人源腺病毒伴随病毒Ⅱ型单克隆抗体Fab段及其全抗体的研制

目的 运用噬菌体表面表达技术，获得基因工程抗腺病毒伴随病毒抗体Fab、IgG。方法 从酶联免疫吸附试验阳性的人外周血中分离淋巴细胞。提取总RNA，逆转录cDNA，聚合酶链反应扩增人IgG Fab轻、重链基因，利用pComb3载体构建噬菌体抗体库。用纯化的腺病毒伴随病毒为固相抗原筛选抗体Fab段，并在E.coli中进行分泌性表达。通过ELISA和间接免疫荧光试验、筛选和鉴定Fab抗体，并进行序列测定。然后将其重链和轻链基因克隆到全抗体表达载体pAC-L-Fc上。转染昆虫sf-9细胞，利用杆状病毒，昆虫细胞系统实现全抗体的分泌型表达，免疫沉淀试验鉴定它的抗蛋白。结果 分离到一株Fab克隆AAVs-31，腺病毒伴随病毒抗原和抗Fab抗体直接ELISA检测阳性，间接免疫荧光试验呈阳性，序列分析结果表明是一新的序列，所获得的基因为人源IgG Fab基因。由Gamma链和Kappa链组成。表达的全抗体ELISA反应、免疫荧光反应和间接免疫荧光试验均为阳性，免疫沉淀试验结果显示它结合病毒颗粒，不结合任一VP1、VP2、VP3单独衣鞘蛋白。结论 用噬菌体表面表达技术首次获得了抗腺病毒伴随病毒Ⅱ型单克隆抗体Fab段，并在真核系统中表达了其全抗体，它们识别的可能是衣鞘蛋白组装时形成的表位。     

Infestation of sand lizards (<Emphasis Type="Italic">Lacerta agilis</Emphasis>) resident in the Northeastern Poland by <Emphasis Type="Italic">Ixodes ricinus</Emphasis> (L.) ticks and their infection with <Emphasis Type="Italic">Borrelia burgdorferi</Emphasis> sensu lato

Sand lizards (Lacerta agilis) were trapped and examined for ticks from May to September in 2002 and 2003 in Northeastern Poland. A total of 233 Ixodes ricinus (L.) ticks (76 larvae and 157 nymphs) was found on 31 of 235 captured lizards (13.2%). The tick infestation is relatively low compared to that of mammals and passerine birds from the same area (Siński et al. 2006, Gryczyńska et al. 2002). Tick infestation depended on the month of capture, being the highest in spring. In autumn no ticks were recorded on any of the captured lizards. The oldest lizards carried the highest number of ticks but no differences related to sex of the host were found. All the collected ticks were analysed by PCR for the presence of Borrelia burgdorferi sensu lato, the etiological agents of Lyme disease. Spirochetes were detected in 11 out of 233 (4.7%) ticks tested. Genetic analysis confirmed that the spirochetes are members of the Borrelia afzelii, B. garinii and B. burgdorferi sensu stricto genospecies. Mixed infection were not detected. The prevalence of infection was analysed in relation to months of the capture, age and sex of the lizards, but differences were not statistically significant. The obtained results suggest that lizards are probably not B. burgdorferi reservoirs, but further studies are required to confirm this.     

Wilms tumor in patients with osteopathia striata with cranial sclerosis

Germline pathogenic variants in AMER1 cause osteopathia striata with cranial sclerosis (OSCS: OMIM 300373), an X-linked sclerosing bone disorder. Female heterozygotes exhibit metaphyseal striations in long bones, macrocephaly, cleft palate, and, occasionally, learning disability. Male hemizygotes typically manifest the condition as fetal or neonatal death. Somatically acquired variants in AMER1 are found in neoplastic tissue in 15–30% of patients with Wilms tumor; however, to date, only one individual with OSCS has been reported with a Wilms tumor. Here we present four cases of Wilms tumor in unrelated individuals with OSCS, including the single previously published case. We also report the first case of bilateral Wilms tumor in a patient with OSCS. Tumor tissue analysis showed no clear pattern of histological subtypes. In Beckwith–Wiedemann syndrome, which has a known predisposition to Wilms tumor development, clinical protocols have been developed for tumor surveillance. In the absence of further evidence, we propose a similar protocol for patients with OSCS to be instituted as an initial precautionary approach to tumor surveillance. Further evidence is needed to refine this protocol and to evaluate the possibility of development of other neoplasms later in life, in patients with OSCS.Subject terms: Genetics, Clinical genetics, Disease genetics     

Interaction between KIR3DL1 and HLA-B*57 supertype alleles influences the progression of HIV-1 infection in a Zambian population

KIR and HLA loci are both highly polymorphic, and some HLA class 1 products bind and trigger cell-surface receptors specified by KIR genes. We examined whether KIR genes act in concert with HLA-B locus to control HIV-1 infection in a sample of Zambian patients. DNA samples from 88 Zambian patients with HIV-1 were examined. Patients were classified as either slow progressors (SP; n = 54) or rapid progressors (RP; n = 34) to AIDS. All were typed for HLA-B and KIR genes. Our results reveal an association between B*57 supertype (B*57s, which includes B*57 and B*58 alleles) and delayed progression to AIDS (p = 0.0007 by pc = 0.015; OR = 5.25). We also observed an increase incidence of Bw4-I80 in patients with slow progression (p = 0.001 by pc = 0.003, OR = 5). This increase was found to be secondary to B*57s. The presence of both KIR3DL1 and B*57S has a significant effect on progression to AIDS (p = 0.0008; OR = 5.61). B*57s genotypes with another HLA-B allele different from those in the trans position, which also had a specificity different to Bw4-I80 (Bw4-T80 or Bw6), was also greater in the SP than in the RP group (p = 0.00003; OR = 10.11). The presence of the inhibitory allele KIR3DL1 in combination with the HLA-B*57s alleles that contain the Bw4-I80 epitope, has a highly protective effect against progression to AIDS in Zambian patients.     

Differences in gallstone structure in primary common bile duct lithiasis and gallbladder lithiasis

Summary Some differences between gallbladder lithiasis and primary common bile duct lithiasis are described. Microbiological cultures and biochemical analyses were carried out on the bile of two groups of patients: 27 suffering from gallbladder and 5 from primary common duct lithiasis. The microstructure and composition of gallstones were also examined by polarized light microscopy and X-ray diffraction. Women predominated in gallbladder lithiasis but not in primary common duct lithiasis group (P<0.05) and body weight was higher in the former group (P<0.02). Primary common duct lithiasis patients had a higher, although not significant, incidence of duodenal diverticulosis (P=0.15), and a higher incidence ofE. coli-positive cultures in bile (P<0.001). No significant difference in the biochemical composition of the bile was found between the groups. Brown pigment stones predominated in primary common duct lithiasis, while cholesterol stones did in gallbladder and secondary common duct lithiasis (P<0.0001). Stones formed in the gallbladder generally show linear, radial growths of cholesterol crystals, while those from the common duct present a polystratified, concentric deposition of microgranules composed mainly of pigmentary salts.These differences should be taken into account as additional criteria in the differential diagnosis between primary and secondary common duct lithiasis, as the classical criteria for diagnosing of the former greatly underestimate its actual incidence. The distinction between primary and secondary common duct lithiasis is of practical significance, since each entity requires different treatment.Abbreviations CBD common bile duct - CBDL common bile duct lithiasis - ERCP endoscopic retrograde cholangiopancreatography - GBL gallbladder lithiasis - HDL high density lipoproteins - PCBDL primary common bile duct lithiasis - SCBDL secondary common bile duct lithiasis - SGOT serum glutamic-oxalacetic transaminase - SGPT serum glutamic-pyruvic transaminase     

N-methylated beta-carbolines protect PC12 cells from cytotoxic effect of MPP+ by attenuation of mitochondrial membrane permeability change

Opening of the mitochondrial permeability transition pore has been recognized to be involved in cell death. The present study investigated the effect of beta-carbolines (harmaline and harmalol) on the MPP(+)-induced change in the mitochondrial membrane permeability and cell death in differentiated PC12 cells. beta-Carbolines and antioxidants (superoxide dismutase, catalase, ascorbate or rutin) prevented the loss of cell viability in PC12 cells treated with 250 microM MPP(+), while the effects of N-acetylcysteine and dithiothreitol were not observed. beta-Carbolines reduced the condensation and fragmentation of nuclei caused by MPP(+) in PC12 cells. beta-Carbolines alone did not exhibit a significant cytotoxic effect on PC12 cells. beta-Carbolines (50 microM) inhibited the decrease in mitochondrial transmembrane potential, cytochrome c release, activation of caspase-3, formation of reactive oxygen species (ROS) and depletion of GSH caused by MPP(+) in PC12 cells. beta-Carbolines reduced the hydrogen peroxide- or SIN-1-induced cell death in PC12 cells. The results suggest that beta-carbolines may attenuate the MPP(+)-induced viability loss in PC12 cells by inhibition of change in the mitochondrial membrane permeability and by antioxidant effect.     

4NQO诱发A／J系小白鼠肺腺癌的研究

目的：通过A／J系小鼠肺腺癌模型,探讨肺肿瘤细胞的起源和恶性转化过程。方法：应用4－氮喹啉1－氧化物（4QO）化学物质为促癌剂,建立A／J系列小鼠肺癌的模型。观察诱发肿瘤时间、肿瘤数目及与类型的关系。结果：RNQO诱发5个月时,共发现肺肿瘤17处,其中肺泡上皮增生（AP）6处。肺泡腺瘤（AA）9处,乳头状腺瘤（PA）2处;诱发7个月时共发现肺肿瘤19处,AP为7处,AA为11处,PA为1处,诱发9个月时共发现肺肿瘤27处,AP为5处,AA为17处,PA为3处,发现乳头状腺癌（PC）2处。结论：4NQO能诱发A／J系小鼠肺腺癌。诱发肿瘤的时间长短与发生肿瘤的组织学类型有密切关系。     

Risk factors of uterine contraction after ureteroscopy in pregnant women with renal colic

International Urology and Nephrology - Ureteroscopy is widely applied in pregnant women with renal colic, but such patients are easy to experience uterine contraction after surgery. There are many...     

不停跳冠状动脉旁路移植术中应用MostCare/PRAM监测系统进行血流动力学监测的临床研究

目的探究Most Care/PRAM系统监测下不停跳冠状动脉旁路移植术(OPCABG)患者术中血流动力学变化情况和预后分析。方法纳入2016年10月至2017年1月安贞医院89例OPCABG患者,其中男53例、女36例,年龄(60.50±8.40)岁。记录术中血流动力学变化情况。按是否发生心肌梗死、低心排血量等严重循环不良事件,分为平稳组和严重循环不良事件组,进行相关分析。结果手术全程监测完整血流动力学数据患者65例,开胸前和关胸后被动抬高试验的每搏量(SV)升高均值分别为23.00%±3.20%和29.40%±3.70%。麻醉、开胸、应用肝素时、搭桥中、应用鱼精蛋白时、关胸和术毕7个时间段,SV明显下降,外周血管阻力指数(SVRI)持续显著增加,最大压力梯度(d P/d T)和心脏循环效率(CCE)在麻醉后明显下降,搭桥时下降到最低,其后逐渐升高;每搏量变异率(SVV)和脉压变异率(PPV)在麻醉后下降,开胸后一直升高。89例患者发生严重循环不良事件共9例,其中4例死亡。严重循环不良事件组术中基础SVRI、SVV和PPV均显著高于平稳组(P<0.05),CCE、d P/d T和SV差异无统计学意义(P>0.05)。术中基础SVRI、CCE、d P/d T、SVV、PPV和SV均值与预后指标均无明显相关性。结论OPCABG术中易出现血流动力学的改变,因此,OPCABG术中宜应用Most Care/PRAM仪进行血流动力学监测,并及时纠正血流动力学异常。