基于全自动显微镜的图像新技术研究

针对显微图像分析、识别需要的全自动控制显微镜,研究开发其相应的图像处理算法。提出了能量谱自动聚焦评价函数算法、自适应选取基准图的图像拼接算法、改进Laplacian算子的多层聚焦图像叠合算法等。该算法模块已成功地应用于所开发的CMIAS显微医学图像分析系统中,取得了满意的应用效果。     

材料显微组织中的晶界识别与提取

本文针对材料制备、加工和热处理过程中涉吸材料显微组织晶界结构的识别与提取问题，采用晶界特征提取、晶界断点修复等形态学图像处理方法，获得了晶界特征和晶粒几何形态学信息，并建立了相应的图像定量分析技术及数据库。     

肝组织免疫组化病理图像的自动分析

本文介绍一个肝组织免疫组化病理图像自动分析系统。该系统由病理图像自动分割、分类识别和定量分析等部分组成。它可定量检测出肝炎病毒抗原含量指数 ,有助于病理医生研究诊断。     

基于图像增强技术的钢铁材料微观组织图像识别的研究

本文采用图像识别技术对钢铁材料在不同状态下形成的不同类型的微观组织照片进行了分析,研究了不同图像增强算法和机器学习算法对钢铁显微组织识别精度的影响.采用图像增强技术将原始图片质量提高,并采用图像特征提取技术获得图像特征.采用随机森林、支持向量机、集成树机器学习算法建立了基于图像特征的钢铁组织分类器.其中基于随机森林算法...     

基于组合BP神经网络的皮肤肿瘤目标识别

本文对皮肤肿瘤目标识别技术进行研究。首先利用阈值分割方法对皮损区域进行分割;然后,依据皮肤肿瘤早期诊断ABCD准则,对皮损区域提取了颜色、纹理和形状等特征,并基于相关性分析对所提取的特征进行优选;最后采用组合BP神经网络模型实现了皮肤肿瘤目标的分类识别。本文方法在黄色人种皮肤镜图像上进行实验,结果表明,该方法具有更高的分类精度,敏感度和特异度分别达到了93.3%和96.7%,识别结果令人满意。     

基于BP神经网络的结核杆菌目标识别

结核杆菌的有效识别是实现结核杆菌显微图像自动分析的关键。论文针对萋尔-尼尔逊方法染色的显微图像,对结核杆菌的提取和识别进行研究。算法首先基于形态学top-hat变换对图像进行增强并进行阈值分割,然后对结核杆菌进行特征分析,定义并提取了包括颜色、形状和边界不规则性等在内的10个特征,最后运用BP神经网络进行识别。实验结果表明,该方法能够有效检测并识别结核杆菌。选取1 200个目标进行七折交叉验证,识别准确率达到98%。     

基于国人皮肤镜黑素细胞肿瘤图像的智能化分类与识别研究

目的建立国人皮肤黑素细胞肿瘤(MT)图像智能化分类与识别方法。方法采用皮肤镜法获取MT图像信息,提出自生成神经网络的自适应聚类分割与特征提取算法,定量分析MT的形状不对称(AY)、形状偏心率(EY)、边界凹陷率(BDR)、过渡区辐射不均匀度(URTA)、颜色多样性(CD)、纹理相关性(TC)六个特征,结合组合神经网络分类器对MT的良、恶性进行分类与识别,经病理验证与统计学分析。结果 642幅MT图像,其中良性82.4%;恶性17.6%;URTA、TC、BDR、CD灵敏度86.73%～95.58%,特异度97.3%～100%;AY和EY灵敏度41.59%～47.78%,特异度69.91%～76.99%;对MT的良、恶性分类与识别,准确率达93.65%;分类符合率经χ2检验均有显著性差异(χ2=4.51,P<0.05)。结论皮肤镜MT图像分析法,可有效实现MT良、恶性分类与自动识别,为解决国人皮肤恶性黑素瘤的智能化识别瓶颈问题奠定了基础。     

金相学和材料显微组织定量分析技术

扼要但比较系统地介绍了材料显微组织几何形态的定量表征与分析技术及其标准化、显微组织仿真及设计、以及金相研究时应注意的材料显微组织的若干特性等内容。对金相学、材相学、体视学、图像分析、虚拟金相学、显微组织仿真及其相互关系亦予以扼要讨论。本论文还给出了一系列金相观测的标准名称以及利用体视学和图像分析方法进行材料显微组织或非金属夹杂物定量分析的标准的例子供查阅、应用。     

利用小波尺度共生矩阵和灰度共生矩阵的SAR图像分类

SAR图像包含有相干斑噪声,传统的方法不能很好地对SAR图像进行分类,本文将反映图像纹理的动态和静态信息特征相结合,提出了一种新的SAR图像分类方法。实验结果证明该方法可以得到较好的分类结果。     

显微图像分析系统测量细胞核DNA含量误差的研究

检测显微图像分析系统测量细胞核ＤＮＡ含量的误差对于判断肿瘤细胞ＤＮＡ倍体分析的准确性具有重要意义。本文检测了ＴＩＧＥＲ显微图像分析仪的系统噪声、聚焦变化、光源亮度与光谱差异、背景光强度分布不均匀的测量误差。结果显示，（１）尽管实现了Ｉｏ和Ｉ在同一空间位置的测量，但是，它们仍给测量结果引入了测量误差，以背景光强度分布不均导致ＩＯＤ的测量误差最大；（２）应根据目标细胞的大小和密度，选择适当的物镜倍率，样品制备应使ＡＯＤ达到一定的密度值以上，才能最大限度地降低测量误差；（３）鸡红细胞核是综合检测和评价显微图像分析系统测量细胞光密度性能的良好生物学标准。     

Monoclonal antibodies against excretory/secretory antigens of Angiostrongylus cantonensis

In the present study, four murine monoclonal antibodies (MAbs) were generated against the excretory/secretory (ES) products of Angiostrongylus cantonensis adult worms; two represented IgG1 and two represented IgM MAbs, and they were designated 12D5, 15F8, 21B7 and 14G10, respectively. Immunoblotting revealed that all of the MAbs predominantly recognized a 98?kDa antigen in the ES products of A. cantonensis adult worms, and no cross reactions were found with the whole worm antigens of some other common parasites, namely, Schistosoma japonicum, Clonorchis sinensis, Paragonimus westermani, Ascaris lumbricoides, Trichinella spiralis, Anisakis sp., Echinococcus granulosus, Taenia solium, and Spirometra erinacei. Immunolocalization showed that all of the four MAbs reacted with the cuticle of the adult parasite, the external surface of its intestinal canal and reproductive organs, and its egg and first-stage larvae in the lungs of rats experimentally infected with A. cantonensis. The generation and characterization of four specific MAbs against A. cantonensis ES antigens provide foundation for the development of specific immunological diagnostic techniques for human infections with A. cantonensis.     

Proteases in the schistosome life cycle: a paradigm for tumour metastasis

Summary Cancers and parasites have a number of properties in common, particularly those that relate to their respective capacities to evade host defence mechanisms. This review highlights the similarities between metastatic tumours and schistosomes in particular, and describes the role that proteases may have in the migration, growth, survival and transmission of the different stages of the schistosome life-cycle in the vertebrate host. An elastase-like serine protease of schistosome larvae has been particularly well characterized, and its substrate profile and other properties are indicative of a role in facilitating migration of the parasite through skin tissue early after infection. The primary structures of a cathepsin B-like enzyme, and a putative haemoglobinase found in adult worms have also recently been derived, these enzymes being responsible for degradation of haemoglobin in erythrocytes upon which the adults feed. Adult schistosome worms reside and produce eggs intravascularly, and the processes that mediate the extravasation and subsequent migration of the egg through host tissue are dependent on both blood platelets and the immune response. Fibrino(geno)lytic enzymatic activity that is present in the egg could modulate the thrombogenic potential that eggs might have as a result of their capacity to cause platelet aggregation in vitro and in vivo. The roles of other proteases and peptidases that have been found in schistosome larvae, worms and eggs are less clear. Some of these enzymes may modulate immunological and haemostatic defence mechanisms and thus prolong survival of the parasite, and the consequences of the interactions between schistosomes and host protease inhibitors could also be immune modulatory.     

Elevated expression of phosphatidylserine in the outer membrane leaflet of human tumor cells and recognition by activated human blood monocytes

We determined whether the presence of phosphatidylserine (PS) in the outer membrane leaflet of human tumor cells correlated with their recognition by activated human monocytes. Three tumorigenic cell lines, A375 melanoma and A431 and Colo-16 carcinomas, and a normal human epidermal keratinocyte line (NHEK) were incubated with monocytes activated to the tumoricidal state by gamma-interferon and lipopolysaccharide. Activated human monocytes bound to and lysed all tumorigenic targets, while the nontumorigenic NHEK were neither bound nor killed. Semiquantitative analysis of PS in the outer leaflet of the cells revealed that the tumorigenic cells expressed 3-7-fold more PS than did the nontumorigenic NHEK. To determine whether enhanced PS expression on the tumor cells was responsible for their recognition by activated monocytes, NHEK were supplemented with exogenously supplied analogues of PS and phosphatidylcholine. PS-labeled NHEK but not phosphatidylcholine-labeled nor control NHEK bound to activated human monocytes. These results suggest a role for PS in monocyte recognition of tumor cells.     

Prospects for oocyte banking and in vitro maturation

There is little debate about the desirability of human oocyte ("egg") banking but plenty of discussion about its prospects. Egg banking is needed by young cancer patients before they undergo potentially sterilizing treatment and is a desirable alternative to in vitro fertilization and embryo cryopreservation. However, egg banking is inefficient-oocytes are sensitive to chilling, often fail to survive freeze-thawing, and are susceptible to cytoskeletal damage and aneuploidy. Currently, even the most optimistic success rates offer patients only a slim chance of pregnancy if few oocytes are available. Ultra-rapid freezing with vitrification may offer advantages over conventional equilibrium cooling protocols and needs to be investigated further. Likewise, freezing immature oocytes followed by in vitro maturation offers practical and theoretical advantages, but this method is still inefficient. Nevertheless, all these technologies are improving, and egg banking will eventually become an option for patients seeking fertility preservation.     

Production of monoclonal antibodies recognizing human hair follicle keratinocytes

To raise new antibodies against the cells contained in human hair follicles, we immunized mice with a mixture of primary cultured human embryogenic keratinocytes thought to contain very immature keratinocytes, as antigen. Using a monoclonal antibody (MAb) producing technique, we obtained two MAbs, which recognize a specific portion of the hair follicle. MAb-8G2 was specific to the companion layer between the inner and outer root sheath of the follicle. This pattern of recognition was quite similar to the expression pattern of K6hf protein, a human hair follicle-specific type II cytokeratin. MAb-sDP showed unique recognition. Immunohistochemistry indicated that MAb-sDP specifically recognized the cells surrounding the dermal papilla (DP). These cells are possibly the germ line cells of hair matrix.     

Cisplatin-DNA damage recognition proteins in human tumour extracts.

Enhanced repair of DNA adducts may be a cause of cis-diamminedichloroplatinum(II) resistance in solid malignancies. Binding of specific damage recognition proteins to the sites of DNA damage may be involved in the initial steps of DNA repair, or alternatively may block access of repair proteins to damaged DNA. Proteins which bind specifically to CDDP-modified DNA were identified in cell extracts from human ovarian carcinoma cell lines by two assays, the gel mobility shift assay and the southwestern blot. In the first assay, proteins complexed with CDDP-modified oligonucleotide and produced two retarded bands, B1 and B2. The B2 complex was partially purified from an ovarian cell extract by anion exchange FPLC, and was shown to bind to DNA damaged by CDDP but not by transDDP or UV irradiation. Using the southwestern blot, proteins of 97, 48, and 25 kD were identified; each of these bound to CDDP-modified but not undamaged oligonucleotide. The partially purified B2 protein fraction contained both the 97 and the 25 kD damage recognition proteins. A human ovarian carcinoma cell line selected in vitro for CDDP-resistance (OV1P/DDP), which is 5-fold more resistant to CDDP than the parental line (OV1P), showed an increase in binding of the 97 and 48 kD damage recognition proteins compared with the parental line. Twelve ovarian cell lines differed by up to 3-fold in their expression of these proteins, but there was no correlation between the amount of damage recognition protein in a cell extract and the cellular sensitivity to CDDP. Damage recognition proteins were also demonstrated in extracts prepared from biopsies of human ovarian, cervical, and testicular malignancies, but there was no apparent difference in the binding activity in extracts from tumours of different CDDP-sensitivity. The functional role of these damage recognition proteins remains to be established.     

Possible relationship between glycosphingolipids and the formation of metastasis in certain human experimental tumors

Two tumors, human sarcoma #1 (HS #1) and human epidermoid carcinoma #3 (HEp #3), were cultured on the chorioallantoic membrane of chick embryos. Under experimental conditions, HS #1 does not metastasize, whereas HEp #3 metastasizes extensively to chick embryo lungs and other organs. The glycosphingolipid profiles of these tumors were studied and HEp #3 wad found to contain about 2.5-fold less lipid-bound sialic acid per 100 mg of total lipid extracted than did HS #1, due mainly to smaller levels of monosialoganglioside (3.7-fold) and disialoganglioside (3.8-fold) in HEp #3. The total amount of neutral glycosphingolipids was approximately the same in both tumors, but their profiles differed. Treatment of these tumors with 6,7,8,9-tetrahydro-1-mercapto-1,2,4-triazolo-[4,3-a]quinazolin-5-ol (2.5 mg/egg/tumor) completely inhibited the formation of metastases in HEp #3 and increased the total content of lipid-bound sialic acid in the tumor by 63% (hematoside, monosialoganglioside, and disialoganglioside by 71, 99, and 67%, respectively). No change was seen in the content of lipid-bound sialic acid in HS #1. Treatment of HEp #3 with a smaller dose of te quinazolinol derivative (1.25 mg/egg) caused an average of 88% inhibition of metastasis, with a 37% increase in lipid-bound sialic acid. Another compound, 2,5-diphenylthiazolo-[5,4-d]thiazole (500 microgram/egg), completely inhibited the formation of metastasis and caused a substantial increase in the amount of lipid-bound sialic acid (77%). The data showed the existence of a correlation between the level of gangliosides in HEp #3 and the ability of these tumors to metastasize.     

Evaluation of chimpanzee antiserum to human carcinoembryonic antigen

An adult chimpanzee (Pan troglodyte) with an endogenous circulating carcinoembryonic antigen (CEA) level of 60 ng/ml was immunized s.c. with human CEA. After 1 year of immunizations, the anti-human CEA antibody titer had plateaued. This chimpanzee antiserum demonstrated high avidity specific recognition of human CEA and showed ionic strength effects for CEA recognition similar to those previously described for goat and baboon anti-CEA antisera. Radioimmunoassay of 93 human plasma samples for CEA content using chimpanzee anti-CEA versus Roche goat anti-CEA antisera gave essentially identical results (R = 0.985). Endogenous CEA in chimpanzee blood was very poorly identified by chimpanzee anti-human CEA antisera compared to Roche goat antisera. Column chromatography of human and chimpanzee CEA in the presence of chimpanzee anti-CEA antibody showed only reactivity for the human CEA. In addition, chimpanzee antiserum had only minimal blocking effect on the binding of either goat or baboon antiserum to human CEA. We conclude from these studies that chimpanzee anti-human CEA antiserum recognized a determinant(s) on human CEA which was different from these recognized by goat or baboon antiserum to human CEA and this determinant(s) was poorly represented on chimpanzee CEA. In contrast, the human CEA determinant(s) recognized by baboon and goat anti-CEA antiserums were readily detected on chimpanzee (CEA).     

Inhibition of proliferation and induction of differentiation of glioma cells with Datura stramonium agglutinin

We found that a lectin, Datura stramonium agglutinin, induced irreversible differentiation in C6 glioma cells. The differentiated cells had long processes, a low rate of proliferation and a high content of glial fibrillary acidic protein. When the medium was replaced with Datura stramonium agglutinin-free medium after 1 h, cell proliferation continued to be inhibited. Experiments with several other lectins indicated that both recognition of linear N-acetyllactosamine repeats and recognition of multiantennary units of cell-surface glycans were required for the inhibition of C6 proliferation. Proliferation of four human glial tumour cells was also inhibited by Datura stramonium agglutinin. Further, these differentiated human glial tumour cells had long processes and a high content of glial fibrillary acidic protein similar to differentiated C6 glioma cells. Taken together, these observations suggest that Datura stramonium agglutinin may be useful as a new therapy for treating glioma without side effects.