Os autores são gratos a Dr. Ana Cotrim, para consulta técnica e Dr. Monzur Murshed para o equipamento. Este trabalho foi financiado em parte pelos Institutos Canadenses de Pesquisa em Saúde.

1. Descrição processuais <ol><li> Por via subcutânea injetar 1 ml / g de peso corporal de 0,5 monohidratada sulfato mg / ml atropina (Cat # 11330, Fluka, EUA) em cada um mouse para evitar que a secreção salivar de interromper o movimento das infusões. </li><li> Anestesiar cada rato com um peso corporal mL / g de ketamina a 100 mg / ml e 20 mg / ml solução xilazina (Phoenix Scientific, AR, EUA) injetou intra-peritoneal (IP) </li><li> Posicione o mouse sobre uma plataforma de plástico feito sob medida na posição ventral: os incisivos superiores estão bloqueados em um fio de metal, e os incisivos inferiores são viciadas em um fio elástico, a fim de manter a boca aberta. </li><li> Use uma seringa de insulina com uma agulha de calibre 29 (Cat # 600145, Tyco Healthcare, MA, EUA), colocado dentro de um tubo de polietileno 0,58 mm de diâmetro (Cat # 427410, Becton Dickinson, MD, EUA; Fig. 2). O tubo de polietileno inserido é 5 milímetros sobre a ponta de uma agulha de 29 gauge. A agulha é usada como um guia rígido para mover o tubo de polietileno em duto de Wharton. O tubo de polietileno é inserido 3-5 mm dentro do duto. Note que apenas o tubo de polietileno entra no ducto da glândula salivar, enquanto a agulha permanece fora do duto. Injetar 50uL de filtrado 0,4% solução de azul de tripano mancha (Cat # 15250, Gibco, EUA) na glândula submandibular via duto de Wharton, que está localizado logo abaixo da língua. A infusão é entregue de forma constante com pressão positiva sobre o pistão da seringa; o dedo permanece no pistão, mesmo depois de todos os infusão tenha sido entregue à glândula, a fim de evitar um refluxo da infusão intra-oral. A solução de azul de tripano é usado no vídeo como um corante para demonstrar a técnica. </li><li> epois a injeção da solução de azul de tripano, o rato foi sacrificado antes da dissecção. A glândula salivar é cirurgicamente expostas, a fim de determinar quanto das infusões realmente permaneceu na glândula. </li></ol> 2. Resultados representativos: Após as infusões foram injetados, o rato foi dissecado para expor cirurgicamente a glândula salivar pós-canulados. Como mostrado na imagem-2, a coloração escura do ducto de Wharton fornece evidências de que a solução de azul de tripano foi infundida com sucesso. <img src="/files/ftp_upload/3074/3074fig1.jpg" alt="Figura 1" /> Figura 1. Canulação da glândula submandibular do rato. <img src="/files/ftp_upload/3074/3074fig2.jpg" alt="Figura 2" /> Figura 2. Tubo de polietileno é envolto em torno da ponta de uma agulha de 29 gauge. A agulha da seringa é usada como um guia rígido para mover o tubo de polietileno em duto de Wharton. O tubo de polietileno está inserido (canulados) mm 3-5 no interior do duto. Note que apenas o tubo de polietileno entra no ducto da glândula salivar, enquanto a agulha permanece fora do duto.

<ol>
	<li>Fox, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acquired+salivary+dysfunction.+Drugs+and+radiation.">Acquired salivary dysfunction. Drugs and radiation.</a> Ann N Y Acad Sci. 842, 132-137 (1998).</li><li>Bhide, S. A., Miah, A. B., Harrington, K. J., Newbold, K. L., Nutting, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiation-induced+xerostomia:+pathophysiology,+prevention+and+treatment.">Radiation-induced xerostomia: pathophysiology, prevention and treatment.</a> Clin Oncol (R Coll Radiol). 21, 737-744 (2009).</li><li>Redman, R. S., Ball, W. D., Mezey, E., Key, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dispersed+donor+salivary+gland+cells+are+widely+distributed+in+the+recipient+gland+when+infused+up+the+ductal+tree.">Dispersed donor salivary gland cells are widely distributed in the recipient gland when infused up the ductal tree.</a> Biotech Histochem. 84, 253-260 (2009).</li></ol>

Todos os experimentos foram aprovados pelo comitê de ética da Universidade e estavam de acordo com as diretrizes do Canadian Council on Animal Care.

A dificuldade em utilizar este método de canulação é evitar que o refluxo da solução injetada. Mesmo se a solução azul de tripano foi temporariamente migraram para a glândula salivar com sucesso, a secreção de saliva pode interferir com as infusões e causar refluxo da solução de azul de tripano para fora da glândula. Como precaução, além de usar atropina para inibir a secreção de saliva, a cânula de polietileno deve permanecer no duto de Wharton eo pistão da seringa realizada durante 60 segundos ap...

<table border="1"><tbody><tr><td> Nome do reagente </td><td> Tipo </td><td> Companhia </td><td> Número de catálogo </td><td> Comentários (opcional) </td></tr><tr><td> Atropina </td><td> Droga </td><td> Fluka </td><td> 11330 </td><td> Para inibir a secreção de saliva </td></tr><tr><td> Ketamina </td><td> Droga </td><td> Phoenix Scientific </td><td></td><td> Sedação </td></tr><tr><td> Xilazina </td><td> Droga </td><td> Phoenix Scientific </td><td></td><td></td></tr><tr><td> Tubos de polietileno (0,58 mm de diâmetro) </td><td> Tubo </td><td> Intramedic PE-10, Becton Dickinson Diagnostic Systems </td><td> 427410 </td><td> Para inserir mais de ponta da agulha para evitar lesão tecidual durante a inserção no ducto de Wharton. </td></tr><tr><td> Seringa de insulina Ultra Comfort (com uma agulha de 29G) </td><td> Seringa </td><td> Tyco Healthcare </td><td> 600145 </td><td></td></tr><tr><td> 4% azul de tripano </td><td> Mancha </td><td> Gibco </td><td> 15250 </td><td></td></tr></tbody></table>

cannulation of the mouse submandibular salivary gland via the wharton's duct

Hipofunção da glândula salivar é grave freqüentemente encontrados em pacientes com síndrome de Sjögren e aqueles que recebem a irradiação terapêutica em sua cabeça e pescoço para tratamento de câncer. Ambos os grupos de sintomas dos pacientes experiência como xerostomia (boca seca), disfagia (deficiente mastigação e deglutição), grave cárie dentária, alteração do paladar, oro-faringe infecções (candidíase), dor, mucosite e desconforto. Uma abordagem inovadora da medicina regenerativa para o tratamento da glândula salivar hipo-função é especulado no RS Redman, E Mezey et al 2009:. Células-tronco pode ser depositado diretamente pela canulação para a glândula como um método potente em reviver as funções do deficiente órgão. Presumivelmente, a migração de células-tronco estrangeiros irão se diferenciar em células glandulares funcionar como parte da glândula salivar anfitrião. Além disso, esta técnica de canulação é um método de entrega expeditos e eficazes para a aplicação clínica de transferência de genes. Aqui nós ilustrar as etapas envolvidas na realização do procedimento canulação do mouse glândula salivar submandibular via duto de Wharton (Fig. 1). C3H camundongos (Charles River, Montreal, Quebec, Canadá) são utilizados para este experimento, que foram mantidos sob condições convencionais limpa no centro de recursos McGill University animal. Todos os experimentos foram aprovados pelo comitê de ética da Universidade e estavam de acordo com as diretrizes do Canadian Council on Animal Care. Para este experimento, uma solução de azul de tripano é infundida a glândula através da abertura do ducto de Wharton, utilizando uma seringa com uma agulha de insulina 29-gauge encerrado dentro de um tubo de polietileno. Posteriormente, o mouse é dissecado para mostrar que as infusões migraram para a glândula com sucesso.

Watch this Scientific Journal Video about Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct at JoVE.com

Cannulation of the Mouse Submandibular Salivary Gland via the Wharton&#039;s Duct

Um protocolo para a canulação do mouse glândula salivar submandibular via ducto de Wharton, é descrito. Para este experimento, a solução de azul de tripano é usado como um tintureiro para demonstrar como esta técnica efetivamente oferece infusões na glândula alvo, e sugerir a confiabilidade dessa nova abordagem como uma potencial terapia medicamentosa / célula clínicos para a regeneração das glândulas salivares.

Canulação da glândula submandibular do rato através do duto salivar de Wharton

Severe salivary gland hypofunction is frequently found in patients with Sj&ouml;gren's syndrome and those who receiving therapeutic 
irradiation in their ...

cannulation-mouse-submandibular-salivary-gland-via-wharton-s

Research

JoVE Journal

Medicine

Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct

25.7K visualizações.  McGill University. Um protocolo para a canulação do mouse glândula salivar submandibular via ducto de Wharton, é descrito. Para este experimento, a solução de azul de tripano é usado como um tintureiro para demonstrar como esta técnica efetivamente oferece infusões na glândula alvo, e sugerir a confiabilidade dessa nova abordagem como uma potencial terapia medicamentosa / célula clínicos para a regeneração das glândulas salivares.

Vídeo: Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct

Collecting Saliva and Measuring Salivary Cortisol and Alpha-amylase in Frail Community Residing Older Adults via Family Caregivers

Salivary measures have emerged in bio-behavioral research that are easy-to-collect, minimally invasive, and relatively inexpensive biologic markers of stress. This article we present the steps for collection and analysis of two salivary assays in research with frail, community residing older adults-salivary cortisol and salivary alpha amylase. The field of salivary bioscience is rapidly advancing and the purpose of this presentation is to provide an update on the developments for investigators interested in integrating these measures into research on aging. Strategies are presented for instructing family caregivers in collecting saliva in the home, and for conducting laboratory analyses of salivary analytes that have demonstrated feasibility, high compliance, and yield quality specimens. The protocol for sample collection includes: (1) consistent use of collection materials; (2) standardized methods that promote adherence and minimize subject burden; and (3) procedures for controlling certain confounding agents. We also provide strategies for laboratory analyses include: (1) saliva handling and processing; (2) salivary cortisol and salivary alpha amylase assay procedures; and (3) analytic considerations.

We demonstrate: (1) procedures for collection of salivary samples in cognitive impaired older adults by family caregivers in the home setting, (2) procedures for measuring stress activity via salivary cortisol and alpha amylase, and (3) representative profiles. Protocols that allow researchers to study stress-linked processes advance our understanding of biological sensitivity and susceptibility.

Salivary measures have emerged in bio-behavioral research  that are easy-to-collect, minimally invasive, and relatively inexpensive  biologic markers of ...

Medicina

Surgical Injury to the Mouse Pancreas through Ligation of the Pancreatic Duct as a Model for Endocrine and Exocrine Reprogramming and Proliferation

Expansion of pancreatic beta cells in vivo or ex vivo, or generation of beta cells by differentiation from an embryonic or adult stem cell, can provide new expandable sources of beta cells to alleviate the donor scarcity in human islet transplantation as therapy for diabetes. Although recent advances have been made towards this aim, mechanisms that regulate beta cell expansion and differentiation from a stem/progenitor cell remain to be characterized. Here, we describe a protocol for an injury model in the adult mouse pancreas that can function as a tool to study mechanisms of tissue remodeling and beta cell proliferation and differentiation. Partial duct ligation (PDL) is an experimentally induced injury of the rodent pancreas involving surgical ligation of the main pancreatic duct resulting in an obstruction of drainage of exocrine products out of the tail region of the pancreas. The inflicted damage induces acinar atrophy, immune cell infiltration and severe tissue remodeling. We have previously reported the activation of Neurogenin (Ngn) 3 expressing endogenous progenitor-like cells and an increase in beta cell proliferation after PDL. Therefore, PDL provides a basis to study signals involved in beta cell dynamics and the properties of an endocrine progenitor in adult pancreas. Since, it still remains largely unclear, which factors and pathways contribute to beta cell neogenesis and proliferation in PDL, a standardized protocol for PDL will allow for comparison across laboratories.

This protocol describes an injury model involving the surgical ligation of the pancreatic duct in the adult mouse pancreas, resulting in severe injury that establishes an environment that allows beta cell neogenesis and proliferation. This model can be used as a tool to study mechanisms involved in beta cell formation.

Lesão cirúrgica para o mouse Pâncreas através da ligadura da pâncreas Duct como um modelo para endócrina e exócrina Reprogramação e Proliferação

Expansion of pancreatic beta cells in vivo or ex vivo, or generation of beta cells by differentiation from an embryonic or adult stem cell, can provide ...

Retroductal Submandibular Gland Instillation and Localized Fractionated Irradiation in a Rat Model of Salivary Hypofunction

Normal tissues that lie within the portals of radiation are inadvertently damaged. Salivary glands are often injured during head and neck radiotherapy. Irreparable cell damage results in a chronic loss of salivary function that impairs basic oral activities, and increases the risk of oral infections and dental caries. Salivary hypofunction and its complications gravely impact a patient's comfort. Current symptomatic management of the condition is ineffective, and newer therapies to assuage the condition are needed. 
Salivary glands are exocrine glands, which expel their secretions into the mouth via excretory ducts. Cannulation of these ducts provides direct access to the glands. Retroductal delivery of a contrast agent to major salivary glands is a routine out-patient procedure for diagnostic imaging. Using a similar procedure, localized treatment of the glands is feasible. However, performing this technique in preclinical studies with small animals poses unique challenges. In this study we describe the technique of retroductal administration in rat submandibular glands, a procedure that was refined in Dr. Bruce Baum's laboratory (NIH)1, and lay out a procedure for local gland irradiation.

Salivary gland hypofunction, a major adverse effect of head and neck radiotherapy diminishes a patient's quality of life. The demonstration of efficacy of new therapies in animal models is a prerequisite before clinical transition. This protocol describes retroductal administration and local irradiation of rat submandibular glands.

Retroductal submandibular Glândula instilação e localizada fracionado irradiação em um modelo de rato de salivar Hipofunção

Normal tissues that lie within the portals of radiation are inadvertently damaged. Salivary glands are often injured during head and neck radiotherapy. ...

Pancreatic Duct Infusion: An Effective and Selective Method of Drug and Viral Delivery

The pancreas is a bifunctional organ with both endocrine and exocrine components. A number of pathologies can afflict the pancreas, including diabetes, pancreatitis, and pancreatic cancer. All three of these diseases mark active areas of study, not only to develop immediate therapy, but also to better understand their pathophysiology. There are few tools to further these areas of study. Pancreatic duct infusion is an important technique that can allow for lineage tracing, gene introduction, and cell line-specific targeting. The technique requires the intricate dissection of the second portion of the duodenum and ampulla, followed by the occlusion of the bile duct and the cannulation of the pancreatic duct. Although the technique is technically challenging at first, the applications are myriad. Ambiguity in the specifics of the procedure between groups highlighted the need for a standard protocol. This work describes the expression of a green fluorescent protein (GFP) within the pancreas after the pancreatic duct infusion of a viral vector expressing GFP versus a sham surgery. The infusion and therefore expression is specific to the pancreas, without expression present in any other tissue type.

Pancreatic duct infusion is an important technique that can allow for lineage tracing, gene introduction, and cell line-specific targeting. A pancreatic duct infusion technique for drug and viral delivery to pancreatic cells is presented here.

Infusão do ducto pancreático: um método eficaz e seletivo de medicamentos e entrega viral

The pancreas is a bifunctional organ with both endocrine and exocrine components. A number of pathologies can afflict the pancreas, including diabetes, ...

Murine Salivary Functional Assessment via Pilocarpine Stimulation Following Fractionated Radiation

Hyposalivation is commonly observed in the autoimmune reaction of Sj&#246;gren's syndrome or following radiation injury to the major salivary glands. In these cases, questions remain regarding disease pathogenesis and effective interventions. An optimized technique that allows functional assessment of the salivary glands is invaluable for investigating exocrine gland biology, dysfunction, and therapeutics. Here, we present a step by step approach to performing pilocarpine stimulated saliva secretion, including tracheostomy and the dissection of the three major murine salivary glands. We also detail the appropriate murine head and neck anatomy accessed during these techniques. This approach is scalable, allowing for multiple mice to be processed simultaneously, thus improving the efficiency of the work flow. We aim to improve the reproducibility of these methods, each of which has further applications within the field. In addition to saliva collection, we discuss metrics for quantifying and normalizing functional capacity of these tissues. Representative data are included from submandibular glands with depressed salivary gland function 2 weeks following fractionated radiation (4 doses of 6.85 Gy).

We present a detailed approach to performing saliva collection, including murine tracheostomy and the isolation of three major salivary glands.

Murino salivar avaliação funcional através de pilocarpina estimulação seguinte fracionada de radiação

Hyposalivation is commonly observed in the autoimmune reaction of Sj&#246;gren's syndrome or following radiation injury to the major salivary glands. In ...

Radiation Treatment of Organotypic Cultures from Submandibular and Parotid Salivary Glands Models Key In Vivo Characteristics

Hyposalivation and xerostomia create chronic oral complications that decrease the quality of life in head and neck cancer patients who are treated with radiotherapy. Experimental approaches to understanding mechanisms of salivary gland dysfunction and restoration have focused on in vivo models, which are handicapped by an inability to systematically screen therapeutic candidates and efficiencies in transfection capability to manipulate specific genes. The purpose of this salivary gland organotypic culture protocol is to evaluate maximal time of culture viability and characterize cellular changes following ex vivo radiation treatment. We utilized immunofluorescent staining and confocal microscopy to determine when specific cell populations and markers are present during a 30-day culture period. In addition, cellular markers previously reported in in vivo radiation models are evaluated in cultures that are irradiated ex vivo. Moving forward, this method is an attractive platform for rapid ex vivo assessment of murine and human salivary gland tissue responses to therapeutic agents that improve salivary function.

Using three-dimensional organotypic cultures to visualize morphology and functional markers of salivary glands may provide novel insights into the mechanisms of tissue damage following radiation. Described here is a protocol to section, culture, irradiate, stain, and image 50&#8211;90 &#956;m thick salivary gland sections prior to and following exposure to ionizing radiation.

Tratamento de radiação de culturas organotypic de glândulas salivares submandibulares e parótidas modelos chave in vivo características

Hyposalivation and xerostomia create chronic oral complications that decrease the quality of life in head and neck cancer patients who are treated with ...

A Mouse Model for Chronic Pancreatitis via Bile Duct TNBS Infusion

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. Several rodent models have been developed to study CP, of which the bile duct 2,4,6 -trinitrobenzene sulfonic acid (TNBS) infusion model replicates the features of neuropathic pain seen in CP. However, bile duct drug infusion in mice is technically challenging. This protocol demonstrates the procedure of bile duct TNBS infusion for generation of a CP mouse model. TNBS was infused into the pancreas through the ampulla of Vater in the duodenum. This protocol optimized drug volume, surgical techniques, and drug handling during the procedure. TNBS-treated mice showed features of CP as reflected by bodyweight and pancreas weight reductions, changes in pain-associated behaviors, and abnormal pancreatic morphology. With these improvements, mortality associated with TNBS injection was minimal. This procedure is not only critical in generating pancreatic disease models but is also useful in local pancreatic drug delivery.

Chronic pancreatitis (CP) is a disease characterized by inflammation and fibrosis of the pancreas, often associated with intractable abdominal pain. This article focuses on refining the technique to generate a mouse model of CP via bile duct infusion with 2,4,6 -trinitrobenzene sulfonic acid (TNBS).

Um modelo de rato para pancreatite crônica através da infusão TNBS do ducto biliar

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. ...

Establishment of a Mouse Severe Acute Pancreatitis Model using Retrograde Injection of Sodium Taurocholate into the Biliopancreatic Duct

The prevalence of acute pancreatitis (AP), especially severe acute pancreatitis (SAP), is increasing in younger age groups annually. However, there is a lack of effective treatments in the current clinical practice. With the easy accessibility of transgenic and knockout strains and their small size, which allows minimal doses of drugs required for in vivo evaluation, a well-established experimental model in mice is preferred for AP research. Moreover, SAP induced through sodium taurocholate (TC) is currently one of the most widely used and best characterized models. This model has been investigated for novel therapies and possible molecular events during the process of AP. Here, we present the generation of an AP mouse model using sodium taurocholate and a simple homemade microsyringe. Moreover, we also provide the methodology for the subsequent histology and serological testing.

A mouse model of severe acute pancreatitis is described herein. The procedure presented here is very rapid, simple, and accessible, thereby potentially allowing the study of the molecular mechanisms and different therapeutic interventions in acute pancreatitis in a convenient way.

Estabelecimento de um modelo de pancreatite aguda grave do rato usando injeção retrógrada de taurocholate de sódio no ducto biliopancreático

The prevalence of acute pancreatitis (AP), especially severe acute pancreatitis (SAP), is increasing in younger age groups annually. However, there is a ...

Establishment of a Simple and Effective Rat Model for Intraoperative Parathyroid Gland Imaging

Parathyroid gland (PG) identification is a critical unmet need in thyroidectomy. The identification of the PG is challenging in thyroid surgery as it is similar in color to the thyroid gland. The lack of effective animal models in preclinical research is a severe limitation for the development of PG identification techniques. This protocol allows for the establishment of a simple and effective rat model for PG identification. In this model, black iron oxide nanoparticles (IONPs) are injected locally in the thyroid gland and rapidly diffuse within the thyroid gland but not the PG. A negatively stained PG and a positively stained thyroid gland can be easily identified by the naked eye without requiring external microscopes. The position of the PG can be identified by increasing the color contrast between the thyroid gland and the PG, based on the color of the black IONPs. This rat model is low-cost and convenient for PG identification, and the IONPs are a novel PG contrast agent.

To date, the development of parathyroid gland (PG) identification methods is limited by the lack of animal models in preclinical research. Here, we establish a simple and effective rat model for intraoperative PG imaging and evaluate its effectiveness by using iron oxide nanoparticles as a novel PG contrast agent.

Estabelecimento de um Modelo de Rato Simples e Eficaz para Imagem Intraoperatória da Glândula Paratireoide

Parathyroid gland (PG) identification is a critical unmet need in thyroidectomy. The identification of the PG is challenging in thyroid surgery as it is ...

Laparoscopic Common Bile Duct Exploration in Patients with a Previous History of Biliary Tract Surgery

For recurrent choledocholithiasis, abdominal adhesions in previous surgeries lead to changes in anatomical structures, and a secondary injury occurs easily when performing another operation for laparoscopic common bile duct exploration (LCBDE), which was once considered a relative contraindication. In view of the limitations of the current surgical technique, this study summarized the surgical approaches and crucial anatomical landmarks for reoperation for LCBDE. Four general surgical approaches were proposed to expose the common bile duct, including the ligamentum teres hepatis approach, the anterior hepatic duodenal ligament approach, the right hepatic duodenal ligament approach, and the hybrid approach. Additionally, this study highlighted seven crucial anatomical landmarks: the parietal peritoneum, the gastrointestinal serosa, the ligamentum teres hepatis, the inferior margin of the liver, the gastric antrum, the duodenum, and the hepatic flexure of the colon, which were helpful to safely separate abdominal adhesions and expose the common bile duct. Moreover, to shorten the time of choledocholithotomy, a sequential method was innovatively applied for the removal of the stones in common bile duct. Mastering the above surgical approaches, including identifying crucial anatomical landmarks and adopting the sequential method will improve the safety of reoperation for LCBDE, shorten the operation time, promote the fast recovery of patients, reduce postoperative complications, and contribute to the popularization and application of this technique.

Programmed surgery based on surgical methods and anatomical markers helps shorten the operation time, reduce complications, and improve the safety of the surgery. This study investigated and summarized the surgical methods and anatomic markers of laparoscopic common bile duct exploration in patients with a history of biliary tract surgery.

Exploração Laparoscópica do Ducto Biliar Comum em Pacientes com História Prévia de Cirurgia das Vias Biliares

For recurrent choledocholithiasis, abdominal adhesions in previous surgeries lead to changes in anatomical structures, and a secondary injury occurs ...