Fine-needle aspiration technique under endoscopic ultrasound guidance:A technical approach for RNA profiling of pancreatic neoplasms

BACKGROUND Early diagnosis of pancreatic ductal adenocarcinoma(PDAC)has been a longstanding challenge.The prognosis of patients with PDAC depends on the stage at diagnosis.It is necessary to identify biomarkers for the detection and differentiation of pancreatic tumors and optimize PDAC sample preparation procedures for DNA and RNA analysis.Most molecular studies are done using paraffin-embedded blocks;however,the integrity of DNA and RNA is often compromised in this format.Moreover,RNA isolated from human pancreatic tissue samples is generally of low quality,in part,because of the high concentration of endogenous pancreatic RNAse activity present.AIM To assess the potential of endoscopic ultrasound-guided fine-needle aspiration(EUS-FNA)to obtain specimens from pancreatic neoplasms for subsequent RNA molecular profiling,including next-generation sequencing(NGS).METHODS Thirty-four EUS-FNA samples were included in this study:PDAC(n=15),chronic pancreatitis(n=5),pancreatic cysts(n=14),mucinous cysts(mucinous cystic neoplasia/intraductal papillary mucinous neoplasia)n=7,serous cystic neoplasms n=5,and pseudocysts n=2.Cyst material consisted of cyst fluid and cyst wall samples obtained by through-the-needle biopsy(TTNB).Samples were stored at -80℃ until analysis.RNA purity(A260/230,A260/280 ratios),concentration,and integrity(RIN)were assessed.Real-time polymerase chain reaction was conducted on all samples,and small RNA libraries were prepared from solid mass samples.RESULTS RNA was successfully extracted from 29/34(85%)EUS-FNA samples:100% pancreatic adenocarcinoma samples,100% chronic pancreatitis samples,70% pancreatic fluid cyst samples,and 50%TTNB samples.The relative expression of GAPDH and HPRT were obtained for all successfully extracted RNA samples(n=29)including lowquality RNA specimens.Low concentration and nonoptimal RIN values(no less than 3)of RNA extracted from EUS-FNA samples did not prevent NGS library preparation.The suitability of cyst fluid samples for RNA profiling varied.The quality of RNA extracted from mucinous cyst fluid had a median RIN of 7.7(5.0-8.2),which was compatible with that from solid neoplasms[6.2(0-7.8)],whereas the quality of the RNA extracted from all fluids of serous cystic neoplasms and TTNB samples had a RIN of 0.CONCLUSION The results demonstrate the high potential of EUS-FNA material for RNA profiling of various pancreatic lesions,including low-quality RNA specimens.

Role of macroscopic on-site evaluation of endoscopic ultrasoundguided fine-needle aspiration/biopsy:Results of a multicentric prospective study

BACKGROUND The concept of macroscopic on-site evaluation(MOSE)was introduced in 2015 when the endoscopist observed better diagnostic yield when the macroscopically visible core on MOSE was superior to 4 mm.Recent studies suggest that MOSE by the endoscopist may be an excellent alternative to rapid on-site evaluation,and some classi-fications have been published.Few studies have assessed the adequacy of histologic cores in MOSE during endoscopic ultrasound-guided fine-needle aspiration/biopsy(EUS-FNA/FNB).AIM To evaluate the performance of MOSE during EUS-FNA/FNB.METHODS This multicentric prospective study was conducted in 16 centers in 3 countries(Egypt,Iraq,and Morocco)and included 1108 patients with pancreatic,biliary,or gastrointestinal pathology who were referred for EUS examination.We prospectively analyzed the MOSE in 1008 patients with available histopathological reports according to 2 classifications to determine the adequacy of the histological core samples.Data management and analysis were performed using a Statistical Package for Social Sciences(SPSS)version 27.RESULTS A total of 1074 solid lesions were biopsied in 1008 patients with available cytopathological reports.Mean age was 59 years,and 509 patients(50.5%)were male.The mean lesion size was 38 mm.The most frequently utilized needles were FNB-Franseen(74.5%)and 22 G(93.4%),with a median of 2 passes.According to 2 classifications,618 non-bloody cores(61.3%)and 964 good samples(95.6%)were adequate for histological evaluation.The overall diagnostic yield of cytopathology was 95.5%.The cytological examination confirmed the diagnosis of malignancy in 861 patients(85.4%),while 45 samples(4.5%)were inconclusive.Post-procedural adverse events occurred in 33 patients(3.3%).Statistical analysis showed a difference between needle types(P=0.035)with a high sensitivity of FNB(97%).The analysis of the relationship between the MOSE-score and the final diagnosis showed a significant difference between the different scores of the MOSE(P<0.001).CONCLUSION MOSE is a simple method that allows endoscopists to increase needle passes to improve sample quality.There is significantly higher FNB sensitivity and cytopathology diagnostic yield with good MOSE cores.

Endoscopic ultrasound-guided fine-needle aspiration cytology diagnosis of solid pseudopapillary tumor of the pancreas: A case report and literature review

We describe the clinical, imaging and cytopathological features of solid pseudopapillary tumor of the pancreas (SPTP) diagnosed by endoscopic ultrasound- guided (EUS-guided) fine-needle aspiration (FNA). A 17-year-old woman was admitted to our hospital with complaints of an unexplained episodic abdominal pain for 2 mo and a short history of hypertension in the endocrinology clinic. Clinical laboratory examinations revealed polycystic ovary syndrome, splenomegaly and low serum amylase and carcinoembryonic antigen (CEA) levels. Computed tomography (CT) analysis revealed a mass of the pancreatic tail with solid and cystic consistency. EUS confirmed the mass, both in body and tail of the pancreas, with distinct borders, which caused dilation of the peripheral part of the pancreatic duct (major diameter 3.7 mm). The patient underwent EUS-FNA. EUS-FNA cytology specimens consisted of single cells and aggregates of uniform malignant cells, forming microadenoid structures, branching, papillary clusters with delicate fibrovascular cores and nuclear overlapping. Naked capillaries were also seen. The nuclei of malignant cells were round or oval, eccentric with fine granular chromatin, small nucleoli and nuclear grooves in some of them. The malignant cells were periodic acid Schiff (PAS)-Alcian blue (+) and immunocytochemically they were vimentin (+), CA 19.9 (+), synaptophysin (+), chromogranin (-), neuro-specific enolase (-), a1- antitrypsin and a1-antichymotrypsin focal positive. Cytologic findings were strongly suggestive of SPTP. Biopsy confirmed the above cytologic diagnosis. EUS- guided FNA diagnosis of SPTP is accurate. EUS findings,cytomorphologic features and immunostains of cell block help distinguish SPTP from pancreatic endocrine tumors, acinar cell carcinoma and papillary mucinous carcinoma.

Rapid on-site evaluation of endoscopic-ultrasound-guided fine-needle aspiration diagnosis of pancreatic masses

Endoscopic ultrasound (EUS) has become an essential tool for the study of pancreatic diseases. Specifically, EUS plays a pivotal role evaluating patients with a known or suspected pancreatic mass. In this setting, differential diagnosis remains a clinical challenge. EUS-guided fine-needle aspiration (FNA) and fine-needle biopsy (FNB) have been proven to be safe and useful tools in this setting. EUS-guided FNA and FNB, by obtaining cytological and/or histological samples, are able to diagnose pancreatic lesions with high sensitivity and specificity. In this context, several methodological features, trying to increase the diagnostic yield of EUS-guided FNA and FNB, have been evaluated. In this review, we focus on the role of rapid on-site evaluation (ROSE). From data reported in the literature, ROSE may increase diagnostic yield of EUS-FNA specimens by 10%-30%, and thus, diagnostic accuracy. However, we should point out that many recent studies have reported adequacy rates of &#x0003e; 90% without ROSE, indicating that, perhaps, at high-volume centers, ROSE may not be indispensable to achieve excellent results. The use of ROSE can be considered important during the learning curve of EUS-FNA, and also in hospital with diagnostic accuracy rates &#x0003c; 90%.

Endoscopic ultrasound-guided fine-needle aspiration cytology in pancreaticobiliary carcinomas:diagnostic efficacy of cell-block immunocyto-chemistry

BACKGROUND： Endoscopic ultrasound-guided fine-needle aspiration cytology was demonstrated to be a useful tool for the diagnosis and staging of pancreaticobiliary neoplastic le- sions. Nonetheless, the diagnostic value of this procedure may be limited by low cellularity of the specimen, contamination of intestinal cells and unfeasibility of ancillary immunocy- tochemical procedures. The present study was to evaluate its usefulness in the diagnosis of neoplastic lesions.

Incidents and adverse events of endoscopic ultrasoundguided fine-needle aspiration for pancreatic cystic lesions

AIM To evaluate the diagnostic value and safety mainly regarding incidents of endoscopic ultrasound-guided fine-needle aspiration(EUS-FNA) for pancreatic cystic lesions(PCLs).METHODS A total of 150 consecutive patients with suspected PCLs were prospectively enrolled from April 2015 to November 2016. We finally enrolled 140 patients undergoing EUS-FNA. We compared the diagnostic accuracy of EUS-FNA and pathological diagnosis, which is regarded as the gold standard, for PCLs. Patients undergoing EUS-FNA at least 1 wk preoperatively were monitored for incidents and adverse events to evaluate its safety.RESULTS There were 88(62.9%) women and 52(37.1%) men among 140 patients, with a mean age of 50.1(± 15.4) years. There were 67 cysts located in the head/uncinate of the pancreas and 67 in the body/tail, and 6 patients had at least 1 cyst in the pancreas. There were 75 patients undergoing surgery and 55 undergoing EUS-FNA with interval at least 1 wk before other operations, with 3 patients undergoing the procedure twice. The accuracy of EUS-FNA in differentiating benign and malignant lesions was 97.3%(73/75), while the accuracy of characterizing PCL subtype was 84.0%(63/75). The incident rate was 37.9%(22/58), whereas only 1 AE was observed in 58 cases.CONCLUSION EUS-FNA is effective and safe for diagnosis of PCLs, however procedure-related incidents are common. Caution should be taken in patients undergoing EUSFNA.

Slow-pull and different conventional suction techniques in endoscopic ultrasound-guided fine-needle aspiration of pancreatic solid lesions using 22-gauge needles

AIM To evaluate the cytological diagnostic capacity and sample quality of the slow-pull technique and compare them with different suction techniques.METHODS From July 2010 to December 2015, 102 patients with pancreatic solid lesions who underwent endoscopic ultrasound-guided fine-needle aspiration(EUS-FNA) with 22-gauge needles were retrospectively evaluated. EUS-FNA diagnosis was based on a cytological examination, and final diagnosis was based on a comprehensive standard of cytological diagnosis, surgical pathology and clinical or imaging follow-up. Cytological specimens were characterized for cellularity and blood contamination. The cytological diagnostic capacity and sample quality of the slow-pull technique and suction techniques with 5-m L/10-m L/20-m L syringes were analyzed.RESULTS Of all of the EUS-FNA procedures, the slow-pull technique and suction techniques with 5-m L/10-m L/20-m L syringes were used in 31, 19, 34 and 18 procedures, respectively. There were significant differences between these four suction techniques in terms of cytological diagnostic accuracy(90.3% vs 63.2% vs 58.8% vs 55.6%, P = 0.019), sensitivity(88.2% vs 41.7% vs 40.0% vs 36.4%, P = 0.009) and blood contamination(score ≥ 2 for 29.0% vs 52.6% vs 70.6% vs 72.2%, P = 0.003). The accuracy and sensitivity of the slow-pull technique were significantly higher than those of the suction techniques using 5-m L(P = 0.03, P = 0.014), 10-m L(P = 0.005; P = 0.006) and 20-mL syringes(P = 0.01, P = 0.01). Blood contamination was significantly lower in the slow-pull technique than in the suction techniques with 10-m L(P = 0.001) and 20-mL syringes(P = 0.007).CONCLUSION The slow-pull technique may increase the cytological diagnostic accuracy and sensitivity with slight blood contamination during EUS-FNA when using 22-gauge needles for solid pancreatic masses.

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.

Value of thyroglobulin combined with ultrasound-guided fine-needle aspiration cytology for diagnosis of lymph node metastasis of thyroid carcinoma

BACKGROUND Surgery for thyroid carcinoma offers a good prognosis;however,cervical lymph node metastasis may occur in the early stage.An effective diagnostic method can accurately guide clinical surgical planning and the scope of lymph node dissection,ultimately improving patient prognosis.AIM To explore the diagnostic value of fine-needle aspiration of thyroglobulin(FNATg)combined with ultrasound(US)-guided fine-needle aspiration cytology for cervical lymph node metastasis in thyroid carcinoma.METHODS We enrolled 209 pathologically confirmed thyroid carcinoma patients who visited our hospital between Jan 2017 and Dec 2020.Patients were tentatively diagnosed with cervical lymph node enlargement using preoperative US.They underwent US-guided fine-needle aspiration cytology and FNA-Tg.The value of single and combined application of the two methods for the diagnosis of cervical lymph node metastasis was calculated.The factors affecting FNA-Tg for diagnosis were analyzed using univariate and multivariate methods.RESULTS FNA-Tg values were significantly higher among patients with positive cervical lymph node metastasis.The sensitivity and specificity of US-guided fine-needle aspiration cytology,FNA-Tg,and US-guided fine-needle aspiration cytology+FNA-Tg were 85.48%and 90.59%,83.06%and 87.06%,and 96.77%and 91.76%,respectively.The area under the receiver operating characteristic curve for USguided fine-needle aspiration cytology,FNA-Tg,and the two combined,was 0.880,0.851,and 0.943,respectively.A long diameter/short diameter ratio<2,an insufficient number of acquired cells,a low serum thyroglobulin level,and an absence of typical metastatic US features increased the risk of cervical lymph node metastasis in thyroid carcinoma patients misdiagnosed using FNA-Tg.CONCLUSION The diagnostic value of FNA-Tg for detecting cervical lymph node metastasis is not high;however,combined with US-guided fine-needle aspiration cytology,it is significantly improved.

Statins decrease the risk of acute pancreatitis after endoscopic ultrasound fine-needle aspiration of pancreatic cysts

Background: Basic and clinical studies suggest that statins may prevent and even ameliorate acute pan- creatitis. The present study was to evaluate whether statin decreases the risk of acute pancreatitis in patients undergoing endoscopic ultrasound-guided ne-needle aspiration of pancreatic cysts. Methods: Out of 456 patients with pancreatic cysts referred to our center between 2006 and 2018, 365 were nally included in analyses: 86 were treated with statins and 279 were not at the time of endo- scopic ultrasound ne-needle aspiration. We compared the acute pancreatitis incidence between the two groups, and we also compared other complications such as bleeding and infections. Results: Median age was 64 years [interquartile range (IQR) 62 69] and median cyst size was 24mm (IQR, 21 29). The most frequent histology was intraductal papillary mucinous neoplasm (45.3% and 42.3% in the two groups, respectively;P =0.98). All 13 patients experiencing post-endoscopic ultrasound acute pancreatitis were from the control group (4.7%), of which 3 were classi ed as severe pancreatitis. None of statin users developed post-procedural acute pancreatitis (odds ratio: 0.15;95% con dence interval: 0.03 0.98;P=0.03). No difference was registered with regard to severe pancreatitis and other complications. Conclusions: Statins exert a bene cial role in preventing acute pancreatitis in patients with pancreatic cysts undergoing endoscopic ultrasound-guided ne-needle aspiration. If con rmed in prospective trials, our ndings may pave the way to an extensive use of statins as prophylactic agents in pancreatic inter- ventional endoscopy.

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.

Fine-needle aspiration cytology of an intrathyroidal nodule diagnosed as squamous cell carcinoma: A case report

BACKGROUND Both squamous cell carcinoma(SCC)and papillary thyroid carcinoma(PTC)are common malignant tumors in the neck.However,seldom has SCC of the thyroid been diagnosed.Further,cytological features of SCC and PTC have rarely been reported.The significance of fine-needle aspiration cytology(FNAC)in the diagnosis of neck masses has been established.Herein,we present an exceedingly rare case of an intrathyroidal SCC diagnosed using FNAC,along with its cytological features.CASE SUMMARY A 66-year-old man presented with a left-sided neck mass.Ultrasound examination showed an ill-defined nodule.The appearance was hypoechoic with a few hyperechoic spots.FNAC of the left thyroid nodule was performed.A cellular smear was obtained,and it showed a large number of neoplastic cells with rich cytoplasm and poor cell adhesion.Tumor cell nuclei showed coarse nuclear chromatin and a few enlarged prominent nucleoli.An increased nuclear/cytoplasm ratio was observed.Thus,malignancy was diagnosed without a confirmed tumor type.Percutaneous tumor biopsy was performed to make a definite diagnosis.The tumor cells showed typical squamous cell characteristics.CONCLUSION Head and neck SCC and PTC have different cytologies.Measures are needed to ensure accurate diagnosis using FNAC.

Simultaneous multiple primary malignancies diagnosed by endoscopic ultrasound-guided fine-needle aspiration:A case report

BACKGROUND Multiple primary malignancies(MPMs)refer to more than one primary malignancy in the same or separate organs of the same patient,and MPMs are considered when different histological characteristics are detected in epidemiological studies.Herein,we report a case presumed to be primary pancreatic cancer with multiple liver metastases by positron-emission tomography/computed tomography(PET/CT)and confirmed to be synchronous liver and pancreatic MPMs by endoscopic ultrasound-guided fine-needle aspiration(EUSFNA).CASE SUMMARY A 50-year-old man was referred to our hospital due to abdominal discomfort for 2 mo.Abdominal CT at a local hospital revealed a pancreatic mass with multiple liver nodules.After being transferred to our hospital,PET/CT confirmed all these lesions to have elevated metabolic activity,and therefore primary pancreatic cancer with multiple liver metastases was considered.EUS-guided liver aspiration unexpectedly found signet-ring cells with a high Ki-67 positive rate(20%),while EUS-guided pancreatic aspiration detected pancreatic neuroendocrine cells with a relatively low Ki-67 positive rate(1%).The final diagnosis from the multidisciplinary team was simultaneous liver and pancreatic MPMs.The patient returned to his local hospital for neoadjuvant chemotherapy and surgery,and he is still alive during the 6-mo postoperative follow-up.CONCLUSION Although rare,MPMs should be considered when treating pancreatic mass with suspected metastatic lesions,and EUS-FNA has proved minimally invasive and accurate.

New treatment for gastric duplication cyst:Endoscopic ultrasonography-guided fine-needle aspiration combined with lauromacrogol sclerotherapy:A case report

BACKGROUND Gastric duplication cysts are very rare disease that are mainly diagnosed by endoscopic ultrasonographic fine-needle aspiration biopsy.In the past,this disease was usually treated with traditional surgery and rarely with minimally invasive endoscopic surgery.However,minimally invasive endoscopic therapy has many advantages,such as no skin wound,organ preservation,postoperative pain reduction,early food intake,fewer postoperative complications,and shorter post-procedure hospitalization.CASE SUMMARY We report a case of endoscopic ultrasonography-guided fine-needle aspiration(EUS-FNA)combined with lauromacrogol sclerotherapy for pyloric obstruction due to gastric duplication cysts.CONCLUSION EUS-FNA combined with lauromacrogol sclerotherapy provides a new option for the treatment of gastrointestinal duplication cysts.

Fine-needle aspiration for periprosthetic fluid removal after implantation of a remote internal-port tissue expander

Background:Use of internal filling ports in tissue expander–based reconstructions are advantageous because of easier self-care,lower infection rates,and fewer instances of capsule formation.The appearance of periprosthetic fluid accumulation after internal-port tissue expander implantation is a common complication that warrants treatment.In this study,we introduced a noninvasive method using fine-needle aspiration(FNA)to remove fluids accumulated after implantation of a remote internal-port tissue expander.Methods:In this study,245 patients who underwent implantation of remote internal-port tissue expanders in our hospital from July 1,2012,to July 1,2019,were included and divided into two groups.In the control group,patients underwent tissue expander implantation before July 1,2016,and large quantities of fluids were removed with surgical aspiration procedures in most cases.In the FNA group,the patients underwent implantation after July 1,2016,and large quantities of fluids were removed first with the FNA procedure.Patients’demographic data,indications for FNA application,and related complications were collected and analyzed.Results:Overall,395 expanders were placed in 245 patients.Postoperative management was similar in both groups.Fluids were managed with 23 expanders in the control group and with 31 expanders in the FNA group.There was no difference in the fluid aspiration rate between the two groups.The surgical aspiration rate was 11.1%(23/208)in the control group.The success rate of FNA was 90.3%(28/31).In the FNA group,the surgical aspiration rate was 1.6%(3/187),which was significantly lower than that in the control group.There were no significant differences in complications between the two groups.Conclusion:FNA can be used for periprosthetic fluid removal after the implantation of a remote internal-port tissue expander in most cases.This method is more convenient and safer than surgical aspiration for the postoperative management of internal-port tissue expander implantation.

Evaluation of clinical relevance of examining K-ras, p16 and p53 mutations along with allelic losses at 9p and 18q in EUS-guided fine needle aspiration samples of patients with chronic pancreatitis and pancreatic cancer

AIM： To establish an optimum combination of molecular markers resulting in best overall diagnostic sensitivity and specificity for evaluation of suspicious pancreatic mass. METHODS： Endoscopic ultrasound （EUS）-guided fine needle aspiration cytology （FNA） was performed on 101 consecutive patients （63 males, 38 females, 60 ± 12 years; 81 with subsequently diagnosed pancreatic cancer, 20 with chronic pancreatitis） with focal pancreatic mass. Samples were evaluated on-site by an experienced cytopathologist. DNA was extracted from Giemsa stained cells selected by laser microdissection and the presence of K-ras, p53 and p16 somatic mutations was tested by cycling-gradient capillary electrophoresis （CGCE） and single-strand conformation polymorphism （SSCP） techniques. In addition, allelic losses of tumor suppressor genes p16 （INK4, CDKN2A） and DPC4 （MADH4, SMAD4） were detected by monitoring the loss of heterozygosity （LOH） at 9p and 18q, respectively. RESULTS： Sensitivity and specificity of EUS-guided FNA were 75% and 85%, positive and negative predictive value reached 100%. The remaining 26% samples were assigned as inconclusive. Testing of molecular markers revealed sensitivity and specificity of 70% and 100% for K-ras mutations （P 〈 0.001）, 24% and 90% for p53 mutations （NS）, 13% and 100% for p16 mutations （NS）, 85% and 64% for aUelic losses at 9p （P 〈 0.001） and 78% and 57% for allelic losses at 18q （P 〈 0.05）. When tests for different molecular markers were combined, the best results were obtained with K-ras ＋ LOH at 9p （92% and 64%, P 〈 0.001）, K-ras ＋ LOH at 18q （92% and 57%, P 〈 0.001）, and K-ras ＋ LOH 9q ＋ LOH 18q （96% and 43%, P 〈 0.001）. When the molecular markers were used as complements to FNA cytology to evaluate inconclusive samples only, the overall sensitivity of cancer detection was 100% in all patients enrolled in the study. CONCLUSION： EUS-guided FNA cytology combined with screening of K-ras mutations and allelic losses of tumor suppressors p16 and DPC4 represents a very sensitive approach in screening for pancreatic malignancy. Molecular markers may find its use particularly in cases where FNA cytology has been inconclusive.

Accuracy of endoscopic ultrasound-guided needle aspiration specimens for molecular diagnosis of non-small-cell lung carcinoma

BACKGROUND Endoscopic ultrasonography-guided fine-needle aspiration(EUS-FNA)and endobronchial ultrasound-guided transbronchial needle aspiration(EBUS-TBNA)are highly sensitive for diagnosing and staging lung cancer.In recent years,targeted therapy has shown great significance in the treatment of non-small cell lung carcinoma(NSCLC).Using these minimally invasive techniques to obtain specimens for molecular testing will provide patients with a more convenient diagnostic approach.AIM To evaluate the feasibility and accuracy of tissue samples obtained using EUSFNA and EBUS-TBNA for molecular diagnosis of NSCLC.METHODS A total of 83 patients with NSCLC underwent molecular testing using tissues obtained from EUS-FNA or EBUS-TBNA at the Tianjin Medical University Cancer Hospital from January 2017 to June 2019.All enrolled patients underwent chest computed tomography or positron emission tomography/computed tomography prior to puncture.We detected abnormal expression of EGFR,KRAS,MET,HER2,ROS1 and anaplastic lymphoma kinase protein.Two patients failed to complete molecular testing due to insufficient tumor tissue.The clinical features,puncture records,molecular testing results and targeted treatment in the remaining 81 patients were summarized.RESULTS In a total of 99 tissue samples obtained from 83 patients,molecular testing was successfully completed in 93 samples with a sample adequacy ratio of 93.9%(93/99).Biopsy samples from two patients failed to provide test results due to insufficient tumor tissue.In the remaining 81 patients,62 cases(76.5%)were found to have adenocarcinoma,11 cases(13.6%)had squamous cell carcinoma,3 cases(3.7%)had adenosquamous carcinoma and 5 cases(6.2%)had NSCLC-not otherwise specified.The results of molecular testing showed EGFR mutations in 21 cases(25.9%),KRAS mutations in 9 cases(11.1%),ROS-1 rearrangement in 1 case(1.2%)and anaplastic lymphoma kinase-positive in 5 cases(6.2%).Twentyfour patients with positive results received targeted therapy.The total effectiveness rate of targeted therapy was 66.7%(16/24),and the disease control rate was 83.3%(20/24).CONCLUSION Tissue samples obtained by EUS-FNA or EBUS-TBNA are feasible for the molecular diagnosis of NSCLC and can provide reliable evidence for clinical diagnosis and treatment.

Gouty Tophus: Cytological Findings in a Case Diagnosed by Fine Needle Aspiration

A case of gouty tophus in the elbow of a 62-year-old male is presented. Fine-needle aspiration (FNA) revealed the presence of acicular crystals, birefringent when viewed under polarized light as is typical for monosodium urate crystals. Histological study of the removed lesion confirmed a typical gouty tophus. We briefly comment the main characteristics of this entity.

Co-Inhibitors of Second Signal of Lymphocyte Response in Human Renal Transplants: PD-L2, GITR, and ILT-2/3/5 Positive Cells from Aspiration Biopsies Associate with Acute Rejection-Freedom

Following organ transplantation, the outcome of the encounter between an APC and a T lymphocyte is strongly dependent on the presence of costimulatory and co-inhibitory molecules, the former associated with allograft rejection and the latter with allograft acceptance. We evaluated the expression of PD-L2, GITR, ILT-2/3/5, and ILT-4 on graft-infiltrating cells procured by Fnab from human KTx under different immunosuppressive regimens. Methods: Fnab biopsies were performed on days 7 or 14 - 30 in stable KTx and on the day of acute rejection diagnosis. Cytopreparations were studied by the enzymatic avidin biotin complex staining. Results: Acute rejection group showed a significant down-regulated expression of PD-L2, GITR, and ILT-2/3/5 as compared to stable group, while for ILT-4 we did not find significant difference. Anti-IL2αR and rapamicyn treatment trend to down-regulate ILT-4 expression, although meaningless. A significantly positive correlation was observed between PD-L2 and GITR expression in Fnab. The PPV for acute rejection diagnosis for both PD-L2 and GITR was clearly above 0.8. Conclusions: Our findings point to an early entrance of cells expressing PD-L2, GITR and ILT-2/3/5 inside human KTx who are going to remain rejection-free. Both PD-L2 and GITR shared a high ability to rule-in and rule-out acute rejection.

Endoscopic Ultrasound-Guided Fine Needle Aspiration Cytology of Gastrointestinal Tract Tumors and Related Organs: Predictive Factors for Accurate Cytopathological Diagnosis

Background: Endoscopic ultrasonography (EUS) is well-established, highly accurate clinical diagnostic test for detection and staging of gastrointestinal tract lesions and related organs. The addition of endoscopic guided fine needle aspiration cytology (EUS-FNAC) has improved the performance characteristics of EUS. Aim: Evaluate the validity of EUS-FNAC in diagnosis of gastrointestinal tract lesions and related organs and assess predictive factors for an accurate EUS-FNAC diagnosis. Methods: Our study included cytological sampling from one hundred sixty-six lesions obtained from gastrointestinal tract and related organs. Factors affecting EUS-FNA accuracy were analyzed. The histopathological results or clinical follow-up were used as the gold standard method. Results: Samples were obtained from: pancreatic masses (n = 80), gastric masses (n = 34), lymph nodes (n = 22), hepatobiliary masses (n = 18) and rectal masses (n = 2). Statistical analysis of sensitivity, specificity, Positive Predictive Value, Negative Predictive Value, and diagnostic accuracy of EUS alone was 80.4%, 51.6%, 75.8%, 76.2% and 75.9% respectively. The sensitivity, specificity, positive predictive value and negative predictive value of EUS-FNAC are 78.8%, 96.8%, 97.6% and 73.2% respectively. Addition of FNAC to EUS improved diagnostic accuracy to 85.5%. Positive statistical association was present between cytological adequacy and adequate cell block preparations, larger lesion size, presence of rapid on site evaluation (ROSE) and obtaining at least two passes from target lesion. Conclusions: EUS is valuable diagnostic and cost effective tool for gastrointestinal tract lesions and related organs when combined with FNAC.