Evaluation of cellular viability by quantitative autoradiographic study of myocardial uptake of a fatty acid analogue in isoproterenol-induced focal rat heart necrosis

Previous studies led us to hypothesize that a fatty acid analogue, 15-p-iodophenyl--methyl pentadecanoic acid (IMPPA or BMIPP), which is taken up but not quickly metabolized by heart cells, would be a more suitable tracer of cellular viability than thallium-201. Biodistribution studies of 1-¹⁴C-IMPPA in conscious, freely moving rats showed that the concentration ratio of radioactivity in the heart with respect to the blood was about 8 for at least 60 min after intravenous administration, permitting its use as a putative tracer in these conscious, freely moving rats. Thereafter, the myocardial uptake of¹⁴C-IMPPA was studied in isoproterenol-treated rats (daily treatment for 10 days in order to induce cardiac hypertrophy and necrotic foci) with respect to control ones. Comparison of myocardial localizations by quantitative autoradiography of the uptake of²⁰¹Tl and¹⁴C-IMPPA with that of triphenyltetrazolium chloride (TTC) staining enabled comparative evaluation of nutritional blood flow, localization and uptake of¹⁴C-IMPPA and necrotic foci size. Distributions of¹⁴C-IMPPA and²⁰¹1 T1 in control rats' hearts were homogeneous, like TTC staining. In infarcted hearts, areas of decreased¹⁴C-IMPPA uptake were nearly the same (100%±5%) as those unstained by TTC. These areas were larger than those showing a decrease in thallium uptake (about 70%±5% of the total scar size). Therefore, IMPPA seems to be a more accurate and sensitive indicator of necrosis localization compared with thallium. It may be a useful agent for assessment of myocardial viability by single photon emission tomography (SPET) imaging.     

p16和p15第二外显子HapⅡ位点在成人急性髓性白血病中的突变

目的　探讨p16和p15基因第二外显子HapⅡ位点在成人急性髓性白血病 (AML)中的突变。方法　用甲基敏感和甲基非敏感限制性内切酶切消化基因组DNA后 ,经PCR扩增和琼脂糖电泳研究 3 1例AML中无p16/p15基因第二外显子缺失的患者中HapⅡ位点的突变情况。结果　在 3 1例成人AML样本中 ,p16E2 的 4个HapⅡ位点和p15E2 的 6个HapⅡ位点均未发生突变。结论　本研究提示p16和p15基因第二外显子中因甲基化而引起的CCGG位点的突变在成人AML患者中不是主要失活方式。     

Synthesis and physico‐chemical properties of peptides in soil humic substances

Abstract: Soil humic substances (HS) are heterologous, polydispersive, and multi‐functional organometallic macromolecules ubiquitous in soils and sediments. They are key players in the maintenance of the belowground ecosystems and in the bioavailability of both organic and inorganic contaminants. It is widely assumed that the peptidic substructures of HS are readily degraded and therefore do not contribute significantly to interactions with contaminants such as toxic metals. To investigate the turnover of humified peptides, laboratory soil aging experiments were conducted with ¹³C‐glucose or ¹⁵N‐nitrate for 8.5 months. Evidence for random‐coil peptidic structures in the labeled HS was obtained from 2‐D nuclear magnetic resonance (NMR), pyrolysis gas chromatography‐mass spectrometry (pyro‐GC‐MS), and circular dichroism data. Interaction of metals with the peptidic carbonyls of labeled HS was rationalized from the solid‐state NMR data. Detailed ¹³C and ¹⁵N labeling patterns of amino acid residues in the acid hydrolysates of HS acquired from NMR and GC‐MS revealed two pools of peptides, i.e. one extant (unlabeled) and the other, newly humified with little isotopic scrambling (fully labeled). The persistence of pre‐existing peptidic structures indicates their resistance to degradation while the presence of fully labeled peptidic amino acids suggests wholesale incorporation of newly synthesized peptides into HS. These findings are contrary to the general notion that humified peptides are readily degraded.     

乳腺肿瘤组织Ha-ras基因蛋白表达与血清CA15-3水平的相关性及其临床价值

用免疫组化ＡＢＣ法和ＩＲＭＡ法分别检测了４２例乳腺癌和３０例良性乳腺病变患者组织中Ｈａ－ｒａｓ基因蛋白（ｒａｓＰ２１）的表达状况及血清ＣＡ１５－３值，结果恶性病变组织中ｒａｓＰ２１呈过度表达，ｒａｓＰ２１的表达与细胞的分化状态密切相关。不同淋巴结转移状况的乳腺癌患者，其血清ＣＡ１５－３差异呈显著性。ＣＡ１５－３水平的变化与ｒａｓＰ２１表达状况显著相关。提示血清ＣＡ１５－３水平为监测乳腺癌病情变化的良好指标，Ｈａ－ｒａｓ癌基因点突变的结果可能导致乳腺癌相关抗原的异常分泌。     

Unusual features in children with inv dup(15) supernumerary marker: a study of genotype-phenotype correlation in Taiwan

Recent molecular cytogenetic studies have elucidated the origin and nature of extra structurally abnormal chromosomes (ESACs) or small supernumerary chromosomes, which are often associated with developmental delay and malformations. We studied the prevalence of inv dup(15) in a nationwide screening programme for mentally retarded children in Taiwan and tried to correlate the genotype and phenotype in those patients. Fluorescence in situ hybridization (FISH) analysis using D15Z, D15Z1, and the cosmids from the Prader-Willi/Angelman syndrome chromosome region (PW/ASCR) was performed on 54 patients (0.45%) with ESACs from 11893 probands within a 5-year period. Of them, inv dup(15) was confirmed in 25 children (46.3%) by FISH analysis. The PW/ASCR probes were used to clarify the size and DNA composition of the markers. Patients with inv dup(15) chromosomes, containing only the heterochromatin or little euchromatin of the proximal 15q (i.e., pter→q11:q11→pter) may have a rather mild or nearly normal phenotype (group 1). Only one patient had some features suggestive of Angelman syndrome, but was considered to be a result of deleted (15)(q12) in the chromosome 15 homologue. Additional copies within D15S11 through GABRB3 (15q11.2-13) resulted in an abnormal phenotype which involved mental and developmental delay but was different from the classical phenotype of PW/AS (groups 2, 3). Signs of autistic behavior did occur in each group. FISH combined with microsatellite analyses showed that the marker was often of maternal origin in de novo cases (n = 12, 86%), or inherited from the mother in only one familial case. Down-inv dup(15) was mentioned in two cases. Unusual features including diaphragmatic eventration, hyperlaxity of joints, arachnodactyly, brain atrophy, epilepsy (particularly infantile spasm), ataxia, genital abnormalities, and cleft lip/palate were noted in those patients. This observation expands the range of phenotypic expression associated with this relatively common ESAC. Conclusion Marked phenotypic diversities exist in children with inv dup(15), dependent upon the size or genetic composition of the markers, degree of mosaicism, parental origin and familial occurrence or not. Patients with a larger inv dup(15) marker chromosome including the PW/ASCR may have a higher risk of abnormalities, but not the typical Prade-Willi/Angelman syndrome phenotype. Received: 11 February 1997 and in revised form: 20 May 1997 / Accepted: 20 May 1997     

IL-15对儿童MDS造血前体细胞bcl-2、bcl-xl mRNA表达的影响

目的 探讨IL-15对骨髓增生异常综合征(MDS)造血细胞bcl-2、bcl—xl mRNA表达的影响，以探索IL-15抑制MDS CD34^ 造血干／祖细胞凋亡的可能机制：方法 采用吸附单克隆抗体的免疫磁珠分离系统，分离纯化17例MDS患儿骨髓CD34^ 细胞，采用RT—PCR检测bcl-2、bcl—xl mRNA表达水平，观察IL-15对它们的影响。结果 IL-15可呈时间与剂量依赖性的增强体外培养的MDS造血前体细胞bcl-2、bcl—xl基因mRNA的表达。结论 IL-15可能为抑制MDS CD34^ 造血干／祖细胞凋亡的作用机制之一。     

甲硫腺苷磷酸化酶在非小细胞肺癌中转录表达

目的 探讨甲硫腺苷磷酸化酶(MTAP)及其连锁基因p14，p15和p16在非小细胞肺癌中的转录表达变化情况。方法 采用RT-PCR方法检测15例新鲜肺癌标本及相应癌旁组织中MTAP及其边锁基因p14、p15和p16mRNA的表达情况。结果 MTAP在73．3％的肿瘤标本中不表达或转录水平降低5倍以上，而MTAP在癌旁对照组织中的表达率却高达93．3％；此外，MTAP在肺腺癌和鳞癌中的低表达现象无显著性差异，但在中／低分化肺癌中低表在显著高于高于分化癌。对7例标本分析表明，1例P15基因在正常组织及癌瘤中都高水平表达，1例均不表达。5例在癌标本中基本不表达．而在正常组织中表达；而p14和p16正常组织中转录表达水平很低，而在配对的7例肺癌组织中却分别有3项高表达。结论 MTPA基因低表达或丢失可能与肺癌的恶性程度密切相关，MTAP的缺失可能独立于p16之外，提示MTAP的低表达或缺失在肿瘤生物学中有其功基础的基础。     

IL-15cDNA转染人喉癌细胞系的建立及其表达的研究

目的建立转人白细胞介素-15(hIL-15)基因的喉癌细胞株,并研究hIL-15的表达情况。方法将携带人白细胞介素-15的质粒pcDNA3.1( )-hIL-15转导入人喉表皮样癌细胞Hep-2中,G418筛选阳性克隆,RT-PCR、Westernblot和ELISA法对hIL-15的表达进行检测。结果IL-15基因成功地转导入Hep-2细胞中并能高效表达。RT-PCR电泳结果显示hIL-15基因在mRNA水平有表达;ELISA法测得106个转染阳性细胞24h内培养上清液中hIL-15的含量为(185.0±12.2)pg/ml,空载体转染及未转染的细胞未检测到hIL-15。结论hIL-15基因成功转染人喉癌细胞系Hep-2细胞且能持续大量表达。     

血液系统疾病中LRP15基因启动子区甲基化状态的研究

目的:探讨LRP15基因启动子区及第一外显子区的甲基化与血液系统疾病的关系和临床意义。方法:采用甲基化特异性PCR方法检测了9例正常人、73例急性白血病患者、8例慢性髓系白血病患者、2例慢性淋巴细胞白血病、12例血液系统良性病患者骨髓LRP15基因启动子区及第一外显子区的甲基化状况;硫化修饰结合限制性内切酶分析及硫化测序验证了一例血液系统良性病患者LRP15基因启动子区及第一外显子区的甲基化。结果:AL患者LRP15基因甲基化阳性率(71.23%)高于正常供者及慢性白血病(10%),差异有统计学意义(P<0.05)。LRP15基因在非恶性血液病患者中甲基化阳性率(55.56%)高于正常供者,硫化修饰结合限制性内切酶分析及硫化测序进一步证实了上述结论。结论:LRP15基因启动子的甲基化与不成熟的白血病细胞有关,与血液系统良性疾病也有一定关系。     

硬膜外麻醉复合全麻对开胸手术患者早期恢复质量的影响

目的 探讨硬膜外腔阻滞复合全麻与肋间神经阻滞复合全麻对开胸手术患者早期恢复质量的影响。方法 选取行开胸手术患者76例,按随机数表法分为两组,硬膜外阻滞复合全麻组（E组）、肋间神经阻滞复合全麻组（C组）,每组38例。E组行硬膜外穿刺置管,术中0.2%罗哌卡因维持;C组行切口及上下肋间神经阻滞,两组全麻诱导及维持一致。采用术后恢复质量量表(quality of recovery QoR-40）对患者进行术前1d、术后3d恢复质量评分;采用视觉模拟评分法（visual analogue scale, VAS）对两组患者进行拔管后4 h、8 h、12 h、24 h安静和活动后的VAS评分;同时记录患者首次下床活动时间及住院时间。结果 E组在术后3dQoR-40各项评分均高于C组,差异有统计学意义（P <0.05）;E组静息和运动状态下术后各时间点VAS评分均低于C组,差异有统计学意义（P <0.05）;E组首次下床活动时间较早,住院时间较C组减少,差异有统计学意义（P <0.05）。结论 硬膜外阻滞复合全身麻醉用于开胸手术患者,不仅镇痛效果良好,还能明显提高术后早期恢复质量,...