巨噬细胞炎性蛋白4的突变和原核表达

目的 探索如何抑制嗜酸细胞的趋化作用，选择β-趋化因子巨噬细胞炎性蛋白4（MIP4）的突变性（Met-MIP4)作为趋化因子受体3的拮抗剂，将Met-MIP4基因在原核细胞中进行表达。方法 设计MIP4基因的PCR引物并进行氨基酸突变，将MIP4N末端的丙氨酸突变为蛋氨酸，以正常人肺酸突变，将MIP4N末端的丙氨酸突变为蛋氨酸。以正常人肺cDNA文库为模板，PCR方法获取Met-MIP4基因，克隆入载体pUC19，测序验证序列已得到突变，将正确的基因插入到GST融合表达载体pGEX-4T中，以IPTG诱导表达。结果 PCR产物为220bp左右的片段，连接入pUC19质粒后测序验证获得正确突变，构建的pGEX-4T融合表达载体在大肠杆菌中表达，经SDS-PAGE凝胶电泳显示有大小约34kU的新生融合蛋白表达。结论 成功突变并克隆了β-趋化因子MIP4基因，SDS-PAGE表明，与GST融合的Met-MIP4突变体已得到表达，为进一步研究其生物学活性奠定了基础。     

Iodogen法标记磁共振单克隆抗体靶向对比剂的实验研究

目的 探讨Iodogen法^131I标记磁共振单克隆抗体靶向对比剂McAb-PMP的方法。方法 采用Iodogen法^131I标记McAb-PMP，并测定标记率，放化纯度，比活度和免疫活性，进行动物免疫显像；结果 标记率为89.7%。放化纯度为3.16mCi/ml，经RM905活度计测定的比活度为89.6μCi/μg。在注射^131I-McAb-PMP后，在结肠癌裸鼠动物模型显像时，24h出现特异性聚集，48h和168h仍然保持聚集。结论 ^131I-McAb-PMP在Iodogen碘化标记后能较好地保持其免疫活性，特异性聚集在肿瘤部位，提示McAb-PMP能够应用于MR成像。     

胸腺切除后血清乙酰胆碱受体抗体改变及与疗效关系

随访２３例因重症肌无力行胸腺切除术的病人。运用灵敏度高、特异性强的ＢＡＳ—ＥＬＩＳＡ法检测手术前、后血清乙酰胆碱受体抗体（ＡｃｈＲ抗体）水平，所得结果均进行统计学分析。２３例病人胸腺切除后，１８例临床症状改善，有效率７８％。术后１４例（６１％）病人血清ＡｃｈＲ抗体水平下降，统计结果表明，手术前后血清ＡｃｈＲ抗体水平差异有显著性（Ｐ＜０．０５），但与症状改善无关（Ｐ＞０．０５）。手术前后血清ＡｃｈＲ抗体水平不影响疗效（Ｐ＞０．０５）。     

链置换式扩增检测羊水中巨细胞病毒DNA

目的：介绍一种简便快速准确检测羊水中ＣＭＶ－ＤＮＡ的改良ＰＣＲ－链置换式扩增用于诊断胎儿先天感染ＣＭＶ。方法，将组成套式ＰＣＲ的外内两对引物按照一定比例（外：内＝１：５０－１００）加在同一试管中一次扩增羊水和胎儿组织中ＣＭＶ－ＤＮＡ。结果：９０例异常孕产史的孕妇羊水检测ＣＭＶ－ＤＮＡ，阳性率为３８．９％（３５／９０），其中合并染色数目异常２例（４７，ＸＹＹ和４７，ＸＸ，＋２１）（已引产）核型及染色     

自体造血干细胞移植治疗恶性血液病

为探讨恶性血液病的有效治疗方法,应用自体骨髓移植(ABMT)38例,自体外周血造血干细胞移植(ABSCT)13例,自体外周血造血干细胞与自体骨髓联合移植15例。治疗白血病54例,恶性淋巴瘤11例,多发性骨髓瘤1例。外周血造血干细胞采用化疗加多抗甲素或C—CSF动员。移植物采用微波、阿克拉霉素净化处理。结果:两组动员方案均有良好动员效果。ABSCT组及联合移植组造血功能恢复比ABMT组快(P<0.05),合并症少。66例中,45例仍呈持续缓解(CCR),中位CCR时间32(5～98)个月,复发21例。3年无病生存率及复发率分别为68.5％及26.1％。结果表明:自体造血干细胞移植是根治恶性血液病的有效手段。ABSCT及联合移植具有造血功能恢复快,合并症少等优点。微波和阿克拉霉素体外净化移植物是一种简便、有效的净化方法。     

A method of cephalometric analysis]

In this paper, 220 children ranging from age 11 to 15 were selected. Each of them received a cephalometric roentgenographic examination. 153 cephalometric items from 13 types of methods which had been widely in use were studied through statistic analysis. By means of "t" test, several items which expressed little difference between the normal occlusion and the malocclusion were eliminated. By using of the cluster analysis we selected one item to represent the items which were with similar effect. Finally, 17 cephalometric items were pick out to provide references for clinical work.     

TREATMENT OF HYALINEMEMBRANE DISEASE BYHUMAN AMNIOTIC FLUIDSURFACTANT

A case of hyaline membrane disease was treated successfully with pulmonary surfactant (PS) isolated from human amniotic fluid. Dosage was 150 mg phospholipid/kg. The exogenous surfactant was instilled into the airway via a tracheal cannula. Clinical symptoms, PO2 and FiO2 improved evidently 24 hours after administration. L/S ratio and phosphatidylglycerol recovered gradually in aspirates. Lung X-ray film manifested "white lung" before instillation of surfactant and showed a striking improvement 3 days after treatment. The duration of mechanical ventilation was 6 days. During the period of recovery complications of patent ductus arteriosus and bacterial pneumonia developed. However, the patient recovered completely and was discharged 32 days after admission.     

The kyphoscoliotic type of Ehlers-Danlos syndrome (type VI): differential effects on the hydroxylation of lysine in collagens I and II revealed by analysis of cross-linked telopeptides from urine

The kyphoscoliotic type of Ehlers-Danlos syndrome (EDS type VIA) (OMIM 225400) is an autosomal recessive connective tissue disorder that results from mutations in the lysyl hydroxylase 1 gene (PLOD1) causing underhydroxylation of lysine residues in tissue collagens, particularly of skin. Previous studies have shown that the pool of collagen cross-linking amino acids, hydroxylysyl pyridinoline (HP) and lysyl pyridinoline (LP) excreted in urine has an abnormally low HP/LP ratio, which is diagnostic of the condition. Here we isolated cross-linked peptides containing these residues from the urine of a child with EDS VIA homozygous for a mutation that results in a stop codon and effective null expression of PLOD1 enzyme activity. Peptides that had originated from bone type I collagen and cartilage type II collagen were identified. A cross-linked N-telopeptide fraction that is derived from bone type I collagen contained only LP, no HP, which means that the helical lysines at residues 930 of alpha 1(I) and 933 of alpha 2(I) of the collagen triple-helix had not been hydroxylated. The equivalent peptide fraction from a normal child's urine gave a ratio of HP to LP of 1.5:1 typical for normal bone collagen. A second cross-linked peptide that is derived from the C-telopeptide domain of cartilage type II collagen showed both HP and LP in a 2:1 ratio, compared with 18:1 for the equivalent peptide from a normal child's urine. The results show that in EDS VIA, bone type I collagen is more markedly underhydroxylated than cartilage type II collagen, at least at those helical sites that form cross-links. The residual fraction of HP found in the urine of EDS VI patients therefore appears to be contributed in significant part by the degradation products of cartilage. Since PLOD1 is null, other PLOD genes must be responsible for the helical hydroxylation activity that results in HP. The presented approach of analyzing urinary cross-linked C-telopeptide fragments of type II collagen may allow the detection of chondrodysplasias due to genetic defects in lysyl hydroxylase isoforms active in cartilage.     

Association of KIR Genotypes and Haplotypes with Susceptibility to Chronic Hepatitis B Virus Infection in Chinese Han Population

Killer immunoglobulin-like receptor （KIR） genes can regulate the activation of NK and T cells upon interaction with HLA class I molecules. Hepatitis B virus （HBV） infection has been regarded as a multi-factorial disorder disease. Previous studies revealed that KIRs were involved in HCV and HIV infection or clearance. The aim of this study was to explore the possibility of the inheritance of KIR genotypes and haplotypes as a candidate for susceptibility to persistent HBV infection or HBV clearance. The sequence specific primer polymerase chain reaction （SSP-PCR） was employed to identify the KIR genes and pseudogenes in 150 chronic hepatitis B （CHB） patients, 251 spontaneously recovered （SR） controls, and 412 healthy controls. The frequencies of genotype G7 M, FZ1 increased in CHB patients compared with healthy control subjects. The frequency of genotype AH was higher in SR controls than that in both CHB patients and healthy controls. The carriage frequencies of genotype G and AH were higher; while, the frequencies of AF and AJ were lower in SR controls than those in healthy control subjects. The frequency of A haplotype was lower, whereas, the frequency of B haplotype was higher in CHB patients and SR controls than those in healthy controls. In healthy controls, haplotype 4 was found lower compared with that in CHB patients and SR controls and the frequency of haplotype 5 was higher in SR controls than that in other two groups. Based on these findings, it seems that the genotypes M and FZ1 are HBV susceptive genotypes; AH, on the other hand, may be protective genotypes that facilitate the clearance of HBV. It appears that the haplotype 4 is HBV susceptive haplotype, whereas, haplotype 5 may be the protective haplotype that facilitates the clearance of HBV. Cellular ＆ Molecular Immunology. 2008;5（6）：457-463.