIVE: Our objective was to clarify the relationship between interindividual differences in nicotine metabolism and CYP2A6 genetic polymorphism. METHODS: Nicotine was administered to 92 healthy Japanese subjects in the form of 1 piece of nicotine gum to investigate the potency of nicotine metabolism. The cotinine-nicotine ratio of the plasma concentration 2 hours after chewing was calculated as an index of nicotine metabolism. The genotypes of CYP2A6 gene, CYP2A6*1A, CYP2A6*1B, CYP2A6*2, CYP2A6*3, CYP2A6*4, and CYP2A6*5, were determined with polymerase chain reaction-restriction fragment length polymorphism. RESULTS: A large interindividual difference in nicotine metabolism was observed. Allele frequencies of CYP2A6*1A, CYP2A6*1B, and CYP2A6*4 were 42.4%, 37.5%, and 20.1%, respectively. The CYP2A6*2, CYP2A6*3, and CYP2A6*5 alleles were not found. Three subjects genotyped as CYP2A6*4/CYP2A6*4 were completely deficient in cotinine formation. The heterozygotes of the CYP2A6*4 allele tend to show lower capacities for cotinine formation. The subjects with CYP2A6*1A/CYP2A6*1B appeared to have higher capacities of cotinine formation than subjects with CYP2A6*1A/CYP2A6*1A, although the difference was not significant. The probit plot of the cotinine-nicotine ratio was not linear; this possibly indicated the existence of a novel mutation in the CYP2A6 gene genotyped as CYP2A6*1B/CYP2A6*4. CONCLUSIONS: The relationship between interindividual differences in nicotine metabolism and CYP2A6 genetic polymorphism in humans was proved.     

Quantitative abnormalities of lipoprotein particles in multiple myeloma

Serum concentrations of total cholesterol and lipoprotein cholesterol, of apolipoproteins A I and A II and of apolipoprotein A I in lipoprotein particles (Lp A I and Lp A) were determined in 43 patients with multiple myeloma. There were striking alterations in the plasma levels of these analytes relative to normal subjects. We observed a decrease of cholesterol levels in LDL, HDL and HDL3 fractions, and of apolipoproteins A I and A II compared with normal subjects. The HDL2 cholesterol was increased. The decrease of apolipoprotein A II was more prominent than apolipoprotein A I. The decrease of apolipoprotein A I concerns only the A I (Lp A), while the A I (Lp A I) was increased. Most of these modifications were correlated with the monoclonal Ig levels.     

Targeting heterogeneous nuclear ribonucleoparticule A1 and A2 proteins by RNA interference promotes cell death in transformed but not in normal mouse cell lines

The heterogeneous nuclear ribonucleoparticule A1 and A2 proteins can bind to vertebrate single-stranded telomeric sequences. Moreover, changes in the levels of heterogeneous nuclear ribonucleoparticule A1 can influence telomere length in mouse and human cells. We have shown previously that the combined knockdown of A1 and A2 proteins in human transformed cells promotes apoptosis. In contrast, a similar reduction in A1 and A2 expression in normal mortal human cell lines does not induce cell death. Here, we show that a variety of mouse cell lines display a similar behavior on reduction of A1 and A2 protein levels using small interfering RNA. In addition, the expression of the mouse A1 cDNA protects human HeLa cells from apoptosis when human A1 and A2 proteins are targeted by RNA interference. Lastly, we show that knockdown of A1 and A2 expression also impairs the growth of a human transformed cell line that does not express telomerase. These results firmly establish A1 and A2 as proteins required for the viability of transformed murine and human cells, irrespective of the status of telomerase expression or the length of the double-stranded telomeric repeat.     

卡马西平所致药疹与CYP3A4＊18B基因型的关联性初探

目的初步探讨卡马西平所致药疹与CYP3A4＊18B基因型的关系,以寻找预测药疹的分子学手段。方法用PCR-RFLP法检测65例卡马西平所致药疹组患者（重型药疹患者11名,轻型药疹患者54名）、100例卡马西平耐受组患者及100例健康对照组患者的CYP3A4＊18B（intron10,G20338→A）基因多态性。结果卡马西平所致药疹组CYP3A4＊1/CYP3A4＊1（G/G）基因型30例,CYP3A4＊1/CYP3A4＊18B（G/A）基因型30例,CYP3A4＊18B/CYP3A4＊18B（A/A）基因型5例,等位基因A频率为30.8%;卡马西平耐受组G/G基因型51例,G/A基因型42例,A/A基因型7例,等位基因A频率为28%;健康对照组G/G基因型44例,G/A基因型49例,A/A基因型7例,等位基因A频率为31.5%。各组间基因型与等位基因频率差异均无统计学意义。结论由于病例数较少,目前尚没有发现卡马西平药疹与CYP3A4＊18B相关联。     

The genetic and phenotypic basis of blood group A subtypes in Koreans

A serological and genetic study of Korean blood donors with phenotypic group A subtypes was performed. There were 176 donors with phenotypic A subtypes identified. Exons 6 and 7 from 57 representative donors were sequenced. The A(var) allele (784 G > A) was cloned and sequenced, and a family study demonstrating its inheritance and unusual serological characteristics was performed. The A102 allele was the most frequently identified allele in phenotypically A2 (58%, 11/19) and A2B (68%, 17/25) donors. Anti-A1 was rarely present amongst A2 and A2B donors. The family study revealed that the A(var) allele was expressed as phenotype A(weak)B in A(var)/B heterozygote members, but as phenotype O in A(var)/O heterozygotes. The most frequent allele in Korean donors with the A2 phenotype differs from its Caucasian counterpart, as does the frequency of anti-A1. The A(var) allele demonstrates allelic enhancement in A(var)/B heterozygotes.     

Role of Aspergillus lentulus 14-α sterol demethylase (Cyp51A) in azole drug susceptibility

Recent studies have demonstrated that some morphologically atypical Aspergillus fumigatus strains are different species belonging to the section Fumigati. Aspergillus lentulus, one of these sibling species, is increasingly reported in patients under corticosteroid treatment. MICs of most antifungals in clinical use are elevated against A. lentulus, and it shows primary resistance to azole drugs. Two A. lentulus cytochrome P450 14-α sterol demethylases, encoded by A. lentulus cyp51A (Alcyp51A) and Alcyp51B genes, were identified. Targeted cyp51A gene knockout in A. lentulus showed that the intrinsic azole resistance of this species is cyp51A dependent. The Δcyp51A strain was morphologically indistinguishable from the A. lentulus wild-type strain, retaining the ability to cause pulmonary disease in neutropenic mice. The heterologous expression of A. lentulus cyp51A was performed in an A. fumigatus cyp51A-deficient strain, confirming that Cyp51A is responsible for the differences in A. lentulus-azole drug interaction.     

Reversal of galectin-1 gene silencing on resistance to cisplatin in human lung adenocarcinoma A549 cells

This study aims to investigate reversal of Galectin-1 gene silencing on resistance to cisplatin in human lung adenocarcinoma A549 (or A549/DDP) in vivo and in vitro. The stably transfected lentivirus vector was used to silence Galectin-1 in human lung adenocarcinoma cell line A549 and A549/DDP cells and the cell lines were cultured and passaged. RT-PCR and western blot assay were used to test A549, A549/DDP cells, silenced Galectin-1A549 (A549/I) cells, Galectin-1 mRNA and protein expression levels, respectively, in A549/DDP (A549/DDP/I) cells. CCK8 assay was used to measure median inhibitory concentration (IC50) in each group and resistant index of A549/DDP cells and A549/DDP/I cells. Tumor model in nude mice was established by armpit injection of A549, A549/DDP, A549/I, A549/DDP/I cells. Cisplatin was injected intraperitoneally in tumor models and growth of tumor was observed in vivo model. Four weeks later, nude mice were killed and tumor weight and diameter was measured. mRNA and protein expression of Galectin-1 in A549/DDP cells was higher than that in A549 cells. mRNA and protein expression of Galectin-1 in A549/DDP/I cells was lower than that in A549/DDP cells. Moreover, IC50 values ​​and resistance index in A549/DDP cells was higher than that in A549 cells group and IC50 values ​​and resistance index A549/DDP/I cell group were lower than that in A549/DDP cells. Additionally, tumor weight and volume in A549/DDP/I cell group were lower than that in A549/DDP. In conclusion, Galectin-1 gene silencing would improve the sensitivity of A549/DDP cells to cisplatin in vivo and in vitro.     

Effect of omeprazole on gastric adenosine A1 and A2A receptor gene expression and function

Adenosine has been shown to inhibit immunoreactive gastrin (IRG) release and to stimulate somatostatin-like immunoreactivity (SLI) release by activating adenosine A(1) and A(2A) receptors, respectively. Since the synthesis and release of gastrin and somatostatin are regulated by the acid secretory state of the stomach, the effect of achlorhydria on A(1) and A(2A) receptor gene expression and function was examined. Omeprazole-induced achlorhydria was shown to suppress A(1) and A(2A) receptor gene expression in the antrum and corporeal mucosa, but not in the corporeal muscle. Omeprazole treatment produced reciprocal changes in A(1) receptor and gastrin gene expression, and parallel changes in A(2A) receptor and somatostatin gene expression. The localization of A(1) and A(2A) receptors on gastrinsecreting G-cells and somatostatin-secreting D-cells, respectively, suggests that changes in adenosine receptor expression may modulate the synthesis and release of gastrin and somatostatin. Thus, the effect of omeprazole on adenosine receptor-mediated changes in IRG and SLI release was also examined in the vascularly perfused rat stomach. After omeprazole treatment, the A(1) receptor-mediated inhibition of IRG and SLI release induced by N(6)-cyclopentyladenosine (A(1) receptor-selective agonist) was not altered, but the A(2A) receptor-mediated augmentation of SLI release induced by 2-p-(2-carboxyethyl-)phenethylamino-5'-N-ethylcarboxamidoadenosine (A(2A)-selective agonist) was significantly attenuated. These findings agree well with the corresponding omeprazole-induced decrease in antral A(2A) receptor mRNA expression. Overall, the present study suggests that adenosine receptor gene expression and function may be altered by omeprazole treatment. Acid-dependent changes in adenosine receptor expression may represent a novel purinergic regulatory feedback mechanism in controlling gastric acid secretion.     

Adsorption of chain type–specific ABO antibodies on Sepharose‐linked A and B tetrasaccharides

BACKGROUND: Antigen‐specific removal of anti‐A and anti‐B on immunoadsorption columns carrying the blood group A and B trisaccharides is one important component of some protocols used in ABO‐incompatible organ transplantation. Because ABO antibodies exist requiring parts of the core saccharide chain for binding, the anti‐A and ‐B–binding capacity of individual and combined, Sepharose‐linked Types 1 through 4 A and B tetrasaccharides with that of the A and B trisaccharides was compared. STUDY DESIGN AND METHODS: Sepharose‐linked A and B tri‐ and tetrasaccharides were used to adsorb anti‐A and ‐B from pooled blood group O serum. Remaining chain type–specific anti‐A and ‐B were detected and quantified in enzyme‐linked immunosorbent assays using wells coated with neoglycoproteins or recombinant mucins carrying A and B determinants on defined core saccharide chains. RESULTS: Significantly more anti‐A Type 3‐ and 4‐specific immunoglobulin (Ig)G remained after adsorption on the A trisaccharide and the A Type 1 and A Type 2 tetrasaccharide than after adsorption on the A Types 3 and 4 tetrasaccharides. Selective adsorption of chain type–specific IgG anti‐B was detected on Sepharose‐linked B tetrasaccharides. In contrast, there were no chain type–specific IgM anti‐A or ‐B. A combination of the A or B tetrasaccharides adsorbed a larger fraction of the IgG anti‐A and ‐B repertoires than the corresponding trisaccharides. CONCLUSION: There are chain type–specific anti‐A and anti‐B IgG, and an adsorber based on a combination of Types 1 through 4 A or B tetrasaccharides will be a more efficient adsorber than an adsorber based on the A or B trisaccharides.