수술 연습 <ol><li> 복부 CT 촬영에서 쉽게 그리고 대동맥에서 직접 발생하는 권리 간장 동맥의 존재를 증명 간 0.3 D 재건의 왼쪽에있는 담즙 관이 (어두운 색상으로 간 실질 조직 내부에 게재) 광범위하게 확대보실 수 있습니다 더 이상의 혈관 이상 없습니다. </li><li> 간장은 기능 8 세그먼트로 동영상에 표시되며 팽창이 세그먼트 2, 3 (왼쪽 분야)로 제한이라고 생각하고, 복강경 왼쪽 sectoriectomy가 계획된로 나눌 수 있습니다. </li></ol> Intraoperative 절차 <ol><li> 환자는 부정사 위치와 환자의 다리 사이에 외과의 스탠드에 배치됩니다. 동영상과 같이 5 trocars 사용하여 배치됩니다 </li><li> 혈관 클램프는 (프링글의 manouvre) 출혈을 줄이기 위해 포털 깡패에 위치 </li><li> 간 실질 조직의 부서는 세그먼트 III와 IV의 낫 모양의 제품 인대 따라 메스 조합 또는 바이폴라 electrocautery 및 고조파를 사용하여 수행됩니다 </li><li> 일단 작은 꽃자루 세그먼트 III 완전히 그것이 혈관 카트리지 45mm 엔도 GIA를 사용하여 나눌 수 있습니다 해부하는 것입니다. </li><li> 그 절차는 주요 세그먼트 II에 작은 꽃자루는 혈관 카트리지 45mm endoGIA을 사용하여 격리에 도달하고 마지막으로 나누어져 있습니다까지 실질의 추가 부문으로 실시 </li><li> 왼쪽 삼각 인대 지금은 혈관 스테이플러를 사용하여 electrocautery하고 왼쪽 간장 정맥은 격리와 나누어집니다를 사용하여 나누어져 있습니다 </li><li> 간 표면은 출혈에 대한 검사, 아르곤 빔은 간 표면에 적용되고 혈관 클램프가 제거됩니다. 이 시점에서 섬유소 접착제는 이후 수술 담즙 누출의 위험을 줄이기 위해에 적용됩니다. 표본은 endobag에 위치하고 제거 </li></ol>

<ol>
	<li>Nguyen, K. T., Gamblin, T. C., Geller, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopic+liver+resection+for+cancer.">Laparoscopic liver resection for cancer.</a> Future Oncol. 4 (5), 661-670 (2008).</li><li>Gumbs, A. A., Gayet, B., Gagner, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopic+liver+resection:+when+to+use+the+laparoscopic+stapler+device.">Laparoscopic liver resection: when to use the laparoscopic stapler device.</a> HPB (Oxford). 10 (4), 296-303 (2008).</li><li>Pulitan&#242;, C., Catena, M., Arru, M., Guzzetti, E., Comotti, L., Ferla, G., Aldrighetti, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopic+liver+resection+without+portal+clamping:+a+prospective+evaluation.">Laparoscopic liver resection without portal clamping: a prospective evaluation.</a> Surg Endosc. 22 (10), 2196-2200 (2008).</li><li>Yonem, O., Bayraktar, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+characteristics+of+Caroli's+disease.">Clinical characteristics of Caroli's disease.</a> World J Gastroenterol. 13 (13), 1934-1937 (2007).</li></ol>

복강경 수술은 안전하게 간 절제술 모두에 양성 및 악성 질환에도 수행할 수 있습니다. 법으로서 접근법을 사용 외과 외상을 줄이면 게시 수술 합병증 및 사후 수술 입원의 발생을 최소화 할 수 있습니다.

laparoscopic left liver sectoriectomy of caroli's disease limited to segment ii and iii

Caroli의 질병은 내부 우산이끼의 담즙 관의 비정상적인 팽창과 같이 정의됩니다 : 그 발병률은 매우 낮은 (백만 인구의 1)과 가지 경우 대부분의 전체 간은 관심과 간 이식은 선택의 치료입니다. 왼쪽 또는 오른쪽으로 기억 상실증에 국한 팽창의 경우 간 절제술을 수행할 수 있습니다. 많은 년 간 절제술의 표준 접근 방식은 상당한 이후 operatie의 병적 상태에 의해 특징 복부의 큰 절개를 통해 정식으로 개복술되었습니다. 최근 법으로서, 복강경 접근 방법은 모두 양성 및 악성 질환에 대한 간 절제술 가능한 수술 기법으로 제안되었습니다. 법으로서 접근 방식의 주요 장점은 빠른 복구 덜 이후 수술 합병증 수있는 수술 외상의 상당한 감소에 의해 표시됩니다. 이 동영상은 Caroli의 질병의 케이스가 발열과 관련된 복통의 갑작스런 발병을위한 위장 부서에 입원 58년 중년 남성에서 발생 보여줍니다 (&gt; 38C °), 구역질, 오한. 복부 초음파는 담낭이나 일반적인 담즙 덕트의 돌을없이 증거와 내부 간의 왼쪽 sited 담즙 관의 큰 팽창을 보여주었다. 이러한 연구 결과는 복부 고해상도 컴퓨터 tomography을 확인했다. 복강경 왼쪽 sectoriectomy이 계획되었다. 다섯 trocars 30가 ° 광을 사용했습니다, 복강의 탐험은 다른 질병에 대한 adhesions이나 증거를 보여주 없습니다. 간 혈액 유입을 조절하기 위해 혈관 클램프 (프링글의 manouvre) 작은 꽃자루 간장에 배치 되었음 Parenchymal 본부는 5mm 바이폴라 포셉 및 5mm 초음파 해부학자의 결합 사용과 수행됩니다. 심각하게 확대 왼쪽 간장 덕트는 격리와 45mm 내시경 혈관 스테이플러를 사용하여 분할되었다. 간 해부 그러면 45mm 혈관 스테이플러의 추가 카트리지로 나뉘어 있던 메인 왼쪽 포털 지점의 고립까지 계속되었다. 그 시점에서 왼쪽으로 간은 왼쪽 간장 정맥으로 연결된 남아 삼각 인대의 사업부는 monopolar의 후크와 분리된 간장 정맥을 사용하여 수행하고 혈관 스테이플러를 사용하여 분할되었다. Haemostatis는 아르곤 빔 응고의 응용 프로그램에 의해 정제된했고 더 출혈은 심지어 혈관 클램프의 제거 (총 프링글의 시간 27분) 이후에 발표되지 않았습니다. 수술 과정은 간기능 검사의 사건이없는, 최소한의 해발 고도는 이후 수술 1 일에 기록했지만 이후 수술 3 일에 배출에 정상으로 돌아였습니다.

Watch this Scientific Journal Video about Laparoscopic Left Liver Sectoriectomy of Caroli's Disease Limited to Segment II and III at JoVE.com

Laparoscopic Left Liver Sectoriectomy of Caroli&#039;s Disease Limited to Segment II and III

이 동영상은 간암의 세그먼트 II와 III로 제한 Caroli 병을 앓고 53 세의 남자에서 perfomed 복강경 좌측 간 sectoriectomy 사건을 선물한다.

Caroli의 질환의 복강경 왼쪽 간 Sectoriectomy는 세그먼트 II와 III로 제한

Caroli's disease is defined as a abnormal dilatation of the intra-hepatica bile ducts: Its  incidence is extremely low  (1 in 1,000,000 population) and in ...

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Biology

Laparoscopic Left Liver Sectoriectomy of Caroli's Disease Limited to Segment II and III

14.0K 조회수.  University of Insubria. 이 동영상은 간암의 세그먼트 II와 III로 제한 Caroli 병을 앓고 53 세의 남자에서 perfomed 복강경 좌측 간 sectoriectomy 사건을 선물한다.

비디오: Laparoscopic Left Liver Sectoriectomy of Caroli's Disease Limited to Segment II and III

A Matrigel-Based Tube Formation Assay to Assess the Vasculogenic Activity of Tumor Cells

Over the past several decades, a tube formation assay using growth factor-reduced Matrigel has been typically employed to demonstrate the angiogenic activity of vascular endothelial cells in vitro1-5. However, recently growing evidence has shown that this assay is not limited to test vascular behavior for endothelial cells. Instead, it also has been used to test the ability of a number of tumor cells to develop a vascular phenotype6-8. This capability was consistent with their vasculogenic behavior identified in xenotransplanted animals, a process known as vasculogenic mimicry (VM)9. There is a multitude of evidence demonstrating that tumor cell-mediated VM plays a vital role in the tumor development, independent of endothelial cell angiogenesis6, 10-13. For example, tumor cells were found to participate in the blood perfused, vascular channel formation in tissue samples from melanoma and glioblastoma patients8, 10, 11. Here, we described this tubular network assay as a useful tool in evaluation of vasculogenic activity of tumor cells. We found that some tumor cell lines such as melanoma B16F1 cells, glioblastoma U87 cells, and breast cancer MDA-MB-435 cells are able to form vascular tubules; but some do not such as colon cancer HCT116 cells. Furthermore, this vascular phenotype is dependent on cell numbers plated on the Matrigel. Therefore, this assay may serve as powerful utility to screen the vascular potential of a variety of cell types including vascular cells, tumor cells as well as other cells.

A tube formation assay is used to evaluate vascular activity of tumor cells.

종양 세포의 Vasculogenic 활동을 평가 Matrigel 기반 튜브 형성 분석

Over the past several decades, a tube formation assay using growth factor-reduced Matrigel has been typically employed to demonstrate the angiogenic ...

연구

생물학

In vivo Liver Endocytosis Followed by Purification of Liver Cells by Liver Perfusion

The liver is the metabolic center of the mammalian body and serves as a filter for the blood. The basic architecture of the liver is illustrated in figure 1 in which more than 85% of the liver mass is composed of hepatocytes and the remaining 15% of the cellular mass is composed of Kupffer cells (KCs), stellate cells (HSCs), and sinusoidal endothelial cells (SECs). SECs form the blood vessel walls within the liver and contain specialized morphology called fenestrae within in the cytoplasm. Fenestration of the cytoplasm is the appearance of holes (&tilde;100 &mu;m) within the cells so that the SECs act as a sieve in which most chylomicrons, chylomicron remnants and macromolecules, but not cells, pass through to the hepatocytes and HSCs 1 (Fig. 1). Due to the lack of a basement membrane, the gap between the SECs and hepatocytes form the Space of Disse. HSCs occupy this space and play a prominent role in regulation and response to injury, storage of retinoic acid and immunoregulation of the liver 2.
SECs are among the most endocytically active cells of the body displaying an array of scavenger receptors on their cell surface 3. These include SR-A, Stabilin-1 and Stabilin-2. Generally, small colloidal particles less than 230 nm and macromolecules in buffer phase are taken up by SECs, whereas, large particles and cellular debris is endocytosed (phagocytosed) by KCs 4. Thus, the bulk clearance of extracellular material such as the glycosaminoglycans from blood is largely dependent on the health and endocytic functions of SECs 5,6. For example, an increase in blood hyaluronan levels is indicative of liver disease ranging from mild to more severe forms 7.
With the exception of one report 8, there are no immortalized SEC cell lines in existence. Even this immortalized cell line is de-differentiated in that it does not express scavenger receptors that are present on primary SECs (our data, not shown). All cell biological studies must be performed on primary cells obtained freshly from the animal. Unfortunately, SECs dedifferentiate under standard culture conditions and must be used within 1 or 2 days upon isolation from the animal. Differentiation of SECs is marked by the expression of Stabilin-2 or HARE receptor 9 , CD31, and the presence of cytoplasmic fenestration 1. Differentiation of SECs can be extended by the addition of VEGF in culture media or by culturing cells in hepatocyte conditioned medium 10,11.
In this report, we will demonstrate the endocytic activity of SECs in the intact organ using radio-labeled heparin for hyaluronan for the SEC-specific Stabilin-2 receptor. We will then purify hepatocytes and SECs from the perfused liver to measure endocytosis.

In vivo Liver Endocytosis Followed by Purification of Liver Cells by Liver Perfusion

The study of liver sinusoidal endothelial cells (SECs) must be performed with primary cells obtained from the animal as no cell lines exist. This method relies on liver digestion and differential centrifugation for SEC purification for subsequent culturing and experimentation.

간 재관류에 의한 간 세포의 정화를 얹는 생체내 간 Endocytosis에서

The liver is the metabolic center of the mammalian body and serves as a filter for the blood.  The basic architecture of the liver is illustrated in ...

The Use of Ex Vivo Whole-organ Imaging and Quantitative Tissue Histology to Determine the Bio-distribution of Fluorescently Labeled Molecules

Fluorescent labeling is a well-established process for examining the fate of labeled molecules under a variety of experimental conditions both in vitro and in vivo. Fluorescent probes are particularly useful in determining the bio-distribution of administered large molecules, where the addition of a small-molecule fluorescent label is unlikely to affect the kinetics or bio-distribution of the compound. A variety of methods exist to examine bio-distribution that vary significantly in the amount of effort required and whether the resulting measurements are fully quantitative, but using multiple methods in conjunction can provide a rapid and effective system for analyzing bio-distributions.
Ex vivo whole-organ imaging is a method that can be used to quickly compare the relative concentrations of fluorescent molecules within tissues and between multiple types of tissues or treatment groups. Using an imaging platform designed for live-animal or whole-organ imaging, fluorescence within intact tissues can be determined without further processing, saving time and labor while providing an accurate picture of the overall bio-distribution. This process is ideal in experiments attempting to determine the tissue specificity of a compound or for the comparison of multiple different compounds. Quantitative tissue histology on the other hand requires extensive further processing of tissues in order to create a quantitative measure of the labeled compounds. To accurately assess bio-distribution, all tissues of interest must be sliced, scanned, and analyzed relative to standard curves in order to make comparisons between tissues or groups. Quantitative tissue histology is the gold standard for determining absolute compound concentrations within tissues.
Here, we describe how both methods can be used together effectively to assess the ability of different administration methods and compound modifications to target and deliver fluorescently labeled molecules to the central nervous system1.

The Use of Ex Vivo Whole-organ Imaging and Quantitative Tissue Histology to Determine the Bio-distribution of Fluorescently Labeled Molecules

Ex vivo whole-organ imaging is a rapid method for determining the relative concentrations of fluorescently labeled compounds within and between tissues or treatment groups. Conversely, quantitative fluorescence histology, while more labor intensive, allows for the quantification of the absolute tissue levels of labeled molecules.

사용<em&gt; 전의 VIVO</em&gt; 전체 기관 이미징 및 정량 조직 조직학는 형광 표지 된 분자의 생물 분포를 확인하는 방법

Fluorescent labeling is a well-established process for examining the fate of labeled molecules under a variety of experimental conditions both in vitro ...

Protocols for Testing the Toxicity of Novel Insecticidal Chemistries to Mosquitoes

New classes of insecticides with novel modes of action are needed to control insecticide resistant populations of mosquitoes that transmit diseases such as Zika, dengue and malaria. Assays for rapid, high-throughput analyses of unformulated novel chemistries against mosquito larvae and adults are presented. We describe protocols for single point-dose and dose response assays to evaluate the toxicity of small molecule chemistries to the Aedes aegypti vector of Zika, dengue and yellow fever,&#160;the malaria vector, Anopheles gambiae and the northern house mosquito, Culex quinquefasciatus, on contact and via ingestion. As an example, we evaluated the toxicity of amitriptyline, a small molecule antagonist of G protein-coupled receptors, via larval, adult topical and adult blood-feeding assay. The protocols provide a starting point to investigate insecticide potential. Results are discussed in the context of additional experiments to explore product applications and mechanisms for delivery.

Protocols are described for assessing the toxicity of chemistries to immature and adult mosquitoes for development as new classes of larvicides, adulticides and endectocides. The protocols enable high-throughput testing of multiple chemistries at single-point dose and subsequent evaluation via dose response assay to determine toxicity on contact or via ingestion.

모기에 소설 Insecticidal 화학 물질의 독성을 테스트 하기 위한 프로토콜

New classes of insecticides with novel modes of action are needed to control insecticide resistant populations of mosquitoes that transmit diseases such ...

Isolation and Fluorescence Imaging for Single-particle Reconstruction of Chlamydomonas Centrioles

Centrioles are large macromolecular assemblies important for the proper execution of fundamental cell biological processes such as cell division, cell motility, or cell signaling. The green algae Chlamydomonas reinhardtii has proven to be an insightful model in the study of centriole architecture, function, and protein composition. Despite great advances toward understanding centriolar architecture, one of the current challenges is to determine the precise localization of centriolar components within structural regions of the centriole in order to better understand their role in centriole biogenesis. A major limitation lies in the resolution of fluorescence microscopy, which complicates the interpretation of protein localization in this organelle with dimensions close to the diffraction limit. To tackle this question, we are providing a method to purify and image a large number of C. reinhardtii centrioles with different orientations using super-resolution microscopy. This technique allows further processing of data through fluorescent single-particle averaging (Fluo-SPA) owing to the large number of centrioles acquired. Fluo-SPA generates averages of stained C. reinhardtii centrioles in different orientations, thus facilitating the localization of distinct proteins in centriolar sub-regions. Importantly, this method can be applied to image centrioles from other species or other large macromolecular assemblies.

Isolation and Fluorescence Imaging for Single-particle Reconstruction of Chlamydomonas Centrioles

We have developed a strategy to purify and image a large number of centrioles in different orientations amenable for super-resolution microscopy and single-particle averaging.

절연 및 Chlamydomonas Centrioles의 단일 입자 재건을 위한 형광 이미징

Centrioles are large macromolecular assemblies important for the proper execution of fundamental cell biological processes such as cell division, cell ...

Isolation of Human Primary Valve Cells for In vitro Disease Modeling

Calcific aortic valve disease (CAVD) is present in nearly a third of the elderly population. Thickening, stiffening, and calcification of the aortic valve causes aortic stenosis and contributes to heart failure and stroke. Disease pathogenesis is multifactorial, and stresses such as inflammation, extracellular matrix remodeling, turbulent flow, and mechanical stress and strain contribute to the osteogenic differentiation of valve endothelial and valve interstitial cells. However, the precise initiating factors that drive the osteogenic transition of a healthy cell into a calcifying cell are not fully defined. Further, the only current therapy for CAVD-induced aortic stenosis is aortic valve replacement, whereby the native valve is removed (surgical aortic valve replacement, SAVR) or a fully collapsible replacement valve is inserted via a catheter (transcatheter aortic valve replacement, TAVR). These surgical procedures come at a high cost and with serious risks; thus, identifying novel therapeutic targets for drug discovery is imperative. To that end, the present study develops a workflow where surgically removed tissues from patients and donor cadaver tissues are used to create patient-specific primary lines of valvular cells for in vitro disease modeling. This protocol introduces the utilization of a cold storage solution, commonly utilized in organ transplant, to reduce the damage caused by the often-lengthy procurement time between tissue excision and laboratory processing with the benefit of greatly stabilizing cells of the excised tissue. The results of the present study demonstrate that isolated valve cells retain their proliferative capacity and endothelial and interstitial phenotypes in culture upwards of several days after valve removal from the donor. Using these materials allows for the collection of control and CAVD cells, from which both control and disease cell lines are established.

This protocol describes the collection of human aortic valves extracted during surgical aortic valve replacement procedures or from cadaveric tissue, and the subsequent isolation, expansion, and characterization of patient specific primary valve endothelial and interstitial cells. Included are important details regarding the processes needed to ensure cell viability and phenotype specificity.

체외 질병 모델링을 위한 인간 일차 판막 세포의 분리

Calcific aortic valve disease (CAVD) is present in nearly a third of the elderly population. Thickening, stiffening, and calcification of the aortic valve ...

Novel In Vivo Micro-Computed Tomography Imaging Techniques for Assessing the Progression of Non-Alcoholic Fatty Liver Disease

Non-alcoholic fatty liver disease (NAFLD) is a growing global health issue, and the impact of NAFLD is compounded by the current lack of effective treatments. Considerable limiting factors hindering the timely and accurate diagnosis (including grading) and monitoring of NAFLD, as well as the development of potential therapies, are the current inadequacies in the characterization of the hepatic microenvironment structure and the scoring of the disease stage in a spatiotemporal and non-invasive manner. Using a diet-induced NAFLD mouse model, we investigated the use of in vivo micro-computed tomography (CT) imaging techniques as a non-invasive method to assess the progression stages of NAFLD, focusing predominantly on the hepatic vascular network due to its significant involvement in NAFLD-related hepatic dysregulation. This imaging methodology allows for longitudinal analysis of liver steatosis and functional tissue uptake, as well as the evaluation of the relative blood volume, portal vein diameter, and density of the vascular network. Understanding the adaptations of the hepatic vascular network during NAFLD progression and correlating this with other ways of characterizing the disease progression (steatosis, inflammation, fibrosis) using the proposed method can pave the way toward the establishment of new, more efficient, and reproducible approaches for NAFLD research in mice. This protocol is also expected to upgrade the value of preclinical animal models for investigating&#160;the development of novel therapies against disease progression.

Novel In Vivo Micro-Computed Tomography Imaging Techniques for Assessing the Progression of Non-Alcoholic Fatty Liver Disease

Using a diet-induced non-alcoholic fatty liver disease (NAFLD) mouse model, we describe the use of novel in vivo micro-computed tomography imaging techniques as a non-invasive method to assess the progression stages of NAFLD, focusing predominantly on the hepatic vascular network due to its significant involvement in NAFLD-related hepatic dysregulation.

비알코올성 지방간 질환의 진행을 평가하기 위한 새로운 생체 내 미세 컴퓨터 단층 촬영 이미징 기술

Non-alcoholic fatty liver disease (NAFLD) is a growing global health issue, and the impact of NAFLD is compounded by the current lack of effective ...

인간 혈청에서 B형 간염 바이러스 DNA의 검출 및 정량화

Real-Time Polymerase Chain Reaction-Based Detection and Quantification of Hepatitis B Virus DNA

Hepatitis B virus (HBV) is a significant cause of liver disease worldwide. It can lead to acute or chronic infections, making individuals highly susceptible to fatal cirrhosis and liver cancer. Accurate detection and quantification of HBV DNA in the blood are essential for diagnosing and monitoring HBV infection. The most common method for detecting HBV DNA is real-time PCR, which can be used to detect the virus and assess the viral load to monitor the response to antiviral therapy. Here, we describe a detailed protocol for the detection and quantification of HBV DNA in human serum or plasma using an IVD-marked real-time PCR-based kit. The kit uses primers and probes that target the highly conserved core region of the HBV genome and can accurately quantify all HBV genotypes (A, B, C, D, E, F, G, H, I, and J). The kit also includes an endogenous internal control to monitor possible PCR inhibition. This assay runs for 40 cycles, and its cutoff is 38 Ct. For the quantification of HBV DNA in clinical samples, a set of 5 quantification standards is provided with the kit. The standards contain known concentrations of HBV-specific DNA that are calibrated against the 4th WHO International Standard for HBV DNA for the nucleic acid test (NIBSC code 10/266). The standards are used to validate the functionality of the HBV-specific DNA amplification and to generate a standard curve, allowing the quantification of HBV DNA in a sample. HBV DNA as low as 2.5 IU/mL was detected using the PCR kit. The high sensitivity and reproducibility of the kit make it a powerful tool in clinical laboratories, aiding healthcare professionals in effectively diagnosing and managing HBV infections.

저자 스포트라이트: B형 간염 바이러스 검출의 발전과 과제 

Real-time polymerase chain reaction (PCR)-based detection and quantification of hepatitis B virus (HBV) DNA is a sensitive and accurate method for diagnosing and monitoring HBV infection. Here, we present a protocol for HBV DNA detection and the viral load measurement of a sample.