胃癌的自体荧光光谱诊断

目的：评价内镜下激光诱发自体荧光光谱诊断胃癌的临床应用价值。方法：采用氦镉（He-Cd）激光系统（激发波长442nm）对38例经病理活检证实的胃癌和慢性胃炎患者进行自体荧光光谱检测，并作自身对照研究。结果：在内镜下以氦镉激光诱发的胃癌自体荧光光谱含主峰（510nm附近）的3个次峰（590nm、670nm和710nm附近），光谱强度和形态与正常组织有显著差异，以荧光强度（FI）662－677nm和FI701－716nm的积分及FI510nm／FI710nm的比值3个参数作判别分析，自体荧光光谱诊断胃癌的敏感性、特异性及阳性和阴性预测值分别为82.9%、91.5%和87.9%、87.8%，其诊断准确率为87.8%。结论：内镜下对胃癌的自体荧光检测具有实时发现病灶、帮助引导活检的重要作用，有望成为早期胃肠道癌肿诊断的有效辅助工具。     

内镜下激光激发自体荧光诊断食管癌和胃癌的研究

为了探讨内镜下激光激发自体荧光对食管癌和胃癌的诊断意义,采用MJ－LF医用激光荧光快速诊断仪,测定人体食管癌及胃癌恶性肿瘤组织的荧光光谱,与胃、十二指肠溃疡病人和慢性浅表性胃炎病人对照研究。结果显示：32例胃癌病人中有25例在肿瘤组织可测到肿瘤特征峰,阳性率78．1％;15例食管癌病人有11例其癌肿组织可测到肿瘤特征峰,阳性率73．3％。二组平均阳性率76．6％。而110例胃、十二指肠溃疡病人及116例慢性浅表性胃炎病人未能测到肿瘤特征峰。结论：上消化道恶性肿瘤食管癌和胃癌病人内镜下的激光荧光光谱测定有76．6％的病人在630nm和／或690nm波长处可测出肿瘤特征峰。激光荧光光谱的测定对食管癌及胃癌的快速诊断具有一定临床意义。     

内镜下激光荧光检查食管癌胃癌及癌前病变的研究

我们采用氙（ｘ＋ｅ）激光光源激发癌组织或癌前病变组织，使该组织产生自体荧光，并作快速记录，以光谱图显示６３０ｎｍ和／或６９０ｎｍ波长处或附近有特征性峰出现诊断为癌及癌前病变。对象和方法有上消化道症状进行内镜检查并作了激光荧光检查和活检病理检查的５８４...     

体内及体外大肠癌组织激光诱发荧光光谱对比分析

目的：探讨体内外大肠癌组织激光诱发荧光（LIF）光谱的相关性。寻找LIF判断大肠癌的最佳特征参数。方法：10例大肠癌患者在肠镜检查时行LIF检测,随后检测该患者手术切除标本的LIF。 以氮分子激光器（波长337nm)为激发光源激发大肠癌组织,所得荧光进入OMAⅢ分析处理。结果：（1）体内、外癌和正常组织LIF光谱形态相似,体内正常组织和癌组织平均主峰强度低于体外组织近一半。体内、外癌组织主峰强度较正常组织也有显著降低。（2）每例患者正常组织的集成荧光强度均高于相应的癌组织。（3）体内癌组织LIF的主峰波长平均向长波侧移动6nm,体外平均移动3nm,体内正常组织较体正常组织平均向短波侧移动3nm。（4）体内、体外癌组织的LIF主、次峰荧光强度比值均显著高于相应正常组织。结论：体内、外大肠癌组织LIF光谱化规律基本一致。体内LIF强度较体外减低,集成荧光强度或主峰强度辅以主峰波长及主次峰比值可以提高肠镜下LIF实时判别大肠癌的灵敏度。     

胃液固有荧光光谱对胃癌诊断价值的初步研究

目的：显示胃癌患者及非癌患者胃液固有荧光光谱的荧光强度的差异,以期用于胃癌的筛查及诊断。方法：收集202例各种胃内疾病患者的胃液,经1：10释释后,检测激发波长为288nm,发射波长范围为300－800nm的固有荧光光谱。用CART V2．0统计软件建立判别模型,计算先验概率及后验概率。结果：各种胃内疾病患者的胃液固有荧光光谱均有3个峰（发射小分别为320－360nm、576nm及670－690nm),胃癌患者的第一荧光峰（发射波长320－360nm)的强度（P1FI）较其他胃内良性疾病患者明显增强。以P1FI≥111．80作为判别指标,用于胃癌诊断的先验概率的敏感度为91．4％,特异度为83．2％,准确度为84．7％。其后验概率的敏感度为85．7％,特异度为82．6％,准确度为83．1％。结论：胃液固有荧光光谱有希望成为一种胃癌的诊断和筛查方法。     

激光诱导大鼠大肠早癌组织自体荧光光谱特征

目的:探索激光诱导大肠早癌组织自体荧光光谱特征.方法:采用二甲肼ip法建立SD大鼠大肠癌动物模型,对大鼠结肠和直肠可疑病变或肿块部位用370 nm激光照射,诱导组织产生自体荧光并收集光谱后取材,根据病理诊断分为正常组、早期癌组、进展期癌组,分析各组光谱特征.结果:正常64例,早期癌78例,进展期癌84例,共226例.各组光谱在460 nm和505 nm处附近有波峰;正常组460 nm左右波峰对应的波长与早期癌组、进展期癌组对应波长比较差异有显著性(457.66±3.28 vs 467.87±7.71,468.60±4.53,P＜0.05).73%(61/84)的进展期大肠癌组织、69%(54/78)的早期大肠癌组织荧光光谱在635 nm附近有荧光峰,而正常组织缺乏该峰.早期癌组和进展期癌组自体荧光在460nm处的荧光强度比在正常组相应强度弱.早期癌组和进展期癌组I635/I460(1.9507±1.1460,2.1368±1.4721)和I635/I600值(0.4215±0.2582,0.4482±0.2309)分别较正常组I635/I460和I635/I600值大且差异有显著性(0.7494±0.1077,0.1416±0.0439,P＜0.05).结论:激光诱导的鼠大肠正常组织与大肠癌组织(包括早期和进展期大肠癌)自体荧光光谱之间存在差异,自体荧光光谱部分特征可用于大肠癌早期诊断.     

固有荧光癌症诊断仪对胃部病变的诊断价值

目的 探索激光诱发固有荧光光谱诊断仪(IFS-I)对胃癌病变的诊断价值.方法 胃镜下收集66例病例,共74个病灶的固有荧光光谱.将三种光谱特征与病理结果相对照,分析诊断方法 的敏感度、特异度和准确率.结果三种光谱特征中符合任意1条即为阳性的诊断敏感度最高,可用于胃癌和良性增生性病变的普查.在(460±20)nm波长处荧光特征的诊断特异性最高,可用于胃癌的精查.结论激光诱发固有荧光光谱诊断仪在胃癌及癌前病变的普查和胃癌明确诊断方面具有较好的应用前景.     

日本血吸虫成虫自发荧光观察及共聚焦λ扫描分析

目的研究日本血吸虫成虫自发荧光现象及共聚焦λ扫描特点,丰富日本血吸虫生物学和光谱学信息。方法小鼠感染日本血吸虫尾蚴6周后剖杀,收集成虫于无菌生理盐水中并分离雌雄虫体。荧光显微镜下分别以紫外光、蓝光、绿光等不同激发光源激发观察虫体自发荧光;激光共聚焦扫描显微镜下分别以405nm、458nm、476nm、488nm、514nm、543nm、633nm等7种激发光激发雄虫口吸盘部做自发荧光λ扫描分析。结果不同激发光照射下日本血吸虫虫体能发出多种不同颜色的自发荧光,以蓝光激发的黄绿色荧光效果最佳,虫体大体结构清晰;λ扫描分别以405nm、488nm、514nm和543nm激发光激发样品,相应敏感的发射波长分别为490-510nm、550-570nm、560-580nm和590-600nm,其中488nm和514nm激发效果较好。458nm、476nm、633nm激发效果欠佳。结论荧光显微镜下可观察日本血吸虫成虫自发荧光,最佳模式是以蓝光激发观察黄绿色荧光;日本血吸虫雄虫自发荧光光谱较宽,主要分布于500-600nm;488nm和514nm为较适宜的激发波长。     

共聚焦激光显微内镜对胃癌病理分型的诊断价值

目的建立共聚焦激光显微内镜（CLE）对胃癌病理分型的诊断标准,评价CLE对胃癌病理分型的诊断价值。方法分析行CLE检查且经组织病理证实为胃癌的36例患者的共聚焦图像,建立CLE对胃癌病理分型的诊断标准。根据共聚焦图像上腺体结构的有无和微血管形态的改变,CLE可将胃癌分为分化型和未分化型两种病理类型。以组织病理结果作为金标准,回顾性研究CLE对不同胃癌病理分型诊断的敏感性、特异性和准确性。结果CLE对分化型胃癌的诊断敏感性、特异性和准确性为81．8％、93．5％和88．7％,对未分化型胃癌的诊断敏感性、特异性和准确性为85．7％、92．3％和90．5％。结论共聚焦激光显微内镜对分化型和未分化型胃癌的诊断与组织病理学诊断有良好的一致性,提供了一种于内镜检查同时在体诊断胃癌病理分型的新工具。     

胃癌维甲酸信号通路相关基因启动子区协同高甲基化的研究

背景：启动子区高甲基化与胃癌中多种抑癌基因表达沉默密切相关。目的：探讨维甲酸信号通路相关基因维甲酸受体B（RAR13）、细胞维生素A结合蛋白1（CRBP1）和他扎罗汀诱导基因1（TIG1）启动子区高甲基化与胃癌的关系。方法：以甲基化特异性聚合酶链反应（MSP）检测40例胃癌标本、10例正常胃黏膜标本和6株胃癌细胞株的RAR13、CRBPI和TIG1基因启动子区甲基化状态，分析i者甲基化状态的相关性及其与胃癌1晦床病理特征的关系。以逆转录聚合酶链反应（RT—PCR）检测胃癌细胞株RAR13、CRBP1和TIG1mRNA表达。结果：40例胃癌组织的RAR13、CRBPI和TIG1基因甲基化率分别为45．0％、32．5％和57．5％，10例正常胃黏膜组织均未检测到上述基因甲基化（P〈0．05）。胃癌组织中RAR13的甲基化状态与CRBP1和TIG1的甲基化状态显著相关（P〈0．05），但三者的甲基化状态与胃癌临床病理特征无相关性。启动子区高甲基化胃癌细胞株相应基因mRNA表达缺失或减弱。结论：胃癌组织常发生维甲酸信号通路相关基因RAR13、CRBP1和TIG1启动子区高甲基化，高甲基化可能是相应基因转录失活的重要原因。     

Autofluorescence imaging analysis of gastric cancer

AIMS: To investigate the characteristics of gastric cancer in autofluorescence images. METHODS: A double‐channel laser scanning confocal microscope with an argon ion laser (excitation wavelength 488 nm) and helium?neon laser (excitation wavelength 543 nm) were used to detect autofluorescence from 16 gastric cancer tissue specimens and corresponding normal gastric tissue. RESULTS: Autofluorescence from normal gastric tissue produced a green‐colored image. The intensity of red color increased obviously in all gastric cancer tissues (100%) after illumination and the tissues produced a reddish‐brown‐colored image. CONCLUSIONS: A reddish‐brown image is characteristic of autofluorescence in gastric cancer detected by an argon ion laser and helium?neon laser with a double‐channel laser scanning confocal microscope. Autofluorescence imaging analysis is useful in the diagnosis of gastric cancer.     

Diagnosis of gastric cancer by using autofluorescence spectroscopy

AIMS: To evaluate the clinical use of laser‐induced fluorescence (LIF) spectra for diagnosing gastric cancer during gastroscopy. METHODS: A helium?cadmium laser system (excitation wavelength = 442 nm) was used to detect autofluorescence in 38 patients with endoscopically and histologically diagnosed gastric cancer and chronic gastritis. RESULTS: The LIF spectra produced by the helium? cadmium laser from gastric cancer tissue had one major peak (510 nm) and three minor peaks (590, 670, 710 nm). The intensity and shape of the LIF spectra in gastric cancer and normal gastric tissue were significantly different. Using the integral fluorescence intensities (FI) at 662?677 and 701?716 nm and the FI ratio of 510 nm/710 nm as the diagnostic parameters, it was possible to diagnose gastric cancer with a sensitivity of 83%, a specificity of 91%, a positive predictive value of 88% and a negative predictive value of 88%. The diagnostic accuracy was 88%. CONCLUSIONS: Using autofluorescence spectro­scopy to detect gastric cancer allows real‐time detection of gastric cancerous lesions and it serves as a guide for taking biopsy specimens. The technique may become a useful tool in the diagnosis of early gastric cancer.     

Autofluorescence excitation-emission matrices for diagnosis of colonic cancer

AIM: To investigate the autofluorescence spectroscopic differences in normal and adenomatous coionic tissues and to determine the optimal excitation wavelengths for subsequent study and clinical application. METHODS: Normal and adenomatous coionic tissues were obtained from patients during surgery. A FL/FS920 combined TCSPC spectrofluorimeter and a lifetime spectrometer system were used for fluorescence measurement. Fluorescence excitation wavelengths varying from 260 to 540 nm were used to induce the autofluorescence spectra, and the corresponding emission spectra were recorded from a range starting 20 nm above the excitation wavelength and extending to 800 nm. Emission spectra were assembled into a three-dimensional fluorescence spectroscopy and an excitation-emission matrix (EEM) to exploit endogenous fluorophores and diagnostic information. Then emission spectra of normal and adenomatous coionic tissues at certain excitation wavelengths were compared to determine the optimal excitation wavelengths for diagnosis of coionic cancer. RESULTS: When compared to normal tissues, low NAD (P)H and FAD, but high amino acids and endogenous phorphyrins of protoporphyrin IX characterized the high-grade malignant coionic tissues. The optimal excitation wavelengths for diagnosis of coionic cancer were about 340, 380, 460, and 540 nm. CONCLUSION: Significant differences in autofluorescence peaks and its intensities can be observed in normal and adenomatous coionic tissues. Autofluorescence EEMs are able to identify coionic tissues.     

Differences in laser-induced autofluorescence between adenomatous and hyperplastic polyps and normal colonic mucosa by confocal microscopy

Laser-induced autofluorescence has been used to discriminate normal from adenomatous colonic mucosa. However, few studies to date have studied the origin of colonic autofluorescence. Using confocal microscopy (excitation wavelength 488 nm), we have shown that autofluorescence at this wavelength is present predominantly in the lamina propria of normal mucosa but in the epithelium in adenomatous and hyperplastic polyps. The intensity ratio of epithelial cell to lamina propria fluorescence was significantly lower (P<0.0001) in normal mucosa (0.52±0.01) compared with either adenomatous (1.6±0.2) or hyperplastic polyps (1.7±0.15). However, the ratios were not significantly different between hyperplastic and adenomatous polyps. Thus, confocal microscopy enables the detection of the sites of autofluorescence within colonic mucosa and the quantitation of differences in fluorescence between different tissue types.     

Characterization of tissue autofluorescence in Barrett''s esophagus by confocal fluorescence microscopy

High grade dysplasia and early cancer in Barrett's esophagus can be distinguished in vivo by endoscopic autofluorescence point spectroscopy and imaging from non-dysplastic Barrett's mucosa. We used confocal fluorescence microscopy for ex vivo comparison of autofluorescence in non-dysplastic and dysplastic Barrett's esophagus. Unstained frozen sections were obtained from snap-frozen Barrett's esophagus biopsy samples and scanned with confocal fluorescence microscopy (458 nm excitation; 505-550 nm [green] and > 560 nm [red] emission). Digital micrographs were taken from areas with homogenous and specific histopathology. Visual inspection and statistical analysis were used to evaluate the image datasets. Dysplastic and non-dysplastic Barrett's esophagus epithelia fluoresced mainly in the green spectrum and the main sources of autofluorescence were the cytoplasm and lamina propria. High-grade dysplasia was differentiated from non-dysplastic Barrett's esophagus by microstructural tissue changes. However, there were no specific changes in either the locations or average intensities of intrinsic green and red autofluorescence at the epithelial level that could differentiate between dysplastic and non-dysplastic Barrett's esophagus epithelia, ex vivo. Detectable differences in autofluorescence between BE and dysplasia/cancer in vivo are probably not caused by specific changes in epithelial fluorophores but are likely due to other inherent changes (e.g. mucosal thickening and increased microvascularity) attenuating autofluorescence from the collagen-rich submucosa. Furthermore, confocal fluorescence microscopy provides 'histology-like' imaging of Barrett's tissues and may offer a unique opportunity to exploit microstructural tissue changes occurring during neoplastic transformation for in vivo detection of high-grade dysplasia in Barrett's patients using newly developed confocal fluorescence microendoscopy devices.     

Autofluorescence endoscopy images of pancreas cancer: report of a case

This is the first report of the observation of pancreas cancer with an autofluorescence endoscopic imaging system (excitation: 437 nm). A case of intraductal papillary adenocarcinoma of pancreas was presented. After pancreatectomy, the resected pancreas was used to test the endoscope (16Fr) in the pancreatic duct. The normal pancreatic duct was seen as light blue and the protruding cancerous lesion was observed as a dark red image. In previous studies, cancerous lesions of the gastrointestinal tract, bronchial tree and bile duct also appeared dark red when examined by autofluorescence endoscopy. In the pancreatic duct, the cancer lesion was also detected as dark red color.     

Evaluation of in vivo endoscopic autofluorescence spectroscopy in gastric cancer

BACKGROUND: The aim of this study was to evaluate light-induced autofluorescence spectroscopy for the in vivo diagnosis of gastric cancer. METHODS: A total of 344 endogenous fluorescence spectra were obtained from normal (164) and cancerous gastric mucosa (180) in 15 patients with pure adenocarcinoma and in 16 patients with gastric cancer containing signet-ring cells. A special light source capable of delivering either white or violet-blue light for the excitation of tissue autofluorescence via the endoscope was used. Endogenous fluorescence spectra emitted by the tissue were collected with a fiberoptic probe and analyzed with a spectrograph. RESULTS: Gastric adenocarcinoma exhibits specific changes in the emitted fluorescence spectra as compared with normal gastric mucosa. By algorithmic classification of the spectra, a sensitivity of 84%, specificity of 87%, a likelihood ratio for a positive test of 6.5 and for a negative test of 0.18 were obtained for the diagnosis of pure adenocarcinoma of the stomach. However, gastric cancer with signet-ring cells exhibits great variation in emitted autofluorescence spectra as compared with normal mucosa. The sensitivity for the diagnosis of all carcinomas containing signet-ring cells was 55%, specificity 85%, the likelihood ratio for a positive test was 3.7 and for a negative test, 0.53. The diagnostic value decreases with increasing numbers of signet-ring cells and tumor grade. CONCLUSIONS: Light-induced autofluorescence spectroscopy is a new and promising bio-optical technique for the endoscopic in vivo diagnosis of gastric adenocarcinoma. The poor diagnostic accuracy for signet-ring cell carcinoma may be explained by the diffuse and frequent submucosal growth of this tumor and the presence of collagen fibers.     

用激光扫描共聚焦显微镜的软件进行消化学分子共存分析

目的　如何应用激光扫描共聚焦显微镜的Ｌａｓｅｒｓｈａｒｐ 软件进行分子共存分析．方法　使用ＢｉｏＲａｄ 公司的Ｌａｓｅｒｓｈａｒｐ 软件分析肝癌和胃组织细胞中不同分子的共存．结果　仔细进行样本制备和图象采集才能得到分子共存的可靠结果． 所有抗体应严格设立对照,确保抗体的特异性,并无交叉反应． 同时进行两个分子荧光标记,应使用间接荧光标记技术． 荧光素的发射波长应无重叠,如用Ａｌｅｘａ４８８ 和丽丝胺罗丹明或Ｔｅｘａｓ Ｒｅｄ ． 发射滤光片应达到可最大采集,同时又避免与其他荧光素发射光谱的相互渗透． 通常应用的一对荧光素是ＦＩＴＣ 和ＴＲＩＴＣ 或ＦＩＴＣ 和Ｔｅｘａｓ Ｒｅｄ ． 用抗体和共聚焦显微镜进行共存研究时根据激光光源的差别分别选用不同的荧光素． 当荧光素的发射光谱存在相互渗透时,应分别采集不同荧光素标记的图象． 使用氪／ 氩激光进行图象采集时,注意仔细平衡激发强度,即使在同时激发的情况下,也可得到充分光谱分离的图象．结论　在图象采集时避免光谱渗透,并从所有图象中去除背底染色后,Ｌａｓｅｒｓｈａｒｐ 共存分析软件包可提供正确的共存系数．