联合培养的成纤维细胞对平滑肌细胞增殖和形态的影响

目的:观察体外联合培养的成纤维细胞对平滑肌细胞形态和增殖的影响,为研究成纤维细胞和平滑肌细胞生物学特性和相互关系以及体外构建含此两种细胞的组织工程皮肤提供理论和实践基础。方法:在体外分别分离培养成纤维细胞和平滑肌细胞并常规传代培养。模拟组织工程皮肤的结构及成纤维细胞和平滑肌细胞间的相互影响途径,按照不同比例(平滑肌细胞:成纤维细胞=1:1,1:2,1:3)建立以胶原凝胶为基质的成纤维细胞与平滑肌细胞联合培养模型。应用荧光标记、组织学观察和MTT法对平滑肌细胞的形态和代谢进行研究。结果:联合培养模型法可以有效地建立起成纤维细胞和平滑肌细胞的相互影响模式。1:1组的成纤维细胞对联合培养的平滑肌细胞增殖有促进作用,1:3组的促增殖作用则不明显。结论:1:1联合培养的成纤维细胞对平滑肌细胞增殖有促进作用,对它们体外培养和扩增以及相互关系的研究对阐明这两种细胞组合作为真皮替代物种子细胞具有重要的意义。     

Three-dimensional recording of the human face with a 3D laser scanner.

Three-dimensional recording of the surface of the human body or of certain anatomical areas has gained an ever increasing importance in recent years. When recording living surfaces, such as the human face, not only has a varying degree of surface complexity to be accounted for, but also a variety of other factors, such as motion artefacts. It is of importance to establish standards for the recording procedure, which will optimise results and allow for better comparison and validation. In the study presented here, the faces of five male test persons were scanned in different experimental settings using non-contact 3D digitisers, type Minolta Vivid 910). Among others, the influence of the number of scanners used, the angle of recording, the head position of the test person, the impact of the examiner and of examination time on accuracy and precision of the virtual face models generated from the scanner data with specialised software were investigated. Computed data derived from the virtual models were compared to corresponding reference measurements carried out manually between defined landmarks on the test persons' faces. We describe experimental conditions that were of benefit in optimising the quality of scanner recording and the reliability of three-dimensional surface imaging. However, almost 50% of distances between landmarks derived from the virtual models deviated more than 2mm from the reference of manual measurements on the volunteers' faces.     

Modified FMCW system for non-contact sensing of human respiration

Abstract

Small displacement detection capability becomes an important key for developing non-contact sensor for human respiration based on radar system. Frequency modulated continuous wave (FMCW) radar has been widely studied and applied for many applications. With respect to the conventional perspective, large bandwidth is needed for detecting the small displacement related to the human respiration. Meanwhile, extracting the respiration pattern from Doppler response has a drawback in identifying the small displacement location. In this paper, the modification on FMCW system was proposed for obtaining the capability in detecting the human respiration and its distance from the radar. Detecting the phase value of the low pass filter (LPF), output from conventional FMCW was investigated and applied as a modification concept. The pattern, rate and amplitude of respiration are extracted from phase detector output. Beat frequency detection is still elaborated for synthesising the reference signal for phase detection. The result shows that the modified FMCW system proposed the capability of detecting the rate and amplitude respiration and location of the target.     

胎鼠脊髓神经干细胞与颌下腺颗粒曲管细胞联合培养的研究

目的研究颌下腺颗粒曲管细胞(GCT细胞)对脊髓神经干细胞(NSCs)增殖分化的影响。方法取胎鼠脊髓NSCs原代和传代培养,取成年雌、雄大鼠GCT细胞原代和传代培养,实验分为3组:NSCs和GCT细胞联合培养组;雌、雄大鼠GCT细胞培养上清液与NSCs培养液混合后培养NSCs;NSCs培养液组为对照组;免疫荧光细胞化学方法进行细胞鉴定,MTT法对比各组NSCs增殖活力。结果NSCs与GCT细胞联合培养7 d后Nestin阳性神经球数多于单独培养,雄性大鼠GCT细胞混合培养液组NSCs增殖活力最强,对照组NSC最弱,雌性混合培养液组介于两者之间。鉴定细胞球Nestin阳性,各组均有少量贴壁分化细胞。结论大鼠GCT细胞分泌物促进NSCs增殖,抑制分化,雄性大鼠GCT细胞作用比雌性大鼠显著。     

交感神经节在共同培养的心肌细胞诱导下神经元的生长和迁移

目的：探讨联合培养中大鼠心肌细胞对交感神经节神经元的生长和迁移的影响。方法：在体外建立大鼠交感神经节与心肌细胞联合培养的模型。用倒置相差显微镜观察交感神经节与心肌细胞联合培养不同时间的活细胞生长状况,计数神经纤维束的数目。并用Holmes还原银染色法计数神经元迁移的数目。结果：联合培养中由交感神经节组织块长出的神经纤维束数目、直径大于5μm的神经纤维束的数目以及由交感神经节组织块迁出的神经元的数目均较单纯交感神经节组织块培养的明显增加。结论：联合培养中分散的心肌细胞诱导培养的交感神经节神经元神经突起的生长和神经元向周围的迁移。     

非接触标测和频谱分析迷走神经介导的心房颤动

目的： 对在体犬迷走神经介导的心房颤动（房颤）进行非接触标测和频谱分析，以探讨其发生和维持机制。方法: 测定8只犬基础情况及双侧迷走神经刺激时心房有效不应期及其离散度，非接触标测和频谱分析房颤时左、右房的电活动。结果: 迷走神经刺激与基础情况相比，左、右心房有效不应期缩短，但有效不应期离散度增大仅见于左房。迷走神经刺激时房颤易诱发和维持，房颤显示反复有序的激动经优先传导路径传播仅见于左房；频谱分析显示左房的主导频谱高于右房［（12.5±1.5)Hz vs (9.3 ±1.2) Hz，P<0.05］。停止迷走神经刺激，左、右房房颤频谱降低［（9.2±0.5)Hz vs (8.5±0.6)Hz， P>0.05］，房颤自发终止。结论: 左、右房电生理特性改变、激动模式差异以及频谱梯度提示迷走神经介导的房颤发生和维持依赖于左房。     

Infertility: Enhancement of outcome from intracytoplasmic sperm injection: does co-culture or assisted hatching improve implantation rates?

In two separate prospectively randomized trials, intracytoplasmicsperm injection (ICSI) cycles were studied in a controlled mannerto monitor the effects of either bovine oviductal epithelialcell co-culture (n = 119) or assisted hatching by zona drilling(n = 100). In the first study, immediately following ICSI, alleggs were placed directly either onto partial monolayers ofbovine oviductal cells or into regular culture medium. Althoughthe embryo developmental rate was apparently compromised inpart by the presence of the co-culture cells, ultimately therewere no significant differences in either the viable pregnancyrate (31.6% co-culture versus 29.0% control) or the embryonicimplantation rate (11.4% co-culture versus 13.6% control). Assistedhatching also had no significant impact on ICSI cycle outcomein terms of either the viable pregnancy rate (30.0% assistedhatching versus 32.0% control) or the embryonic implantationrate (8.5% assisted hatching versus 13.5% control). However,in female patients aged 235 years, assisted hatching appearedto convey a marginally significant benefit in terms of boththe viable pregnancy rate (35.5% assisted hatching versus 11.1%control) and the embryonic implantation rate (103% assistedhatching versus 3.1 % control). It seems that the overall improvementof ICSI cycle outcome cannot be achieved by the general applicationof either co-culture or assisted hatching. Nevertheless, itis possible that there remain specific patient groups that mightbenefit from selected use of either of these modalities.     

Fertilization and early embryology: Effects of embryo density and co-culture of unfertilized oocytes on embryonic development of in-vitro fertilized mouse embryos

We have evaluated the effects of embryo density and the co-cultureof unfertilized (degenerating) oocytes on the development ofin-vitro fertilized (IVF) mouse embryos. In experiment 1, groupsof one, five, 10 or 20 zygotes were cultured in 20 µldrops of modified human tubal fluid (HTF) medium for 168 h at38.7°C in 5% CO₂ and 95% air. As the embryo density increased,significantly (P < 0.05) higher rates of embryos reachedhatched blastocyst stage. In addition, the time required forhatching after IVF was significantly (P < 0.05) shortenedby the increase in embryo density. In experiment 2, 10 IVF zygoteswere cultured with or without 10 unfertilized (degenerating)oocytes in 20 µl drops of HTF medium. The rates of IVFembryos that developed to morula, blastocyst, expanded blastocystand hatched blastocyst stages were decreased significantly (P< 0.01) by culturing embryos with unfertilized oocytes comparedwith culturing embryos alone. In experiment 3, groups of oneor 10 IVF zygotes or 10 IVF zygotes plus 10 unfertilized oocyteswere cultured in 20 µl drops of HTF medium and the numberof cells per blastocyst was examined at 120 h after IVF. Increasingembryo density resulted in a significant (P < 0.05) increasein the number of cells per blastocyst. In contrast, the cellnumber of IVF embryos that developed to blastocyst decreasedsignificantly (P < 0.05) when they were cultured with unfertilizedoocytes. The results suggest that in-vitro development of IVFmouse embryos is enhanced by increasing embryo density and isimpaired by co-culture with unfertilized (degenerating) oocytes.     

IL-18在体外培养系统中诱导肿瘤快速杀伤效应及肿瘤特异性CTL

目的应用rhlL-18在体外培养系统(Coculture system in vitro，CCs)中诱导快速肿瘤杀伤效应及诱导肿瘤特异性细胞毒性T淋巴细胞(Cytotoxic T Lymphocyte，CTL)。方法 采用StemSep^TM免疫磁性细胞分离法分离人外周血NK细胞、T细胞及树突细胞(DCs)，流式细胞仪分析细胞表型，125I-UdR标记的细胞毒实验检测杀伤活性，ELISA方法检测IFN-7的产生量。结果 在CCs中，rhIL-18诱导出快速肿瘤杀伤效应，这种杀伤效应无抗原特异性、不受MHC限制，DCs和T细胞的存在与否对其无明显影响。在同一培养系统中，肿瘤抗原存在的条件下，96h后，rhIL-18能够诱导并促进CTL介导的肿瘤特异性杀伤效应。结论 rhIL-18能够在体外培养系统中相继诱导肿瘤快速杀伤效应及肿瘤特异性CTL。     

Ultrastructure of preimplantation human embryos co-cultured with human ampullary cells

Ova with two pronuclei were co-cultured with established human ampullary cell lines and various stages of preimplantation embryonic development were monitored by Nomarski optics and then assessed by transmission electron microscopy (TEM). Fifteen embryos ranging from the 2-cell stage to blastocyst hatching were examined for normal and abnormal features. Their ultrastructure was similar to that of embryos cultured in Whittingham's T6 medium, reported previously. Seven embryos were evidently morphologically normal and showed good organization of fine structure. Most cellular organelles underwent progressive changes during early development. There was evidence of enhanced embryonic genome activation at the 8-cell stage. Invariably, all embryos had few too many fragments, some internalized, which were later segregated into the blastocoele or found outside the trophoblast of the late morula and blastocysts. Six grossly 'normal' embryos assessed by Nomarski had multiple nuclei of various dimensions, which highlights the subjectivity of embryo assessment in the IVF laboratory. Incomplete incorporation of chromatin into nuclei and formation of micronuclei were evident in some blastomeres. The results are discussed in relation to early embryonic loss, prevalent in IVF. Significant events reported include the detection of centrioles at the 8-cell stage, cavitation of the early blastocyst and the initiation of blastocyst hatching visualized by TEM.