球结膜图像的血管识别

目的：在人体微循环系统中,球结膜图像对血压的测定以及未来血管疾病发展有着良好的预测性。通过显微镜观察,虽可以清晰地看到球结膜图像,但是想直接从球结膜图像中识别出球结膜血管却较为困难,本文希望对球结膜图像进行边缘检测,以利于血管的准确识别。方法：采集24位彩色显微图像,对其进行灰度化处理及图像滤波后,利用M-H算子进行边缘检测。结果：处理后的显微图像可以非常完美的识别多级血管。结论：可见M-H算子适宜于进行球结膜图像的边缘检测。     

眼底图像中硬性渗出物检测算法

目的利用眼底图像中硬性渗出物(hard exudates,HE)的亮度与边缘特征,提出一种基于Canny边缘检测算法与形态学重构相结合的HE自动检测方法,以解决目前算法灵敏度低、检测结果中视盘和血管的干扰等问题,对糖尿病视网膜病变(diabetic retinopathy,DR)的自动筛查具有重要意义。方法检测算法包括4个步骤。步骤一,图像预处理,主要包括RGB通道选取、基于形态学的图像对比度增强。步骤二,视网膜图像关键结构的消除,利用基于Gabor滤波的血管分割方法,消除血管边缘对HE检测的影响。将本文视杯分割算法应用在眼底图像红色通道上实现视盘自动分割,消除视盘及其边缘对HE检测的影响。步骤三,利用改进的Canny边缘检测算法和形态学重构方法对HE进行提取。步骤四,基于形态学的图像后处理,消除眼底图像边缘部分假阳性区域。最后利用该算法测试公开数据库中的40幅图像(35幅HE病变图像,5幅正常图像)。结果该算法对基于病变的灵敏性(sensitivity,SE)和阳性预测值(positive predictive value,PPV)分别为93.18%和79.26%,基于图像的灵敏性、特异性(specificity,SP)和准确率(accuracy,ACC)分别为97.14%、80.00%和95.00%。结论与其他方法对比,基于Canny边缘检测算法与形态学重构相结合的HE自动检测算法具有较好的可行性。     

Cell contour extraction on multi-threshold images.

A procedure for the extraction of cell and nuclear contours from digital images is presented. The procedure is simple and suitable for efficient implementation in an interactive environment where the user selects a region of interest containing the cell to be segmented. Segmentation is facilitated by multi-thresholding where the threshold values are determined by an iterative algorithm using the mean square error criteria to cluster the image pixel values. Edges are detected on the threshold images by employing a binary morphological filter and the desired contours are selected by considering closed contours in the edge image. The cell region is first extracted by performing segmentation, edge detection and contour extraction on the region of interest. The nuclear region is subsequently extracted by reapplication of the algorithms within the extracted cell region. Segmentation examples are presented.     

定量血管内光学相干层析成像的研究进展

血管内光学相干层析成像(IVOCT)是目前分辨率最高的血管内成像技术,可对冠状动脉血管腔及管壁内膜下病变进行快速、清晰的成像。仅根据组织结构的层析图像无法精确识别粥样硬化斑块成分(如钙化、纤维化、脂质和混合斑块),需要形态结构之外的生理信息的对比机制,获得具有临床诊断价值的组织参数,即定量IVOCT(qIVOCT)。本文针对根据IVOCT原始背向散射信号和灰阶图像定量测量血管壁组织的光学特性参数、弹性参数和血流动力学参数的研究现状进行归纳和总结,分析目前存在的问题,展望可能的发展方向。     

Serotonin- and somatostatin-positive goblet cell carcinoid of the duodenum

In the duodenum, mixed exocrine-endocrine tumors exhibiting both neuroendocrine and glandular differentiations [cf. appendiceal goblet cell carcinoids (GCCs)] are rare. We present a Japanese case with a duodenal GCC that was found during pathologic examination of a gastrectomy specimen removed for gastric mucosal cancer. The tumor was widely distributed within both the first portion of the duodenum and the gastric antrum, although mucosal involvement was observed only in the duodenum. The tumor cells formed solid nests, trabeculae, or tubules, and some displayed a goblet cell appearance. They were immunoreactive against antibodies for both serotonin and somatostatin, and showed an argentaffin reaction (similar to a "midgut" enterochromaffin cell carcinoid). Ultra-structurally, the tumor cells had an amphicrine nature. Physicians encounter GCC in the duodenum only rarely, and its discovery may be incidental. Its diagnosis will be challenging and will require careful clinical and pathologic examinations.     

一种基于相位相关法及数学形态学方法的眼底血管图像自动拼

Precise mosaic techniques are essential for quantitative evaluation of retinal images to make early detection of fundus anomalies feasible. Opening of a gray-scale image by a gray-scale structuring element(SE) can generate a background image. Image mosaic was achieved by subtraction this background image from the original image and then applying a phase correlation method to find translation difference. Because of the accuracy characteristics of the phase correlation method and the speed of the FFT hardware, this new algorithm can work very fast and accurately, compared to conventional techniques. The method was also characterized by an outstanding robustness against correlated noise and disturbances, such as those encountered with nonuniform illumination. The results confirm the robustness of the chosen approach.     

Automatic analysis of image of surface structure of cell wall-deficient EVC

Some computer applications for cell characterization in medicine and biology, such as analysis of surface structure of cell wall-deficient EVC (El Tor Vibrio of Cholera), operate with cell samples taken from very small areas of interest. In order to perform texture characterization in such an application, only a few texture operators can be employed: the operators should be insensitive to noise and image distortion and be reliable in order to estimate texture quality from images. Therefore, we introduce wavelet theory and mathematical morphology to analyse the cellular surface micro-area image obtained by SEM (Scanning Electron Microscope). In order to describe the quality of surface structure of cell wall-deficient EVC, we propose a fully automatic computerized method. The image analysis process is carried out in two steps. In the first, we decompose the given image by dyadic wavelet transform and form an image approximation with higher resolution, by doing so, we perform edge detection of given images efficiently. In the second, we introduce many operations of mathematical morphology to obtain morphological quantitative parameters of surface structure of cell wall-deficient EVC. The obtained results prove that the method can eliminate noise, detect the edge and extract the feature parameters validly. In this work, we have built automatic analytic software named "EVC.CELL".     

Morphological evidence of neutrophil-tumor cell phagocytosis (cannibalism) in human gastric adenocarcinomas

The phenomenon of neutrophil-tumor cell emperipolesis or phagocytosis has been documented by light microscopy in various human carcinomas, but little is known about the cellular pathological processes and the morphological changes involved. In an attempt to clarify the nature of this phenomenon, the authors' ultrastructural studies on the relationships among neutrophils and tumor cells in human gastric carcinomas are reviewed and analyzed. At the electron microscopy level, apoptotic neutrophils were found within vacuoles of adenocarcinoma cells in 2 cases. They showed either early apoptotic morphology with perinuclear chromatin aggregation but cytoplasm integrity or late apoptotic morphology with uniform, collapsed nucleus and tightly packed cytoplasmic granules. A light microscopy review of 200 cases of resected gastric carcinomas identified 22 cases (11%) that were characterized by neutrophil-tumor cell phagocytosis (cannibalism). TUNEL staining confirmed the presence of apoptotic neutrophils within the cytoplasm of the tumor cells. This study provides light and electron microscopic evidence of apoptotic neutrophils phagocytosed by gastric adenocarcinoma cells. The morphological features of neutrophil-tumor cell phagocytosis (cannibalism) would suggest a particular mechanism of tumor-immune escape in human gastric carcinoma.     

基于超声影像组学的原发性肝细胞癌分级预测

目的:针对原发性肝细胞癌（HCC）肿瘤分级预测难题，提出一种基于灰阶超声成像的影像组学预测模型。方法:首先，由超声医生对肿瘤区域进行手动分割，其次，采用影像组学方法对肿瘤区域提取形状、一阶统计、纹理特征，计算特征间Pearson相关系数剔除冗余特征，最后通过单变量分析筛选得到特征子集，采用LASSO构建HCC分级预测模型；利用留一法计算模型的受试者操作特性曲线下的面积（AUC）评估模型对HCC分级的预测能力。结果:利用43例经手术病理证实的HCC患者的灰阶超声图像构建HCC分级预测模型，所建模型由6个与分级高度相关的影像特征组成，模型具有较强的预测能力（AUC=0.76）。结论:基于灰阶超声成像的影像特征与HCC分级高度相关，所建影像组学模型能够较好地预测HCC分级。     

白介素32在胃癌细胞中的表达及意义

目的观察白介素32（IL-32）在胃癌细胞株中的表达情况,探讨其在胃癌发生、发展中的意义。方法采用逆转录PCR、real-time PCR及Western blot法分别从基因和蛋白水平上检测IL-32在正常胃上皮细胞GES-1以及胃癌细胞株MKN-45、KATO中的表达情况。结果逆转录PCR及real-time PCR结果均显示胃癌细胞IL-32 mRNA表达水平明显高于正常胃上皮细胞,逆转录PCR灰度比值（目的 /内参）为：GES-1（0.34±0.09）、MKN-45（0.79±0.11）、KATO（0.90±0.17）,差异有统计学意义（P〈0.05）。real-time PCR Ct值显示：MKN-45的IL-32基因表达量是GES-1的（5.34±1.09）倍,KATO是GES-1的（4.07±1.69）倍,差异均有统计学意义（P〈0.05）。Western blot结果显示IL-32在蛋白表达水平的结果与前一致,灰度比值（目的 /内参）为：GES-1（0.30±0.10）、MKN-45（0.92±0.32）、KATO（0.86±0.15）,差异有统计学意义（P〈0.05）。结论IL-32在胃癌细胞株中呈高表达,推测其可能在胃癌的发生、发展中起一定作用。     

基于形态学去噪的兔眼相干光断层成像中角膜形态自动识别

目的对兔眼相干光断层成像(optical coherence tornography,OCT)中的角膜边缘识别进行研究,以期从不具有较高清晰度的图像中自动获得角膜边缘以及相关形态学参数。方法首先利用Otsu算法对兔眼OCT图像进行二值化,并以半径为5像素的圆盘形结构元素进行形态学去噪运算消除内部伪边缘,通过Canny算子获得角膜边缘图像;然后使用筛选过错误边缘点后的其余边缘点进行边缘的二次曲线一般方程的曲线拟合,求出各点曲率半径;最后用克朗巴哈系数法与人工描绘的边缘获得的相应点的曲率半径进行一致性检验。结果基于形态学去噪的Canny算法获得了较为连续平滑的角膜内外边缘曲线,与手工标点拟合曲线求出的曲率半径值具有较高的一致性(Cronbach’sα=0.953,P0.05)。结论基于形态学去噪的Canny算法可以较为准确地对OCT图像中的兔眼角膜的形态进行识别。     

Morphological Evidence of Neutrophil-Tumor Cell Phagocytosis (Cannibalism) in Human Gastric Adenocarcinomas

The phenomenon of neutrophil-tumor cell emperipolesis or phagocytosis has been documented by light microscopy in various human carcinomas, but little is known about the cellular pathological processes and the morphological changes involved. In an attempt to clarify the nature of this phenomenon, the authors' ultrastructural studies on the relationships among neutrophils and tumor cells in human gastric carcinomas are reviewed and analyzed. At the electron microscopy level, apoptotic neutrophils were found within vacuoles of adenocarcinoma cells in 2 cases. They showed either early apoptotic morphology with perinuclear chromatin aggregation but cytoplasm integrity or late apoptotic morphology with uniform, collapsed nucleus and tightly packed cytoplasmic granules. A light microscopy review of 200 cases of resected gastric carcinomas identified 22 cases (11%) that were characterized by neutrophil-tumor cell phagocytosis (cannibalism). TUNEL staining confirmed the presence of apoptotic neutrophils within the cytoplasm of the tumor cells. This study provides light and electron microscopic evidence of apoptotic neutrophils phagocytosed by gastric adenocarcinoma cells. The morphological features of neutrophil-tumor cell phagocytosis (cannibalism) would suggest a particular mechanism of tumor-immune escape in human gastric carcinoma.     

Methods for preparing tissue microarray slides using xenografts with different levels of HER2 expression to standardize HER2 detection

Gene amplification and protein overexpression of human epidermal growth factor receptor type 2 (HER2) are specific targets for HER2-targeting drugs in breast, gastric, salivary gland, and colorectal cancers. The histopathological determination of HER2 status is crucial for treatment, highlighting the importance of improving HER2 detection accuracy in clinical practice. We prepared tissue microarray (TMA) slides for use as control slides for the standardization of gastric HER2 testing. Four human gastric cancer cell lines with HER2 scores of 3+, 2+, 1+, and 0 were xenografted in NOG mice. The TMA slides were constructed using samples from three different areas in these tumors. Staining properties were determined using six clinical kits for HER2. In TMA, HER2-positive tumors with HER2 scores of 3+ and 2+ showed good staining with all diagnostic kits, and the tissue images were similar to those of clinical samples. Xenograft tumor slides could potentially be used as external controls to standardize staining conditions for a variety of kits and may improve the accuracy of HER2 detection in clinical practice.     

Analysis and recognition of touching cell images based on morphological structures

Automated analysis and recognition of cell-nuclear phases using fluorescence microscopy images play an important role for high-content screening. A major task of automated imaging based high-content screening is to segment and reconstruct each cell from the touching cell images. In this paper we present new useful method for recognizing morphological structural models of touching cells, detecting segmentation points, determining the number of segmented cells in touching cell image, finding the related data of segmented cell arcs and reconstructing segmented cells. The conceptual frameworks are based on the morphological structures where a series of structural points and their morphological relationships are established. Experiment results have shown the efficient application of the new method for analysis and recognition of touching cell images of high-content screening.     

Ultrastructural Descriptions of Heterotypic Aggregation between Eosinophils and Tumor Cells in Human Gastric Carcinomas

A histological variant of gastric adenocarcinoma, characterized by an intense tumor-associated tissue eosinophilia (TATE), has been occasionally reported in the literature. The purpose of this ultrastructural study was to determine the interactions between frequently occurring eosinophils and tumor cells in gastric carcinoma characterized by TATE. Fresh tumor tissue of 92 gastric carcinomas was processed for both light and electron microscopic examination. Intense TATE was found in 7 out of 92 (7.6%) gastric carcinomas (6 of intestinal-type and 1 of diffuse-type). Electron microscopy, selectively performed in 7 cases with intense TATE, revealed eosinophils, singly or in groups, in contact with damaged or necrotic tumor cells. Activated eosinophils showing piecemeal degranulation were also found in intimate contact with viable tumor cells, characterized by plasma membrane caveolar invaginations. The authors regard this close morphological relationship as in vivo evidence for possible cross-talk between eosinophil and viable tumor cell, a conclusion that has already been drawn from experimental studies, but until now inadequately supported by ultrastructural observations in a human tumor.     

Ultrastructural descriptions of heterotypic aggregation between eosinophils and tumor cells in human gastric carcinomas

A histological variant of gastric adenocarcinoma, characterized by an intense tumor-associated tissue eosinophilia (TATE), has been occasionally reported in the literature. The purpose of this ultrastructural study was to determine the interactions between frequently occurring eosinophils and tumor cells in gastric carcinoma characterized by TATE. Fresh tumor tissue of 92 gastric carcinomas was processed for both light and electron microscopic examination. Intense TATE was found in 7 out of 92 (7.6%) gastric carcinomas (6 of intestinal-type and 1 of diffuse-type). Electron microscopy, selectively performed in 7 cases with intense TATE, revealed eosinophils, singly or in groups, in contact with damaged or necrotic tumor cells. Activated eosinophils showing piecemeal degranulation were also found in intimate contact with viable tumor cells, characterized by plasma membrane caveolar invaginations. The authors regard this close morphological relationship as in vivo evidence for possible cross-talk between eosinophil and viable tumor cell, a conclusion that has already been drawn from experimental studies, but until now inadequately supported by ultrastructural observations in a human tumor.     

Molecular identification of main cellular lineages as a tool for the classification of gastric cancer

Gastric carcinomas (GC) are heterogeneous tumors comprising variable amounts of cells of different lineage phenotype, including gastric mucous cells (surface--SMC or gland--GMC) and intestinal cells (IC). The evaluation of tumor behavior has classically depended on strictly morphological classifications of tumors. Microsatellite instability (MSI) is frequently detected in GC, but whether MSI affects all gastric cellular lineages or exclusively occurs in unique cellular lineages in GC is not known. The aims of this study were to test a combination of anti-mucin antibodies to classify gastric cancer into predominant cell lineage phenotype and to determine whether MSI in GC is associated with particular cellular tumor phenotypes. Fifty-five GC were immunophenotyped with antibodies specific for SMC, GMC, or IC. DNA was extracted from tumor and non-neoplastic gastric tissues and amplified with 5 microsatellite markers. A mixed cellular pattern was the most frequent phenotype of GC (61%) and was seen in both glandular (63%) and diffuse (58%)-type tumors. No significant difference in the rate of MSI was found in tumors with predominant gastric, intestinal or mixed phenotype. However, tumors with null or low-level expression of cellular lineage differentiation markers displayed MSI more frequently than tumors with high-level expression (40% v 20%). In conclusion, different gastric carcinoma cell lineage patterns can be easily identified with the 3 immunohistochemical markers used in this study. The 3 main cellular lineage components of gastric cancer can be similarly affected by microsatellite instability, consistent with the notion that MSI is an early event in gastric carcinogenesis.     

Detection of disseminated tumor cells in neuroblastoma: 3 log improvement in sensitivity by automatic immunofluorescence plus FISH (AIPF) analysis compared with classical bone marrow cytology

The sensitive detection of bone marrow involvement is crucial for tumor staging at diagnosis and for monitoring of the therapeutic response in the patient's follow-up. In neuroblastoma, only conventional cytomorphological techniques are presently accepted for the detection of bone marrow involvement, yet since the therapeutic consequences of the bone marrow findings may be far-reaching, the need for highly reliable detection methods has become evident. For this purpose, we developed an automatic immunofluorescence plus FISH (AIPF) device which allows the exact quantification of disseminated tumor cells and the genetic verification in critical cases. In this study, the power of the immunofluorescence technique is compared with conventional cytomorphology. 198 samples from 23 neuroblastoma patients (stages 4 and 4s) at diagnosis and during follow-up were investigated. At diagnosis, 45.6% of the samples (26 of 57) which were positive by AIPF investigation were negative by cytomorphology. During follow-up, 74.2% (49 of 66) of AIPF-positive samples showed no cytological signs of tumor cell involvement. False negative morphological results were found in up to 10% of tumor cell content. A tumor cell infiltrate below 0.1% was virtually not detectable by conventional cytomorphology. Using the sensitive immunofluorescence technique, the analysis of only two instead of four puncture sites did not lead to false negative results. Thus, the immunofluorescence technique offers an excellent tool for reliable detection and quantification of disseminated tumor cells at diagnosis and during the course of the disease.     

基于抑制性突触多层神经元群放电编码的图像边缘检测

图像边缘检测所获得的目标轮廓细节质量,对于后续图像分析或理解过程具有重要作用。提出一种基于视觉机制的图像边缘检测新方法:构建抑制性突触连接的多层神经元群,在待检测图像的激励下,分析7像素×7像素视觉感受野窗口内互连接神经元的脉冲放电过程,记录发放时刻进行次序编码;同时考虑神经元之间的侧向抑制作用,在选择注意机制作用下获得增强图像;之后利用Log Gabor滤波器模拟视觉系统中的方向选择特性,获取8个方向的滤波结果,经过输出层神经元群的融合处理并经灰度映射到0～255的范围后获得边缘图像。对含有丰富边缘细节特性的24幅菌落图像进行处理,其处理结果的ROC评价指标均值为0.698 4,优于PCNN法的0.659 3;从评价指标的均方差来看,该结果具有更好的一致性。另外,从信息熵评价指标来看,该方法同样具有一定的优势,能够有效提取图像的边缘信息,而且也能反映更多层次的图像细节。所提出的方法为利用视觉生理特性进行图像处理提供了崭新而有效的思路。     

Gallbladder shape extraction from ultrasound images using active contour models

Gallbladder function is routinely assessed using ultrasonographic (USG) examinations. In clinical practice, doctors very often analyse the gallbladder shape when diagnosing selected disorders, e.g. if there are turns or folds of the gallbladder, so extracting its shape from USG images using supporting software can simplify a diagnosis that is often difficult to make. The paper describes two active contour models: the edge-based model and the region-based model making use of a morphological approach, both designed for extracting the gallbladder shape from USG images. The active contour models were applied to USG images without lesions and to those showing specific disease units, namely, anatomical changes like folds and turns of the gallbladder as well as polyps and gallstones. This paper also presents modifications of the edge-based model, such as the method for removing self-crossings and loops or the method of dampening the inflation force which moves nodes if they approach the edge being determined. The user is also able to add a fragment of the approximated edge beyond which neither active contour model will move if this edge is incomplete in the USG image. The modifications of the edge-based model presented here allow more precise results to be obtained when extracting the shape of the gallbladder from USG images than if the morphological model is used.