无菌包的灭菌标记应贴在封口处

无菌包的灭菌标记应贴在封口处吴涛《基础护理学》一书中介绍供应室工作时，要求灭菌物品和未灭菌物品，必须严格分开放置，并作出标记。要求无菌包有标记，主要是指要标出灭菌时间和已灭菌的标记。目前，供应室无菌包的标记多采用“三Ｍ高压灭菌指示胶带”（以下简称胶带...     

淋巴显象剂硫化锑胶体冻干药盒的研制

＾９９ｍＴｃ标记的硫化锑胶体是目前性能最优的淋巴显象剂，但因无标记药盒供应，使推广应用受限。我们研制了一种硫化锑胶体冻干药盒，有效保存期达一年以上，复溶性能良好，颗粒均匀（２０￣４０ｎｍ），标记率约９４％，标记稳定性大于２４小时，经临床试用，其淋巴显象效果良好。     

5-IAF标记兔心肌肌钙蛋白C的荧光特性

背景：心肌肌钙蛋自在检测心肌梗死的诊断中有极其重要的临床意义。目的：将荧光标记物5-iodoaccetamidofluorescein（5-IAF）应用于心肌肌钙蛋白，观察其是否可用作与收缩调节蛋白之间相互作用的研究。设计、时间及地点：随机对照实验，于2002—01／2005—12在广州医学院人体解剖教研室完成。材料：成年兔，由广州医学院实验动物中心提供。方法：利用反转录一聚合酶链反应技术制备兔心肌肌钙蛋白C的基因片段，应用基因重组技术将兔心肌肌钙蛋白C基因片段克隆到pET表达载体，根据PCR定点诱变的方法，取得突变型的兔cTnC（C35S），分别将5-IAF和2-（4’-（iodoacetamido）anilino）naphthalene-6sulfonicacid（IAANS）标记在兔肌钙蛋白C（C35s），并进行恒态荧光测定和时间-分辨荧光测定。主要观察指标：兔cTnC（C35S），IAF及cTnC（C35S）-IAANS的荧光特性。结果：5-IAF标记的兔肌钙蛋白C在波长491nm处进行激发，其发射峰值出现在波长520nm处，加入饱和的镁离子可导致荧光强度减少35％，继续用饱和的钙离子滴定，其荧光强度再减少35％，而且波峰向蓝光方向移动3nm。IAANS标记的cTnC（C35S）在加进镁离子和钙离子后引起的荧光反射的跃迁，并不出现在用5-IAF标记的cTnC上。然而，不管使用哪一种荧光标记物，钙引起cTnC的构象变化这个作用并没有改变。钙滴定实验表明，两种荧光标记物标记的荧光发射结合参数是一致的。结论：5-IAF标记可以应用于心肌肌钙蛋白与其他收缩调节蛋白之间相互作用的研究。     

CM-DiI标记大鼠骨髓间充质干细胞的体外体内的示踪观察

目的建立骨髓间充质干细胞（BMSCs）的一个简单、有效的标记和示踪方法并对BMSCs移植治疗心肌梗死作初步观察，为进一步的研究提供依据。方法应用CM—DiI细胞标记液标记第三代大鼠BMSCs，应用荧光显微镜对标记的细胞进行体外示踪观察；8只SD大鼠，制作急性心肌梗死模型，将已标记CM—DiI的BMSCs经心肌注射途径移植给急性心肌梗死模型大鼠，在移植后1周、2周、3周、4周分别取心肌组织行冰冻切片，并在荧光显微镜下观察移植细胞；于移植后4周应用免疫组织化学法检测移植细胞心肌缝隙连接蛋白43（connexin 43）的表达。结果CM-DiI标记大鼠BMSCs效率达100％，标记后BMSCs生长和增殖特性未受影响，形态无改变，经过传代培养14d仍然保持较清晰的红色荧光。已标记细胞在大鼠心梗模型心肌内移植后1周、2周、3周、4周，心肌组织冰冻切片可找到移植细胞，CM—DiI与BMSCs结合稳定，荧光标记效果维持一个月无明显衰退，免疫组化切片显示移植细胞少量表达connexin43。结论CM—DiI细胞标记液标记BMSCs，对细胞生长特性无影响，操作简单，标记效果好，BMSCs移植到梗死心肌4周后仍存活，并少量表达心肌特异性蛋白connexin43。CM—DiI标记细胞法可用于进一步的BMSCs移植治疗心肌梗死的研究。     

药物介入及运动试验心肌断层显的临床应用

１１０例心绞痛及６０例心肌梗塞患者行药物介入及运动试验＾９９ｍＴｃ－ＭＩＢＩ心肌断层显象。结果表明，心肌断层显象诊断心绞痛及心肌梗塞的阳性率、可定位病灶节段数较ＥＣＧ均高，是一种有价值的玩创伤性诊断方法。但其诊断下壁、后壁心肌缺血及梗塞时要注意假阳性。     

药物介入及运动试验心肌断层显象的临床应用

110例心绞痛及60例心肌梗塞患者行药物介入及运动试验~(99m)Tc-MIBI心肌断层显象。结果表明,心肌断层显象诊断心绞痛及心肌梗塞的阳性率、可定位病灶节段数较ECG均高,是一种有价值的无创伤性诊断方法。但其诊断下壁、后壁心肌缺血及梗塞时要注意假阳性。     

加标记的心脏磁共振成像研究进展

加标记的心脏磁共振影像(tagged MRI)是20世纪80年代末期出现的一种医学成像技术.标记线的引入,为使用非介入方式研究心肌运动提供了新的方法.如何有效地获取和使用tagged MRI提供的信息成为当前医学影像领域的一个研究热点.作者结合近年来在tagged MRI所做的研究工作,从信息处理角度,对tag线跟踪、心脏MRI分割方法、左心室的形状恢复与运动重建、心肌形变应变研究和验证方法等进行了论述,并就该领域今后的研究方向提出了建议.     

混合抗体标记法在免疫组化技术中的应用及价值

免疫组化双标记染色技术是在一张组织切片上同时标记两种不同的抗体，可同步检测和观察两种不同抗原的表达情况，通常需要专用的试剂盒，而且操作过程复杂，染色时间长，即耗时又费力。我们用p63＋34βE12、p63＋SMA混合抗体法分别标记前列腺基底细胞及乳腺组织中的肌上皮细胞，既增加了单抗体标记法的敏感性，又达到了双标记法的染色效果，是一种简便易行、经济实用的免疫组化标记技术。     

流式细胞术检测胎儿红细胞的影响因素探讨

目的 探讨流式细胞术检测胎儿红细胞的影响因素。方法45名健康产妇脐血样本分别用同血型健康人外周血调整为5种浓度，使用抗胎儿血红蛋白（HbF）单克隆抗体，用直接标记和间接标记2种方法分别测定HbF阳性细胞比例．将样本放置24h、48h和7211后再次测定。结果以Fetal Cell Count TM Kit Ⅱ双色标记碳酸酐酶和HbF抗体[碳酸酐酶-异硫氰酸荧光素（CA-FITC）、HbF-藻红蛋白（PE）]为标准，5种浓度的直接标记结果均高于间接标记（P〈0．05），阳性细胞数低时差异有统计学意义（P〈0．01）。样本放置48h直接标记结果未见明显下降，间接标记结果下降明显，放置72h直接标记结果也有明显下降（P〈0．05）。结论利用流式细胞仪检测母血中胎儿红细胞含量受到标记方法、样本放置时间等因素的影响。较好的方法是采用直接标记法，染色完毕立即检测，最好不要超过3d，样本必须避光保存。     

羧基荧光素琥珀酰亚胺酯标记肿瘤细胞及其在细胞毒检测中的价值

目的探讨化学染料羧基荧光素二乙酸盐琥珀酰亚胺酯（CFDA-SE）标记肿瘤细胞的最佳条件及其在细胞毒检测中的价值。方法用不同浓度的CFDA-SE标记K562（人红白血病细胞）、YAC-1（小鼠淋巴瘤细胞）、A375（人乳腺癌细胞）、MCF-7（人黑色素瘤细胞）不同肿瘤细胞系，并检测其0—24h的荧光强度，观察荧光强度变化的时间动力学，选择CFDA-SE标记细胞的最佳浓度；CFDA-SE标记细胞后孵育时间为1—6h，再用DNA染料碘化丙啶（PI）标记，流式细胞仪分析CFDA-SE和PI双标记细胞，并计算细胞死亡率，研究CFDA-SE对细胞的毒性。CFDA-SE标记时间分别为5、6、7、8、10、15min，测定标记不同时间的荧光强度和细胞的死亡率，选择最佳标记时间；用CFSE在最佳条件下标记靶细胞进行细胞毒检测实验。结果对于不同肿瘤细胞系，CFDA-SE标记的最佳浓度不同；CFDA-SE对细胞没有毒性，死亡率低于5％；最佳标记时间为8min。人外周血单个核细胞和BALB／c鼠脾淋巴细胞对肿瘤细胞K562、YAC-1均表现出杀伤活性，杀伤百分率随效靶比和共孵育时间的增加而增加。最佳效靶比为50：1—25：1，共孵育时间为2—4h。结论CFDA-SE具有标记细胞稳定，能用于细胞培养时间较长的研究，不影响细胞功能，适用于流式细胞仪检测细胞毒实验的优点，是一种很好的标记细胞的荧光素染料。     

Pathology associated with a human case of rabies in the United Kingdom caused by European bat lyssavirus type-2

Human infection with the rabies-related virus European bat lyssavirus type-2 (EBLV-2) has only been reported on two occasions. Here we report the pathology observed within spinal cord and visceral tissues associated with EBLV-2 infection for the first time. Neuronal labelling with an anti-rabies nucleocapsid monoclonal antibody was observed and appeared indistinguishable from the labelling reported from human infection with rabies virus.     

转导野生型p53基因提高K562细胞对紫外线诱导的细胞凋亡的敏感性

目的：观察野生型ｐ５３基因转染的Ｋ５６２细胞对紫外线诱导其细胞凋亡的影响。方法：应用基因转染技术将重组野生型ｐ５３的逆转录病毒载体转导ｐ５３蛋白缺失的Ｋ５６２细胞，用紫外线照射Ｋ５６２ｐ５３细胞不同时间后再培养不同时间，用ＤＮＡ片段化和细胞ＤＮＡ原位末端标记技术检测细胞凋亡情况。结果：紫外线照射Ｋ５６２ｎｅｏ和Ｋ５６２ｐ５３细胞８分钟后再培养７～１２小时，Ｋ５６２ｎｅｏ和Ｋ５６２ｐ５３两组细胞的凋亡差异显著。结论：野生型Ｐ５３蛋白在Ｋ５６２细胞表达后，能加速紫外线诱导的Ｋ５６２细胞凋亡，为临床应用ｐ５３进行基因治疗提供了有意义的实验基础     

Labelling of olfactory ensheathing cells with micron‐sized particles of iron oxide and detection by MRI

A crucial issue in transplant‐mediated repair of the damaged central nervous system (CNS) is serial non‐invasive imaging of the transplanted cells, which has led to interest in the application of magnetic resonance imaging (MRI) combined with designated intracellular magnetic labels for cell tracking. Micron‐sized particles of iron oxide (MPIO) have been successfully used to track cells by MRI, yet there is relatively little known about either their suitability for efficient labelling of specific cell types, or their effects on cell viability. The purpose of this study was to develop a suitable MPIO labelling protocol for olfactory ensheathing cells (OECs), a type of glia used to promote the regeneration of CNS axons after transplantation into the injured CNS. Here, we demonstrate an OEC labelling efficiency of >90% with an MPIO incubation time as short as 6 h, enabling intracellular particle uptake for single‐cell detection by MRI without affecting cell proliferation, migration and viability. Moreover, MPIO are resolvable in OECs transplanted into the vitreous body of adult rat eyes, providing the first detailed protocol for efficient and safe MPIO labelling of OECs for non‐invasive MRI tracking of transplanted OECs in real time for use in studies of CNS repair and axon regeneration. Copyright © 2012 John Wiley & Sons, Ltd.     

In vitro and in vivo studies using 111 indium oxine, 111 indium oxine sulfate and 99m Tc oxine in erythrocyte labeling

The optimal conditions for red blood cell labelling using 111indium oxine, 111indium oxine sulphate and 99mTc oxine were established both in vitro as well as in vivo. The coagulant had no effect on labelling efficiency. Other variables such as the incubation time, temperature, duration, cell number and concentration of the complex exert a significant influence on labelling efficiency. Labelling efficiency of red blood cells is very high also under non-optimum conditions as compared with other cells (leucocytes, platelets).     

The Reliability of the Scholander-Roughton Method of Microdetermination of the Oxygen Content of Blood,Using Aerosol and Saponin as the Hemolytic Agents

The application of neutron activation paper chromatography has been shown to be most suitable for the analysis of phospholipids in clinical research where labelling of phospholipids with P³² is not possible and only a minute amount of sample is available.

This technique made it possible for the first time to analyze phospholipids in human liver biopsy samples and a comparison was made between human liver and plasma phospholipids. Much less phosphatidylethanola-mine and phosphoinositide were found in human plasma.

Further evidence was presented for the presence of lysolecithin in human plasma.     

Optimisation of the Synthesis and Cell Labelling Conditions for [89Zr]Zr-oxine and [89Zr]Zr-DFO-NCS: a Direct In Vitro Comparison in Cell Types with Distinct Therapeutic Applications

Background

There is a need to better characterise cell-based therapies in preclinical models to help facilitate their translation to humans. Long-term high-resolution tracking of the cells in vivo is often impossible due to unreliable methods. Radiolabelling of cells has the advantage of being able to reveal cellular kinetics in vivo over time. This study aimed to optimise the synthesis of the radiotracers [⁸⁹Zr]Zr-oxine (8-hydroxyquinoline) and [⁸⁹Zr]Zr-DFO-NCS (p-SCN-Bn-Deferoxamine) and to perform a direct comparison of the cell labelling efficiency using these radiotracers.

Procedures

Several parameters, such as buffers, pH, labelling time and temperature, were investigated to optimise the synthesis of [⁸⁹Zr]Zr-oxine and [⁸⁹Zr]Zr-DFO-NCS in order to reach a radiochemical conversion (RCC) of >95 % without purification. Radio-instant thin-layer chromatography (iTLC) and radio high-performance liquid chromatography (radio-HPLC) were used to determine the RCC. Cells were labelled with [⁸⁹Zr]Zr-oxine or [⁸⁹Zr]Zr-DFO-NCS. The cellular retention of ⁸⁹Zr and the labelling impact was determined by analysing the cellular functions, such as viability, proliferation, phagocytotic ability and phenotypic immunostaining.

Results

The optimised synthesis of [⁸⁹Zr]Zr-oxine and [⁸⁹Zr]Zr-DFO-NCS resulted in straightforward protocols not requiring additional purification. [⁸⁹Zr]Zr-oxine and [⁸⁹Zr]Zr-DFO-NCS were synthesised with an average RCC of 98.4 % (n = 16) and 98.0 % (n = 13), respectively. Cell labelling efficiencies were 63.9 % (n = 35) and 70.2 % (n = 30), respectively. ⁸⁹Zr labelling neither significantly affected the cell viability (cell viability loss was in the range of 1–8 % compared to its corresponding non-labelled cells, P value > 0.05) nor the cells’ proliferation rate. The phenotype of human decidual stromal cells (hDSC) and phagocytic function of rat bone-marrow-derived macrophages (rMac) was somewhat affected by radiolabelling.

Conclusions

Our study demonstrates that [⁸⁹Zr]Zr-oxine and [⁸⁹Zr]Zr-DFO-NCS are equally effective in cell labelling. However, [⁸⁹Zr]Zr-oxine was superior to [⁸⁹Zr]Zr-DFO-NCS with regard to long-term stability, cellular retention, minimal variation between cell types and cell labelling efficiency.

     

Quantification of feto-maternal haemorrhage (FMH) by flow cytometry: anti-fetal haemoglobin labelling potentially underestimates massive FMH in comparison to labelling with anti-D

Many centres now routinely use flow cytometry to quantify feto-maternal haemorrhage (FMH). However, which flow cytometric method is the most accurate in quantifying FMH is currently unknown. An audit of clinical results in which FMH had been estimated by both directly conjugated monoclonal anti-D and anti-fetal haemoglobin (HbF) labelling suggested that the anti-HbF labelling method may underestimate massive FMH in comparison to labelling with anti-D. Subsequent to this audit, 46 samples of adult D-negative blood were spiked with varying amounts of D-positive cord blood (0.05-10% fetal cells per sample), and the number of fetal cells present was quantified by both labelling methods. The percentage of fetal cells detected by anti-D was not significantly different to the estimated percentage of fetal cells added to each sample (P = 0.636). However, anti-HbF labelling significantly underestimated the percentage of fetal cells present (P = 0.0001). In comparison to anti-D, the percentage of fetal cells detected by anti-HbF was also significantly lower (P < 0.0001). The difference in fetal cell detection between anti-D and anti-HbF labelling was only apparent in the spiked samples containing > or =1% fetal cells per sample. In samples containing < or =0.6% fetal cells, no significant difference in the detection of fetal cells between anti-D and anti-HbF labelling was observed (P = 0.11). To allow adequate immunoprophylaxis in D-negative mothers with massive FMH, we recommend that anti-D labelling should be used in the routine flow cytometric estimation of FMH.     

Observations on the Chromium Labelling of ACD-Stored and Previously Frozen Red Cells

Chromium labelling characteristics of both ACD-stored and previously frozen red cells were evaluated. The chromium uptake of previously frozen red cells processed by agglomeration was inversely related to the hemoglobin level of the suspending fluid. Ascorbic acid was not needed for the labelling of previously frozen, agglomerated red cells.
Cellular injury, as measured by increase in supernatant hemoglobin during post-thaw storage at 4 C, occurred with the agglomerated, previously frozen red cells when: (1) Na₂ EDTA was present in the glycerolizing solution; (2) the disaggregation of the agglomerated red cell mass was carried out with 75 rather than 250 ml of isotonic saline; and (3) the storage temperature of the glycerolized red cells was interrupted for one week with a storage interim at either 4 C or −20 C.
By use of a phthalate ester technic, red cells were separated into three fractions on the basis of cellular density. Preferential chromium labelling of red cells was noted: the lightest fraction contained significantly more radioactivity than the heaviest fraction.     

Erythrocyte life span in growing swine as determined by glycine-2-C14

Red blood cell survival studies were performed on five normal growing swine by following the C¹⁴-specific activity of hemoglobin and heme after the administration of glycine-2-C¹⁴. The erythrocytes of normal growing swine appear to be destroyed both by a random and an age-dependent process. Random destruction accounts for the larger portion of the cells which are destroyed. The "mean" red cell survival time was 62 days. This represents the interval from the time of incorporation of 50 per cent of the maximal amount of labelling achieved to the time when the level had decreased once more to the 50 per cent amount. The " ‘corrected’ average potential life span" of the red cells was 86 ± 11.5 days. This figure was obtained by subtracting the number of days required to attain 80 per cent of the maximal labelling from the average survival time of red cells destroyed by an age-dependent process as distinguished from random destruction.