激光捕获显微切割制备垂体腺瘤蛋白质组学研究样品

目的 探讨应用免疫组织化学导航激光捕获显微切割(LCM)技术获取高同质性泌乳素垂体腺瘤细胞及正常垂体泌乳素细胞的方法及可行性.方法 冰冻泌乳素腺瘤标本及对照正常垂体常规制备8μm冰冻覆膜切片,采用两步法免疫组织化学方法染色;应用LCM技术切割腺瘤内及正常对照垂体内泌乳素细胞;选择性获取同质泌乳素细胞和配对正常细胞.结果 60 min内完成免疫组织化学染色,细胞显色较理想.采用4μm紫外线激光可以准确、快速地分离片状甚至单个泌乳素分泌细胞;无间质细胞污染,细胞形态保持完好.结论 应用LCM技术可以成功获垂体瘤蛋白组学研究中的异质性问题,所获组织同质性高、操作过程稳定是垂体瘤蛋白质组学研究较有价值的样品制备方法.     

Laser Doppler vibrometry: a new tool for diagnosing hearing loss with an intact eardrum

The diagnosis of hearing loss with an intact eardrum frequently requires an entire battery of hearing tests, without the guarantee of an exact diagnosis. The techniques frequently provide only orientation for it, without establishing the site of the lesion and the etiology of the hearing loss. Laser Doppler vibrometry is a new technique, which has recently proved capable, of partially resolving this problem. The method is based on the study of the sound-induced vibration of the eardrum in humans in vivo, using a laser Doppler vibrometer. The method proved to be useful in the diagnosis of the pathology of the middle ear sound transmission system, avoiding the need for exploratory tympanotomy. Called "laser-audiometry", the method promises to become a new diagnostic tool for hearing impairment.     

激光捕获显微切割技术在神经科学研究中的应用进展

背景 激光捕获显微切割技术(laser capture microdissection,LCM)是一项在显微镜直视下应用激光技术从组织中分离同质细胞,进行后续研究的新技术.目的 现就LCM的基本原理和分类、在神经科学研究的近期应用、主要进展及未来发展作一综述.内容 近年来随着样本切片制备、染色、核酸提取和扩增等辅助技术的改进,该技术在神经科学的研究中得到广泛应用,且表明其具有显著优势.趋势 针对神经科学研究的特点,进一步探索和改进相关技术,促进LCM技术在神经科学研究领域中的应用.     

Gastrointestinal tonometry: a new tool for the anesthesiologist

Intestinal hypoperfusion is among the factors implicated in sepsis and multiorgan failure. Splanchnic blood flow may be sacrificed to maintain supply to vital organs, even when hemodynamic alterations are minor. The sensitivity of invasive hemodynamic monitoring for detecting intestinal hypoperfusion is low. This paper aims to review current knowledge about indirect measurement of splanchnic perfusion by way of gastrointestinal tonometry. We review the pathophysiology of ischemic intestinal lesions, the basis for gastrointestinal tonometry, and the method. Finally we discuss clinical applications (early diagnosis of ischemic colitis and ischemia of the flap after esophageal reconstruction, weaning from mechanical ventilation, abdominal compartment syndrome, liver transplant, heart surgery, prognostic factors and care of the critically ill patient). An adequate understanding of this monitoring technique and management of information it provides can give an early warning of the intestinal hypoperfusion that precedes other serious systemic complications.     

Interphase cytogenetics: a new tool for the study of genetic changes in brain tumors.

Interphase cytogenetics is the application of nonradioactive in situ hybridization with chromosome-specific DNA probes to interphase nuclei. In this study, interphase cytogenetics was used to investigate 66 primary brain tumors (33 gliomas, 30 meningiomas, and three medulloblastomas) for numerical chromosomal aberrations of chromosomes 1, 6, 7, 10, 11, 17, 18, X, and Y. Of the 33 gliomas (17 astrocytomas grades II, III, and IV, five oligoastrocytomas, seven oligodendrogliomas, and four ependymal tumors), 22 were near diploid, while the remaining 11 showed a significant triploid or tetraploid component. The predominant specific aberrations in gliomas were an over-representation of chromosome 7 (13 cases) and an under-representation of chromosome 10 (16 cases). These changes were observed in grade III and grade IV astrocytomas, as well as in oligodendrogliomas. Other frequent numerical changes were a gain of chromosome 17 (six cases) and a loss of chromosome 18 (seven cases). This loss of chromosome 18 seemed relatively specific for gliomas with an oligodendroglial component (six cases). Only two of 33 gliomas displayed no genetic abnormality with the probes used. Seven patients with astrocytomas died of their brain tumor during the clinical follow-up period. Their astrocytomas did not show a different chromosomal constitution compared to the other gliomas. For the meningiomas, the probe panel was extended with a probe specific for chromosome 22. Loss of chromosome 22 was obvious in 21 of the 30 meningiomas, and was the sole abnormality in 11 meningiomas; in the other 10, this loss was associated with other chromosomal changes. Five of these tumors with additional aberrations were recurrent or atypical meningiomas. It is suggested that interphase cytogenetics can contribute to a better understanding of the biological behavior of these tumors and possibly result in better insights into prognosis and strategies for therapy.     

Localization of postresection EGF receptor expression using laser capture microdissection

Background/Purpose: Epidermal growth factor (EGF) and its receptor (EGFR) are key components in the genesis of adaptation after small bowel resection (SBR). Within intestinal homogenates, EGFR expression is increased after SBR; however, the exact cells responsible for altered EGFR expression are unknown. In this study, laser capture microdissection (LCM) microscopy was used to elucidate the specific cellular compartment(s) responsible for postresection changes in EGFR expression. Methods: Male ICR mice underwent a 50% proximal SBR or sham operation. After 3 days, frozen sections were taken from the remnant ileum. Individual cells from villi, crypt, muscularis, and mesenchymal compartments were isolated by LCM. EGFR mRNA expression for each cell compartment was quantified using real-time polymerase chain reaction (PCR). Results: EGFR expression was increased after SBR within the crypt (2-fold) and muscularis compartments (3-fold). There were no changes detected after SBR in the villus tips or mesenchymal compartments. Conclusions: Increased expression of EGFR in crypts directly correlates with the zone of cell proliferation and supports the hypothesis that EGFR signaling is crucial for the mitogenic stimulus for adaptation. The finding of increased EGFR expression in the muscular compartment is novel and may implicate a role for EGFR as a mediator of the muscular hyperplasia seen after massive SBR. J Pediatr Surg 38:440-445.     

Integrated medical records for surgical services: a new tool for a new hospital model

According to a modern definition, the patient's medical record is a tool shared by health-care professionals whose purpose is to support the planning, providing and documenting of patient-centred care. The requirements and regulations of medical records, which date back to the sixties, need now be adapted to more recent organisational models based on different levels of care intensity and strongly integrated patient-oriented care approaches designed to create connectivity, alignment and collaboration within and between different professional fields. On this basis, the SS. Cosma and Damiano Hospital Chief Physician endorsed a multiprofessional project providing for the complete reorganisation of surgical activities according to the above-mentioned approach and, at the same time, for the elaboration of an "integrated" medical record, capable of overcoming the traditional separation between medical and nursing documents. This paper presents the results of a long period of complex teamwork consisting in revising and sharing a new medical record model based on principles of professional integration and patient-centredness in health-care provision.     

Computerized contrast angiosonography: a new diagnostic tool for the urologist?

OBJECTIVES: To evaluate the diagnostic potential of echo-enhanced ultrasonography (US) for depicting the vascularization pattern of renal cell carcinoma (RCC), and calculating the first-pass effect using harmonic imaging, against that obtained by triphasic helical computed tomography (CT). PATIENTS AND METHODS: Sixty patients with surgically confirmed RCC underwent US using B-mode and power Doppler methods with or without an intravenous microbubble echo-enhancing agent. After depicting and defining the tumour extent by B-mode US, the first-pass effect/enhancement by the echo-enhancing agent within the lesion, and that of a reference area of unaffected renal cortex, were recorded on-line by calculating the mean pixel intensity. Time-intensity curves, i.e. the rise time and gradient of both the suspected tumour and reference areas, were constructed. RESULTS: Using B-mode US, the extent of all tumours was delineated (mean tumour size 3.8 cm, SD 0.6). After applying the microbubble agent all tumours were enhanced, whereas the perfusion was decreased (in 48%), increased (in 16%) or similar (in 36%) compared with the cortical reference area. Using the Hounsfield classification, these results correlated well with the hypo/hypervascularity shown on CT. CONCLUSION: Ultrasonography has considerable potential in diagnosing RCC, if combined with echo-enhancing methods, harmonic imaging and computer-based calculation of tumour vascularization. Dynamic US studies should provide a diagnostic yield similar to that of CT.