抗HBsAg Fab片段和α-干扰素融合蛋白的克隆及表达

目的 为深入研究乙型肝炎的生物导向治疗，构建以人抗乙型肝炎表面抗原（ＨＢｓＡｇ）抗本Ｆａｂ片段国导向作用，αＡ－干扰素（ＩＦＮ－αＡ）为治疗作用的融合蛋白Ｆａｂ／ＩＦＮ－αＡ原核表达载体ｐＨＳ／ＩＦＮ－αＡ。方法 用ＰＣＲ体外扩增目的基因ＩＦＮ－αＡ，并在其ＤＮＡ两端引入酶切点及５′端人工合成的Ｌｉｎｋｅｒ，重组人抗ＨＢｓＡｇ缺本Ｆａｂ片段功体ｐＨＳ相应酶切位点，酶切位点，酶切和ＤＮＡ测序鉴定筛选     

实时超声造影和融合影像导航下微波消融与手术治疗小肝癌患者疗效比较*

目的 研究实时超声造影和融合影像导航技术下微波消融与手术治疗小肝癌患者的疗效。方法 2013年12月～2016年12月我院收治的142例原发性肝癌患者被随机分为两组,每组71例。对观察组患者实施实时超声造影和融合影像导航下微波消融治疗,给予对照组实施外科手术切除治疗。采用酶速率法测定血清α-L-岩藻糖苷酶(AFU)水平,采用ELISA法测定血清甲胎蛋白(AFP)水平,采用免疫放射法测定血清糖类抗原19-9(CA19-9)水平。结果 在术后3个月,观察组肿瘤完全消融率为93.8%,对照组完全切除率达100.0%(P>0.05);两组血清AFP、AFU和CA19-9水平比较差异无统计学意义(P＞0.05);术后,观察组肺部感染、胆漏、皮肤切口感染、腹腔或胸腔积液等并发症发生率为4.2%,显著低于对照组的25.3%(P＜0.05);随访3～36个月(中位随访18个月),观察组累积生存率为73.2%(52/71),对照组为71.8%(51/71),两组差异无统计学意义(Log-rank值=0.086,P=0.763)。结论 在实时超声造影联合融合影像导航技术支持下行微波消融治疗小肝癌患者,疗效与手术切除相当,但并发症更少,值得临床积极开展和验证。     

MRI and dual-energy CT fusion anatomic imaging in Ru-106 ophthalmic brachytherapy

PurposeBrachytherapy with Ru-106 is widely used for the treatment of intraocular tumors, and its efficacy depends on the accuracy of radioactive plaque placement. Ru-106 plaques are MRI incompatible and create severe metal artifacts on conventional CT scans. Dual-energy CT scans (DECT) may be used to suppress such artifacts. This study examines the possibility of creating fusion images from MRI scans (preoperatively) and DECT scans (with the plaque in place) as a tool for confirming the anatomic accuracy of plaque placement.Methods and MaterialsSix patients with intraocular lesions (5 with choroidal melanoma and 1 with a retinal vasoproliferative lesion) were included. Fusion images of preoperative MRI scans and DECT scans with the plaque in place were created with the Demo version of the ImFusion suite (ImFusion GmbH, Munchen Germany). Clearance margins between the tumor and plaque edge in axial, transverse, and coronal planes as well as the elevation of the posterior plaque edge from the sclera were recorded and associated with the location of the lesion.ResultsPlaque-tumor clearance margins for transverse, sagittal, and coronal planes were higher for anteriorly located lesions (5.13 mm ± 0.11 [5.0–5.2], 5.10 mm ± 0.26 [4.9–5.4], and 5.33 mm ± 0.45 [4.9–5.8] respectively) than for posteriorly located lesions (4.16 mm ± 1.44 [2.5–5.1], 4.13 mm ± 1.42 [2.5–5.1], and 4.2 mm ± 1.21 [2.8–5.0], respectively). The elevation of the posterior plaque edge from the sclera was 0.33 mm ± 0.28 [0–0.5] and 0.63 mm ± 0.60 [0.7–1.2] for posterior and anterior lesions, respectively.ConclusionsFusion images between DECT and MRI scans may be used as a tool to confirm the accuracy of Ru-106 plaque placement in relation with the intraocular tumors in ophthalmic brachytherapy.     

A rare case of clear cell sarcoma with 4 types of EWSR1-ATF1 fusions detected not in primary site but in metastatic site

Clear cell sarcoma is a unique tumor which has EWSR1-ATF1 or EWSR1-CREB1 fusion. Several patterns of EWSR1-ATF1 fusion are observed in clear cell sarcoma. Since type 5–7 fusions were reported recently, they are classified as type 1–7. We examined EWSR1-ATF1 and EWSR1-CREB1 fusions in a single case of clear cell sarcoma with lung metastasis in a 36-year-old Japanese man. As a result, we found only type 1 EWSR1-ATF1 fusion in the primary site, but 4 types of EWS-ATF1 fusion (type 1, 2, 5, 6) were detected in the metastatic site. These 4 types of fusion were completely identical to the recent report, but the case had the same fusion patterns in both primary and metastatic sites. In our case, increased splicing activity in the EWSR1-ATF1 fusion might be acquired at the metastatic site. There is another possibility that metastasis might develop through the increased splicing activity in the fusion.     

Is instrumented lateral lumbar interbody fusion superior to stand-alone lateral lumbar interbody fusion for the treatment of lumbar degenerative disease? A meta-analysis

The purpose of this meta-analysis was to compare the fusion rate and outcomes directly between patients who underwent stand-alone lateral lumbar interbody fusion (LLIF) and LLIF with supplemental posterior instrumentation. A comprehensive literature search was performed for relevant studies using PubMed, EMBASE, Web of Science, and Cochrane Library. The stand-alone and instrumented LLIF were compared by the fusion rate, the radiographic parameters, the cage subsidence rate, the clinical outcomes, the complication rate, and the reoperation rate. A total of 13 studies comprising 1090 patients with lumbar degenerative disease (LDD) were included. There was no significant statistical difference in the complication rate, and there was no significant clinical difference in the improvement of clinical outcomes at the last follow-up between patients who underwent stand-alone and instrumented LLIF. Nevertheless, lower fusion rate (RR, 0.92; 95% CI 0.87 to 0.98, P = 0.006), inferior restoration of disk height (WMD, −0.68; 95% CI −1.04 to −0.32, P < 0.001) and segmental lordosis (WMD, −1.28; 95% CI −2.30 to −0.27, P = 0.013), higher cage subsidence rate (RR, 1.68; 95% CI 1.36 to 2.07, P < 0.001), and higher reoperation rate (RR, 2.12; 95% CI 1.02 to 4.43, P = 0.045) were observed in the stand-alone group. Both stand-alone and instrumented LLIF were effective in improving the clinical outcomes of patients with LDD. However, the stand-alone LLIF was associated with lower fusion rate, inferior maintenance of indirect decompression, and higher reoperation rate due to high-grade cage subsidence. For patients with risk factors of high-grade cage subsidence, the LLIF with posterior instrumentation may be the better choice.     

癌胚抗原启动子调控的白细胞介素-2/干扰素β融合基因的原核及真核表达质粒的构建

目的 根据细胞因子间协同作用特点,构建人白细胞介素-2（IL-2）与干扰素（IFN）β的融合基因,并用癌胚抗原（CEA）基因启动子调控其在结肠癌细胞株中的靶向表达.方法 分别用原核及真核表达载体,检测融合蛋白的表达情况.PCR、RT-PCR法扩增得到CEA启动子、IL-2 cDNA、IFNβ cDNA,应用原核表达质粒pGEX-5X-1、真核表达质粒pcDNA3.1/HisA、绿色荧光蛋白质粒pEGFP-C1,构建出原核表达质粒和真核表达质粒;SDS-PAGE电泳检测原核表达;用脂质体法将真核表达质粒转入结直肠癌细胞株Lovo（CEA高表达）、HT-29（CEA低表达）和对照Hela细胞（CEA阴性）,转染后分别以流式细胞仪、荧光显微镜、Western印迹等方法检测肿瘤细胞凋亡情况及蛋白表达情况.结果融合基因可在大肠杆菌及肿瘤细胞内表达融合蛋白;对肿瘤细胞有明显的杀伤作用;由CEA启动子调控的融合基因在Lovo细胞的表达高于HT-29细胞,在CEA阴性的Hela细胞中几乎不表达.结论 CEA启动子可靶向性调控融合基因在肿瘤细胞的表达,具有协同作用的细胞因子融合蛋白可能有更强的抗肿瘤作用.     

同种异体骨与自体骨填充椎间融合修复脊髓型颈椎病的比较

背景:目前用于颈椎前路的填充材料较多,以自体骨、同种异体骨多见,但自体骨取材引发供骨区出血、感染、术后疼痛等并发症被临床工作者逐渐重视。同种异体骨具有良好的生物相容性及安全性,可作为一种比较理想的融合器填充材料。目的:对比异体骨与自体骨填充椎间融合器结合钢板内固定在脊髓型颈椎病患者颈椎前路融合中的临床修复效果。方法:2012年1月至2013年6月对44例脊髓型颈椎病共44节段实施前路椎间盘摘除、椎间融合联合钢板内固定治疗,取颈前斜切口2.0-3.0 cm,切除椎间盘及后纵韧带,尽量保留终板。其中24例取自体髂骨填充椎间融合器,20例采用同种异体骨填充,以JOA评分、Odom’s疗效评定标准及X线射片评定两组疗效。结果与结论:两组患者均获得12-18个月随访。两组间术后并发症及JOA评分差异无显著性意义。临床疗效评定(Odom’s标准)优良率,同种异体骨组高于自体骨组,差异有显著性意义(P<0.05)。术后3,6个月同种异体骨组融合率较自体骨组低,12个月时两组融合率均为100%。影像学结果提示,术后3,6,12个月同种异体骨组与自体骨组融合节段椎间隙高度、前凸Cobb角之间差异无显著性意义(P>0.05)。提示同种异体骨作为椎间融合器填充材料,融合率与自体髂骨相似,并可维持颈椎生理曲度及恢复椎间隙高度,可作为理想的骨填充材料应用于颈椎前路手术。