EUS-FNB
Dotaz
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- Klíčová slova
- EUS (endoskopická ultrasonografie, endosonografie), EUS - navigovaná antitumorózní terapie), EUS-CPB (EUS - navigovaný blok celiakálního plexu), EUS-CDS (EUS - navigovaná choledochoduodenostomie), EUS-CPN (EUS - navigovaná celiakální neurolýza), EUS-FNI (EUS - navigovaná tenkojehlová injekční terapie), EUS-FNB (EUS - navigovaná tenkojehlová biopsie), EUS-RFA (EUS - navigovaná radiofrekvenční ablace), EUS-TCB (EUS - navigovaná trucut biopsie), EPGS (endoskopická pseudocystogastrostomie),
- MeSH
- diagnostické techniky gastrointestinální * přístrojové vybavení trendy využití MeSH
- endosonografie * metody přístrojové vybavení využití MeSH
- lidé MeSH
- nemoci jater diagnóza prevence a kontrola terapie MeSH
- nemoci slinivky břišní diagnóza prevence a kontrola terapie MeSH
- nemoci trávicího systému * diagnóza prevence a kontrola terapie MeSH
- nemoci žaludku diagnóza prevence a kontrola terapie MeSH
- nemoci žlučníku diagnóza prevence a kontrola terapie MeSH
- statistika jako téma MeSH
- Check Tag
- lidé MeSH
We compare two types of pancreatic carcinoma samples obtained by EUS-guided fine needle biopsy (EUS-FNB) in terms of the success rates and clinical validity of analysis of two most commonly investigated DNA/RNA pancreatic cancer markers, KRAS mutations and miR-21 expression. 118 patients with pancreatic ductal adenocarcinoma underwent EUS-FNB. The collected sample was divided, one part was stored in a stabilizing solution as native aspirate (EUS-FNA) and second part was processed into the cytological smear (EUS-FNC). DNA/RNA extraction was followed by analysis of KRAS mutations and miR-21 expression. For both sample types, the yields of DNA/RNA extraction and success rates of KRAS mutation and miRNA expression were evaluated. Finally, the resulting KRAS mutation frequency and miR-21 prognostic role were compared to literature data from tissue resections. The overall amount of isolated DNA/RNA from EUS-FNC was lower compared to the EUS-FNA, average yield 10 ng vs 147 ng for DNA and average yield 164 vs. 642 ng for RNA, but the success rates for KRAS and miR-21 analysis was 100% for both sample types. The KRAS-mutant detection frequency in EUS-FNC was 12% higher than in EUS-FNA (90 vs 78%). The prognostic role of miR-21 was confirmed in EUS-FNC (p = 0.02), but did not reach statistical significance in EUS-FNA (p = 0.06). Although both types of EUS-FNB samples are suitable for DNA/RNA extraction and subsequent DNA mutation and miRNA expression analysis, reliable results with clinical validity were only obtained for EUS-FNC.
- MeSH
- biopsie tenkou jehlou pod endosonografickou kontrolou MeSH
- cytodiagnostika metody MeSH
- DNA analýza MeSH
- duktální karcinom slinivky břišní diagnóza MeSH
- fixace tkání metody MeSH
- lidé středního věku MeSH
- lidé MeSH
- mikro RNA analýza MeSH
- mutace MeSH
- nádorové biomarkery analýza MeSH
- nádory slinivky břišní diagnóza MeSH
- odběr biologického vzorku metody MeSH
- protoonkogenní proteiny p21(ras) genetika MeSH
- senioři MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
The European Society of Gastrointestinal Endoscopy (ESGE) has recognized the need to formalize and enhance training in diagnostic endoscopic ultrasound (EUS). This manuscript represents the outcome of a formal Delphi process resulting in an official Position Statement of the ESGE and provides a framework to develop and maintain skills in diagnostic EUS. This curriculum is set out in terms of the prerequisites prior to training; the recommended steps of training to a defined syllabus; the quality of training; and how competence should be defined and evidenced before independent practice. 1: Trainees should have achieved competence in upper gastrointestinal endoscopy before training in diagnostic EUS. 2: The development of diagnostic EUS skills by methods that do not involve patients is advisable, but not mandatory, prior to commencing formal training in diagnostic EUS. 3: A trainee's principal trainer should be performing adequate volumes of diagnostic EUSs to demonstrate maintenance of their own competence. 4: Training centers for diagnostic EUS should offer expertise, as well as a high volume of procedures per year, to ensure an optimal level of quality for training. Under these conditions, training centers should be able to provide trainees with a sufficient wealth of experience in diagnostic EUS for at least 12 months. 5: Trainees should engage in formal training and supplement this with a range of learning resources for diagnostic EUS, including EUS-guided fine-needle aspiration and biopsy (FNA/FNB). 6: EUS training should follow a structured syllabus to guide the learning program. 7: A minimum procedure volume should be offered to trainees during diagnostic EUS training to ensure that they have the opportunity to achieve competence in the technique. To evaluate competence in diagnostic EUS, trainees should have completed a minimum of 250 supervised EUS procedures: 80 for luminal tumors, 20 for subepithelial lesions, and 150 for pancreaticobiliary lesions. At least 75 EUS-FNA/FNBs should be performed, including mostly pancreaticobiliary lesions. 8: Competence assessment in diagnostic EUS should take into consideration not only technical skills, but also cognitive and integrative skills. A reliable valid assessment tool should be used regularly during diagnostic EUS training to track the acquisition of competence and to support trainee feedback. 9: A period of supervised practice should follow the start of independent activity. Supervision can be delivered either on site if other colleagues are already practicing EUS or by maintaining contacts with the training center and/or other EUS experts. 10: Key performance measures including the annual number of procedures, frequency of obtaining a diagnostic sample during EUS-FNA/FNB, and adverse events should be recorded within an electronic documentation system and evaluated.
- MeSH
- biopsie tenkou jehlou pod endosonografickou kontrolou MeSH
- endosonografie metody MeSH
- gastrointestinální endoskopie * výchova MeSH
- kurikulum * MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Evropa MeSH
The European Society of Gastrointestinal Endoscopy (ESGE) has recognized the need to formalize and enhance training in endoscopic retrograde cholangiopancreatography (ERCP) and endoscopic ultrasound (EUS). This manuscript represents the outcome of a formal Delphi process resulting in an official Position Statement of the ESGE and provides a framework to develop and maintain skills in ERCP and EUS. This curriculum is set out in terms of the prerequisites prior to training; recommended steps of training to a defined syllabus; the quality of training; and how competence should be defined and evidenced before independent practice. 1: Trainees should be competent in gastroscopy prior to commencing training. Formal training courses and the use of simulation in training are recommended. 2: Trainees should keep a contemporaneous logbook of their procedures, including key performance indicators and the degree of independence. Structured formative assessment is encouraged to enhance feedback. There should be a summative assessment process prior to commencing independent practice to ensure there is robust evidence of competence. This evidence should include a review of a trainee's procedure volume and current performance measures. A period of mentoring is strongly recommended in the early stages of independent practice. 3: Specifically for ERCP, all trainees should be competent up to Schutz level 2 complexity (management of distal biliary strictures and stones > 10 mm), with advanced ERCP requiring a further period of training. Prior to independent practice, ESGE recommends that a trainee can evidence a procedure volume of > 300 cases, a native papilla cannulation rate of ≥ 80 % (90 % after a period of mentored independent practice), complete stones clearance of ≥ 85 %, and successful stenting of distal biliary strictures of ≥ 90 % (90 % and 95 % respectively after a mentored period of independent practice). 4: The progression of EUS training and competence attainment should start from diagnostic EUS and then proceed to basic therapeutic EUS, and finally to advanced therapeutic EUS. Before independent practice, ESGE recommends that a trainee can evidence a procedure volume of > 250 cases (75 fine-needle aspirations/biopsies [FNA/FNBs]), satisfactory visualization of key anatomical landmarks in ≥ 90 % of cases, and an FNA/FNB accuracy rate of ≥ 85 %. ESGE recognizes the often inadequate quality of the evidence and the need for further studies pertaining to training in advanced endoscopy, particularly in relation to therapeutic EUS.
1: ESGE recommends that, where there is a suspicion of eosinophilic esophagitis, at least six biopsies should be taken, two to four biopsies from the distal esophagus and two to four biopsies from the proximal esophagus, targeting areas with endoscopic mucosal abnormalities. Distal and proximal biopsies should be placed in separate containers.Strong recommendation, low quality of evidence. 2: ESGE recommends obtaining six biopsies, including from the base and edge of the esophageal ulcers, for histologic analysis in patients with suspected viral esophagitis.Strong recommendation, low quality of evidence. 3: ESGE recommends at least six biopsies are taken in cases of suspected advanced esophageal cancer and suspected advanced gastric cancer.Strong recommendation, moderate quality of evidence. 4: ESGE recommends taking only one to two targeted biopsies for lesions in the esophagus or stomach that are potentially amenable to endoscopic resection (Paris classification 0-I, 0-II) in order to confirm the diagnosis and not compromise subsequent endoscopic resection.Strong recommendation, low quality of evidence. 5: ESGE recommends obtaining two biopsies from the antrum and two from the corpus in patients with suspected Helicobacter pylori infection and for gastritis staging.Strong recommendation, low quality of evidence. 6: ESGE recommends biopsies from or, if endoscopically resectable, resection of gastric adenomas.Strong recommendation, moderate quality of evidence. 7: ESGE recommends fine-needle aspiration (FNA) and fine-needle biopsy (FNB) needles equally for sampling of solid pancreatic masses.Strong recommendation, high quality evidence. 8: ESGE suggests performing peroral cholangioscopy (POC) and/or endoscopic ultrasound (EUS)-guided tissue acquisition in indeterminate biliary strictures. For proximal and intrinsic strictures, POC is preferred. For distal and extrinsic strictures, EUS-guided sampling is preferred, with POC where this is not diagnostic.Weak recommendation, low quality evidence. 9: ESGE suggests obtaining possible non-neoplastic biopsies before sampling suspected malignant lesions to prevent intraluminal spread of malignant disease.Weak recommendation, low quality of evidence. 10: ESGE suggests dividing EUS-FNA material into smears (two per pass) and liquid-based cytology (LBC), or the whole of the EUS-FNA material can be processed as LBC, depending on local experience.Weak recommendation, low quality evidence.
Background/Objectives: Although the overall survival prognosis of patients in advanced stages of pancreatic ductal adenocarcinoma (PDAC) is poor, typically ranging from days to months from diagnosis, there are rare cases of patients remaining in therapy for longer periods of time. Early estimations of survival prognosis would allow rational decisions on complex therapy interventions, including radical surgery and robust systemic therapy regimens. Understandably, there is great interest in finding prognostic markers that can be used for patient stratification. We determined the role of various KRAS mutations in the prognosis of PDAC patients using biopsy samples and circulating tumor DNA. Methods: A total of 118 patients with PDAC, clinically confirmed by endoscopic ultrasound-guided fine-needle aspiration biopsy (EUS-FNB), were included in the study. DNA was extracted from cytological slides following a standard cytology evaluation to ensure adequacy (viability and quantity) and to mark the tumor cell fraction. Circulating tumor DNA (ctDNA) was extracted from plasma samples of 45 patients in stage IV of the disease. KRAS mutations in exons 12 and 13 were detected by denaturing capillary electrophoresis (DCE), revealing a minute presence of mutation-specific heteroduplexes. Kaplan-Meier survival curves were calculated for individual KRAS mutation types. Results:KRAS mutations were detected in 90% of tissue (106/118) and 44% of plasma (20/45) samples. All mutations were localized at exon 2, codon 12, with G12D (GGT > GAT) being the most frequent at 44% (47/106) and 65% (13/20), followed by other types including G12V (GGT > GTT) at 31% (33/106) and 10% (2/20), G12R (GGT > CGT) at 17% (18/106) and 10% (2/20), G12C (GGT/TGT) at 5% (5/106) and 0% (0/20) and G12S (GGT/AGT) at 1% (1/106) and 5% (1/20) in tissue and plasma samples, respectively. Two patients had two mutations simultaneously (G12V + G12S and G12D + G12S) in both types of samples (2%, 2/106 and 10%, 2/20 in tissue and plasma samples, respectively). The median survival of patients with the G12D mutation in tissues was less than half that of other patients (median survival 101 days, 95% CI: 80-600 vs. 228 days, 95% CI: 184-602), with a statistically significant overall difference in survival (p = 0.0080, log-rank test), and furthermore it was less than that of all combined patients with other mutation types (101 days, 95% CI: 80-600 vs. 210 days, 95% CI: 161-602, p = 0.0166). For plasma samples, the survival of patients with this mutation was six times shorter than that of patients without the G12D mutation (27 days, 95% CI: 8-334 vs. 161 days, 95% CI: 107-536, p = 0.0200). In contrast, patients with detected KRAS G12R in the tissue survived nearly twice as long as other patients in the aggregate (286 days, 95% CI: 70-602 vs. 162 days, 95% CI: 122-600, p = 0.0374) or patients with other KRAS mutations (286 days, 95% CI: 70-602 vs. 137 days, 95% CI: 107-600, p = 0.0257). Conclusions: Differentiation of specific KRAS mutations in EUS-FNB and ctDNA (above all, the crucial G12D and G12R) is feasible in routine management of PDAC patients and imperative for assessment of prognosis.
- MeSH
- biopsie tenkou jehlou pod endosonografickou kontrolou * MeSH
- cirkulující nádorová DNA genetika krev MeSH
- dospělí MeSH
- duktální karcinom slinivky břišní * genetika patologie krev MeSH
- lidé středního věku MeSH
- lidé MeSH
- mutace * MeSH
- nádorové biomarkery genetika MeSH
- nádory slinivky břišní * genetika patologie mortalita MeSH
- prognóza MeSH
- protoonkogenní proteiny p21(ras) * genetika MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- tekutá biopsie metody MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Cíl práce: Práce se zabývá klinickou částí výzkumu lidských embryonálních kmenových buněk (hESC). Cílem projektu je vznik somatických buněčných typů použitelných ve vývoji léčiv, regenerativní medicíně a buněčné terapii. Výhledem je umožnit cílenou terapii dosud nevyléčitelné nemoci. Pluripotentní hESC mají neomezenou kapacitu pro sebeobnovu. Této vlastnosti se využívá v terapii, kdy jsou z hESC vytvořeny chybějící nebo poškozené buňky v lidském těle. Je zájem vytvořit hESC linie v klinické kvalitě, použitelné v předklinických a klinických studiích. Metodika: Vytvoření hESC musí respektovat legislativu ČR a EU. Podmínkou bylo vypracovat informovaný souhlas obou dárců pro darovaná vyřazená embrya, která nejsou vhodná pro léčbu oplodněním in vitro dle směrnice 2004/23/ES. Centrum asistované reprodukce (CAR) FN Brno se podílí na odběru oocytů, kultivaci a kryokonzervaci embryí, komunikaci s klienty a zajišťování informovaných souhlasů dárců embryí. Byl vypracován předávací protokol a metodika předávání rozmrazených embryí s originálním číselným kódem. Před předáním embryí na spoluautorské pracoviště – Centrum buněčného a tkáňového inženýrství (CTEF) ICRC FN u sv. Anny – je provedeno jejich rozmrazení, v případě potřeby dokultivování do stadia blastocysty, a následně je proveden asistovaný hatching. Výsledky: V období leden 2018 až červenec 2020 bylo obesláno 138 vybraných vhodných klientek na dárcovství, z nichž 52 nereagovalo, 19 ukončilo a 29 prodloužilo skladování embryí. Pouze 38 klientek, tj. 27,5 %, souhlasilo s jejich využitím na přípravu hESC. Ve stejném období probíhala osobní komunikace s vhodnými klienty CAR a bylo získáno dalších 17 dárců embryí. Celkem bylo získáno 160 embryí od 55 dárkyň ve věku 26–42 let. Nejčastěji byla embrya zamrazena ve stadiu blastocysty (53 embrií – 33,1 %) a moruly (74 embrií – 46,3 %). Z 29 geneticky vyšetřených embryí je 5 euploidních (17,2 %), 2 mozaiky a 22 aneuploidních nebo s translokací či přenašečů s monogenní vadou. Závěr: Byl vypracován a Etickou komisí LF MU a FN Brno schválen informovaný souhlas, bylo vybráno a zajištěno 160 darovaných embryí. Je vypracován předávací protokol a metodika předávání. Plán předávání rozmrazených anonymizovaných embryí zahrnuje cca 5 rozmrazených blastocyst týdně s provedeným asistovaným hatchingem. Po předání embryí na CTEF probíhá izolace embryoblastu s následnou kultivací. Ustanovené buněčné linie hESC musí splnit specifikovaná kritéria bezpečnosti, stability a pluripotence. Věříme, že v souladu s plánem projektu získáme nejméně tři linie hESC v klinické kvalitě, poprvé vytvořené v ČR, respektující požadavky na léčivé přípravky Advanced Medicinal Therapy Products (AMTP).
Objective: The work deals with a clinical part of human embryonic stem cell (hESC) research. The aim of the project is the differentiation of somatic cell types, useful in drug development, regenerative medicine and cell therapy. The aim of this work is to enable targeted therapy of yet incurable diseases. The pluripotent hESCs have unlimited self-renewal capacity. This ability is used in therapy to create missing or damaged cells in the human body. It is of interest to develop clinical-grade hESC lines useful in preclinical and clinical studies. Methods: The derivation of the hESC must respect the legislation of the Czech Republic and the EU. The aim was to develop an informed consent of both donors for donated discarded embryos that are not suitable for treatment by in vitro fertilization according to Directive 2004/23/EC. The FNB‘s Center for Assisted Reproduction (CAR) participates in oocyte collection, cultivation and cryopreservation of embryos, communication with clients and ensuring the informed consent of embryo donors. A transport protocol and a methodology for handing over the thawed embryos with the original numerical code were developed. Before the embryos are handed over to the ICRC co-author‘s workplace (CTEF), they are thawed and, if necessary, recultivated to the blastocyst stage; afterwards, assisted hatching is performed. Results: In the period from January 2018 to July 2020, 138 selected suitable clients were asked for donations, with 52 not responding, 19 terminating and 29 extending the embryo storage. Only 38 clients, i.e. 27.5%, agreed with the usage of their embryos for the preparation of hESCs. In the same period, personal communication with suitable CAR clients took place and another 17 embryo donors were obtained. A total of 160 embryos were obtained from 55 donors aged 26 to 42 years. The embryos were most often frozen in the blastocyst (53 embryos – 33.1%) and morula (74 embryos – 46.3%) stages. Of the 29 genetically examined embryos, only 5 are euploid (17.2%), 2 are mosaic and 22 are aneuploid or with translocations or carriers with a monogenic defect. Conclusion: We have an informed consent prepared and approved by the Ethics Committee of the Masaryk University and the University Hospital Brno; 160 donated embryos have been selected and secured. A transport protocol and handover methodology are developed. The plan for the transfer of thawed anonymized embryos in the first phase, October – December 2020, includes approximately 5 thawed blastocysts per week with assisted hatching. After their transfer to the CTEF, the embryoblast will be isolated with subsequent cultivation. The established hESCs must meet the specified criteria of safety, stability and pluripotency. We believe that, in accordance with the project plan, we will obtain at least 3 clinical-grade hESC lines, the first created in the Czech Republic, respecting the requirements for Advanced Medicinal Therapy Products (AMTP).
- Klíčová slova
- výběr embryí,
- MeSH
- blastocysta MeSH
- buněčná a tkáňová terapie MeSH
- embryoblast MeSH
- lidé MeSH
- lidské embryonální kmenové buňky MeSH
- nakládání s embryem * MeSH
- vitrifikace MeSH
- získávání tkání a orgánů MeSH
- Check Tag
- lidé MeSH
