同步辐射光源相衬成像技术在形态学研究中的应用

目的探讨同步辐射相位衬度成像在形态学研究中的应用前景。方法以北京同步辐射装置（BSRF）4W1A束线提供的硬X线,采用衍射增强成像（DEI）和类同轴成像法对经10％福尔马林固定的正常猪、大鼠、小鼠的肝、肾、肺、骨等组织进行成像,并与成像样品的相邻组织及成像后原位组织切片的光镜染色结果进行对比分析。结果衍射增强成像显示大鼠肝脏血管的逐级分支,走行方向清楚,最细可分辨直径为30μm的血管;猪肝脏中央静脉、肝小叶分界隐约可见;大鼠肾脏皮、髓质分界清楚,弓形动脉、静脉及髓质区集合管、乳头管结构清晰可见,最细可分辨直径为30μm的管道结构;小鼠肺支气管树分支清楚,重叠肺泡隐约可见。类同轴成像大鼠股骨头关节软骨及骨小梁结构清楚。结论利用同步辐射相位衬度成像方法,样品无需制片处理即可观察到光镜及其他影像学技术尚不能直接观察到的脉管系统模式结构及其与周围组织的结构关系,此外还可以观察到常规X线成像方法无法观察到的软骨影像,比较适合形态学研究。     

67例未分化结缔组织病患者尿隐血和尿红细胞位相临床追踪观察

目的:追踪观察未分化结缔组织病(undifferentiated connective tissue disease,UCTD)患者尿隐血和尿红细胞位相的动态变化。方法:选取67例UCTD患者,每3个月进行中段尿尿液分析和尿红细胞位相检查,追踪1年,观察各指标变化情况。结果:一年后,67例UCTD患者尿隐血阳性由纳入观察时的48例(71.64%)上升到55例(82.09%),尿红细胞位相异常(畸形红细胞>80%)者由55例(82.09%)上升到60例(89.55%)。尿蛋白阳性者由11例(16.42%)上升到17例(25.37%)。67例患者5次检查尿隐血阳性率及尿红细胞位相异常阳性率均高于尿蛋白阳性率,差异有统计学意义(χ2分别为9.52,12.36,P值均<0.05)。结论:UCTD患者有尿隐血和尿红细胞位相异常,轻微但缓慢加重,可能与潜在肾损害有关。     

1，25－双羟维生素D_3对培养人角朊细胞增殖，细胞形态及HLA－DR，ICAM－1表达的影响

用体外培养人角朊细胞研究了１．２５－双羟维生素Ｄ_３（１，２５（ＯＨ）_２Ｄ_３）对角朊细胞增殖，细胞形态及ＨＬＡ－ＤＲ，ＩＣＡＭ－１分子表达的影响，培养液中加入１，２５（ＯＨ）_２Ｄ_３可使细胞增殖明显抑制。用流式细胞仪测定，１，２５（ＯＨ）_２Ｄ_３处理６天后角朊细胞体积较未处理的明显变大。用免疫组化染色显示１，２５（ＯＨ）_２Ｄ_３对经ＩＦＮ－γ和ＴＮＦａ刺激的角朊细胞ＨＬＡ－ＤＲ和ＩＣＡＭ－１分子表达无明显影响。     

Interneurons in the tritocerebrum of the crayfish

In isolated head preparations of the freshwater crayfish Orconectes limosus 268 local and projecting interneurons with branches in the tritocerebrum have been penetrated with glass microelectrodes and characterized for their sensory inputs. Using 3 criteria (sensory modality, site of receptors, response type of interneurons), the interneurons found were divided into 16 classes. The interneurons were either unimodal mechanoreceptive (89%) or bimodal (9% responding to mechanical and chemical stimuli, 2% responding to mechanical and visual stimuli). No trimodal interneurons were found. Within each modality the neurons received mostly bilateral input (70% of all interneurons responding to antennal stimulation, 84% of all chemosensitive interneurons). If the input was lateralized it was more often ipsilateral. The types of interneuronal responses evoked by sensory stimulation were: neurons that were exclusively excited (84%), those that were exclusively inhibited (10%), those that were excited or inhibited depending on the modality or laterality of the stimulus (6%), those showing long lasting excitatory aftereffects (3%), and those showing excitation or inhibition upon identical stimulation depending on the state of the neurons while being stimulated (1%). Interneurons that responded to mechanical antennal stimulation responded best either to low (10 Hz) or to high (100 Hz) stimulus frequencies. Six neurons responded best to a certain phase relationship between the movements of both antennal flagellae.     

利用小鼠胚胎干细胞贴壁制备拟胚体的新方法研究

目的为改进拟胚体制备过程中分化不同步等问题,探讨建立小鼠胚胎干细胞贴壁制备拟胚体的新方法。方法沈阳军区总医院心内科于2004年10月至2006年1月对小鼠进行研究,当小鼠胚胎干细胞R1生长至70%～80%亚融合时,将其消化成单细胞,以1×106的细胞数接种到直径100mm组织培养皿中,用含低浓度白血病抑制因子(LIF)1μg/L的胚胎干细胞培养液连续贴壁培养3d后,收集贴壁细胞团继续悬浮培养。对悬浮培养的拟胚体及其冰冻切片行形态学观察,对悬浮及贴壁分化的拟胚体行甲胎蛋白(AFP)、血小板内皮细胞粘附分子-1(PECAM-1)及神经丝蛋白68KD(NF-68)免疫荧光染色。结果形态学观察用本法得到的拟胚体大小均一,分化同步,能展示从简单拟胚体到成熟囊性拟胚体的典型发育过程。免疫荧光染色显示,AFP、PECAM-1及NF-68三种标志物表达均为阳性。结论与传统方法相比,该法操作简便、拟胚体形成率高、分化阶段同步,具有分化为3个胚层来源细胞的能力,可作为胚胎早期发育和胚胎干细胞分化等研究的理想工具。     

藤梨根提取物对大肠癌LoVo细胞增殖的抑制作用及诱导凋亡的影响

目的:研究藤梨根提取物(ethanol extract from radix of actinidia chinensis,EERAC)对人大肠癌LoVo细胞增殖和凋亡的影响.方法:提取藤梨根抗癌有效活性成分(EERAC),按浓度分为4处理组(10、40、160、320mg/L)和空白对照组(0mg/L).各实验组经作用24、48、72h后,进行一般形态学和AO/EB荧光染色观察;MMT法检测细胞增殖的抑制情况;免疫组织化学(immunohistochemistry,IHC)法测定LoVo细胞中凋亡相关基因Bcl-2、Bax、Caspase-3的蛋白表达变化.结果:与空白对照组比较,一般形态学显示EERAC处理组能使细胞密度减低,增殖变慢;细胞逐渐变大,细胞间接触变松,胞浆中颗粒增多,细胞脱壁现象和周围碎片增多;荧光染色观察可见处理组细胞呈橙红色荧光,细胞核出现碎片状或固缩状的凋亡特征学形态改变,凋亡现象与EERAC的浓度呈正相关性;MTT法检测显示,EERAC处理组对LoVo细胞的最佳作用时间为72h,最大抑制率为79.48%,具有浓度和时间的依赖性(P<0.01);IHC检测结果显示EERAC作用LoVo细胞24h后,Bcl-2表达明显减弱,Bax、Caspase-3表达水平明显增高,Bcl-2/Bax比值下降,差异具有统计学意义(P<0.05),其效应与浓度相关.结论:EERAC具有明显抑制LoVo细胞增殖的作用,其机制可能与降低Bcl-2表达,上调Bax、Caspase-3的表达水平,激活线粒体凋亡途径有关.     

髂前上棘、耻骨联合及股骨头中心的影像定位与体表关系

目的通过X线骨盆平片测量,计算股骨头中心与这两点的关系。分析其特点,找到一个简便、相对准确的体表确定股骨头运动中心的方法。方法门诊选取健康男女性各20人﹙无腰髋症状及病史﹚,得到年龄、身高、体重。摄标准骨盆正位片,拍照前体表找到髂前上棘、定位,于其上放置参照物。使用测量软件测量骨盆宽度、高度、髂耻线及股骨头中心数据。将各项参数收集后进行统计学分析。结果股骨头中心距髂耻连线距离平均为2.72cm左右。股骨头中心在髂耻连线的投影点均位于髂耻连线中点的外侧,距离髂耻中点1.08cm左右。结论股骨头运动中心是髋膝关节置换及骨折手术的重要标志,通过髂前上棘、耻骨联合等体表标志确定股骨头中心简便易行、值得推广,可作为临床手术定位的辅助。     

水牛痒螨和兔痒螨的形态学观察

通过光镜及扫描电镜对采自四川西部的水牛痒螨和兔痒螨进行了形态学观察。水牛痒螨幼虫大小为(283·1±32·6)μm×(202·3±42·5)μm,表纹多为横纹,假头鄂基及腹面第1、2对足基部为竖纹或未形成明显表纹;若虫大小为(392·6±101·2)μm×(299·4±73·0)μm,腹面第4对足之间具有角质窝,雌若虫尾部具有1对性乳突;雌螨大小为(498·5±88·4)μm×(357·9±61·9)μm,产卵孔和阴道分别位于虫体腹面中部和末端,肛门位于虫体背面末端;雄螨大小为(388·4±78·8)μm×(305·0±54·3)μm,尾部有瘤状突起、性吸盘和生殖孔。兔痒螨和水牛痒螨各期形态相似。兔痒螨幼虫大小为(279±60·64)μm×(180±56·37)μm;若虫大小为(496±76·67)μm×(356±41·73)μm;雌螨大小为(624·78±76·69)μm×(400·87±59·84)μm;雄螨大小为(435±64·68)μm×(330±39·72)μm。2种痒螨若虫腹中部均观察到数量不等的角质窝,首次观察到2种痒螨成虫第1对足跗节上具有一圆形突起、足末端分两节的吸盘吸柄和雌螨第4对足附近的角质窝。     

The influences of muscle fibre proportions and areas upon EMG during maximal dynamic knee extensions

This study is an investigation of the relationship between muscle morphology and surface electromyographic (EMG) parameters [mean frequency of the power spectrum (MNF), signal amplitude (root mean square, RMS) and the signal amplitude ratio (SAR; i.e. the ratio between the RMS level during the passive part of the contraction cycle and the RMS level during the active part of the contraction cycle)] during 100 maximal dynamic knee extensions at 90° · s⁻¹. Each contraction cycle comprised of 1 s of active knee extension and 1 s of passive knee flexion. The surface EMG was recorded from the vastus lateralis muscle. Twenty clinically healthy subjects participated in the study, and muscle biopsy samples of the vastus lateralis were obtained from 19 of those subjects. The relationships between muscle morphology and EMG were investigated at three stages of the test: initially, during the fatigue phase (initial 40 contractions), and at the endurance level (the final 50 contractions). Major findings on correlations are that SAR and MNF tended to correlate positively with the proportion of type 1 fibres, and RMS correlated positively with the proportion of type 2 muscle fibres. The muscle fibre areas showed little correlation with the EMG variables under investigation. The results of the present study showed that the three EMG variables of a dynamic endurance test that were investigated (RMS, MNF and SAR) were clearly correlated with the proportions of the different fibre types, but only to a small extent with fibre areas. These findings contradict some of the theoretical models of the EMG, especially for parameters in the frequency domain. Accepted: 17 June 1999     

不同类型异基因造血干细胞移植后的急性白血病患者骨髓形态学比较

目的观察单体型相合造血干细胞移植及全相合异基因造血干细胞移植后骨髓形态学差异及造血重建情况。方法在235名实施异基因造血干细胞移植患者中,单体型相合120名,采用外周血造血干细胞联合骨髓移植;115名全相合的患者,采用单纯的外周血造血干细胞移植。在造血干细胞移植成功出层流室+30 d行骨髓穿刺抽吸涂片观察骨髓形态学差异,并观察造血重建情况。结果 235名患者均成功重建造血。全相合患者白细胞及血小板重建的时间分别14.7 d(12～26 d)及20.6 d(13～32 d);单体型相合造血干细胞移植患者白细胞及血小板重建的时间分别12 d(10～21 d)及18 d(15～30 d);2者之间的差异具有统计学意义(P0.05)。2组患者造血干细胞移植后骨髓全部完全缓解。在骨髓增生程度、粒红比例和细胞形态等方面有差异,但差异无统计学意义(P0.05)。结论单体型相合造血干细胞移植较全相合移植造血重建早,移植后骨髓形态学有差异,但差异无统计学意义。