B组轮状病毒WH-2株vp7基因的原核表达与抗体的制备

目的：在大肠杆菌中表达人B组轮状病毒WH-2株VP7蛋白并制备其兔抗血清。方法：根据B组轮状病毒（GBRV）WH-2株vp7基因的全序列设计引物,用PCR的方法扩增得到vp7基因的编码区。将其克隆到原核GST融合表达载体pGEX-KG内,转化大肠杆菌E.coliDH5α,IPTG诱导表达人B组轮状病毒WH-2株VP7蛋白,经SDS-PAGE分离纯化表达的蛋白免疫新西兰兔,制备人B组轮状病毒WH-2株VP7蛋白抗血清。结果：经鉴定确认,vp7基因以正确的方式插入到载体中,此重组表达载体经IPTG诱导后,可表达相对分子量为53.4 kDa的GST-VP7融合蛋白。制备的抗血清经同样诱导表达的表达载体pGEX-KG表达产物吸收后1：500倍稀释后用Western Blot分析,与53.4 kDa的GST-VP7融合蛋白获得特异性显色信号。结论：人B组轮状病毒WH-2株VP7蛋白成功在大肠杆菌中GST融合表达,所表达的蛋白和制备的抗体不但为研究结构与功能提供了物质基础,也为GBRV所引起的疾病预防、诊断和治疗等流行病学研究和临床诊断奠定了基础,具有重要实际应用价值。     

Expression of the cell surface antigen detected by the monoclonal antibody A7 in pancreatic carcinoma cell lines

In a previous study, we used a murine monoclonal antibody, A7, against human colon carcinoma as a drug-carrier to treat colorectal cancer.¹ In the present study, we found that MAb A7 also reacted immunohistochemically with 73% of human pancreatic carcinoma cell lines, with the A7 antigen mainly being detected on the cell surface. However, the A7 antigen was found in only 9% of the spent media of these human pancreatic carcinoma cell lines by ELISA. On the other hand, the positive incidence of CA19-9, POA, ferritin, CEA, DU-PAN-2 and SLX in those spent media was 100%, 64%, 64%, 55%, 55% and 36%, respectively. These results suggest that the A7 antigen may only rarely be shed into the sera of pancreatic cancer patients, in which case MAb A7 could be a suitable drug-carrier in targeting chemotherapy for pancreatic cancer patients.     

人牙本质涎蛋白在COS-7细胞中的表达

目的 :构建人牙本质涎蛋白 (humandentinsialoprotein ,hDSP)真核表达载体 ,用瞬时系统表达目的基因。方法 :将hDSP全长基因定向克隆入真核表达质粒载体pcDNA3 ,构建pcDNA3 hDSP重组质粒 ,脂质体转染法转染COS 7细胞 ,48～ 72h收集细胞及培养上清 ,Westernblot和免疫组化检测表达产物。结果 :酶切鉴定表明重组表达载体构建成功 ;Westernblot检测显示在 60 0 0 0处出现一条清晰的特异性免疫阳性条带 ;免疫组化染色显示细胞胞质内有大量棕黄色颗粒 ,进一步证明该转染方法可行 ,转染效率较高。结论 :hDSP全长基因可以在COS 7细胞获得瞬时高效表达。     

8-硝基白杨素的合成及其抗肿瘤作用

目的　探讨 8-硝基白杨素的抗肿瘤活性。方法　荷Lewis肺癌雄性小鼠 5 0只 ,随机分为5组 (NS对照组 ,环磷酰胺组 ,低、中、高剂量 8-硝基白杨素组 ) ,每组 1 0只 ,均采用腹腔注射给药。末次给药 2 4h后 ,颈椎脱位处死小鼠 ,观察各组瘤重减轻情况及转移结节数。结果　低、中、高剂量 8-硝基白杨素均能明显抑制肿瘤的生长和肿瘤转移结节的数 (阳性药对照组和各处理组与对照组相比 ,P均<0 .0 1 ) ,高剂量 ( 5 0mg kg) 8-硝基白杨素与 1 0 0mg kg的环磷酰胺的抗肿瘤作用相当 (P >0 .0 5 )。结论　 8-硝基白杨素作为一种新黄酮衍生物 ,具有良好的抗肿瘤作用。     

A clinical and genetic study of 56 Saudi Wilson disease patients: identification of Saudi-specific mutations

Wilson disease (WD) is a hereditary disorder, with recessive transmission and genetic heterogeneity. Several mutations of ATP7B, the gene underlying WD, were reported in many ethnic groups. In this study, mutation screening in ATP7B of 56 Saudi Arabian WD patients was undertaken. The clinical data of all patients were recorded. The entire ATP7B coding sequence, including intron-exon boundaries were screened for mutation by the polymerase chain reaction (PCR)-based mutation detection technique and DNA sequencing. Thirty-nine patients were symptomatic at presentation and 17 subjects were pre-symptomatic siblings of affected patients. Fourteen patients had neurological, 11 patients had mixed (hepatic and neurological), and 14 patients had hepatic presentations. Family history suggestive of WD was present in 72% of cases and 68% had consanguineous parents. Genetic analysis showed disease-causing mutations in three exons (exons 8, 19 and 21) of the ATP7B gene in 28 patients (50%). Mutations in exons 21 (18 cases) and 19 (one case) were unique for Saudis. This large series of Saudi patients with WD has shown wide variability in the genomic substrate of WD. There is no correlation between genotype and clinical presentation.     

共刺激分子B7在移植免疫中的研究现状及展望

B7家族成员是最重要的共刺激分子,其中对B7-1、B7-2分子的研究最为深入,近年来,该家族中B7-H1、B7-H2、B7-H3、B7-DC、B7-H4等新成员也被陆续发现,其作用机制主要是为T细胞活化提供所需的正性和负性共刺激信号,通过复杂的网络平衡作用决定最终的免疫效应。随着器官移植的迅速发展,诱导对移植物的特异性耐受成为解决移植排斥的重要途径,正性共刺激通路B7CD28通路、B7-H2ICOS通路、B7-H3通路的过度活化,以及负性共刺激通路B7CT-LA-4通路、B7-H1、B7-DCPD-1通路、B7-H4BTLA通路的发现,在移植免疫中意义尤为突出。     

Microvascular decompression for hemifacial spasm: postoperative neurologic follow-up and evaluation of life quality

Microvascular decompression (MVD) is an effective and safe treatment in hemifacial spasm (HFS). Postoperative evaluations are usually made by neurosurgeons. Follow-up studies performed by neurologists and postoperative quality of life (QoL) investigations are lacking. All 25 HFS patients operated with MVD in our centre between 2000 and 2004 were evaluated with the recently validated HFS-7 scheme, extended with the item 'sleep disturbance due to HFS' (HFS-8). The patients underwent a careful neurological examination median 3 years after the operation. The evaluation focused on clinical aspects, changes in blood pressure and time until observable effect of MVD. The evaluation of HFS-7 questionnaire and the extended form (HFS-8) showed significant improvement in QoL after MVD. Neurological outcome was in almost all cases excellent or good. Eleven (44%) patients had no neurological deficits at all. Only one patient had serious complications with ipsilateral facial palsy, deafness, balance problems and vertigo. The other patients had minor neurological findings or symptoms. Eighteen (72%) patients experienced early effect within 3 months after MVD; seven (28%) patients had late effect between 6 and 14 months. Median age of the patients with late effect (62.6 years) was significantly higher than in those with early effect (52.7 years).     

血管紧张素(1-7)和依那普利拉对烧伤血清离体灌注大鼠心脏功能的影响

Objective To investigate effects of angiotcnsin ( 1-7 ) [ Ang( 1-7 ) ] and enalaprilat on function of isolated rat heart perfused by burn serum. Methods Eighty SD rats were used to prepare burn serum. Hearts of another 24 SD rats were isolated to reproduce Langendorff perfusion model. The rat hearts were divided into different groups with different perfusion fluids as K-H buffer group, K-H buffer containing 20% burn serum group ( burn serum group) , K-H buffer containing 20% burn serum and 2 μg/mL enala-prilat group (enalaprilat group), and K-H buffer containing 20% burn serum and 1 nmol/mL Ang(1-7) group [ Ang(1-7) group]. The rat hearts were perfused for 30 mins with each of above-mentioned fluids in different groups. Then left ventrieular systolic pressure (LVSP) , left ventricular end diastolic pressure ( LV-EDP) , ± dp/dt max, coronary flow(CF) , level of creatine kinase (CK) and lactate dehydrogenase (LDH) in respective coronary effluent were determined. Results Compared with LVSP ( 11.2 ± 1.0 kPa, 1 kPa = 7.5 mm Hg), +dp/dt max (642±53 kPa/s), -dp/dt max (380±61 kPa/s) and CF level in K-H buffer group, CF, LVSP(5.9±0.8, 8.0±1.1, 8.9±1.3 kPa, respectively), +dp/dt max(275±37, 454±48, 479±63 kPa/s, respectively), - dp/dt max ( 135±35, 219±47, 277±58 kPa/s, respectively ) of burn serum group, those levels in Ang(1-7) group, and enalaprilat group were decreased obviously(P<0.05 or P <0.01 ), but LVEDP, level of CK and LDH in coronary effluent were increased. Compared with those parameters in burn serum group, CF, LVSP, ±dp/dt max of Ang(1-7) group and enalaprilat group were increased obviously(P<0.05 or P<0.01), and LVEDP, level of CK and LDH in coronary effluent were decreased obviously(P<0.01). Conclusions Ang(1-7) and enalaprilat can effectively improve left ventricular function of isolated rat heart perfused by burn serum and mitigate myocardial injury.