Effectiveness of Histopathological Examination of Ultrasound-guided Puncture Biopsy Samples for Diagnosis of Extrapulmonary Tuberculosis

Objective To evaluate the diagnostic value of histopathological examination of ultrasound-guided puncture biopsy samples in extrapulmonary tuberculosis(EPTB).Methods This study was conducted at the Shanghai Public Health Clinical Center.A total of 115patients underwent ultrasound-guided puncture biopsy,followed by MGIT 960 culture(culture),smear,Gene Xpert MTB/RIF(Xpert),and histopathological examination.These assays were performed to evaluate their effectiveness in diagnosing EPTB in comparison to two different diagnostic criteria:liquid culture and composite reference standard(CRS).Results When CRS was used as the reference standard,the sensitivity and specificity of culture,smear,Xpert,and histopathological examination were(44.83%,89.29%),(51.72%,89.29%),(70.11%,96.43%),and(85.06%,82.14%),respectively.Based on liquid culture tests,the sensitivity and specificity of smear,Xpert,and pathological examination were(66.67%,72.60%),(83.33%,63.01%),and(92.86%,45.21%),respectively.Histopathological examination showed the highest sensitivity but lowest specificity.Further,we found that the combination of Xpert and histopathological examination showed a sensitivity of 90.80%and a specificity of 89.29%.Conclusion Ultrasound-guided puncture sampling is safe and effective for the diagnosis of EPTB.Compared with culture,smear,and Xpert,histopathological examination showed higher sensitivity but lower specificity.The combination of histopathology with Xpert showed the best performance characteristics.

Percutaneous ultrasound-guided coaxial core needle biopsy for the diagnosis of multiple splenic lesions: A case report

BACKGROUND The overlap of imaging manifestations among distinct splenic lesions gives rise to a diagnostic dilemma.Consequently,a definitive diagnosis primarily relies on his-tological results.The ultrasound(US)-guided coaxial core needle biopsy(CNB)not only procures sufficient tissue to help clarify the diagnosis,but reduces the incidence of puncture-related complications.CASE SUMMARY A 41-year-old female,with a history of pulmonary tuberculosis,was admitted to our hospital with multiple indeterminate splenic lesions.Gray-scale ultrasono-graphy demonstrated splenomegaly with numerous well-defined hypoechoic ma-sses.Abdominal contrast-enhanced computed tomography(CT)showed an en-larged spleen with multiple irregular-shaped,peripherally enhancing,hypodense lesions.Positron emission CT revealed numerous abnormal hyperglycemia foci.These imaging findings strongly indicated the possibility of infectious disease as the primary concern,with neoplastic lesions requiring exclusion.To obtain the precise pathological diagnosis,the US-guided coaxial CNB of the spleen was ca-rried out.The patient did not express any discomfort during the procedure.CONCLUSION Percutaneous US-guided coaxial CNB is an excellent and safe option for obtaining precise splenic tissue samples,as it significantly enhances sample yield for exact pathological analysis with minimum trauma to the spleen parenchyma and sur-rounding tissue.

Comparison of Cognitive Registration Transrectal Ultrasound-Guided Targeted Biopsy of Prostate to Systematic 12-Core Biopsy: A Retrospective, Multicentre Study

Introduction: Prostate cancer (PCa) is the third most prevalent cancer among Malaysian males, often diagnosed at advanced stages, leading to suboptimal outcomes. While transrectal ultrasound-guided systematic biopsy (TRUS-SB) is the primary diagnostic method, prebiopsy multiparametric magnetic resonance imaging (mpMRI) is gaining popularity in identifying suspicious lesions. This study addresses the lack of comprehensive investigations into the efficacy of cognitive registration TRUS targeted biopsy (COG-TB) compared to conventional TRUS-SB, considering the resource limitations of the Malaysian healthcare system. Materials and Methods: A retrospective cohort study was conducted in two Malaysian healthcare facilities. 116 adult patients with a prostate-specific antigen (PSA) level of more than 4 ng/mL who underwent both COG-TB and TRUS-SB between October 2020 and March 2022 were included. Primary outcomes were cancer detection rate and histopathological outcomes, including Gleason score. Results: COG-TB showed a higher overall cancer detection rate (50%) compared to TRUS-SB (44%). Clinically significant cancer detection rates were similar between COG-TB and TRUS-SB (37.1%). Further analysis revealed that both COG-TB and TRUS-SB detected clinically significant cancer in 30.2% of patients, did not detect it in 56.0%, and had conflicting findings in 16 patients (p Conclusion: COG-TB and TRUS-SB have comparable detection rates for clinically significant prostate cancer, with COG-TB showing a higher tendency to detect insignificant prostate cancer. Further studies comparing these methods are warranted.

Hemobilia and other complications caused by percutaneous ultrasound-guided liver biopsy

Hemobilia accounts for approximately 3%of all major percutaneous liver biopsy complications,and rarely results from arterioportal fistula.We report a patient who suffered from four complications over 11 d after ultrasound-guided percutaneous liver biopsy:hemobilia,acute pancreatitis,acute cholecystitis,and multiple stomach ulcers.Digital subtraction angiography was done after consultation with doctors,and showed obvious arteriovenous fistula of the right liver.The hepatic artery was selected and embolized by spring orbs.The active bleeding was stopped after embolization of the hepatic artery.The patient was discharged home on day 12 after embolization and remained well.

Endoscopic ultrasound-guided tissue acquisition for the diagnosis of focal liver lesion

In patients with liver tumors,the histopathology examination can assist in diagnosis,staging,prognosis,and therapeutic management strategy.Endoscopic ultrasound(EUS)-guided tissue acquisition using fine needle aspiration(FNA)or more newly fine needle biopsy(FNB)is a well-developed technique in order to evaluate and differentiate the liver masses.The goal of the EUS-FNA or EUS-FNB is to provide an accurate sample for a histopathology examination.Therefore,malignant tumors such as hepatocarcinoma,cholangiocarcinoma and liver metastasis or benign tumors such as liver adenoma,focal hyperplastic nodular tumors and cystic lesions can be accurately diagnosed using EUS-guided tissue acquisition.EUS-FNB using 19 or 22 Ga needle provide longer samples and a higher diagnostic accuracy in patients with liver masses when compared with EUS-FNA.Few data are available on the diagnostic accuracy of EUS-FNB when compared with percutaneously,ultrasound,computer tomography or transjugulary-guided liver biopsies.This review will discuss the EUS-guided tissue acquisition options in patients with liver tumors and its efficacy and safety in providing accurate samples.The results of the last studies comparing EUS-guided liver biopsy with other conventional techniques are presented.The EUS-guided tissue acquisition using FNB can be a suitable technique in suspected liver lesions in order to provide an accurate histopathology diagnosis,especially for those who require endoscopy.

Endoscopic ultrasound-guided through-the-needle microforceps biopsy and needle-based confocal laser-endomicroscopy increase detection of potentially malignant pancreatic cystic lesions:A single-center study

BACKGROUND Currently,there is insufficient data about the accuracy in the diagnosing of pancreatic cystic lesions(PCLs),especially with novel endoscopic techniques such as with direct intracystic micro-forceps biopsy(mFB)and needle-based confocal laser-endomicroscopy(nCLE).AIM To compare the accuracy of endoscopic ultrasound(EUS)and associated techniques for the detection of potentially malignant PCLs:EUS-guided fine needle aspiration(EUS-FNA),contrast-enhanced EUS(CE-EUS),EUS-guided fiberoptic probe cystoscopy(cystoscopy),mFB,and nCLE.METHODS This was a single-center,retrospective study.We identified patients who had undergone EUS,with or without additional diagnostic techniques,and had been diagnosed with PCLs.We determined agreement among malignancy after 24-mo follow-up findings with detection of potentially malignant PCLs via the EUSguided techniques and/or EUS-guided biopsy when available(EUS malignancy detection).RESULTS A total of 129 patients were included, with EUS performed alone in 47/129. In 82/129 patients,EUS procedures were performed with additional EUS-FNA (21/82), CE-EUS (20/82), cystoscopy(27/82), mFB (36/82), nCLE (44/82). Agreement between EUS malignancy detection and the 24-mo follow-up findings was higher when associated with additional diagnostic techniques thanEUS alone [62/82 (75.6%) vs 8/47 (17%);OR 4.35, 95%CI: 2.70-7.37;P < 0.001]. The highestmalignancy detection accuracy was reached when nCLE and direct intracystic mFB were bothperformed, with a sensitivity, specificity, positive predictive value, negative predictive value andobserved agreement of 100%, 89.4%, 77.8%, 100% and 92.3%, respectively (P < 0.001 comparedwith EUS-alone).CONCLUSIONThe combined use of EUS-guided mFB and nCLE improves detection of potentially malignantPCLs compared with EUS-alone, EUS-FNA, CE-EUS or cystoscopy.

Endoscopic ultrasound-guided fine-needle aspiration biopsy-Recent topics and technical tips

Endoscopic ultrasound-guided fine-needle aspiration biopsy (EUS-FNA) is a useful procedure that enables reliable pathological diagnoses of pancreatobiliary diseases, subepithelial lesions, and swollen lymph nodes. In recent years, a pathological diagnosis based on EUS-FNA has made it possible to provide accurate treatment methods not only in these fields, but also in respiratory organs and otorhinolaryngology. This review discusses the latest topics pertaining to EUS-FNA as well as procedural tips.

Septic shock due to Granulicatella adiacens after endoscopic ultrasound-guided biopsy of a splenic mass:A case report

BACKGROUND Endoscopic ultrasound-guided fine needle aspiration or biopsy(EUS-FNA or FNB)has become a popular method for diagnosing various lesions of the gastrointestinal tract and surrounding tissue due to the accuracy and safety.To the best of our knowledge,no case report of severe infection after EUS-FNB of a solid lesion in the spleen has been described.Herein,we report a rare case of septic shock after EUS-FNB of a splenic mass.CASE SUMMARY A 45-year-old male patient presented to the outpatient clinic due to an incidentally detected splenic mass.A definitive diagnosis could not be established based on the abdominal magnetic resonance imaging.EUS of the spleen showed a 6 cm-sized,relatively well-demarcated,heterogeneous mass,and EUS-FNB with a 22G needle was performed.Ten days after the procedure patient developed septic shock and a splenic abscess was identified.Blood culture revealed growth of Granulicatella adiacens.After the treatment with antibiotics the patient underwent surgical resection,and the pathological examination showed diffuse large B-cell lymphoma.The patient received chemotherapy and he is in complete remission.CONCLUSION Infection of a splenic mass after EUS-FNB is a rare complication and prophylactic antibiotics might be considered.

New Wireless Handheld Ultrasound-Guided Vacuum-Assisted Breast Biopsy (VABB) Devices: An Important Innovation in Breast Diagnosis

Vacuum-assisted breast biopsy (VABB) plays a fundamental role in the preoperative assessment of breast lesions pro- viding large histology samples that are useful to define diagnoses and biological parameters to guide treatment planning. This technique has been used in our institute since 2000 and two new wireless handheld ultrasound-guided VABB de- vices have been introduced since May 2012. In this report we analyze our experience with these revolutionary devices which are able to provide the option of single-insertion contiguous tissue samples respectively with a 13/14-gauge aperture. Our initial experience on 75 lesions shows that these devices are safe, fast, procedurally advantageous for operators and well accepted by patients. Finally VABB procedures can markedly reduce the need for surgical biopsy.

Staging the axilla in women with breast cancer: the utility of preoperative ultrasound-guided needle biopsy

Preoperative staging of the axilla in women with invasive breast cancer using ultrasound-guided needle biopsy(UNB) identifies approximately 50% of patients with axillary nodal metastases prior to surgical intervention. Although moderately sensitive, it is a highly specific staging strategy that is rarely falsely-positive, hence a positive UNB allows patients to be triaged to axillary lymph-node dissection(ALND) avoiding potentially unnecessary sentinel node biopsy(SNB). In this review, we extend our previous work through an updated literature search, focusing on studies that report data on UNB utility. Based on data for 10,934 breast cancer patients, sourced from 35 studies, a positive UNB allowed triage of 1,745 cases(simple proportion 16%) to axillary surgical treatment: the utility of UNB was a median 19.8% [interquartile range(IQR) 11.6%-26.7%] across these studies. We also modelled data from a subgroup of studies, and estimated that amongst patients with metastases to axillary nodes, the odds ratio(OR) for high nodal disease burden for a positive UNB versus a negative UNB was 4.38 [95% confidence interval(95% CI): 3.13, 6.13], P<0.001. From this model, the estimated proportion with high nodal disease burden was 58.9%(95% CI: 50.2%, 67.0%) for a positive UNB, whereas the estimated proportion with high nodal disease burden was 24.6%(95% CI: 17.7%, 33.2%) if UNB was negative. Overall, axillary UNB has good clinical utility and a positive UNB can effectively triage to ALND. However, the evolving landscape of axillary surgical treatment means that UNB will have relatively less utility where surgeons have modified their practice to omission of ALND for minimal nodal metastatic disease.

The Diagnostic Significance of Ultrasound and Ultrasound-guided Coarse Needle Biopsy in Breast Cancer in Pregnancy

Objective:To explore the significance of ultrasound and ultrasound-guided coarse needle biopsy in the clinical diagnosis of breast cancer patients in pregnancy.Methods:A total of 10 patients with breast cancer in pregnancy were selected to study from October 2018 to October 2019 in our hospital,who were given preoperative ultrasound results and ultrasound-guided coarse-needle biopsy histopathology,and the results were analyzed.Results:All the 10 patients showed thickening of the mammary gland body,appeared to pick up disorderly situation,most of the dilated catheter,including 8 patients with solid hypoechoic mass,irregular shape,and“crab foot”edge burr.The corresponding aspect ratio value was greater than 1.The other 2 patients presented with cystic mixed masses with poor boundary definition,irregular shape,and mostly accompanied by posterior echo enhancement.Conclusion:Simple ultrasound diagnosis for patients with breast cancer during pregnancy has some errors,and should be used in combination with ultrasound-guided coarse needle biopsy operation.This method has a high clinical diagnosis rate and significant clinical application value,so it is worth promoting.

Diagnosis and excision of breast multi-focal lesions by ultrasound-guided vacuum-assisted biopsy:a comparative evaluation

Objective To compare and evaluate the efficacy of diagnosis and excision for appropriately selected breast multi-focal lesions and solitary lesion by ultrasound-guided vacuum-assisted biopsy(UGVAB).Methods Among 392 appropriately selected patients,187 patients with multi-focal lesions and 205 patients with solitary lesion were treated by the 8-gauge UGVAB from May 2007 to June 2009.All lesions were removed as completely as possible.The patients with benign pathology underwent physical and ultrasound examinations at one week and 6 months after procedure.Results During the procedure,only three patients had vasovagal syncope and twenty others complained of other intraoperative discomfort.An accurate pathological diagnosis was obtained in all lesions.There was no apparent false-negative result among the 696 lesions with benign pathology at a follow-up of 6 months after procedure.The rates of malignant or premalignant pathology,postoperative complications and residual lesions in patients with multi-focal lesions were higher than those in patients with solitary lesion.If each lesion was considered as a subject of study,there was no significant difference between the two groups.Conclusion UGVAB is an effective method for diagnosis and excision of appropriately selected breast multi-focal lesions and can be used routinely.

The efficacy and safety of endoscopic ultrasound-guided fine-needle biopsy in gallbladder masses

Background: Endoscopic ultrasound-guided fine-needle biopsy(EUS-FNB) is a widely used modality for acquiring various target samples, but its efficacy in gallbladder masses is unknown. The aim of this retrospective study was to evaluate the efficacy and safety of EUS-FNB in patients with gallbladder masses. Methods: The study samples were composed of patients from March 2015 to July 2019 who needed to identify the nature of gallbladder masses through EUS-FNB. The outcomes of this study were the adequacy of specimens, diagnostic yields, technical feasibility, and adverse events of the EUS-FNB in gallbladder masses. Results: A total of 27 consecutive patients with a median age of 58 years were included in this study. The 22-gauge FNB needle was feasible in all lesions. The median follow-up period of the patients was 294 days. The specimens sufficient for diagnosis account for 89%(24/27) and 93%(25/27) in cytology and histology, respectively. The overall diagnostic yields for malignancy showed the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 95.45% [95% confidence interval(CI): 75.12%-99.76%], 100%(95% CI: 46.29%-10 0%), 10 0%(95% CI: 80.76%-100%), 83.33%(95% CI: 36.48%-99.12%), and 96.30%(95% CI: 80.20%-99.99%), respectively. The subgroup analysis revealed that FNB could obtain sufficient specimens and high diagnostic yields in both gallbladder mass < 20.5 mm group and ≥20.5 mm group. One patient experienced mild abdominal pain after the procedure and recovered within one day. Conclusions: EUS-FNB is a reasonable diagnostic tool for the pretreatment diagnosis of patients with gallbladder masses, especially for patients who may miss the opportunity of surgery and need sufficient specimens to identify the pathological type so as to determine chemotherapy regimens. Further largescale studies are needed to confirm our conclusion.

Result of 10 Years of Experience in Echo-Guided Kidney Biopsy Punctures in the Nephrology Department of the Martigues Hospital Center, France

Introduction: Percutaneous renal biopsy (PRB) is an essential procedure for the diagnosis and therapeutic management of many primary or secondary nephropathies. Objectives: To identify the indications, to determine the profile of the diagnosed nephropathies and to evaluate the short-term complications related to the practice of echo-guided PRB at the Martigues hospital center. Methodology: This was a retrospective and descriptive study carried out on the records of patients who underwent echo-guided native kidney biopsy from January 1, 2010 to December 31, 2019 in the nephrology department of the Martigues Hospital. Results: The analysis of 123 cases of echo-guided PRB involved 76 men and 47 women with a sex ratio of 1.6. The mean age was 55.92 ± 17.80 with age extremes of 16 and 87 years. Glomerular syndromes were the main indication with 42 cases of nephrotic syndrome (34.1%), 15 cases of nephritic syndrome (12.2%), 11 cases of rapidly progressive glomerulonephritis syndrome (8.9%), and 6 cases of recurrent macroscopic hematuria syndrome (4.9%). The histological findings were 47 cases of primary glomerular lesions (38.3%), 32 cases of nephroangiosclerosis lesions (26%), 24 cases of secondary glomerulopathy (19.5%), 9 cases of interstitial nephritis (7.3%), 2 cases of myelomatous nephropathy (1.6%), and 9 cases (7.3%) of unclassified histological lesions. Twenty-two hypertensive patients (40.7%) had nephroangiosclerosis lesions (p = 0.001). The follow-up was simple in 119 patients (96.7%). Macroscopic hematuria was noted in 4 patients (3.3%). It was associated with a perirenal hematoma in 2 patients including 1 transfused case. Conclusion: Our data provide an important contribution to the understanding of the prevalence and clinical presentation of renal diseases in the nephrology department of the Martigues hospital center.

Comprehensive review on endoscopic ultrasound-guided tissue acquisition techniques for solid pancreatic tumor

Pancreatic ductal adenocarcinoma is speculated to become the second leading cause of cancer-related mortality by 2030,a high mortality rate considering the number of cases.Surgery and chemotherapy are the main treatment options,but they are burdensome for patients.A clear histological diagnosis is needed to determine a treatment plan,and endoscopic ultrasound(EUS)-guided tissue acquisition(TA)is a suitable technique that does not worsen the cancer-specific prognosis even for lesions at risk of needle tract seeding.With the development of personalized medicine and precision treatment,there has been an increasing demand to increase cell counts and collect specimens while preserving tissue structure,leading to the development of the fine-needle biopsy(FNB)needle.EUS-FNB is rapidly replacing EUS-guided fine-needle aspiration(FNA)as the procedure of choice for EUS-TA of pancreatic cancer.However,EUS-FNA is sometimes necessary where the FNB needle cannot penetrate small hard lesions,so it is important clinicians are familiar with both.Given these recent developments,we present an up-to-date review of the role of EUS-TA in pancreatic cancer.Particularly,technical aspects,such as needle caliber,negative pressure,and puncture methods,for obtaining an adequate specimen in EUS-TA are discussed.

Overlooked risk for needle tract seeding following endoscopic ultrasound-guided minimally invasive tissue acquisition

Endoscopic ultrasound-guided minimally invasive tissue acquisition can be performed by two approaches as follows: Endoscopic ultrasound-guided fineneedle aspiration(EUS-FNA) and endoscopic ultrasound-guided fine-needle biopsy(EUS-FNB). These have been evolved into leading approaches and widely used for the histological diagnosis of tumors in the gastrointestinal tract and adjacent organs. However, the role of EUS-FNA and EUS-FNB in disease diagnosis and evaluation remains controversial. Although the incidence of surgery-associated complications remains low, the consequences of needle tract seeding can be serious or even life-threatening. Recently, increasing case reports of needle tract seeding are emerging, especially caused by EUS-FNA. This complication needs serious consideration. In the present work, we integrated these case reports and the related literature, and summarized the relevant cases and technical characteristics of needle tract seeding caused by EUS-FNA and EUSFNB. Collectively, our findings provided valuable insights into the prevention and reduction of such serious complication.

Influence of the safety and diagnostic accuracy of preoperative endoscopic ultrasound-guided fine-needle aspiration for resectable pancreatic cancer on clinical performance

AIM: To evaluate the safety and diagnostic accuracy of endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) in a cohort of pancreatic cancer patients.

Peritoneal dissemination of pancreatic cancer caused by endoscopic ultrasound-guided fine needle aspiration:A case report and literature review

BACKGROUND Endoscopic ultrasound-guided fine needle aspiration(EUS-FNA)is a biopsy technique widely used to diagnose pancreatic tumors because of its high sensitivity and specificity.Although needle-tract seeding caused by EUS-FNA has been recently reported,dissemination of pancreatic cancer cells is generally considered to be a rare complication that does not affect patient prognosis.However,the frequency of dissemination and needle-tract seeding appears to have been underestimated.We present a case of peritoneal dissemination of pancreatic cancer due to preoperative EUS-FNA.CASE SUMMARY An 81-year-old man was referred to the Department of Surgery of our hospital in Japan owing to the detection of a pancreatic mass on computed tomography during medical screening.Trans-gastric EUS-FNA revealed that the mass was an adenocarcinoma;hence laparoscopic distal pancreatectomy with lymphadenectomy was performed.No intraoperative peritoneal dissemination and liver metastasis were visually detected,and pelvic lavage cytology was negative for carcinoma cells.The postoperative surgical specimen was negative for carcinoma cells at the dissected margin and the cut end margin;however,pathological findings revealed adenocarcinoma cells on the peritoneal surface proximal to the needle puncture site,and the cells were suspected to be disseminated via EUSFNA.Hence,the patient received adjuvant therapy with S-1(tegafur,gimeracil,and oteracil potassium);however,computed tomography performed 5 mo after surgery revealed liver metastasis and cancerous peritonitis.The patient received palliative therapy and died 8 mo after the operation.CONCLUSION The indications of EUS-FNA should be carefully considered to avoid iatrogenic dissemination,especially for cancers in the pancreatic body or tail.

Three‑dimensional Modeling and Simulation of Muscle Tissue Puncture Process

Needle biopsy is an essential part of modern clinical medicine.The puncture accuracy and sampling success rate of puncture surgery can be effectively improved through virtual surgery.There are few three-dimensional puncture(3D)models,which have little significance for surgical guidance under complicated conditions and restrict the development of virtual surgery.In this paper,a 3D simulation of the muscle tissue puncture process is studied.Firstly,the mechanical properties of muscle tissue are measured.The Mooney-Rivlin(M-R)model is selected by considering the fitting accuracy and calculation speed.Subsequently,an accurate 3D dynamic puncture model is established.The failure criterion is used to define the breaking characteristics of the muscle,and the bilinear cohesion model defines the breaking process.Experiments with different puncture speeds are carried out through the built in vitro puncture platform.The experimental results are compared with the simulation results.The experimental and simulated reaction force curves are highly consistent,which verifies the accuracy of the model.Finally,the model under different parameters is studied.The simulation results of varying puncture depths and puncture speeds are analyzed.The 3D puncture model can provide more accurate model support for virtual surgery and help improve the success rate of puncture surgery.

Strategic insights into the cultivation of pancreatic cancer organoids from endoscopic ultrasonography-guided biopsy tissue

BACKGROUND The frequent suboptimal efficacy of endoscopic ultrasound-guided fine-needle biopsy(EUS-FNB)to culture pancreatic cancer(PC)organoids(PCOs)poses a major challenge in the advancement of personalized medicine for advanced PC.AIM To explore how to obtain appropriate puncture tissues from EUS-FNB and optimize the strategy for efficiently constructing PCOs,providing an efficient tool for the advancement of personalized medicine.METHODS Patients who underwent EUS-FNB for the diagnosis of PC tissue were prospectively enrolled.We refined the endoscopic biopsy procedures and organoid cultivation techniques.All tissue specimens verified by on-site pathological assessment were cultured in a semi-suspended medium in a microfluidic environment.We assessed differences in PCOs cultured beyond and below five generations examining patient demographics,specimen and organoid attributes,and the sensitivity of organoids to a panel of clinical drugs through cell viability assays.RESULTS In this study,16 patients with PC were recruited,one sample was excluded because onsite cytopathology showed no tumor cells.Successful organoid generation occurred in 93.3%(14 of 15)of the EUS-FNB specimens,with 60%(9 of 15)sustaining over five generations.Among these patients,those with a history of diabetes,familial cancer,or larger tumors exhibited enhanced PCO expandability.The key factors influencing longterm PCOs expansion included initial needle sample quality(P=0.005),rapid initiation of organoid culture postisolation(P≤0.001),and high organoid activity(P=0.031).Drug sensitivity analysis revealed a partial response in two patients following therapeutic intervention and surgery and stable disease in four patients,indicating a moderate correlation between organoid response and clinical outcomes.CONCLUSION Optimal initial needle sampling,rapid and precise biopsy sample processing,process isolated samples as soon as possible,and sufficient cellular material are crucial for successful cultivating PCOs.High organoid activity is an important factor in maintaining their long-term expansion,which is essential for shortening the time of drug sensitivity analysis and is the basis of PC research.