Detection Of Bladder Cancer Research Articles

P133 - Bladder cancer detection in urine using DNA methylation markers: A prospective preclinical validation

P133 - Bladder cancer detection in urine using DNA methylation markers: A prospective preclinical validation

UBC®Rapid Is Sensitive in Detecting High-Grade Bladder Urothelial Carcinoma and Carcinoma in situ in Asian Population

Objectives: Bladder cancer is a common type of malignancy. UBC®Rapid is a novel immunoassay detecting urine cytokeratin 8/18 protein. Studies have shown good accuracy of UBC®Rapid in detecting bladder cancer. UBC®Rapid has however to date not been evaluated in Asian patients. We evaluated UBC®Rapid in detecting Asian bladder cancer, together with urine cytology. Methods: In total, 112 patients were recruited from National University Hospital Singapore and 103 patients were included in this study, comprising 49 patients with bladder urothelial carcinoma (UC), 21 patients with bladder benign lesions, and 33 patients with normal bladder. All the bladder cancer and benign lesions were confirmed by histology. Voided urine was collected for UBC®Rapid test. The results were compared with urine cytology. Results: The bladder UC group had remarkably higher UBC®Rapid value than the control groups. The sensitivity, specificity, positive, and negative predictive value of UBC®Rapid in detecting bladder UC were 53%, 85.5%, 76.5%, and 66.8%, respectively. Those of urine cytology were 40.8%, 96.3%, 90.9%, and 64.2%, respectively. Adding UBC®Rapid to urine cytology increased sensitivity to 57.1% but decreased specificity to 81.8%. UBC®Rapid was sensitive in detecting high-grade bladder UC (61.1%) and carcinoma in situ (CIS) (72.7%), as compared to urine cytology for bladder UC (55.6%) and CIS (54.5%), respectively. Conclusion: UBC®Rapid is sensitive in detecting high-grade bladder UC and CIS in Asian population. It may be useful as an adjunct test to achieve better detection of bladder cancer.

Prospective assessment of VI-RADS score in multiparametric MRI in bladder cancer: accuracy and the factors affecting the results.

PURPOSE We aimed to investigate the accuracy of Vesical Imaging - Reporting and Data System (VI-RADS) in the detection of muscle-invasive bladder cancer (MIBC) and to determine which factors affect the results of this scoring system. METHODS A prospective data analysis of 80 patients who were detected to have bladder tumor was performed between March 2019 and October 2020. VI-RADS scoring was used to determine the probability of muscle invasion. The scores were compared with pathological results to evaluate the accuracy of the VI-RADS scoring system. Interobserver agreement was assessed by VI-RADS scoring of 20 randomly chosen patients by a different experienced radiologist. RESULTS Using the VI-RADS scoring system, the sensitivity, specificity, positive predictive value, and negative predictive value of multiparametric magnetic resonance imaging (mpMRI) were 87.5%, 87.5%, 63.6%, and 96.6%, respectively. The interobserver agreement expressed as the interclass correlation coefficient (ICC) was 0.72 (95% CI: 0.44-0.84, P < .001). In addition, the flat appearance of the tumor was an important factor affecting the accuracy of the VI-RADS score (odds ratio: 5.3 [95% CI: 1.1-27.0] and relative risk: 1.87 [95% CI: 1.24-2.82]). CONCLUSION The mpMRI, used in conjunction with VI-RADS, has proven to be an effective imaging method for detecting muscle invasion in cases of bladder cancer. VI-RADS scoring system can distinguish whether there is a muscle-invasive and non-muscle invasive bladder cancer with acceptable accuracy. In addition, the flat appearance of the tumor is an important entity that can affect the accuracy of the VI-RADS scoring system.

Application of surface plasmon resonance imaging biosensors for determination of fibronectin, laminin-5 and type IV collagen in serum of transitional bladder cancer patients

The urothelial basement membrane (UBM) contains type IV collagen, laminin-5, and fibronectin. Urothelial neoplasm can break through the UBM underlying the urothelium to invade the lamina propria. Conceptually, all bladder cancer staging over Ta (T1–T4) may demonstrate disturbances in the UBM structure, as well as alterations in the serum concentrations of the studied components. The aim of this study was to determine the blood serum concentration of collagen IV, laminin-5 and fibronectin in bladder cancer patients. Quantification of their concentrations and correlation with various clinicopathological parameters may be useful for making more accurate predictions and identifying high-risk patients. The study included 96 patients with bladder cancer confirmed by transurethral resection or cystectomy and 26 patients with diagnosed cystitis chronica or BPH (benign prostate hyperplasia). Collagen IV, laminin-5 and fibronectin were detected using Surface Plasmon Resonance Imaging biosensors. Significant differences in blood serum concentrations of the studied biomarkers were observed between the control group and bladder cancer patients, as well as between nonmuscle-invasive and muscle-invasive groups. ROC analysis gave satisfactory results for differentiation between the control group and bladder cancer group (AUC 0.92–0.99), with lower values only for collagen IV between nonmuscle-invasive and muscle-invasive patients (AUC 0.71), and a statistically insignificant difference for laminin-5. Laminin-5 concentration was more closely correlated to tumour grade, size and recurrence rate; fibronectin to tumour stage, size and morphology; and collagen IV to tumour stage, grade and recurrence rate. The relations between serum concentrations of the presented biomarkers of the urothelial basement membrane may be useful for bladder cancer detection and for determination of the tumour stage, hence simplifying the making of therapeutic decisions.

Confocal Laser Endomicroscopy for Bladder Cancer Detection: Where Do We Stand?

Introduction: Confocal laser endomicroscopy (CLE) is a relatively new technology that allows for a real-time in situ microscopic characterization of tissue lesions, being able to discriminate between low- and high-grade ones. After a first period of slow diffusion caused by technological limitations and elevated costs, CLE applications are rapidly spreading in different branches of medicine, and there is mounting evidence of its advantages for the management of different tumors such as bladder cancer (BCa), from both a diagnostic and a clinical point of view. In this systematic review (SR), we evaluate the state-of-the-art CLE for BCa management. Material and methods: We performed an SR and quality assessment analysis of the current literature in this regard following the PRISMA guidelines. All data were independently verified by two different authors and discrepancies were solved by a third author. Moreover, a quality-assessment analysis according to QUADAS-2 criteria was performed to evaluate the studies selected for SR. Results: A total of 158 articles were retrieved; of which 79 were rejected and 38 were removed as duplicates. After article selection, seven prospective studies were assessed for data extraction. These accounted for 214 patients overall, with a correspondence rate between CLE and histopathological examination ranging from 54.6 to 93.6%. Regarding quality assessment, three out five prospective studies have at least a high risk of bias in one QUADAS-2 domain, whereas the applicability always has a low risk of bias. Conclusion: Despite actual technical limitations, the preliminary results of this appealing technology are encouraging and should prompt further investigations.

Multicentric validation of diagnostic tests based on BC-116 and BC-106 urine peptide biomarkers for bladder cancer in two prospective cohorts of patients.

Non-invasive urine-based biomarkers can potentially improve current diagnostic and monitoring protocols for bladder cancer (BC). Here we assess the performance of earlier published biomarker panels for BC detection (BC-116) and monitoring of recurrence (BC-106) in combination with cytology, in two prospectively collected patient cohorts. Of the 602 patients screened for BC, 551 were found eligible. For the primary setting, 73 patients diagnosed with primary BC (n = 27) and benign urological disorders, including patients with macroscopic haematuria, cystitis and/or nephrolithiasis (n = 46) were included. In total, 478 patients under surveillance were additionally considered (83 BC recurrences; 395 negative for recurrence). Urine samples were analysed with capillary electrophoresis-mass spectrometry. The biomarker score was estimated via support vector machine-based software. Validation of BC-116 biomarker panel resulted in 89% sensitivity and 67% specificity (AUCBC-116 = 0.82). A diagnostic score based on cytology and BC-116 resulted in good (AUCNom116 = 0.85) but not significantly better performance (P = 0.5672). A diagnostic score including BC-106 and cytology was evaluated (AUCNom106 = 0.82), significantly outperforming both cytology (AUCcyt = 0.72; P = 0.0022) and BC-106 (AUCBC-106 = 0.67; P = 0.0012). BC-116 biomarker panel is a useful test for detecting primary BC. BC-106 classifier integrated with cytology showing >95% negative predictive value, might be useful for decreasing the number of cystoscopies during surveillance.

Clinical utility of vesical imaging-reporting and data system (VI-RADS) in non-muscle invasive bladder cancer (NMIBC) patients candidate for en-bloc transurethral resection: A prospective study.

To assess accuracy of vesical imaging-reporting and data system (VI-RADS) 5-point score in detection of muscle invasive bladder cancer and avoiding second look transurethral resection of the tumors (TURBT). Additionally, to assess safety and efficacy of bipolar en-block transurethral urethral resection of bladder tumor. Patients with bladder mass up to 5 cm were included in the study. VI-RADS 5-point score was done preoperative for all cases and postoperatively before second look TURBT. Patients were followed up for 12 months. In all, 80 cases were eligible for the study. Preoperative VI-RADS score at cutoff of 3 had sensitivity of 89.3 %, specificity 83.3 %, postive predective value (PPV) 92.6 %, negative predictive value (NPV) 76.9 %, accuracy of 87.5 %, while at cutoff 2 sensitivity was 82.1%, specificity 91.7%, PPV 95.8%, NPV 68.8%, accuracy of 85.0%. Operative time 28.8 ± 9.4 minutes, hemoglobin drop 0.3 ± 0.05 g/dl, catheterization time 2.8 ± 0.8 days, hospital stay 1.4 ± 0.4 days. No complications occurred. Recurrence in field of resection 3.75%. Detrusor muscle was available in 76 cases (95%). Postoperative VI-RADS score at cutoff of 3 had sensitivity of 78.6%, specificity 77.8%, PPV 84.6%, NPV 70.0%, accuracy of 78.3%. At cutoff 2 VI-RADS score sensitivity was 71.4%, specificity 77.8%, PPV 83.3%, NPV 63.6%, accuracy of 73.9%. VI-RADS 5-point score showed high sensitivity and specificity in preoperative discrimination of non-muscle invasive bladder cancer (NMIBC) from muscle invasive bladder cancer cases and in avoiding unnecessary second look TURBT. Bipolar en-block TURBT technique is both safe and efficacious in resecting NMIBC cases with low recurrence rate.

Urine exosomes as biomarkers in bladder cancer diagnosis and prognosis: From functional roles to clinical significance.

Bladder cancer is one of the top ten most common cancers and top ten causes of cancer death globally. 5-year survival rates have decreased in Australia from 66% to 55% in the past three decades. The current gold standard for diagnosis is cystoscopy. However, cystoscopies are an invasive and health-resource intensive procedure which has sub-optimal sensitivity for flat lesions such as CIS (carcinoma in situ) and low specificity for differentiating inflammation from cancer - hence requiring biopsies under anesthesia. Frequent and life-long surveillance cystoscopy is required for most patients since there are high rates of progression and local recurrence in high-risk non-muscle invasive cancer (NMIBC) as well as poor outcomes associated with delayed detection of muscle-invasive bladder cancer (MIBC). There is an unmet need for a non-invasive test to provide better discrimination and risk-stratification of bladder cancer which could aid clinicians by improving patient selection for cystoscopy; enhanced risk stratification methods may guide the frequency of surveillance cystoscopies and inform treatment choices. Exosomes, which are nano-sized extracellular vesicles containing genetic material and proteins, have been shown to have functional roles in the development and progression of bladder cancer. Exosomes have also been demonstrated to be a robust source of potential biomarkers for bladder cancer diagnosis and prognosis and may also have roles as therapeutic agents. In this review, we summarize the latest evidence of biological roles of exosomes in bladder cancer and highlight their clinical significance in bladder cancer diagnosis, surveillance and treatment.

BladMetrix: a novel urine DNA methylation test with high accuracy for detection of bladder cancer in hematuria patients

BackgroundCystoscopy is the gold standard for bladder cancer detection, but is costly, invasive and has imperfect diagnostic accuracy. We aimed to identify novel and accurate DNA methylation biomarkers for non-invasive detection of bladder cancer in urine, with the potential to reduce the number of cystoscopies among hematuria patients.ResultsBiomarker candidates (n = 32) were identified from methylome sequencing of urological cancer cell lines (n = 16) and subjected to targeted methylation analysis in tissue samples (n = 60). The most promising biomarkers (n = 8) were combined into a panel named BladMetrix. The performance of BladMetrix in urine was assessed in a discovery series (n = 112), consisting of bladder cancer patients, patients with other urological cancers and healthy individuals, resulting in 95.7% sensitivity and 94.7% specificity. BladMetrix was furthermore evaluated in an independent prospective and blinded series of urine from patients with gross hematuria (n = 273), achieving 92.1% sensitivity, 93.3% specificity and a negative predictive value of 98.1%, with the potential to reduce the number of cystoscopies by 56.4%.ConclusionsWe here present BladMetrix, a novel DNA methylation urine test for non-invasive detection of bladder cancer, with high accuracy across tumor grades and stages, and the ability to spare a significant number of cystoscopies among patients with gross hematuria.

Untargeted ultra-high-resolution mass spectrometry metabolomic profiling of blood serum in bladder cancer

Bladder cancer (BC) is a common urological cancer of high mortality and recurrence rates. Currently, cystoscopy is performed as standard examination for the diagnosis and subsequent monitoring for recurrence of the patients. Frequent expensive and invasive procedures may deterrent patients from regular follow-up screening, therefore it is important to look for new non-invasive methods to aid in the detection of recurrent and/or primary BC. In this study, ultra-high-performance liquid chromatography coupled with ultra-high-resolution mass spectrometry was employed for non-targeted metabolomic profiling of 200 human serum samples to identify biochemical signatures that differentiate BC from non-cancer controls (NCs). Univariate and multivariate statistical analyses with external validation revealed twenty-seven metabolites that differentiate between BC patients from NCs. Abundances of these metabolites displayed statistically significant differences in two independent training and validation sets. Twenty-three serum metabolites were also found to be distinguishing between low- and high-grade of BC patients and controls. Thirty-seven serum metabolites were found to differentiate between different stages of BC. The results suggest that measurement of serum metabolites may provide more facile and less invasive diagnostic methodology for detection of bladder cancer and recurrent disease management.

Detecting and monitoring bladder cancer with exfoliated cells in urine.

Current methods for the diagnosis and monitoring of bladder cancer are invasive and have suboptimal sensitivity. Liquid biopsy as a non-invasive approach has been capturing attentions recently. To explore the ability of urine-based liquid biopsy in detecting and monitoring genitourinary tumors, we developed a method based on promoter-targeted DNA methylation of urine sediment DNA. We used samples from a primary bladder cancer cohort (n=40) and a healthy cohort (n=40) to train a model and obtained an integrated area under the curve (AUC) > 0.96 in the 10-fold cross-validation, which demonstrated the ability of our method for detecting bladder cancer from the healthy. We next validated the model with samples from a recurrent cohort (n=21) and a non-recurrent cohort (n=19) and obtained an AUC > 0.91, which demonstrated the ability of our model in monitoring the progress of bladder cancer. Moreover, 80% (4/5) of samples from patients with benign urothelial diseases had been considered to be healthy sample rather than cancer sample, preliminarily demonstrating the potential of distinguishing benign urothelial diseases from cancer. Further analysis basing on multiple-time point sampling revealed that the cancer signal in 80% (4/5) patients had decreased as expected when they achieved the recurrent-free state. All the results suggested that our method is a promising approach for noninvasive detection and prognostic monitoring of bladder cancer.

Efficacy of Voided Urine Cytology Compared to Cystoscopic Findings in the Detection of Bladder Cancer- A Pakistani Perspective

Objective: To compare the efficacy of voided urine cytology with findings of cystoscopy and histopathology of biopsy specimens in the diagnosis of bladder cancer.&#x0D; Study Design: Comparative cross-sectional study&#x0D; Place and Duration of Study: Armed forces Institute of Urology (AFIU), Rawalpindi Pakistan from Jan 2019 to Jan 2020.&#x0D; Methodology: All patients presenting to the urology clinic with complaints of haematuria, visible and non-visible, and any radiologic evidence of bladder growth were included in the study after informed consent. Urine cytology was performed for all patients, followed by cystoscopy under anaesthesia, transurethral resection was conducted, and biopsy was taken where needed.&#x0D; Results: 170 patients were included in the study134 (78.8%) were males, while 36 (21.2%) were females. The mean age was 54 ± 9.47 years (range 36 to 73 years). The overall sensitivity of voided urine cytology was 46.7%, while specificity was 79.2%. The positive predictive value was 85.1%, and the negative predictive value was 36.9%.&#x0D; Conclusion: Bladder cancer is a disease which demands an early diagnosis, prompt treatment, and long-term follow-up. Cystoscopy remains the gold standard for this purpose; urine cytology can be used as a supplement as it is non-invasive, more specific and cost-effective.

Targeted elastin-like polypeptide fusion protein for near-infrared imaging of human and canine urothelial carcinoma.

Cystoscopic visualization of bladder cancer is an essential method for initial bladder cancer detection and diagnosis, transurethral resection, and monitoring for recurrence. We sought to develop a new intravesical imaging agent that is more specific and sensitive using a polypeptide based NIR (near-infrared) probe designed to detect cells bearing epidermal growth factor receptors (EGFR) that are overexpressed in 80% of urothelial carcinoma (UC) cases. The NIR imaging agent consisted of an elastin like polypeptide (ELP) fused with epidermal growth factor (EGF) and conjugated to Cy5.5 to give Cy5.5-N24-EGF as a NIR contrast agent. In addition to evaluation in human cells and tissues, the agent was tested in canine cell lines and tissue samples with naturally occurring invasive UC. Flow cytometry and confocal microscopy were used to test cell-associated fluorescence of the probe in T24 human UC cells, and in K9TCC-SH (high EGFR expression) and K9TCC-Original (low EGF expression) canine cell lines. The probe specifically engages these cells through EGFR within 15 min of incubation and reached saturation within a clinically relevant 1 h timeframe. Furthermore, ex vivo studies with resected canine and human bladder tissues showed minimal signal from normal adjacent tissue and significant NIR fluorescence labeling of tumor tissue, in good agreement with our in vitro findings. Differential expression of EGFR ex vivo was revealed by our probe and confirmed by anti-EGFR immunohistochemical staining. Taken together, our data suggests Cy5.5-ELP-EGF is a NIR probe with improved sensitivity and selectivity towards BC that shows excellent potential for clinical translation.

Metabolomic and elemental profiling of blood serum in bladder cancer

Bladder cancer (BC) is one of the most frequently diagnosed types of urinary cancer. Despite advances in treatment methods, no specific biomarkers are currently in use. Targeted and untargeted profiling of metabolites and elements of human blood serum from 100 BC patients and the same number of normal controls (NCs), with external validation, was attempted using three analytical methods, i.e., nuclear magnetic resonance, gold and silver-109 nanoparticle-based laser desorption/ionization mass spectrometry (LDI-MS), and inductively coupled plasma optical emission spectrometry (ICP-OES). All results were subjected to multivariate statistical analysis. Four potential serum biomarkers of BC, namely, isobutyrate, pyroglutamate, choline, and acetate, were quantified with proton nuclear magnetic resonance, which had excellent predictive ability as judged by the area under the curve (AUC) value of 0.999. Two elements, Li and Fe, were also found to distinguish between cancer and control samples, as judged from ICP-OES data and AUC of 0.807 (in validation set). Twenty-five putatively identified compounds, mostly related to glycans and lipids, differentiated BC from NCs, as detected using LDI-MS. Five serum metabolites were found to discriminate between tumor grades and nine metabolites between tumor stages. The results from three different analytical platforms demonstrate that the identified distinct serum metabolites and metal elements have potential to be used for noninvasive detection, staging, and grading of BC.

Analysis of the Relationship between Bladder Cancer Gene Mutation and Clinical Prognosis by High-Throughput Sequencing.

Bladder cancer is one of the most common malignant tumors in urology in China. The analysis of gene mutation in bladder cancer and its relationship with clinical characteristics and prognosis will provide a basis for accurate treatment of bladder cancer. The aim of this study was to analyze the mutations and functional regions of bladder cancer-related genes based on high-throughput sequencing, and to explore the relationship between mutations and clinicopathological features, as well as its prognosis and clinical implication. From April 2020 to October 2020, a total of 47 patients with bladder cancer in the Department of Urology, Affiliated Hospital of Chengde Medical College were studied. Gene sequencing was performed using Nextseq CN500 System, a high-throughput sequencing platform. The results of gene detection were described, and the relationship and clinical value of high frequency mutated genes with clinicopathological features and prognosis were systematically analyzed. A total of 29 mutation genes, 61 exons, and 95 mutation sites were identified in this study. The frequencies of TP53, FGFR3, PIK3CA, ERBB2, MUC4, and KRAS mutation are relatively high, accounting for 59.92 % of the total mutation frequency. The TP53 was significantly associated with muscle invasive bladder cancer, T2 stage, and progression-free survival, while FGFR3 was significantly associated with non-muscle invasive bladder cancer and T1 stage. High-throughput sequencing technology provides a successful approach for detecting bladder cancer gene mutations. The TP53, FGFR3, PIK3CA, ERBB2, MUC4, and KRAS genes have high mutation frequencies in bladder cancer patients. The TP53, FGFR3 and PIK3CA genes may play a predictive role in the prognosis of bladder cancer, which may hold certain guiding significance for in-depth study of the pathogenesis of bladder cancer and the development of targeted therapies.

Role of multiparametric magnetic resonance imaging in the diagnosis and staging of urinary bladder cancer.

ObjectivesTo assess the role of multiparametric magnetic resonance imaging (mp-MRI) in the diagnosis and staging of urinary bladder cancer (BC).Materials and methodsFifty patients diagnosed with bladder masses underwent mp-MRI study. The results of 3 image sets were analyzed and compared with the histopathological results as a reference standard: T2-weighted image (T2WI) plus dynamic contrast-enhanced (DCE), T2WI plus diffusion-weighted images (DWI), and mp-MRI, including T2WI plus DWI and DCE. The diagnostic accuracy of mp-MRI was evaluated using receiver operating characteristic curve analysis.ResultsThe accuracy of T2WI plus DCE for detecting muscle invasion of BC was 79.5% with a fair agreement with histopathological examination (κ = 0.59); this percentage increased up to 88.6% using T2WI plus DWI, with good agreement with histopathological examination (κ = 0.74), whereas mp-MRI had the highest overall accuracy (95.4%) and excellent agreement with histopathological data (κ = 0.83). Multiparametric MRI can differentiate between low- and high-grade bladder tumors with a high sensitivity and specificity of 93.3% and 98.3%, respectively.ConclusionsMultiparametric MRI is an acceptable method for the preoperative detection and accurate staging of BC, with reasonable accuracy in differentiating between low- and high-grade BC.

The use of blue-light cystoscopy in the detection and surveillance of nonmuscle invasive bladder cancer.

Nonmuscle invasive bladder cancer is associated with a high risk of recurrence as well as progression to muscle-invasive disease. Therefore, adequate visualization and identification of malignant lesions as well as complete resection are critical. Traditional white-light cystoscopy is limited in its ability to detect bladder cancer, specifically carcinoma in situ. Blue-light cystoscopy makes use of the intravesical instillation of a heme precursor to differentiate areas of malignancy from normal tissue. A narrative review of the literature on the use of blue-light cystoscopy in bladder cancer was conducted. Blue-light cystoscopy has been shown in several randomized clinical trials to increase detection of Ta, T1, and carcinoma in situ, as well as reduce risk of recurrence at 12 months as compared with traditional white-light cystoscopy. Research into the effects of blue-light cystoscopy on risk of disease progression has produced mixed results, in part due to changing definitions of progression. However, more recent research suggests a correlation with decreased risk of progression. Whereas the use of blue-light was initially limited to rigid cystoscopy in the operating room, results from a recent randomized clinical trial showing enhanced detection of recurrent disease using blue-light in-office surveillance flexible cystoscopy have led to expanded Food and Drug Administration approval. Overall, blue-light cystoscopy offers promise as an enhancement to white-light cystoscopy for the detection of nonmuscle invasive bladder cancer and may yield additional benefits in reducing disease recurrence and progression. Further prospective research is needed to evaluate the true benefit of blue-light cystoscopy in terms of disease progression as well as the cost-effectiveness of this technique.

Glycosylated phospholipid-coated upconversion nanoparticles for bioimaging of non-muscle invasive bladder cancers.

Detection of non-muscle invasive bladder cancer (NMIBC) is crucial to facilitate complete tumor resection, thus improving the survival rate as well as reducing the recurrence frequency and treatment expense. Fluorescence imaging cystoscopy is an effective method for the detection of NMIBC. However, its application is limited as the commonly applied fluorescent agents such as dyes and photosensitizers usually lack specific tumor accumulation and are vulnerable to photobleaching. Furthermore, the broad emission band of conventional fluorescent agents limits their imaging and detection efficacy. To overcome these limitations, upconversion nanoparticles (UCNPs) have been selected as the fluorescent agent, due to their resistance to photobleaching, less background auto-fluorescence, and narrow emission bands. In order to achieve active tumor targeting, the UCNPs are coated with a glycosylated phospholipid layer. The glycosylated phospholipid-coated UCNPs exhibited high selective accumulation in cancer cells over normal cells and enhanced the upconversion luminescence (UCL) (at 540nm and 660nm) from bladder cancer cells under 980nm laser irradiation. Glycosylated phospholipid coating that promotes uptake of UCNPs by cancer cells, and UCL emitted from UCNPs under NIR (980nm) laser irradiation for cancer cell imaging.

Self-calibration SERS sensor with “core-satellite” structure for detection of hyaluronidase activity

Hyaluronidase expression is known to be upregulated in various pathological conditions. A method based on a combination of ratiometric surface-enhanced Raman scattering (SERS) and magnetic separation is described for the determination of hyaluronidase (HAase) activity. Gold nanospheres (AuNPs) functionalized by 4-mercaptophenylboronic acid (4-MPBA) form stable cyclic esters with diol on hyaluronic acid (HA) by the boronic acid group, while Fe3O4@DTNB@Au modified with mercaptoethylamine (MEA) was used as a capture substrate to bind to the carboxyl group on the surface of HA, forming the “Au@4-MPBA@HA/Fe3O4@DTNB@Au@MEA” “core-satellite” structure. When HAase is present, HA is enzymatically disrupted, resulting in the destruction of the “core-satellite” structure, the SERS signal of 4-MPBA is subsequently weakened. The gold shell in the substrate protects the 5,5′-Dithio bis-(2-nitrobenzoic acid) (DTNB) from the external environment, which makes it become an ideal internal standard (IS) molecule for subsequent calibration. Under optimal conditions, the I1075/I1324 varied in the range of 10−3 - 10 U‧mL−1 HAase activity, with a limit of detection (LOD) of 0.32 mU‧mL−1，below the level of HAase in normal human body fluids. This method has been successfully applied to the determination of HAase activity in urine and is expected to provide a new method in disease detection, especially in the non-invasive detection of bladder cancer.

Label-free observation of morphological alteration of irradiated-urothelial bladder carcinoma cells through digital holographic microscopy

Radiotherapy is an effective treatment for certain patients with muscle-invasive bladder cancer and radio-sensitivity detection plays a vital role during bladder cancer radio treatment because radiotherapy responses have profound influences on a patient’s prognosis. Although several potential biomarkers were investigated to assess the radio-response of bladder cancer, studies on detecting radio-sensitivity based on morphological characteristics of cancer cells at the single-cell level are rare. In fact, morphological parameters are vital characteristics of cells that could provide direct information to infer the physiological statuses of cells and evaluate the response of cells to the external stimulations. In this study, digital holographic microscopy was applied to quantify morphological parameters of bladder cancer cells (HT-1376) at the single-cell level and their alterations after exposure to four different radiation doses, i.e., 0 Gy (control), 4, 8, and 12 Gy. Based on the reconstructed phase images, four morphological parameters of cells, namely, cell phase volume (CPV), cell projected area (CPA), cell average phase height (CAPH), and cell maximum phase height (CMPH), were quantitatively calculated. The results show that the change rates of CPV, CAPH, and CMPH were increased with the radiation dose rising, while the change rate of CPA was decreased with the radiation dose increasing. Moreover, the change rates of CPV, CPA, CAPH, and CMPH were different between control group and 12 Gy treated group. The results demonstrate that morphological characteristics have the potential to be utilized to estimate the radio-sensitivity of bladder cancer cells, and it may provide new perspectives to establishing label-free methods to detect radio-sensitivity and guide radiotherapy in bladder cancer.