Comparisons of voided urine cytology, nuclear matrix protein-22 and bladder tumor associated antigen tests for bladder cancer of geriatric male patients in Taiwan, China

Aim： To compare the results of bladder tumor associated antigen （BTA TRAK）, nuclear matrix protein 22 （NMP 22） and voided urine cytology （VUC） in detecting bladder cancer. Methods： A total of 135 elderly male and 50 healthy volunteers enrolled in this study were classified into three groups： （i） 93 patients with bladder cancer; （ii） 42 patients with urinary benign conditions; and （iii） 50 healthy volunteers. BTA TRAK and NMP 22 kits were used to detect bladder cancer. Voided urine cytology was used to compare the sensitivity and specificity of the screening tests. Results： The sensitivity and specificity of cytology, BTA TRAK and NMP 22 were 24% and 97%, 51% and 73%, 78% and 73%, respectively. The level of NMP 22 increased with tumor grading. The BTA TRAK kit has the lowest sensitivity among the screening tests. The NMP 22 with the best sensitivity can be an adjunct to cytology for evaluating bladder cancer. Conclusion： The NMP 22 test has a better correlation with the grading of the bladder cancer than BTA TRAK. As cytology units are typically not available in hospitals or in outpatient clinics, NMP 22 might be a promising tool for screening bladder cancer.

Primary Urethral Malignant Melanoma in an Elder Woman Diagnosed by Urine Cytology

Primary urethral malignant melanoma is an extremely rare malignancy with early metastasis and often delayed diagnosis, resulted in poor prognosis. Literature on this entity, especially with regard to urinary cytology is limited. We report here an extremely rare case of primary urethral malignant melanoma (MM) developed in a 95-year-old Japanese woman. A urinalysis showed macrophages containing brown granular pigments, and the urinary cytology was positive for atypical cells containing brown pigments suggestive of MM. Pathological examination of the biopsy specimen from the tumor revealed MM of the urethra. Immunohistochemically, the tumor cells showed strong cytoplasmic reactivities against antibodies, HMB-45 and Malan-A. Melanoma cells were immunohistochemically positive for c-Kit in their cytoplasm and cell membrane, although mutations in BRAF V600E and N/K-RAS were not detected. The urinary cytology and immunohistochemistry were useful for the rapid and accurate diagnosis for the urethral MM.

Diagnosis of pancreatic cancer by cytology and telomerase activity in exfoliated cells obtained by pancreatic duct brushing during endoscopy

BACKGROUND: Telomerase activity is reported to be specific and frequent in human pancreatic cancer. We conducted this study to assess the usefulness of monitoring telomerase activity in exfoliated cells obtained by pancreatic duct brushing during endoscopic retrograde cholangiopancreatography (ERCP) for the diagnosis of pancreatic cancer. METHODS: Exfoliated cells obtained by pancreatic duct brushing during ERCP from 21 patients (18 with pancreatic cancer, 3 with chronic pancreatitis) were examined. Telomerase activity was detected by polymerase chain reaction and telomeric repeat amplification protocol assay (PCR-TRAP-ELISA). RESULTS: D450 values of telomerase activity were 0.446 +/- 0.2700 in pancreatic cancer and 0.041 +/- 0.0111 in chronic pancreatitis. 77.8% (14/18) of patients with pancreatic cancer had cells with telomerase activity. None of the samples from patients with chronic pancreatitis showed telomerase activity, when the cutoff value of telomerase activity was set at 2.0. Cytological examination showed cancer cells in 66.7% (12/18) of the patients. CONCLUSIONS: Telomerase activity may be an early malignant event in pancreatic cancer development. Cytology and telomerase activity in cells obtained by pancreatic duct brushing may complement each other for the diagnosis of pancreatic cancer.

Genomic characterization of peritoneal lavage cytology-positive gastric cancer

Objective: Positive peritoneal lavege cytology(CY1) gastric cancer is featured by dismal prognosis, with high risks of peritoneal metastasis. However, there is a lack of evidence on pathogenic mechanism and signature of CY1and there is a continuous debate on CY1 therapy. Therefore, exploring the mechanism of CY1 is crucial for treatment strategies and targets for CY1 gastric cancer.Methods: In order to figure out specific driver genes and marker genes of CY1 gastric cancer, and ultimately offer clues for potential marker and risk assessment of CY1, 17 cytology-positive gastric cancer patients and 31matched cytology-negative gastric cancer patients were enrolled in this study. The enrollment criteria were based on the results of diagnostic laparoscopy staging and cytology inspection of exfoliated cells. Whole exome sequencing was then performed on tumor samples to evaluate genomic characterization of cytology-positive gastric cancer.Results: Least absolute shrinkage and selection operator(LASSO) algorithm identified 43 cytology-positive marker genes, while Mut Sig CV identified 42 cytology-positive specific driver genes. CD3G and CDKL2 were both driver and marker genes of CY1. Regarding mutational signatures, driver gene mutation and tumor subclone architecture, no significant differences were observed between CY1 and negative peritoneal lavege cytology(CY0).Conclusions: There might not be distinct differences between CY1 and CY0, and CY1 might represent the progression of CY0 gastric cancer rather than constituting an independent subtype. This genomic analysis will thus provide key molecular insights into CY1, which may have a direct effect on treatment recommendations for CY1and CY0 patients, and provides opportunities for genome-guided clinical trials and drug development.

Learning Discriminatory Information for Object Detection on Urine Sediment Image

In clinical practice,the microscopic examination of urine sediment is considered an important in vitro examination with many broad applications.Measuring the amount of each type of urine sediment allows for screening,diagnosis and evaluation of kidney and urinary tract disease,providing insight into the specific type and severity.However,manual urine sediment examination is labor-intensive,time-consuming,and subjective.Traditional machine learning based object detection methods require hand-crafted features for localization and classification,which have poor generalization capabilities and are difficult to quickly and accurately detect the number of urine sediments.Deep learning based object detection methods have the potential to address the challenges mentioned above,but these methods require access to large urine sediment image datasets.Unfortunately,only a limited number of publicly available urine sediment datasets are currently available.To alleviate the lack of urine sediment datasets in medical image analysis,we propose a new dataset named UriSed2K,which contains 2465 high-quality images annotated with expert guidance.Two main challenges are associated with our dataset:a large number of small objects and the occlusion between these small objects.Our manuscript focuses on applying deep learning object detection methods to the urine sediment dataset and addressing the challenges presented by this dataset.Specifically,our goal is to improve the accuracy and efficiency of the detection algorithm and,in doing so,provide medical professionals with an automatic detector that saves time and effort.We propose an improved lightweight one-stage object detection algorithm called Discriminatory-YOLO.The proposed algorithm comprises a local context attention module and a global background suppression module,which aid the detector in distinguishing urine sediment features in the image.The local context attention module captures context information beyond the object region,while the global background suppression module emphasizes objects in uninformative backgrounds.We comprehensively evaluate our method on the UriSed2K dataset,which includes seven categories of urine sediments,such as erythrocytes(red blood cells),leukocytes(white blood cells),epithelial cells,crystals,mycetes,broken erythrocytes,and broken leukocytes,achieving the best average precision(AP)of 95.3%while taking only 10 ms per image.The source code and dataset are available at https://github.com/binghuiwu98/discriminatoryyolov5.

Measurement of very low-molecular weight metabolites by traveling wave ion mobility and its use in human urine samples

The collision cross-sections(CCS)measurement using ion mobility spectrometry(IMS)in combination with mass spectrometry(MS)offers a great opportunity to increase confidence in metabolite identification.However,owing to the lack of sensitivity and resolution,IMS has an analytical challenge in studying the CCS values of very low-molecular-weight metabolites(VLMs250 Da).Here,we describe an analytical method using ultrahigh-performance liquid chromatography(UPLC)coupled to a traveling wave ion mobility-quadrupole-time-of-flight mass spectrometer optimized for the measurement of VLMs in human urine samples.The experimental CCS values,along with mass spectral properties,were reported for the 174 metabolites.The experimental data included the mass-to-charge ratio(m/z),retention time(RT),tandem MS(MS/MS)spectra,and CCS values.Among the studied metabolites,263 traveling wave ion mobility spectrometry(TWIMS)-derived CCS values(TWCCSN2)were reported for the first time,and more than 70%of these were CCS values of VLMs.The TWCCSN2 values were highly repeatable,with inter-day variations of<1%relative standard deviation(RSD).The developed method revealed excellent TWCCSN2 accuracy with a CCS difference(DCCS)within±2%of the reported drift tube IMS(DTIMS)and TWIMS CCS values.The complexity of the urine matrix did not affect the precision of the method,as evidenced by DCCS within±1.92%.According to the Metabolomics Standards Initiative,55 urinary metabolites were identified with a confidence level of 1.Among these 55 metabolites,53(96%)were VLMs.The larger number of confirmed compounds found in this study was a result of the addition of TWCCSN2 values,which clearly increased metabolite identification confidence.

Landscape of urine biomarkers for bladder cancer:molecular function,cell-of-origin,and bibliometric trend

Background:The effective management of bladder cancer(BCa)depends on the early diagnosis and surveillance.Previous studies have explored numerous urinary molecules as potential biomarkers of BCa.However,the molecular functions and cell-of-origin profiles of these biomarkers are yet to be elucidated.In this study,we aimed to provide a comprehensive overview of the landscape of urinary biomarker genes for BCa.Methods:We conducted an exhaustive literature search in PubMed,through which 555 biomarker genes were identified.We then analyzed the BCa single-cell atlas to infer the cellular origin of these BCa urine biomarker genes and performed functional enrichment analysis to gain insights into the functional molecular implications of these biomarkers.Results:These genes are involved in tumor proliferation,angiogenesis,cellmigration,and cell death and are predominantly expressed in epithelial and stromal cells.Interestingly,our analysis ofmultiomics tumor data revealed a discordance between tissue and urine in terms of differential methylation and RNA expression,suggesting that biomarker discovery for liquid biopsies should ideally begin with the analysis of bodily fluids rather than relying interest and that test strategies incorporating multiple molecular markers represent an ongoing trend.Conclusions:Collectively,our study has built a landscape of BCa urine biomarker genes,uncovered molecular insights into these biomarkers,and revealed the bibliometric trends in this field,which will contribute to the discovery of novel biomarkers in the future.

Impact of hyperthermic intraperitoneal chemotherapy on gastric cancer survival: Peritoneal metastasis and cytology perspectives

BACKGROUND Gastric cancer presenting with peritoneal metastasis is notably associated with diminished survival prospects.The use of cytoreductive surgery in conjunction with hyperthermic intraperitoneal chemotherapy(HIPEC)has been shown to increase survival rates in these patients.Despite these advancements,debates persist regarding the magnitude of survival improvement attributed to this treatment modality.The present investigation examined survival outcomes following HIPEC in individuals diagnosed with gastric cancer and peritoneal metastasis,and it took a comparative analysis of patients exhibiting positive and negative cytological findings.Between April 2013 and March 2020,84 patients with advanced gastric cancer treated at our institution were categorized into three cohorts:HIPEC(20 patients with peritoneal metastasis),cytology-positive(23 patients without peritoneal nodules but with positive wash cytology),and cytology-negative(41 patients with advanced gastric cancer,no peritoneal nodules,and negative wash cytology).The HIPEC cohort underwent gastrectomy with HIPEC,while the cytology-positive and cytology-negative groups received gastrectomy alone.The demographic,pat-hological,and survival data of the groups were compared.RESULTS The HIPEC cohort-predominantly younger females-exhibited relatively extended surgical durations and high blood loss.Nevertheless,the complication rates were consistent across all three groups.Median survival in the HIPEC group was 20.00±4.89 months,with 1-year,2-year,and 3-year overall survival rates of 73.90%,28.70%,and 9.60%,respectively.These figures paralleled the survival rates of the cytology-positive group(52.20%at 1 year,28.50%at 2 years,and 19.00%at 3 years).Notably,47%of patients experienced peritoneal recurrence.CONCLUSION HIPEC may offer a modest improvement in short-term survival for patients with gastric cancer and peritoneal metastasis,mirroring the outcomes in cytology-positive patients.However,peritoneal recurrence remained high.

A Case of Struma Ovarii Diagnosed by Cytology during Laparoscopic Surgery

We report a case of pure struma ovarii tumor diagnosed by cytology during laparoscopic surgery. The patient was a 34-year-old Japanese woman, gravida 1, para 1, who had the left adnexal mass, and was pre-operatively diagnosed as left ovarian endometriotic cyst or mature cystic teratoma by magnetic resonance imaging findings. She underwent laparoscopy, and the content of the left ovarian cystic tumor was found to be yellow gelatinous material, suggesting mature cystic teratoma. The imprint cytology of the tumor showed benign glandular pattern, suggesting struma ovarii. Histopathological findings led us to the diagnosis of pure struma ovarii with positive reactions for thyroglobulin and thyroid transcription factor-1. No metastases or disseminated lesions were detected. The patient has no recurrent signs 7 months after the operation.

A Case of Serous Tubal Intraepithelial Carcinoma Diagnosed by Cytology during the Operation

We experienced a case of serous tubal intraepithelial carcinoma (STIC), which is the earliest morphologically recognizable precursor of pelvic high grade serous carcinoma, diagnosed by cytology during the operation. A 48-year-old Japanese woman visited to our department because of abnormal cytological result and the left adnexal mass on female cancer screening. Pre-operatively, she was diagnosed as cervical intraepithelial neoplasia (CIN) 3, adenomyosis, and the left adnexal cystic lesion with papillary growth. At laparotomy, the left adnexal tumor turned out to be cystic fallopian tube with small papillary growth on the inner surface, but abnormal findings were not present on bilateral ovaries. The cytology of the imprint smears from the papillary projection and ascitic fluid showed positive, suggesting serous adenocarcinoma. Histopathological examination of the lesion revealed the left fallopian tube lesion was STIC with immunohistochemically positive reactivity against p53. No metastases including disseminated lesions were noted. The patient received four courses of systemic chemotherapy, and had no recurrent signs 10 months after the operation.

Effect of vaginal speculum lubrication on cervical cytology and discomfort during smear examination

AIM: To evaluate the role of lubricant gel in the cytology of a Pap smear and whether it results in an improvement in the discomfort experienced by women while undergoing Pap smear screening.METHODS: A total of 151 women were analyzed in the study. After screening for inclusion criteria, a Pap smear was taken with no lubricant in all the women and the discomfort experienced was rated on a visual analogue scale. The women underwent a second Pap smear on the next visit using a lubricant gel and were again rated on a visual analogue scale for the discomfort felt. The pathologist was blinded to the fact of whether the lubricating gel was used.RESULTS: The number of unsatisfactory smears in the no gel group was 3 vs 5 in the gel group, P < 0.05. However, a significant difference(P = 0.00) was observed in the visual analogue pain score in both groups, suggesting that application of lubricant gel over the speculum improves the pain experienced by women. CONCLUSION: Using a small amount of lubricant overthe speculum does not impair cervical cytology but significantly improves the discomfort experienced by women while undergoing a Pap smear.

影响Urine-H-11干化学法检测尿液分析的因素

Urine—H—11尿液试纸是干化学法检测尿液分析的一种方法，利用干化学法进行尿液分析，能够简便，快速为临床诊断、治疗、预防和监测疾病提供可靠的实验依据，但是，Urine—H—11干化学法也受许多因素的影响，而导致结果的准确性，为了使大家更多的了解影响Urine-H—11干化学法检测尿液的各项因素，准确的为临床提供诊断依据，现将有关Urine—H—11干化学法检测尿液的影响因素总结如下：

Detection of Neisseria Gonorrhoeae from Urine with Ligase Chain Reaction

Objective: To evaluate the value of ligase chainreaction(LCR) in the diagnosis of diplococcusgonorrhoeae in urine. Methods: LCR detection of the urine for Neisseriagonorrhoeae and bacteria culture of discharge was per-formed simultaneously to 276 patients with urethritisor cervicitis seeking treatment in sex transmitted dis-eases (STDs) outpatient clinic. For specimens withdiscordant results, polymerase chain reaction wasconducted. The purpose was to detect the respectivesensitivity and specificity of bacteria culture and LCR. Results: 24 of 276(8.7%) patients had positive LCRresults and 21 of 276(7.6%) were positive for culture.5 specimens had discordant results from LCR andbacteria culture. The sensitivity and specificity of LCRin the diagnosis of gonorrhoeae were 92.3% and100% respectively. Conclusion: This study showed that LCR had ahigher sensitivity and specificity for the diagnosis ofgonorrhoeae from urine.

Comparison of brush and basket cytology in differential diagnosis of bile duct stricture at endoscopic retrograde cholangiopancreatography

BACKGROUND： A previous report has identified a significantly higher sensitivity of cancer detection for dedicated grasping basket than brushing at endoscopic retrograde cholangiopancreato- graphy （ERCP）. This study aimed to compare the diagnostic accuracy of Geenen brush and Dormia basket cytology in the differential diagnosis of bile duct stricture. METHOD： The current study enrolled one hundred and fourteen patients who underwent ERCP with both Geenen brush and Dormia basket cytology for the differential diagnosis of bile duct stricture at our institution between January 2008 and December 2012. RESULTS： We adopted sequential performances of cytologic samplings by using initial Geenen brush and subsequent Dormia basket cytology in 59 patients and initial Dormia basket and subsequent Geenen brush cytology in 55 patients. Presampling balloon dilatations and biliary stentings for the stricture were performed in 17 （14.9%） and 107 patients （93.9%）, respectively. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of Geenen brush cytology for the diagnosis of malignant bile duct stricture were 75.0%, 100.0%, 100.0%, 66.7% and 83.3%, respectively, and those of Dormia basket cytology were 64.5%, 100.0%, 100.0%, 58.5% and 76.3%, respectively （P=0.347 and 0.827 for sensitivity and accuracy, respectively）. The good and excellent cellular yields （≥grade 2） were obtained by Geenen brush and Dormia basket cytology in 88 （77.2%） and 79 （69.3%） patients, respectively.CONCLUSION： The sensitivity, specificity and accuracy of biliary sampling with a Dormia basket are comparable to those with conventional Geenen brush cytology in the detection of malignant bile duct stricture.

Determination of urine catecholamines and metanephrines by reversed-phase liquid chromatography-tandem mass spectrometry

The measurement of urine catecholamine and metanephrine concentrations is important for biochemical screening and diagnosis of pheochromocytoma.The goal of this work was to develop a simple liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for determining catecholamines and metanephrines in urine to replace an existing liquid chromatographic method using electrochemical detection.Urine samples were prepared using Oasis weak-cation-exchange cartridges.The eluate was analyzed on an Agilent ZORBAX Eclipse Plus Phenyl-Hexyl column in 3 min.Adrenaline,noradrenaline,dopamine,metanephrine,normetanephrine,and their deuterated internal standards were monitored in positive electrospray ionization mode by multiple reaction monitoring(MRM).No evidence of ion suppression was observed.The assay was linear up to 5μmol/L for adrenaline,5μmol/L for noradrenaline,6.1μmol/L for dopamine,5.6μmol/L for metanephrine,and 34.6μmol/L for normetanephrine,with lower limits of quantification of 5,5,12,6 and 7nmol/L,respectively.The intra-day and inter-day precisions for all analytes ranged from 0.59%to 4.64%and1.98%to 4.80%,respectively.External quality assurance samples were assayed and showed excellent agreement with the target values.This simple method provides an improved assay for determining urine catecholamines and metanephrines.

A Study on Bisphenol A,Nonylphenol,and Octylphenol in Human Urine amples Detected by SPE-UPLC-MS

Objective To establish a comprehensive analytical method based on SPE‐UPLC‐MS for the simultaneous determination of bisphenol A （BPA）, nonylphenol （NP）, and octylphenol （OP） in urine samples. Methods Sixty urine samples collected from healthy subjects were analyzed for BPA, NP, and OP concentrations. The samples were de‐conjugated by adding β‐glucuronidase and sulfatase. After the enzymatic treatment, the samples were subjected to the OASIS HLB column solid phase extraction cartridges so as to be cleaned and concentrated. The UPLC separation was performed on a Acquity UPLCTM BEH C18 column （2.1×100 mm, 1.7 μm） with a gradient elution system of methanol‐water as the mobile phase. Triple‐quadrupole mass spectrometry analyzer was used for the qualitative and quantitative analysis of UPLC‐MS/MS system. Results The limit of detection of BPA, NP, and OP was 0.10, 0.10, and 0.15 ng/mL, respectively. The recoveries of BPA, NP and OP were 80.1%‐108%, 81.3%‐109%, and 81.5%‐98.7%, respectively. Among the 60 urine samples, BPA was detected in 8 samples at the level of 0.297‐32.7ng/mL, NP was detected in 29 samples at the level of 1.69‐27.8 ng/mL, and OP was detected in 17 samples at the level of 0.407‐11.1 ng/mL. Conclusion The method is simple with high sensitivity and selectivity, and is suitable for the determination of BPA, NP, and OP in urine. As shown by our analysis , BPA, NP, and OP appear to be prevalent in human urine. This is particularly true for NP. The results from our study is therefore valuable for future studies to assess the exposure to BPA, NP, and OP in the general population.

Simultaneous determination of fluorine and iodine in urine by ion chromatography with electrochemical pretreatment

A new method for the simultaneous determination of fluorine and iodine in urine by ion chromatography （IC） with electrochemical pretreatment has been developed. The pretreatment was performed in a novel electrochemical oxidationneutralization device （EOND）, in which iodide of the sample was oxidized to iodate and the alkaline digestion sample solution was neutralized. Under the optimized conditions, the limits of detection （LOD, S/N = 3） were 2.5 μg/L for fluoride and 20 μg/L for iodate, respectively. The recoveries were in the range of 93-102% for fluoride and 86-98% for iodate.

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.

Urea hydrolysis and recovery of nitrogen and phosphorous as MAP from stale human urine

Laboratory-scale tests for magnesium ammonium phosphate（MAP）precipitation following urea hydrolysis of human urine were conducted using orthogonal experiment design.The effects of initial pH,temperature and the volumetric ratios of stale urine to fresh urine,on urea hydrolysis in urine were studied to determine the final hydrolysis time to recover most nitrogen from separated human urine by MAP.With a volumetric ratio of stale to fresh urine>10% and at temperature≥20℃,urea hydrolysis could be completed in two days.Alkaline pH inhibited urea hydrolysis progress.The final pH values were all around 9.0 following urine hydrolysis,while the suspension pH might act as an indicator to detect the start and extent of urea hydrolysis.Over 95% of ammonium nitrogen and over 85% of phosphorus from hydrolyzed urine as MAP precipitate were obtained using MgCl;·6H;O and Na;HPO;·12H;O as precipitation agents at pH 8.5,molar ratio of Mg;:NH;-N:PO;-P at（1.2-1.3）:1:1,mixing speed of 120 r/min,and precipitation time and reaction time of 3 h and 15 min,respectively.The precipitate has a structure resembling pure MAP crystal.