| ObjectiveLaryngeal cancer is one of the common malignant tumors in Otolaryngology-Head and Neck Surgery, surgical operation is the primary measurement of laryngeal carcinoma at present. As the irregular morphous, the complex structure and 3D feature of the growth of tumor infiltration, surgical eradication of the primary laryngeal lesion called for an accurate grasp of the scope of tumor infiltration, spatial 3D structure and the scientifical surgical resectional scope. Positive margin is one of the major surgical operation failures. Master of carcinoma infiltration is not only the premise of precisely defined scope of surgical margin, but also the effective means of reducing positive margin. Precise surgical margin determines the survival rate of laryngeal cancer patients, the preserve retention of anatomical structure and reconstruction of laryngeal function. In this study, based on 3D-Doctor 3.5 software and 3D reconstruction techniques, serial slice of the normal 3D reconstruction of laryngeal specimens were produced in the experiment one, to establish a whole database of serial tissue sections of normal larynx, to ensure the normal throat samples of correct understanding of morphological structure; experiment 2 by the comparison between serial slices and MSCT and 3D reconstruction for laryngeal imaging provide anatomical basis for guiding the clinical application of MSCT in the throat; serial slices of laryngeal biopsy specimens and 3D reconstruction were completed. Observation of 3D morphology of laryngeal cancer invasion and growth characteristics were finished, visualization of human organs was used in laryngeal morphology study on laryngeal cancer through a comprehensive understanding of spatial structure, familiarity with the invasive growth of laryngeal features and determination of the laryngeal operation of 3D cutting margin, so that partial laryngectomy will be more reasonable to guide the clinical treatment of laryngeal cancer surgery.Methods:1. Serial slices of 30 (21male,9female) normal whole laryngeal specimen from the north region of China were produced, thickness of slice is 5-15μm, the slice was selected every 300μm, HE stained. The photos of serial slices were acquired by professional miscrospur photograph system. A whole database of serial tissue sections of normal larynx was established. Comparative study was executed between general specimens and the virtual specimens after 3D reconstructions with 3D-Doctor 3.5 software in the graphic workstation of INTEL XEON 5472 (double route). Four reference diameters of clinical laryngeal invasion pathways were selected and the corresponding morphological parameters were measured.2. MSCT images of 30 volunteers (12male,18female) normal necks were acquired and 3D reconstruction of MSCT images were accomplished by 3D-Doctor 3.5 software in the professional graphics workstation of INTEL XEON 5472 (double route), and measurement data of the serial slices and MSCT images were compared with the 3D reconstruction of serial sections, which the measurements parameters are same to that of experiment one.3.3 specimens with en bloc surgical resection in laryngeal carcinoma patients were collected, which including supraglottic type, glottic type and straddled glottic type, T3, T4) were cut along the long axis of tumor specimens so as to produce serial sections, slice images were obtained by professional microspur photograph system, and virtual model of 3D reconstruction of laryngeal carcinoma were accomplished by 3D-Doctor 3.5 software in the professional graphics workstation of INTEL XEON 5472 (double route). Compared with the image of preoperative MSCT, the serial sections of laryngeal cancer and its histological features of infiltration characteristics were grasped.Results:1.1 Difference with the previous measurements studies, the attachment of vocal cord is above 0.718mm±0.487,95%confidence interval for the 0.369mm-1.066mm. The distances from the attachment of vocal cord in thyroid carrlage to the superior notch and the distance to inferior notch and their rate of the gender are no significant difference (P>0.05), except that, the other measurements data are significant difference on the gender (P<0.05).1.2 The measuring data of general specimens are more than that of the virtual specimens of serial slices, but the difference is not significant (P>0.05).1.3 Arytenoid cartilage lie in the posterior 1/2-2/3 of thyroid cartilage. In addition to the depth of vocal cord attached (in the vocal cords vocal cords attached to the edge of the end of the distance) to 1.665mm-1.736mm, muscular process to the thyroid cartilage endosteum distance 3.651mm-3.715mm, the rest of the indicators are gender differences (P<0.05).1.4 Measurements of pre-epiglottic and paraglottic space:pre-epiglottic space in the sagittal plane of the area 80.930mm2±8.911, in the adjacent area of the median sagittal plane 84.757 mm2±15.482. Paraglottic space after vocal cord level in the area of section 172.400 mm2±6.968, the middle point in the film by the vocal cord on coronal section area of 248.751 mm2±5.543, in the arytenoid cartilage posterior coronal section area of 221.893 mm2±9.473, notch between the ladle Department coronal section area of 186.678 mm2±13.249.The area measurements of laryngeal spaces in the different section provide an anatomic reference data for clinicians on the infiltration of laryngeal cancer.1.5 Measurements of four reference diameters on common infiltration paths of laryngeal cancer: the distance of the stem end of the epiglottis to the anterior commissure was males 5.122mm±1.031, females 4.051mm±0.944; the distance of the epiglottic end of hyoepiglottic ligament to ventricular bands was male 23.807±3.747, female 20.638mm±2.748; the distance of midpoint of aryepiglottic fold to ventricular bands was male 21.708mm±4.724, female to 18.423mm±4.245; the distance of vocal cord to the outer edge of thyroarytenoid muscle was male 3.322±0.724, female 2.371mm±1.032; the distance of vocal cord to endosteum of thyroid carlilage lamina was male 7.245±1.524, female 6.833mm±1.425. Gender differences in all measured parameters were significant (P<0.05).1.6 Transection position of vocal process:Because the distance of the posterior end of vocal process to the attachment edge of vocal cord was 1.665mm-1.736mm, the length of vocal process was male 5.194mm, female 4.092mm. Therefore, transaction position at the anterior 1/3 and 1/2 of vocal process is more appropriate.1.7 Thyroid cartilage fenestration improvements, main points:(1) determine the window center of a circle:Using the midpoint of the posterior border of thyroid cartilage center of the circle to the midpoint of posterior margin of the thyroid cartilage to the muscular process of arytenoid cartilage from the projection point for the radius, the thyroid cartilage marker board to do the first arc; muscle process as center of a circle, with half of the distance between top and bottom radius of the thyroid cartilage board to do the second arc; two windows shall be centered on the intersection curve;. (2) from the outside layer to remove the thyroid cartilage window, hard to be modest, avoid cartilage fracture; (3) along the lower edge of the windows or cricoarytenoid muscle to determine muscle process. Operation Notes:(1) to avoid the external branch of superior laryngeal nerve injury. (2) to avoid injury pyriform sinus mucosa.2. MSCT image of the thyroid cartilage, epiglottic cartilage and cricoid cartilage showed clearly, arytenoid cartilage showed incomplete, the structure in the laryngeal space can not be displayed. The virtual model of MSCT reconstruction were measured and compared with serial slices of the laryngeal cartilage was no significant difference (P> 0.05). Using 3D-Doctor 3.5 software, cutting 3D reconstruction of MSCT virtual model,6 cutting plane were obtained as the same level of serial slices, the area of pre-epiglottic space and paraglottic space were compared between MSCT and serial slices, the results there was no significant difference (P>0.05). In the common ways of laryngeal cancer infiltration, four reference diameters measurements were compared between tissue sections and MSCT, the results showed:the distance of vocal cords to the thyroid cartilage endosteum of MSCT less than that of serial slices, the other parameters of MSCT are more than serial slices, but there were all no significant difference (P>0.05).3. Serial sections of the cross-section observation of laryngeal carcinoma were observed, boundaries were distinguished clearly between tumor and normal tissue. In the cross-section aspect, type of tumor was round, irregular-shaped and so on, the color of the tumor is darker than the normal, and the tumor showed substantial part of the heterogeneity, with some regions more dense in some regions, loose, color rendering differences, but the larger is the tumor,the more obvious kinds of phenomena are not homogeneous. Laryngeal space shape is irregular, tumor growth is the spatial 3D growth and no direction, which can be invasivetowords the growth of any organization around in the 3D reconstruction model of laryngeal carcinoma.Conclusion1. The data sets of serial slices of whole normal larynx were established. The virtual specimens of 3D reconstruction are identified with the real general specimen basically, and provided the anatomical data fo clinical applications and imaging.2. MSCT in excess of its fine structure shows poor resolution, the details of serial slices are more full and clear than MSCT 3D reconstruction. Based on the human laryngeal serial slices,3D reconstruction is apt to medical teaching and research. MSCT and its 3D reconstruction is apt to clinical applications and can be used as clinical diagnosis and imaging examination of the auxiliary tool.3. The virtual model of Laryngeal resection specimens of 3D reconstruction of serial sections provide a visual image of the tumor 3D graphics, help clinicians are familiar with the spatial laryngeal morphology and growth characteristics of infiltration, improve diagnosis and treatment of laryngeal carcinoma.Creative point:1. Complete firstly the serial slices of larynx, and establish the database of tissue slices of larynx.2. Using professional graphics workstation processing tissue slices, to accomplish virtual 3D reconstruction of laryngeal tissue sections. From multiple perspectives, multiple levels, visually display morphological characteristics of the organizational structure of larynx, which is better than the 3D reconstruction of MSCT, is more effective in guiding the clinics, and is helpful to determine the 3D cutting edge of laryngeal surgery.
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