中国汉族人群凝血因子Ⅸ基因限制性片段长度多态性的研究

为探讨中国人群中凝血因子Ⅸ（FⅨ）基因SalⅠ，NruⅠ和MseⅠ酶切位点的限制性片段长度多态性（RFLP），采用聚合酶链反应（PCR）方法，以三代内相互间无亲缘关系的正常汉族人为研究对象，对其FⅨ基因内含子1第192位以及5′端－793位核苷酸A和G的频率和5′端－698位核苷酸T和C的频率进行了分析，其中对FⅨ－192共分析了214条X染色体，对FⅨ－793共分析了210条X染色体，对FⅨ－698共分析了206条X染色体。结果表明，FⅨ基因内含子1第192位A和G的频率分别为0．878和0．122，杂合体频率为0．213；－793位核苷酸A和G的频率分别为0．552和0．448，杂合体频率为0．494，FⅨ－698位核苷酸T和C的频率分别为0．311和0．689，杂合体频率为0．429。结论提示：FⅨ基因SalⅠ,NruⅠ和MseⅠ酶切位点的RFLP有利于我国血友病B的携带检测和产前诊断。     

Abnormalities in Monocyte Recruitment and Cytokine Expression in Monocyte Chemoattractant Protein 1–deficient Mice

Monocyte chemoattractant protein 1 (MCP-1) is a CC chemokine that attracts monocytes, memory T lymphocytes, and natural killer cells. Because other chemokines have similar target cell specificities and because CCR2, a cloned MCP-1 receptor, binds other ligands, it has been uncertain whether MCP-1 plays a unique role in recruiting mononuclear cells in vivo. To address this question, we disrupted SCYA2 (the gene encoding MCP-1) and tested MCP-1–deficient mice in models of inflammation. Despite normal numbers of circulating leukocytes and resident macrophages, MCP-1^−/− mice were specifically unable to recruit monocytes 72 h after intraperitoneal thioglycollate administration. Similarly, accumulation of F4/80+ monocytes in delayed-type hypersensitivity lesions was impaired, although the swelling response was normal. Development of secondary pulmonary granulomata in response to Schistosoma mansoni eggs was blunted in MCP-1^−/− mice, as was expression of IL-4, IL-5, and interferon γ in splenocytes. In contrast, MCP-1^−/− mice were indistinguishable from wild-type mice in their ability to clear Mycobacterium tuberculosis. Our data indicate that MCP-1 is uniquely essential for monocyte recruitment in several inflammatory models in vivo and influences expression of cytokines related to T helper responses.     

跟骨定量超声骨质测量参数与骨密度及骨组织形态计量学指标的关系

目的:分析跟骨定量超声骨质测量中各参数与骨密度及形态计量学指标的相关性。方法:选择2004-01/2005-12广州市第六人民医院和中山大学三院骨科小腿以上截肢患者38例,将其跟骨定量超声测定的超声振幅衰减平均值与健康青年人骨峰值进行比较,>-2.5 SD者为骨量正常组(12例),<-2.5 SD者为骨质疏松组(26例)。分别进行跟骨定量超声、双能X线骨密度测量仪及骨形态计量学测量,应用直线相关分析法分析跟骨定量超声测定中各参数与骨密度及骨组织形态计量学各指标的相关性。结果:38例全部进入结果分析。①骨质疏松组跟骨超声振幅衰减平均值和骨硬度指数值均小于骨量正常组(P<0.01)。②骨量正常组跟骨骨密度值显著高于骨质疏松组[(352±16),(233±14)mg/cm2,P<0.01]。③骨量正常组跟骨平均骨小梁间距或弥散度低于骨质疏松组而松质骨体积高于骨质疏松组(P<0.05)。④超声振幅衰减平均值和骨硬度指数与骨密度呈直线正相关(r=0.814,0.326,P<0.01,0.05)。⑤超声传播速度与骨小梁游离末端、平均骨小梁间距呈直线负相关(r=-0.688,-0.712,P<0.01),与小梁间连点数、松质骨体积呈直线正相关(r=0.672,0.794,P<0.01);骨硬度指数与平均骨小梁间距呈直线负相关(r=-0.358,P<0.05),与松质骨体积呈直线正相关(r=0.513,P<0.01)。结论:跟骨定量超声测量中,超声振幅衰减平均值能较好地反映骨的密度,超声传播速度能较好地反映骨的质量,而骨硬度指数能较综合地反映骨强度的改变。     

Ultrasonic enhancement of gene transfection in murine melanoma tumors

The enhancement of gene transfection by ultrasound (US) was evaluated in vitro and in vivo using the B16 mouse melanoma model. Cultured cells were either exposed in suspensions in vitro or implanted subcutaneously in female C57BL/6 mice for 10–14 days and, subsequently exposed, in vivo. For comparison to results with a luciferase plasmid, a reporter plasmid for green fluorescent protein (GFP) was used to evaluate transfection efficiency. US was supplied by a system, similar to a Dornier HM-3 lithotripter, that produced shock waves (SW) of 24.4 MPa peak positive and 5.2 MPa peak negative pressure amplitudes at the focus. The plasmids were mixed with the suspensions to achieve 20 μL mL⁻¹, or were injected intratumorally to provide 0.2 mg DNA per mL of tumor. Acoustic cavitation was promoted by retaining 0.2 mL of air in the 1.2-mL exposure chambers in vitro and by injecting air at 10% of tumor volume in vivo. In vitro, cell counts declined to 5.3% of shams after 800 SW exposure, with 1.4% of the cells expressing GFP after 2 days of culture. In vivo, 2 days after 400 SW exposure, viable-cell recovery from excised tumors was reduced to 4.2% of shams and cell transfection was enhanced by a factor of about 8, reaching 2.5% of cell counts (p < 0.005 in t-test). These results show that strong tumor ablation induced by US shock wave treatment can be coupled with simultaneous enhancement of gene transfection.     

Lentiviral Vectors and Adeno‐Associated Virus Vectors: Useful Tools for Gene Transfer in Pain Research

Pain, especially chronic pain, has always been a heated point in both basic and clinical researches since it puts heavy burdens on both individuals and the whole society. A better understanding of the role of biological molecules and various ionic channels involved in pain can shed light on the mechanism under pain and advocate the development of pain management. Using viral vectors to transfer specific genes at targeted sites is a promising method for both research and clinical applications. Lentiviral vectors and adeno‐associated virus (AAV) vectors which allow stable and long‐term expression of transgene in non‐dividing cells are widely applied in pain research. In this review, we thoroughly outline the structure, category, advantages and disadvantages and the delivery methods of lentiviral and AAV vectors. The methods through which lentiviral and AAV vectors are delivered to targeted sites are closely related with the sites, level and period of transgene expression. Focus is placed on the various delivery methods applied to deliver vectors to spinal cord and dorsal root ganglion both of which play important roles in primary nociception. Our goal is to provide insight into the features of these two viral vectors and which administration approach can be chosen for different pain researches. Anat Rec, 301:825–836, 2018. © 2017 The Authors. The Anatomical Record published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists.     

不同手术方式对前列腺增生患者生存质量的影响

目的比较不同手术方式对前列腺增生患者术后生存质量的影响。方法对采用开放性前列腺切除(OP)和经尿道电汽化切割(TUEVAP)治疗的83例前列腺增生(BPH)患者的国际前列腺症状评分(IPSS)和生存质量进行评价。结果两组患者手术时间、术中出血、前列腺切除量有显著性差异(P<0.05);术后并发症的发生和再手术率无显著性差异(P>0.05)。两组患者手术后IPSS评分和生存质量均较术前明显改善。其中TUEVAP组改善更为明显(P<0.05)。结论在BPH治疗过程中,必须结合术者的操作经验、设备状况和BPH患者的自身条件选择手术方式,不同的手术方式对手术病人生存质量将会有不同程度的影响。     

免疫磁性分选系统大量分选CD34+细胞的实验研究

CD3 4 细胞的分离纯化在自体外周血干细胞、异基因骨髓移植 /外周血干细胞移植及干细胞研究中具有重要的应用价值。为了摸索大量分选CD3 4 细胞的方法 ,本研究应用Isolex3 0 0i磁性分选系统富集CD3 4 细胞 ,采用流式细胞术监测分选前后细胞表面标志 ,经CD3 4 细胞体外增殖实验及克隆形成实验验证分选获得的CD3 4 细胞生物学活性。结果显示 ,所完成的 5例次自体外周血富集CD3 4 细胞时 ,先收获单个核细胞约 ( 3 .5 -6.0 )× 10 10 ,CD3 4 细胞占单个核细胞百分率为 ( 0 .5 5 - 1.2 ) % ;收获的CD3 4 阳性细胞总数为 ( 2 - 3 )× 10 8,纯度为 ( 75 - 85 ) % ,回收率为 ( 4 0 - 65 ) %。体外实验表明 ,在SCF IL 3 FL TPO EPO存在下 ,经过 3 - 4天培养 ,可扩增 2 - 3倍 ;经CFU GM、BFU E集落形成实验显示具有形成集落的能力 ,证实分选后细胞具有造血祖细胞生物活性。结论 :应用Isolex3 0 0iCD3 4 细胞分选仪可以高效大量富集CD3 4 细胞 ,适于临床应用。     

中频交变磁场照射后小鼠的行为学变化

目的:目前电磁辐射对生物体行为学方面的研究还很薄弱,设立不同磁场的强度和不同照射的周期,观察中频领域磁场照射对小鼠的自主活动和学习记忆的影响。方法:实验于2007-05-10/06-15在清华大学医学院和中国医学科学院药用植物研究所完成。①实验材料:磁场发生装置:中频交变磁场发生装置由清华大学工程物理系医学物理与工程研究所自主研发,可产生频率为40kHz,场强为28.8A/m,28.8kA/m的中频交变磁场。自主活动测试箱:中国医学科学院药用植物研究所提供,为一可封闭的金属箱,内置摄像头。4只黑色塑料测试桶分别置于箱子四角。水迷宫测试箱:中国医学科学院药用植物研究所提供。②实验动物及方法:将80只小鼠随机分为不同场强照射组和对照组。强磁场照射1组:11.6kA/m/40kHz,1h/d,连续照射7d。强磁场照射2组:11.6kA/m/40kHz,2h/d,连续照射7d。强磁场照射3组:11.6kA/m/40kHz,2h/d连续照射15d。弱磁场照射组:28.8kA/m/40kHz,2h/d,连续照射7d。对照组除了未放入磁场照射,其他条件与照射组一致。实验过程中对动物处置符合动物伦理学要求。③实验评估:观察其一般活动的改变,并采用自主活动测试箱和水迷宫测试箱进行测试,观察各组小鼠的自主活动和学习记忆的改变。结果:①一般行为观察:与对照组小鼠对比,经过中频交变磁场照射过的各组小鼠活跃度减低,毛色较差。强磁场照射3组(145G,2h/d,连续15d)于第11天和13天分别死亡1只。②自主活动检测结果:强磁场照射2,3组小鼠的运动路程、运动速度、运动时间明显低于对照组(P<0.05)。③水迷宫测试结果:在学习记忆检测阶段撤掉平台,发现各组动物2min内穿越平台原来所在象限的次数没有明显差别(P>0.05)。结论:中频交变磁场照射会给小鼠的自发活动造成一定影响,对学习记忆没有影响。     

两种微创治疗下肢静脉曲张的观察护理

目的观察微创方法治疗下肢静脉曲张患者的护理体会。方法本组115例下肢静脉曲张病人用微创治疗（激光腔内闭合治疗55例，射频腔内闭合治疗60例），采用小切口大隐静脉高位结扎、血管腔内原位闭合，属支及交通支电凝、结扎观察术后效果、并发症等。结果所有病人下肢肿胀、酸痛、乏力在手术后1个月逐步消失；皮肤色素沉着在术后1个月减轻，3个月逐步恢复。并发症的发生，激光和射频治疗组分别为：16.36％（9／55），16．67％（10／60）；全部病例无下肢深静脉血栓形成和切口感染。小腿皮肤麻木3～6个月逐步恢复，其余并发症在1个月内均恢复。结论微创治疗具有明显的优越性，但与传统手术一样，同样可能发生一些并发症，因此术后要严密观察和精心护理，积极配合医师完成整个治疗过程。     

应用28通道近红外光谱分析恢复过程中老年期抑郁症患者前额叶的激活特征

目的：认知功能障碍是老年期抑郁症的可逆性症状，试验以认知功能研究为突破口，采用28通道的近红外光谱技术分析处于恢复过程中的老年期抑郁症患者在执行词语流畅性测验时前额叶的激活特点，探讨老年期抑郁症的发病机制。 方法：①分组：患者组为2006～03／12首都医科大学附属北京安定医院收治的12例处于恢复过程中的老年期抑郁症患者，入组时符合DSM-4有关重度和中度抑郁发作或复发的标准诊断，首次发病年龄≥60岁，均服用SSRIs类抗抑郁药物治疗。对照组为12例性别、年龄和教育程度与患者组相匹配的健康志愿者。所有受试者排除脑器质性病变：②试验方法：所有受试者接受近红外光谱测查，在28通道的CW5（TechEnInc．American）NIRS系统上完成。通过近红外光谱系统监测被试者在执行词语流畅性测验任务过程中，前额叶氧合血红蛋白和脱氧血红蛋白浓度的变化，从而反映前额叶认知任务相关的激活效应。采用组块设计的词语流畅性测验作为认知激活任务，任务范式包括词语重复任务和词语流畅性任务两种试验。 结果：24例受试者均进入结果分析。①在健康老年人和处于恢复过程中的老年期抑郁症患者，词语流畅性测验能够激活双侧前额叶．未发现半球单侧化激活效应。②处于恢复过程中的老年期抑郁症患者在词语流畅性测验任务过程中，左侧前额叶激活效应较对照组减弱。③尽管在健康老年人和处于恢复过程中的老年期抑郁症患者，近红外光谱显示了不同的激活模式，但词语流畅性测验的成绩不存在显著性的组间差异（P〉0．05）。 结论：处于恢复过程中的老年期抑郁症患者存在词语流畅性测验相关的功能性左侧前额叶功能减退，微血管功能失调和多巴胺系统异常也许在老年期抑郁症的病理生理学机制中起重要作用。