乙型肝炎病毒全基因组PCR扩增产物的直接测序

目的建立乙型肝炎病毒全基因组PCR扩增产物直接测序的方法并对序列进行分析。方法PCR法扩增乙型肝炎病毒全长基因,PCR扩增产物纯化后直接进行DNA序列测定;将已测定序列的PCR扩增产物用上述方法进行再测序以评价该方法的保真性;应用进化树分析及对齐比较等方法分析HBV基因型和YMDD变异情况。结果20例标本中有18例扩增阳性并得到全长基因组序列;保真性分析表明,本法引起的人为核苷酸突变率约为1‰;序列分析表明,18例标本中有10例为基因型B、8例为基因型C;2例出现耐拉米夫定的YMDD变异,均为基因型C。结论乙型肝炎病毒全基因组PCR扩增产物直接测序,方法简便,可满足临床和科研的需要。     

2D舌侧自锁托槽简易间接粘结技术的临床应用

目的：探索2D舌侧自锁矫治技术（2dimensional lingual brackets self-ligating）托槽简易间接粘结方法在成人正畸临床中的应用效果。方法：先用光固化复合树脂将2D舌侧自锁托槽在水平与垂直方向上准确粘接于石膏模型的牙齿舌侧面,再转移粘结于患者牙齿舌侧面。结果：9例2D舌侧自锁矫治技术简易间接托槽方法粘结的成人患者,平均年龄（34．2±14．7）岁,矫治后牙位在水平与垂直方向上均较直接粘结者理想,托槽二次粘结发生率低,且由于首次粘结的粘结剂尚有补偿牙齿舌侧面形态变异的作用,故避免了由此导致的矫治后前牙牙根转距的偏差。另外,经首次粘结后的托槽底板与牙齿舌侧面的适合性更好,使粘结至牙面时更易找到正确位置。结论：简易托槽间接粘结法可有效克服2D舌侧自锁托槽直接粘结时的各种误差,对患者开口度等主观方面的依从性要求相对较低、并可缩短托槽直接粘结的椅旁操作时间,使托槽粘结位置更准确,有利于提高2D舌侧自锁矫治技术的临床矫治效果。     

重视急性肾损伤的临床研究

急性肾损伤(AKI)发病率和死亡率在世界范围内逐年增加。2012年改善全球肾脏病预后组织(KDIGO)指南明确了新的AKI定义与分期。AKI的新定义给基础研究和常规临床实践带来了巨大的变化,提高了AKI诊断的灵敏度,降低了漏诊率,提高了预测危重患者死亡的能力,临床实用性更强。在此基础上,中国开展了几个大型的、多中心AKI流行病学调查,初步阐明了中国AKI流行病学现状和危险因素。在急性肾损伤的治疗方面,更加强调肾脏的保护与支持,越来越认识到早期干预、液体管理、营养支持以及肾脏替代治疗(RRT)时机的综合管理对AKI预后影响的重要性。     

慢病毒介导CCL5-siRNA对乳腺癌细胞生物学行为的影响

Objective To study the effect of lentivirus-mediated CCL5-RNAi on the biological behaviors of human breast cancer cells. Methods CCL5-specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector, pGCSIL-GFP. Human high-metastatic breast cancer cells, MDA-MB-231, were infected by CCL5-siRNA recombinant lentivirus, which was set as KD group. Cells infected with CCL5-NC was as NC group, and cells cultured was as CON group. The expression of CCL5 mRNA and protein in MDA-MB-231 cells was detected by RT-PCR and western blot, respectively. Cell growth suppression and cell cycle was observed by MTT assay and fluorescence activated cell sorting (FACS). Colony formation and migration ability were determined by colony-rorming assay and Boyden chamber method. Results After infection of CCL5-siRNA recombinant lentivirus, the expression level of CCL5 mRNA and protein in MDA-MB-231 cells as well as the colony formation and migration ability decreased significantly, but cell's proliferation was not affected obviously. Compared with MDA-MB-231 (0.88± 0.15) and MDA-MB-231/CCL5-NC (1.00±0.07) cells, the expression of CCL5 mRNA in MDA-MB-231/ CCL5-siRNA decreased to 0.18±0.03, P<0.01. Compared with MDA-MB-231/CCL5-NC (1.82±0.18) cells, the expression of CCL5 protein in MDA-MB-231/CCL5-siRNA decreased to 0.33±0.13, P <0.01. Colony-forming assay and Boyden chamber method showed that the colony formation and migration ability of MDA-MB-231/CCL5-siRNA decreased markedly (P<0.05). The clone count in KD group was (0.33± 0.10), which was a significant decrease from (0.97±0.09) (NC group) and (1.04±0.07) (CON group), P<0.05. The number of cells that migrated through the chamber membrane of KD group (38± 15) was less than that of NC group (77±11, P <0.05) and CON group (69±9, P <0.05). However, MTT assay and FACS revealed that the proliferation of MDA-MB-231/CCL5-siRNA was not different from MDA-MB-231/CCL5-NC and MDA-MB-231 (P>0.05), the proliferation index (PI) of group KD, NC and CON were (0.48±0.02), (0.44±0.05) and (0.47±0.02) respectively. The difference was not statistically significant by multiple comparison (P>0.05). Conclusion CCL5-specific siRNA can specifically suppress the colony formation and migration of human high-matastatic breast cancer cells.     

Fusion versus Bryan Cervical Disc in two-level cervical disc disease: a prospective,randomised study

In this prospective study, our aim was to compare the functional results and radiographic outcomes of fusion and Bryan Cervical Disc replacement in the treatment of two-level cervical disc disease. A total of 65 patients with two-level cervical disc disease were randomly assigned to two groups, those operated on with Bryan Cervical Disc replacement (31) and those operated on with anterior cervical fusion with an iliac crest autograft and plate (34). Clinical evaluation was carried out using the visual analogue scale (VAS), the Short Form 36 (SF-36) and the neck disability index (NDI) during a two year follow-up. Radiological evaluation sought evidence of range of motion, stability and subsidence of the prosthesis. Substantial reduction in NDI scores occurred in both groups, with greater percent improvement in the Bryan group (P = 0.023). The arm pain VAS score improvement was substantial in both groups. Bryan artificial cervical disc replacement seems reliable and safe in the treatment of patients with two-level cervical disc disease.     

肾细胞癌灌注CT表现与微血管密度及血管内皮生长因子表达的相关性

目的 探讨肾细胞癌的多层螺旋灌注CT表现及其与微血管密度(MVD)、血管内皮生长因子(VEGF)表达的相关性. 方法 经手术病理确诊的肾细胞癌73例,其中透明细胞癌65例、乳头状腺癌3例、嫌色细胞癌5例.术前行多层螺旋灌注CT扫描,分别测量肾癌病灶血容量(BV)、血流量(BF)、平均通过时间(MTT)和毛细血管表面通透性(PS).免疫组化染色检查肾癌组织中MVD及VEGF表达.并与灌注扫描各参数进行相关性分析. 结果 肾癌病灶BV、BF、MTT及PS分别为(17.2±8.3)ml/100g,(262±176)ml·min-1·100g-1、(7.1±3.4)s、(25±13)ml·min-1·100g-1.MVD为7.6～96.3(42.3±21.0);VEGF表达阳性38例(52.1%),其中Ⅰ级24例(32.9%)、Ⅱ级10例(13.7%)、Ⅲ级4例(5.5%).MVD与VEGF无相关性.肾癌病灶MVD与BV、BF、PS呈正相关(P<0.01),与MTT呈负相关(P<0.05);VEGF与所有灌注参数均不相关.结论肾癌灌注CT扫描参数可反映肿瘤组织中的血管生成情况.     

头颈部CTA判断大脑中动脉区脑梗死患者供血动脉狭窄的价值

目的探讨头颈部计算机断层扫描血管成像（CTA）在判断大脑中动脉（MCA）区脑梗死患者血管狭窄程度以及与脑梗死程度的相关I生。方法对68例MCA区脑梗死患者进行头颈部CTA检查,分析受累血管狭窄程度及其与脑梗死程度的关系。结果68例中,颈内动脉（ICA）和MCA均正常者5例,单纯ICA狭窄共16例,其中轻度狭窄2例,中度狭窄4例,重度狭窄5例,闭塞5例;单纯MCA狭窄共41例,其中轻度狭窄6例,中度狭窄12例,重度狭窄18例,MCA闭塞5例;MCA合并ICA狭窄6例。ICA或MCA狭窄程度与脑梗死类型有关,狭窄程度越严重,梗死程度越严重（r=0.832,0．938,P〈0．05）。结论头颈部CTA在判断血管狭窄的同时,可预测患者脑梗死的程度,对脑梗死的临床诊断和治疗方案的选择具有重要的指导价值。     

孕马采尿期养殖成本分析

【目的】本文通过对采集孕马尿(PMU)为主的马养殖经营状况的调研,分析孕马养殖的方式、规模以及成本结构等,提出有效提高孕马养殖效益的途径。【方法】在伊犁地区新源县,本文采用问卷调查和走访典型养殖户的方式,统计归纳数据;采用损益平衡分析法对PMU养殖进行分析研究,确定在不亏损的条件下的最低生产量,发现当损益平衡点Qf=6.1(匹),牧户PMU养殖数量超过6匹时即可营利。【结果】综合分析孕马养殖成本,探索减低孕马养殖成本的方式,增加孕马养殖收入。     

大骨节病儿童手部X线征与年龄的关系

本文分析了2800例4～13岁大骨节病儿童掌指骨X线病变与年龄的关系,结果表明手部掌指骨干骺端X线征随年龄增加而趋于下降,骨端、骨骺、腕骨X线征的类型与检出率则随年龄而呈上升趋势,反映出大骨节病关节病变的范围有随年龄增长而扩大和增重的趋势。     

中国北方汉族男子fms样酪氨酸激酶基因多态性与高住高练低训敏感性的关联性研究

目的:探讨fms样酪氨酸激酶(Flt-1)基因多态性与低氧训练敏感性的关联性。方法:选取70名中国北方平原地区汉族男子进行4周高住高练低训(HiHiLo),方案为每日在低氧环境(O2浓度为14.5%～14.8%,大约相当于海拔3000米)中休息10h,每周进行3次75%VO2max低氧训练(O2浓度为15.4%,大约相当于海拔2500米),其余时间在常氧下训练。测试其HiHiLo前后的VO2max及训练期间的血象指标。用PCR-RFLP法分析Flt-1基因SNP/A193019G多态性。结果:(1)4周HiHiLo后,AA基因型VO2max增加幅度高于AG基因型57.54%,但无显著性差异。(2)在HiHiLo期间(除第23天外),AA基因型Hb的变化幅度始终高于AG基因型,但无显著性差异。结果提示,与AG基因型者相比,AA基因型者可能有对低氧训练更敏感的趋势。