胆汁微泡附加压力及其成核意义的分析

根据压力因素对结晶过程的影响，讨论了胆汁微泡的附加压力及其成核意义；分析了胆固醇单水结晶只从胆汁微泡中生成的原因；以附加压力观点，说明了胆汁微泡聚集、融合现象的原因和意义。     

一种适合于中枢内小核团注射HRP溶液的方法——套管式微玻管注射法

在中枢内小核团注射HRP溶液时,常碰到由于注射量不易控制而造成注入量过多,注射范围过大的问题。我们通过实验试制成套管式微玻管。用这种微玻管注射HRP溶液,设备简单,操作方便,能够较好地控制注入量和注射范围,也能较好地避免针道污染,使向中枢     

胞质内单精子注射与遗传学检测

胞质内单精子注射 (ICSI)为治疗男性不育发挥了重要的作用 ,但是由于男性的遗传因素 ,如染色体异常、CF基因突变、Y染色体微缺失等所引起的男性不育将会给所生后代带来遗传学缺陷。本文综述了 ICSI技术为治疗上述男性不育所要进行的遗传学检测及其存在问题与展望     

小鼠卵细胞质内显微注入单精子受精的实验研究

为了探讨提高小鼠卵细胞质内单精子注入受精率的方法，选取鼠龄１２－１４周的健康昆明白小鼠作为精子和卵子的供体，受用ＩＣＳＩ技术，以受精后二细胞卵裂的形成率为指标，了解不同采卵时间，不同微注射针参数及不同培养液对细胞质单精子注入的影响。结果表明，ｈＣＧ注射后１８－１９ｈ采卵，用针尖内径为４－５μｍ，斜面角度为３５－４０度的微注射针进行ＩＣＳＩ操作受精后卵子置ＣＺＢ中培养可获得较多的２细胞胚，卵裂率明显     

胞质内单精子注射与遗传学检测

胞质内单精子注射（ＩＣＳＩ）为治疗男性不育发挥了重要的作用,但是由于男性的遗传因素,如染色体异常、ＣＦ基因突变、Ｙ染色体微缺失等所引起的男性不育将会给所生后代带来遗传学缺陷。本综述了ＩＣＳＩ技术为治疗上述男性不育所要进行的遗传学检测及其存在的问题与展望。     

利培酮微球注射过程及注射部位疼痛的调查:12周多中心评价

背景:注射用利培酮微球是第一个新型抗精神病药的长效制剂,它的疗效和安全性已经得到证实。但是因其注射部位为臀部深部肌肉注射,可能引起较重的疼痛刺激。目的:明确注射用利培酮微球治疗12周后注射部位疼痛情况的变化,探索注射过程及注射部位疼痛与注射剂量、注射次数的关系。方法:选择符合DSM-IV精神分裂症诊断标准的住院或门诊患者57例,年龄18～65岁,每2周注射一次利培酮微球,可根据病情调整剂量为25,37.5,50mg,同时采用100mm目测类比量表对患者注射过程疼痛和注射部位的疼痛程度进行评价,研究者应用他评量表对受试者疼痛情况进行调查,分析注射剂量、注射次数对注射过程疼痛、注射部位疼痛的影响。结果与结论:利培酮微球注射的不同剂量组间注射过程的疼痛程度差异无显著性意义(F=1.35,P0.05),说明注射过程的疼痛与注射用利培酮微球注射剂量无关;注射部位的疼痛程度与利培酮微球的注射剂量有相关性,50mg组的疼痛程度较37.5,25mg组的注射部位的疼痛程度大,明确临床注射过程中应对高剂量利培酮微球注射患者进行重点的护理干预。     

河南单精子显微注射辅助生殖技术获成功

河南医科大学第一附属医院开展单精子显微注射辅助生殖技术获得成功。据专家介绍，第一代试管婴儿技术主要解决女性因素导致的不孕问题。第二代试管婴儿技术又称单精子显微注射试管婴儿，单精子显微注射是目前国内最先进的辅助生殖技术。实施方案是将一个精子注射入一个成...     

革兰氏阴性杆菌发生过程控制的研究

本研究选择奇异变形杆菌和绿肿杆菌等作为实验体系，从群体发生发展过程的宏观表现上探讨了生物控制的时空表达及非线性问题。结果表明，杆菌发生过程的时相周期性可以表现为空间的波形靶图样，并具有分形的特征。实验结果表明，菌群在生长过程中的代谢产物是富含生物信息的控制中介机制。我们认为，生物系统局部存在着准平衡和准封闭的微生态环境，生物液晶的存在和相变是细菌群体发生过程的重要协同中介因素。     

应用微丸脉冲注射与压电传感器提高放射性核素弹丸注射的安全性

放射性核素动态显像通常要求显像剂以弹丸注射的方法进行静脉注射。由于人工操作存在误差和放射性伤害的限制，使用自动注射装置代替人工进行弹丸注射具备应用潜力。本研究使用可编程注射泵对微丸脉冲注射的效果进行了对比和热实验验证，并使用压电传感器预压法进行整体气泡识别实验和大鼠尾静脉模拟注射验证。结果表明，在相同注射峰值速度下，微丸脉冲冲洗（约83μL/脉冲）的有效冲洗体积相比匀速注射降低了49.65%，相比人工冲洗降低了25.77%。为避免长管道对药液体积的稀释影响，使用压电传感器进行密封预压检测法能够较为精确地预测注射器内100μL以上的气泡。在大鼠尾静脉模拟注射实验中，通过预压100μL生理盐水，将针头置于不同组织时，在1 s内压电传感器反馈了差异较大的压强衰减率，分别为肌肉2.78%、皮下17.28%和静脉54.71%。微丸脉冲注射法和压电传感器密封预压法在提高放射性核素自动弹丸注射的安全性方面具备应用潜力。     

乳酸-羟基乙酸共聚物缓控释微球的制备及突释成因与影响因素

背景：乳酸-羟基乙酸共聚物是一种生物可降解高分子材料,以乳酸-羟基乙酸共聚物为原料制备的载药微球和纳米粒既可提高药物的稳定性,又能实现缓释、控释和靶向释放。 目的：分析乳酸-羟基乙酸共聚物缓控释微球的制备方法以及突释的成因、影响因素和改进方法。 方法：应用计算机检索1990/2010中国期刊全文数据库和PubMed数据库与乳酸-羟基乙酸共聚物缓控释微球的制备及突释联系紧密的文章。 结果与结论：目前乳酸-羟基乙酸共聚物缓释微球制备方法主要有单凝聚法、乳化-固化法、喷雾干燥法。造成其突释的原因首先是药物分子和聚合物分子之间的相互作用太弱,导致药物很容易从微球进入释放递质中,其次是在微球释放初期,药物从微球中的孔洞和缝隙中释放出来导致突释。影响突释程度的具体因素有乳酸-羟基乙酸共聚物的相对分子质量、浓度、微球载药量、主药理化性质、微球制备方法及制备参数等。虽然国内外对突释机制以及控制突释措施的研究都还处于初步阶段,通过对各影响因素加以适当优化与控制,可在一定程度上减少微球的突释率,突释问题应该能够得到解决和控制。     

Comparison of methods for introducing vectors based on bovine papillomavirus-1 DNA into mammalian cells

The intracellular structure of several vectors based on BPV-1 DNA has been analyzed following transfection into mouse C127 cells by the calcium phosphate method or, for the first time, by microinjection directly into the nucleus. It is shown that the method of introduction markedly affects the fate of a BPV-1 based vector. In general, microinjection appears to do little damage to DNA and is more likely to result in a vector replicating extrachromosomally as a monomeric structure of the same size as the input DNA. The method of selection for transformed cells, e.g., focus formation versus resistance to the neomycin analog G418, can also affect the intracellular state of the BPV-1 vector DNA. The nature of the recipient mammalian cell also influences whether a vector can replicate extrachromosomally or whether it integrates. BPV-1 based vectors, which replicated predominantly as multicopy intact extrachromosomal forms in mouse C127 cells, were always found to have integrated at low copy number in mouse LtAp20 cells.     

Infection of human cells with polyoma virus

Human cells which are resistant to infection with polyoma virus (PV) and PV-DNA by the conventional virus-absorption method were infected by microinjection of the virus or the viral-DNA into the cell cytoplasm. Induction of PV-tumor (T) antigen production and synthesis of cellular DNA were found in a large number of the infected cells, but no viral capsid (V) antigen was detected. Among 3 × 10³ virus-injected cells, none developed into a transformed cell colony. Multinucleated human muscle cells (myotubes), in which DNA synthesis and mitosis are irreversibly repressed after microinjection of PV, do not incorporate [³H]thymidine but produce T-antigen.     

Inhibition of SV40-induced cellular DNA synthesis by microinjection of monoclonal antibodies

The region of the SV40 large T-antigen molecule recognized by a panel of monoclonal antibodies has been determined using hybrid Adeno-SV40 viruses, and manual microinjection of cloned deletion mutants. In addition, an investigation was made of how monoclonal antibodies microinjected into the nucleus can affect the ability of the T-antigen coding gene to stimulate cell DNA synthesis. The monoclonal antibody Pab 14, that recognized the -COOH terminal half of large T, was comicroinjected into quiescent cells together with plasmid pCl-1. This plasmid contains only that part of the T-antigen coding gene that extends from nucleotide residue 120, counterclockwise to nucleotide residue 4002, and makes a truncated T antigen 33,000 in molecular weight and missing the last 435 amino acids on the -COOH terminal side. Monoclonal antibody Pab 14 did not inhibit the stimulation of cellular DNA synthesis caused by microinjection of pCl-1, although it did inhibit cell DNA synthesis induced by microinjection of pSV2G, a recombinant plasmid that contains the entire T-antigen coding gene of SV40.     

急性脑缺血损伤大鼠海马神经元谷氨酸转运体的表达

目的：研究急性脑缺血损伤大鼠海马神经元谷氨酸转运体（EAAC1）的表达变化。 方法： 采用EAAC1反义寡核苷酸脑内注射，用插线法建立大鼠局灶性脑缺血模型(MCAO)。运用Western blot法和TTC染色观察缺血区EAAC1表达和梗塞体积；采用RT-PCR 和Western blot法，测定海马EAAC1 mRNA和蛋白在缺血1 h、6 h、24 h的变化。结果： 注射EAAC1反义寡核苷酸组大鼠梗塞体积［(105.67±8.70) mm3］显著小于正义组。缺血1 h大鼠海马EAAC1 mRNA表达（0.963±0.117）与假手术组（0.907±0.113）无明显差异，缺血6h、24h持续高于缺血1 h（分别为1.116±0.104和1.428±0.078）。而海马EAAC1蛋白表达（0.640±0.027）在缺血24 h高于假手术组，缺血1 h和6h EAAC1表达与假手术组比较无显著差异（分别为0.330±0.018、0.330±0.015）。结论： EAAC1可促进缺血脑损伤,在急性脑缺血病理过程中表达增加。     

Pertussis toxin administration increases the expression of proneurotensin and preproenkephalin A mRNAs in rat striatum.

The effect of a unilateral intrastriatal microinjection of pertussis toxin on the expression of proneurotensin and preproenkephalin A mRNAs in the adult rat neostriatum was investigated using a technique of non-radioactive in situ hybridization. Control sham microinjected animals received an equal volume of vehicle only and were processed in parallel with the pertussis toxin-treated rats. All rats were allowed to recover from the stereotaxic surgery for 22 h before being killed and their brains rapidly removed and processed for in situ hybridization using alkaline phosphatase-labelled oligonucleotide probes. In comparison to sham microinjected rats, a single intrastriatal microinjection of pertussis toxin (1 microgram) resulted in a significant increase in the amount of both proneurotensin and preproenkephalin A mRNAs in the ipsilateral neostriatum. For proneurotensin mRNA, this increase was reflected by a substantial increase in the number of mRNA-containing cells detected. Proneurotensin mRNA-containing cells detected in the nucleus accumbens appeared to be unaffected by the intrastriatal pertussis toxin microinjection. In contrast, the significant increase in preproenkephalin A mRNA, when compared to the contralateral uninjected striatum and the ipsilateral striatum of control sham injected rats, was reflected by an increase in the cellular amount of preproenkephalin A mRNA and not by an increase in the number of mRNA-containing cells detected. These results demonstrate that the expression of both proneurotensin mRNA and preproenkephalin A mRNA in the adult rat striatum are rapidly increased in vivo by an intrastriatal microinjection of pertussis toxin.     

Selective immortalization of mammary epithelial cells by microinjection of SV40 DNA into human milk cells

Immortalization of mammary epithelial cells has been obtained by microinjection of SV40 DNA into epithelial cells from primary cultures of rabbit mammary glands. A modification of this technique, allowing selective immortalization of a specific cell type of mammary epithelial cells from human milk by microinjection of SV40 DNA, is described. It includes the selection of luminal epithelial cells from human milk cells after identification of these cells by antibodies directed against specific markers such as intermediate filaments and membrane antigens, and finally, the introduction of SV40 DNA into cell nuclei which will selectively immortalize this restricted cell population. This procedure can be used to derive permanent cell lines with a defined phenotype when only a few cells are available within a heterogeneous cell population.     

Vasopressin release and drinking induced by intracranial injection of angiotensin II in monkey.

The effects of intracerebral injection of angiotensin II (AII) on both water intake and arginine-vasopressin (AVP) release were tested on unanesthetized rhesus monkeys (Macaca mulatta). Injection of 10(-10) mol of peptide was administered with a cannula microinjection system stereotaxically implanted into different diencephalic structures. The preoptic area, anterior part of third ventricle, caudate nucleus, and septum appeared to be the injection sites most effective in eliciting both drinking behavior and AVP release when the animal did not have access to water. On the contrary, when water was presented, AVP release was blocked after AII microinjections in the preoptic area and the third ventricle. No drinking was observed after microinjection in the supraopticus nucleus although AVP release was stimulated. These data suggest that AII might be effective in the regulation of water balance by centrally controlling both the input (drinking) and the output (ADH secretion) of water.     

A hypothalamic alpha-adrenergic mechanism mediating the thermogenic response to electrical stimulation of the lower brainstem in the guinea pig

Summary Electrical stimulation of the lower brain stem (ESLB) at sites presumed to be parts of the ascending noradrenergic system was carried out in unanaesthetized young guinea pigs. At neutral ambient temperature ESLB elicited a thermogenic response resembling that evoked by microinjection of noradrenaline into the hypothalamus. The response consisted of a rise in oxygen uptake (to more than 50% of the resting value) and of body temperatures, especially of the interscapular adipose tissue. In some cases shivering was also evoked. The thermogenic response to ESLB was completely blocked by additional microinjection of an adrenergic alpha-receptor blocker, phentolamine, into the hypothalamic area where the noradrenergic fibres were presumed to terminate. Subsequent intrahypothalamic injection of noradrenaline, which had formerly been shown to restore the decreased responses to repeated ESLB, failed to restore the effectiveness of ESLB after phentolamine. It is concluded that the thermogenic responses to ESLB are mediated by a noradrenergic pathway ascending to the hypothalamus and not by direct stimulation of efferent pathways controlling the peripheral target system. The hypothalamic transmission can be prevented by an alpha-adrenergic blockade.Supported by the Deutsche Forschungsgemeinschaft SFB 122, Projeckt B 1     

Development of a vibratory microinjection method

To reduce cellular damage by pronuclear microinjection and nuclear transfer, we have recently developed a vibratory microinjection method. A micropipette was fixed to a piezoelectric ceramic with a resonance frequency of 70 kHz. When this micropipette was vibrated, it easily entered a mouse-fertilized egg without any sharp depression of the cell body, whereas a sharp, deep depression at the insertion site was observed when the micropipette was not vibrated. A depression rate defined as a rate of a depth of depression over an original cell diameter was utilized as an index of cellular deformation. The depression rates with and without vibration were 11.1 +/- 5.2% (N = 24) and 40.4 +/- 8.8% (N = 16), respectively (P < 0.0001, Student's t-test). In conclusion, the vibratory microinjection method is a new, useful option for gene transfer because it resulted in much less cellular deformation, therefore implicating less cellular damage.     

Regulation mechanism of simian virus 40 late gene expression in primary kidney cells and simian virus 40 transformed 3T3 cells.

Primary and continuous lines of mouse cells are nonpermissive for simian virus 40 (SV40). These cells, infected by the conventional virus absorption method, promote expression of the early but not of the late viral genes. Induction of V-antigen synthesis was found in a large number of primary mouse kidney cells and SV40-transformed $\text{Balb}\text{c}\text{-3T3}$ cells (SV-T2) infected with SV40-DNA component I by the microinjection technique. The proportion of SV40 V-antigen-synthesizing cells is correlated to the quantity of injected DNA molecules in both primary mouse kidney and SV-T2 cells.