胃癌合并慢性病性贫血患者红细胞膜表面CD35和CD58的表达

目的研究细胞因子对正常人和胃癌伴慢性病性贫血(anemia of chronic cliseasis, ACD)的影响.方法采用ELISA方法测定CD35和CD58水平,红细胞酵母花环法测定RBC-C3bRR和RBC-ICR,聚乙醇沉淀比浊法测定CIC,流式细胞术测定CD2.结果胃癌组和正常对照组进行比较RBC-C3b差异有统计学意义,P<0.01;RBC-ICR差异有统计学意义,P<0.05;CD35差异有统计学意义,P<0.01;而CD58虽然低于正常对照组,但差异无统计学意义,P>0.05;CIC测定结果均正常.结论胃癌患者RBC-C3b、CD35和CD58水平下降与胃癌患者具有免疫活性的细胞因子有直接关系.但CD58无统计学意义.     

肺癌患者血浆中APC基因启动子甲基化定量检测研究

背景与目的：APC基因编码的蛋白参与信号转导途径,大量研究证实APC启动子区的高甲基化在肿瘤的发生发展中起重要作用。本研究建立血浆APC基因实时荧光定量甲基化特异性基因扩增（methylation—specific PCR,MSP）检测方法,并对临床肺癌患者血浆进行检测,以确定该方法在肺癌诊断中的应用价值。方法：将APC基因启动子甲基化阳性肺癌细胞株NCI—H460细胞用有限稀释法获取单个集落形成的细胞,以经典的酚一氯仿法提取细胞DNA,并用MSP对APC基因甲基化进行验证,紫外分光光度计定量并以10倍稀释的浓度梯度依次投入到200μL健康人血浆中．得到模拟肺癌患者血浆,利用磁珠核酸提取方法从模拟肺癌及肺癌患者、肺部良性疾病患者和健康对照者血浆中提取DNA,对血浆DNA模板进行亚硫酸氢盐化学修饰;以模拟血浆样品作为标准品,采用外标准曲线法对各种血浆样品中APC甲基化进行定量分析。结果：所建立的实时荧光定量MSP检测方法的线性范围为1．5×10^2-1．5×10^5拷贝／mL。用该方法检测甲基化肿瘤细胞的DNA其最低检测限为1．5×10^2拷贝／mL。78例肺癌患者有40例组织中检测出APC基因甲基化阳性．其中的19例（47．5％）肺癌患者血浆中APC基因启动子甲基化阳性,APC甲基化浓度为1．67×10^2-6．78×10^3拷贝／mL,中位浓度为1．60×10^3拷贝／mL。38例组织阴性的肺癌患者、31例肺部良性疾病患者和23例健康者的血浆APC基因启动子甲基化均为阴性。结论：实时荧光定量MSP检测血浆APC基因甲基化在肺癌诊断方面具有潜在应用价值。     

阻塞性睡眠呼吸暂停综合征免疫网络及其与临床的相关性

目的探讨红细胞表面补体受体Ⅰ型(CR1)活性、红细胞表面CD58表达和血清中可溶性白细胞介素-2受体(SIL-2R)、白细胞介素-6(IL-6)水平在阻塞性睡眠呼吸暂停综合征(OSAS)中的作用、变化规律及其与疾病严重程度的关系。方法采用肿瘤红细胞花环法、流式细胞分析法及酶联免疫吸附法检测20例中、重度OSAS患者(研究组)及15例正常人(对照组)的红细胞CR1活性、CD58表达和血清中SIL-2R、IL-6水平。结果(1)研究组患者肿瘤红细胞花环率(ECR1CaRR)较正常对照组降低(P<0.05);CD58表达明显高于对照组(P<0.01)。(2)研究组患者SIL-2R及IL-6水平均明显高于对照组(P均<0.01)。(3)OSAS患者SIL-2R、IL-6与呼吸紊乱指数(AHI)无相关性(P均>0.05),与动脉血氧饱和度低于90%的时间占总睡眠时间百分比(SIT90%)均呈正相关(P均<0.01)。ECR1CaRR、CD58表达与SIT90%无相关性(P均>0.05)。结论血清中SIL-2R、IL-6水平变化与OSAS患者病情密切相关,可作为临床分度及评价疗效的指标;红细胞免疫功能与细胞因子间存在相互调节作用。     

Epigenetic enhancement of p66Shc during cellular replicative or premature senescence

The cytoplasmic protein p66Shc is expressed in a wide range of cell types, initially believed to be involved in signaling pathways that regulate cell growth and oxidative stress. Here the epigenetic alterations in the promoter of p66Shc were investigated in replicative senescence and in premature senescence induced by hydrogen peroxide in human embryonic pulmonary fibroblast cells. In both cases p66Shc expression was elevated compared to that seen in growing cultures. However, methylation-specific PCR and bisulfite sequencing revealed that the CpG sites were hypermethylated in all cultures. In addition, quantitative chromatin immunoprecipitation showed increased histone H4 acetylation and histone H3 Lys-4 methylation during replicative senescence, while the increased acetylation of histone H3 and H4, as well as increased H3 Lys-4 methylation, was seen in premature senescence persistence. These findings suggest that histone modifications of p66Shc might be the molecular event in cellular senescence. Taken together, the epigenetic enhancement of p66Shc is associated with the specifically increased histone acetylation and methylation, which may contribute to cellular replicative senescence or premature senescence.     

Design and synthesis of novel organometallic complexes using boronated phenylalanine derivatives as potential anticancer agents

Drug design and discovery studies are important because of the prevalence of diseases without available medical cures. New anticancer agents are particularly urgent because of the high mortality rate associated with cancer. A series of mononuclear gold (III) and platinum (II) complexes based on boronated phenylalanine (BPA) were designed and synthesized using 4,4’-dimethyl-2,2’-dipyridyl (L1) or 1,10-phenanthroline-5,6-dion (L2) ligands to obtain promising anticancer drug candidates. Proton nuclear magnetic resonance, infrared, mass spectrometry, and elemental analyses were utilized for chemical characterizations. Cell viability, cancer cell colony formation, endothelial tube formation, and cytoskeleton staining assays were performed using A549 lung adenocarcinoma and human umbilical vein endothelial cells (HUVECs) to investigate preliminary pharmacological activities. L1-based platinum (II) complex (BPA-L1-Pt) was the most promising complex, and has similar activity with the approved chemotherapy drug cis-platinum. Half maximal inhibitory concentration values for BPA-L1-Pt were 9.15 µM on A549s and 16.61 µM on HUVECs; the values for cis-platinum were 5.24 µM on A549s and 23.14 µM on HUVECs. Consequently, further synthesis studies should be performed to boost the cancer cell selectivity feature of BPA by varying metal and ligand types.     

Immunoglobulin-producing cells in CSF and blood from patients with multiple sclerosis and other inflammatory neurological diseases enumerated by Protein-A plaque assay

Using the Protein-A plaque assay, numbers of IgG + IgA + IgM producing cells determined in patients with multiple sclerosis (MS) were 0.1–5% in CSF and 0.1–0.7% in peripheral blood; interestingly, 7 of 11 MS patients had IgM producing cells in CSF. In patients with aseptic meningitis (AM), the corresponding values were 0.04–7.5% in CSF and 0.4–2.4% in peripheral blood. There were more Ig producing cells in peripheral blood from patients with AM and MS than in healthy subjects. cocorrelation between numbers of IgG producing cells in CSF and the concentrations of intrathecally produced IgG (CSF IgG index) was registered in patients with AM: the same was true for IgA. The Protein-A plaque method, adopted for 20 × 10³ lymphocytes, makes possible enumeration of Ig-producing cells in CSF and discrimination among cells secreting different Ig classes, thereby being a powerful tool for studying immune reactions in the CNS-CSF compartment.     

Reflux Control Following Gastroplasty

A Belsey gastroplasty was performed on 135 patients, 132 of whom were available for follow-up. Despite a low incidence (1.5%) of anatomical recurrence, the operation failed to control reflux effectively, and the incidence of continued reflux is 44.6%. Because of this failure to control reflux, a Nissen fundoplication has been added to the gastroplasty tube.In a group of 78 patients, radiological recurrence has occurred in 1 patient, with no patient experiencing symptoms of reflux. Manometric comparison between the Belsey and Nissen gastroplasty shows more effective tone elevation of the high pressure zone and a more effective decrease in disordered motor activity of the lower esophagus.     

An experimental model of hyper-irritability in the trigeminal skin field of the rat.

Cutaneous hyper-irritability was produced in the trigeminal sensory distribution of chronically prepared rats by injecting various central stimulants into the dorsal subarachnoid space overlying caudal medulla. Effective substances were classified into three groups according to the behavioral patterns induced. These were (1) picrotoxin and penicillin G-K, (2) strychnine and brucine, and (3) D,L-homocysteic acid. Of these substances, picrotoxin was the most effective. The hyper-irritability elicited by picrotoxin or penicillin G-K was associated with a relatively localized trigger zone in the face, while that elicited by strychnine or brucine spread beyond the trigeminal region to the first cervical dermatome. D,L-homocysteic acid evoked only spontaneous scratching and vocalization. Picrotoxin-induced hyper-irritability involved facilitation of impulse transmission from cutaneous afferent terminals to second-order neurons in the trigeminal nucleus caudalis.