Project description:Needle knife papillotomy (NKP) is still considered a rescue technique after conventional access failure due to traditional high complication rates, albeit data are maturing for early use of NKP in standard endoscopic retrograde cholangiopancreatography indications. By contrast, in certain settings NKP should be prioritized to a first-class indication, such as in choledochocele management and/or, more often encountered in clinical practice, true papillary stone impaction with or without gallstone pancreatitis. The latter results in prompt stone release; thus, the procedure might become alternatively designated as "needle knife excision."
Project description:BackgroundSome clinically important genetic variants are not easily evaluated with next-generation sequencing (NGS) methods due to technical challenges arising from high- similarity copies (e.g., PMS2, SMN1/SMN2, GBA1, HBA1/HBA2, CYP21A2), repetitive short sequences (e.g., ARX polyalanine repeats, FMR1 AGG interruptions in CGG repeats, CFTR poly-T/TG repeats), and other complexities (e.g., MSH2 Boland inversions).MethodsWe customized our NGS processes to detect the technically challenging variants mentioned above with adaptations including target enrichment and bioinformatic masking of similar sequences. Adaptations were validated with samples of known genotypes.ResultsOur adaptations provided high-sensitivity and high-specificity detection for most of the variants and provided a high-sensitivity primary assay to be followed with orthogonal disambiguation for the others. The sensitivity of the NGS adaptations was 100% for all of the technically challenging variants. Specificity was 100% for those in PMS2, GBA1, SMN1/SMN2, and HBA1/HBA2, and for the MSH2 Boland inversion; 97.8%-100% for CYP21A2 variants; and 85.7% for ARX polyalanine repeats.ConclusionsNGS assays can detect technically challenging variants when chemistries and bioinformatics are jointly refined. The adaptations described support a scalable, cost-effective path to identifying all clinically relevant variants within a single sample.
Project description:Conventional embolization such as coiling of acute upper gastrointestinal bleeding may be rendered impossible or unsuitable due to anatomical and technical factors. Liquid (Onyx) embolization in such situations may prove valuable and life saving, though literature on the subject, particularly of acute upper gastrointestinal bleeding due to duodenal ulcer, is sparse. We present a technically challenging case of acutely rebleeding duodenal ulcer embolized successfully using ethylene polyvinyl alcohol polymer (Onyx).
Project description:CRISPR/Cas9 represents a valuable tool to determine protein function, but technical hurdles limit its use in challenging settings such as cells unable to grow in vitro like primary leukemia cells and xenografts derived thereof (PDX). To enrich CRISPR/Cas9-edited cells, we improved a dual-reporter system and cloned the genomic target sequences of the gene of interest (GOI) upstream of an out-of-frame fluorochrome which was expressed only upon successful gene editing. To reduce rounds of in vivo passaging required for PDX leukemia growth, targets of 17 GOI were cloned in a row, flanked by an improved linker, and PDX cells were lentivirally transduced for stable expression. The reporter enriched scarce, successfully gene-edited PDX cells as high as 80%. Using the reporter, we show that KO of the SRC-family kinase LYN increased the response of PDX cells of B precursor cell ALL towards Vincristine, even upon heterozygous KO, indicating haploinsufficiency. In summary, our reporter system enables enriching KO cells in technically challenging settings and extends the use of gene editing to highly patient-related model systems.
Project description:BackgroundPeriampullary diverticula (PAD) often detected during endoscopic retrograde cholangiopancreatography (ERCP), and ERCP remains the primary approach to treating bile duct stones, and papilla cannulation plays a critical role in the success of ERCP. PAD can reduce the cannulation success rate. Needle knife precut greatly promoted the clinical application of precut incision. However, this approach also increases the risk of various adverse events. The present study aimed to compare the perioperative outcomes of pancreatic duct guide wire-assisted needle knife precut and conventional needle knife precut for PAD with difficult cannulation.MethodsA total of 230 cases of PAD with difficult cannulation of the duodenal papilla diagnosed by ERCP between June 2009 and December 2021 were retrospectively reviewed. The exclusion criteria were set as follows: patients with ERCP history, coagulopathy prothrombin time two times longer or platelet (PLT) count ≤70×109/L, or an inability to tolerate endoscopy due to severe heart/lung diseases. Pancreatic duct guide wire-assisted needle knife precut (Group A) was performed in 135 cases, and conventional needle knife precut (Group B) was performed in 95 cases. All clinical data were analyzed retrospectively. SPSS20.0 statistical software was used for the t-test and analysis of variance. P<0.05 was considered statistically significant.ResultsThe operating time of the needle knife precut was significantly shorter in Group A (18.44±6.65 min) compared with Group B (32.05±13.15 min, P<0.01). Moreover, the success rate of the cannulation was markedly higher in Group A (100%, 135/135) compared with Group B (78.9%, 75/95). Intraoperative complications occurred in 15 (11.1%) and 26 (27.4%) cases in Groups A and B, respectively (P<0.01). Postoperative complications occurred in 10 (7.4%) and 17 (17.9%) cases in Groups A and B, respectively (P<0.01). Our results showed notable differences in the operating time, success rate of cannulation, intraoperative complication rates, and postoperative complication rates between the two approaches.ConclusionsPancreatic duct guide wire-assisted needle knife precut appeared to be a safe and effective modality for PAD with difficult cannulation in the duodenal papilla.
Project description:Interventions: Group A: Tech knife group Group A performs ESD using a Tech knife.
Group B: Dual knife J group Group B performs ESD using a Dual knife.
Primary outcome(s): Resection time
Study Design: Parallel Randomized