The influence of manual segmentation strategies and different phases selection on machine learning-based computed tomography in renal tumors: a systematic review and meta-analysis.

BACKGROUND: Delineating the region/volume of interest (ROI/VOI) and selecting the phases are of importance in developing machine learning (ML). The results will change when choosing different methods of drawing the ROI/VOI and selecting different phases. However, there is no related standard for delineating the ROI/VOI and selecting the phases in renal tumors to develop ML based on computed tomography (CT). METHODS: The PubMed and Web of Science were searched for related studies published until March 1, 2023. Inclusion criteria were studies that developed ML models in renal tumors from CT images. And the binary diagnostic accuracy data were extracted to obtain the outcomes, such as sensitivity (SE), specificity (SP), accuracy (ACC), and area under the curve (AUC). RESULTS: Twenty-three papers were included in the meta-analysis with a pooled SE of 87% (95% CI 85-88%), SP of 82% (95% CI 79-85%), and AUC of 91% (95% CI 89-93%) in phases; a pooled SE of 82% (95% CI 80-84%), SP of 85% (95% CI 83-86%), and AUC of 90% (95% CI 88-93%) in phases combined with delineating strategies, respectively. In all different combinations, the contour-focused and single phase produce the highest AUC of 93% (95% CI 90-95%). In subgroup analyses (sample size, year of publication, and geographical distribution), the performance was acceptable on phases and phases combined strategies. CONCLUSIONS: To explore the effect of manual segmentation strategies and different phases selection on ML-based CT, we find that the method of single phase (CMP or NP) combined with contour-focused was considered a better strategy compared to the other strategies.

Association between NDUFS1 from urinary extracellular vesicles and decreased differential renal function in children with ureteropelvic junction obstruction.

BACKGROUND: Ureteropelvic junction obstruction (UPJO) is the most common cause of pediatric congenital hydronephrosis, and continuous kidney function monitoring plays a role in guiding the treatment of UPJO. In this study, we aimed to explore the differentially expressed proteins (DEPs) in the urinary extracellular vesicles(uEVs) of children with UPJO and determine potential biomarkers of uEVs proteins that reflect kidney function changes. METHODS: Preoperative urine samples from 6 unilateral UPJO patients were collected and divided into two groups: differential renal function (DRF) ≥ 40% and DRF < 40%.We subsequently used data-independent acquisition (DIA) to identify and quantify uEVs proteins in urine, screened for DEPs between the two groups, and analyzed biofunctional enrichment information. The proteomic data were evaluated by Western blotting and enzyme-linked immunosorbent assay (ELISA) in a new UPJO testing cohort. RESULTS: After one-way ANOVA, a P adj value < 0.05 (P-value corrected by Benjamin-Hochberg) was taken, and the absolute value of the difference multiple was more than 1.5 as the screening basis for obtaining 334 DEPs. After analyzing the enrichment of the DEPs according to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment combined with the protein-protein interaction (PPI) network results, we selected nicotinamide adenine dinucleotide-ubiquinone oxidoreductase core subunit S1 (NDUFS1) for further detection. The expression of NDUFS1 in uEVs was significantly lower in patients with DRF < 40% (1.182 ± 0.437 vs. 1.818 ± 0.489, P < 0.05), and the expression level of NDUFS1 was correlated with the DRF in the affected kidney (r = 0.78, P < 0.05). However, the NDUFS1 concentration in intravesical urine was not necessarily related to the change in DRF (r = 0.28, P = 0.24). CONCLUSIONS: Reduced expression of NDUFS1 in uEVs might indicate the decline of DRF in children with UPJO.

The application of machine learning based on computed tomography images in the identification of renal tumors in children.

Background: The clinical manifestations of Wilms tumor and non-Wilms tumor in children are similar, and the only way to confirm the diagnosis is by postoperative pathology. Computed tomography (CT) is one of the main methods for preoperative diagnosis of the two, but it is also difficult to distinguish because it is easily affected by the subjective influence and the experience of the radiologists. Methods: The CT images of 82 children with renal tumors admitted to the Department of Pediatric Urology, Shandong Provincial Hospital from January 2011 to March 2022 were retrospectively analyzed. First, we drew the two-dimensional (2D) region of interest (ROI) of the largest cross-section on the corticomedullary phase (CMP) and nephrogenic phase (NP) images, and extracted seven types of 107 features in the ROI. Then, the texture features with similarity greater than 95% and repetition less than 90% were screened out, and the remaining texture features were further screened by analysis of variance (ANOVA) and recursive feature elimination (RFE). Finally, 15 texture feature were used to build the machine learning (ML) models. We used the synthetic minority oversampling technique (SMOTE) and 10-fold cross-validation to build ML models and verified them in the training, testing, and internal validation sets. The area under the receiver-operating characteristic curve (AUC) and calibration curve were used to evaluate the diagnostic performance. Results: We collected 77 CMP and 81 NP images, which were randomly divided into the training set and the testing set according to the ratio of 7:3. In the internal validation of CMP, the Mean-PCC-ANOVA-5-AE pipeline model achieved the highest AUC 0.792 [95% confidence interval (CI): 0.653-0.930], and its accuracy (ACC), sensitivity (SEN), and specificity (SPE) were 0.833, 0.539 and 0.927, respectively. Correspondingly, in NP, the Mean-PCC-ANOVA-2-LR pipeline model achieved the highest AUC 0.655 (95% CI: 0.485-0.82) in the internal validation. The ACC, SEN, and SPE were 0.696, 0.539, and 0.744, respectively. Conclusions: The ML models based on CT images have good diagnostic efficiency in differentiating Wilms tumors from non-Wilms tumors in children.

Widening of narrow urethral plates with lateral skin in TIP hypospadias repair: single center series.

BACKGROUND: To compare the outcomes of hypospadias repair using tubularized incised plate (TIP) urethroplasty and modified TIP with lateral skin to widen the urethral plate (WTIP). MATERIALS AND METHODS: Data were obtained from pre-pubertal boys who underwent primary hypospadias repair between May 2018 and July 2023. The cases were divided into two groups; one group underwent TIP with urethral plate ≥ 6 mm width and the other group with urethral plate width < 6 mm underwent WTIP. WTIP urethroplasty was performed by widening incisions on the outer margins of the urethral plate to incorporate penile and glandular skin lateral to the urethral plate to facilitate tubularization. Complication rates and urinary functions were compared. RESULTS: A total of 157 patients were enrolled in this study. Eighty-eight cases with narrow urethral plate were subjected to WTIP urethroplasty, and the rest were subjected to TIP urethroplasty. The preoperative glans width in WTIP group was less than that in TIP group (P < 0.001), and 44.3% had midshaft meatus in WTIP group compared to 17.4% in TIP group (P < 0.001). However, the incidences of postoperative complications (17.6% vs. 21.6%, P = 0.550) were not statistically different between the TIP and WTIP groups. In addition, both groups did not differ significantly in postoperative uroflowmetry assessment. CONCLUSIONS: The described technique helps to create an adequately caliber aesthetic neomeatus and facilitates tubularization, especially in hypospadias with a narrow urethral plate. Our data suggest that augmentation of a narrow urethral plate with WTIP has a similar surgical outcome to that of the TIP procedure in patients with a wide urethral plate.

The underlying molecular mechanism of ciliated epithelium dysfunction and TGF-ß signaling in children with congenital pulmonary airway malformations.

The aim of this study was to investigate the variation in gene expression in the complete transcripts of Congenitalpulmonary airwaymalformation (CPAM) of the lung using Next Generation Sequencing (NGS) technology. There were 20 cases involving children with CPAM were used for selection of study sample. NGS was used to establish RNA-Seq libraries for the two groups of samples separately, and both groups were conducted to differential expression analysis and Gene Ontology (GO) functional enrichment analysis. The pathways of the differential genes were analyzed to find the enriched target pathways. A total of 592 genes were expressed with significant differences (CPAM vs. normal tissue, P < 0.05). GO functional analysis of DEGs indicated that abnormal ciliary function played a role in the development of CPAM. Subsequently, analysis of these genes pathways showed the TGF-ß signaling pathway was significantly enriched. Finally, the results of immunohistochemical analysis of some DEGs showed that a significant reduction in the expression of SMAD6, a gene related to the TGF-ß signaling pathway, led to abnormal activation of the pathway. TGF-ß signaling pathway involved in the evolution of the disease obtained by DEGs enrichment pathway analysis. SMAD6, a gene involved in this pathway, might be a potential biomarker for the diagnosis and treatment of CPAM.

Rare pediatric synchronous bilateral testicular germ cell tumors of different pathological types: a case report.

The occurrence of synchronous bilateral testicular germ cell tumors (BTGCTs) of different pathologic histologic types in pediatric patients is rare. We reported a case of a left testicular yolk sac tumor (YST) combined with a right testicular mature teratoma. Left orchiectomy and right testis-sparing surgery were performed. Retroperitoneal recurrence was noted 6 months after surgery. The patient underwent reoperation for the resection of a retroperitoneal mass, which was pathologically diagnosed as a recurrent YST. A full cycle of chemotherapy was then administered. No tumor metastasis or recurrence has yet been detected. We present this new case, and we review the previous literature on synchronous BTGCTs to explore the clinicopathologic features and summarize the diagnostic and therapeutic experience. Radical orchiectomy, as the standard treatment for YSTs, should be considered with caution in patients with bilateral testicular tumors. Rapid intraoperative frozen pathology provides support for timely surgical planning. In patients with intraoperative frozen pathologic specimens suggestive of benign lesions, testis-sparing surgery is the preferred treatment option.

Bilateral Wilms tumor in a patient with a horseshoe and duplex kidney: A case report.

The occurrence of horseshoe kidney with duplex urinary collecting systems is rare. Herein, we report a case of bilateral Wilms tumor (BWT) in a patient with a concurrent horseshoe kidney and left duplex kidney, which had not been previously reported. The patient was treated with neoadjuvant chemotherapy, followed by surgical resection and adjuvant chemotherapy. The tumor recurred 6 months postoperatively. A second resection was performed, followed by the administration of chemotherapy and radiotherapy. The patient passed away 15 months after the initial diagnosis of BWT.

Ureteral dilation recovery after intravesical reimplantation in children with primary obstructive megaureter.

Background: To observe the postoperative recovery following ureteral dilation in primary obstructive megaureter (POM) after ureteral implantation, and evaluate the risk factors affecting ureter diameter resolution. Materials and Methods: A retrospective study was performed in patients with POM who underwent ureteral reimplantation using the Cohen procedure. Patient characteristics, perioperative parameters, and postoperative outcomes were also analysed. A widest ureteral diameter of <7 mm was defined as a normal shape and outcome. Survival time was defined as the time from surgery to ureteral dilation recovery or to the last follow-up. Results: A total of 49 patients (54 ureters) were included in the analysis. The survival time ranged from 1 to 53 months. The shapes of a total of 47 (87.04%) megaureters recovered, and most (29/47) resolutions happened within 6 months after surgery. In the univariate analysis, bilateral ureterovesical reimplantation (p = 0.015), ureteral terminal tapering (p = 0.019), weight (p = 0.036), and age (p = 0.015) were associated with the recovery time of ureteral dilation. A delayed recovery of ureteral diameter was noted in bilateral reimplantation (HR = 0.336, p = 0.017) using multivariate Cox regression. Conclusions: Ureteral dilation in POM mostly returned to normal within six postoperative months. Moreover, bilateral ureterovesical reimplantation is a risk factor for delayed postoperative recovery of ureter dilation in POM.

Laparoscopic extraction of a urethral self-inflicted needle from pelvis in a boy: a case report.

Introduction: Self-insertion of foreign bodies in the urethra is an infrequent occurrence in children, and their management aims to minimize urethral morbidity. Endoscopic removal presents a significant challenge, particularly in boys. Currently, there are few reports on laparoscopic management of urethral foreign bodies that have migrated to the pelvic cavity. Case description: An 11-year-old boy presented to the emergency department with complaints of increased frequency of micturition and dysuria. A sharp sewing needle was discovered lodged in the posterior urethra mucosa during cystoscopy. Attempts to remove the needle using an endoscopic grasping forceps were unsuccessful due to the forceps' weak biting power. During a digital rectal examination, the needle migrated into the pelvic region, wedged between the prostatic urethra and the rectal ampulla. After careful inspection of the peritoneal reflection over the fundus of the bladder, the needle was identified and successfully removed through laparoscopy without any complications. Psychiatric counseling was advised for this patient, who was in good condition during an 8-week follow-up. Conclusions: Our case demonstrates the first recorded use of laparoscopy to remove a self-inserted urethral needle that had migrated into the pelvic region, after failed attempts at endoscopic extraction. Future cases may benefit from considering laparoscopic interventions for similar circumstances.

Outcome of Renal Function after Laparoscopic Pyeloplasty in Children with High-Grade Hydronephrosis.

INTRODUCTION: This study aimed to analyze the changes in differential renal function (DRF) after laparoscopic pyeloplasty in children with high-grade hydronephrosis and factors influencing DRF improvement. METHODS: We reviewed the clinical data of unilateral ureteropelvic junction obstruction (UPJO) patients with SFU grade IV who underwent laparoscopic pyeloplasty between February 2018 and October 2020. The patients were divided into two groups: DRF improvement ≥5% (group 1) and DRF improvement <5% (group 2). Preoperative, operative, and postoperative parameters were included in the statistical analysis in both groups. RESULTS: A total of 29 patients with a median age of 6 months were included. The preoperative DRF in group 1 was significantly lower than that in group 2 (36.97 ± 8.47% vs. 45.19 ± 5.22%, p = 0.004). Logistic regression and receiver operating characteristic analysis showed the preoperative DRF was the predictor for renal function improvement after pyeloplasty (p = 0.021) and had a significant predictive value (p = 0.004). CONCLUSION: Nearly half of the patients with high-grade UPJO had improved renal function at 1 year follow-up after laparoscopic pyeloplasty. Preoperative DRF was a predictor of renal function improvement, and postoperative functional recovery was superior in children with lower preoperative DRF.

Ossifying renal tumor of infancy: a case report.

Ossifying renal tumor of infancy (ORTI) is an extremely rare benign renal solid tumor with typical clinical and pathological features. Most cases are diagnosed in infants that are less than 12 months of age and is more common in males. The first symptom in most patients is painless gross hematuria. Microscopically, the tumor has three main components: an osteoid core, osteoblast-like cells, and spindle cells. We reported a case of a 21-day-old patient diagnosed with ORTI who underwent partial nephrectomy and had good follow-up. The unique features of this case are the strong expression of Wilms Tumor-1 (WT-1) and a high Ki-67 index in the hot spot area. ORTI is considered to have a favorable prognosis. Due to the rarity of WT-1 positivity and high Ki-67 index, we should be highly aware that this patient needs to be followed closely. In addition, we reviewed the available literature on ORTI, with the aim of summarizing the diagnostic and therapeutic experience. The diagnosis needs to be given cautiously on the basis of clinical symptoms, imaging, and pathologic examination. Depending on the location and extent of the tumor, surgery can be performed by partial nephrectomy or nephrectomy to avoid overtreatment.

A novel targeted iron oxide nanocarrier for inhibiting M2-type macrophages in the tumor microenvironment.

Background: Tumor-associated macrophages (TAMs) are vital to the tumor microenvironment. They are classified as antitumor M1-type or protumor M2-type macrophages. M2-type macrophages accumulate in the tumor stroma and are related to poor prognosis. Iron oxide nanoparticles are used as drug delivery vehicles because of the structure of carboxyl groups on their surface and their ability to be easily phagocytosed by macrophages. Aim: The signal transducer and activator of transcription 6 (STAT6) signaling pathway controls M2 macrophage polarization, but the STAT6 signaling pathway inhibitor AS1517499 lacks efficient targeting in vivo. Thus, our study aimed to block the polarization of TAMs to M2-type macrophages. Methods and Material: We used ultrasmall superparamagnetic iron oxide nanoparticles (USPIONs) as drug carriers coated with the STAT6 signaling pathway inhibitors AS1517499 and CD163 monoclonal antibodies to synthesize the targeted nanocomplex AS1517499-USPION-CD163 utilizing the carbodiimide method. Then, we determined its physicochemical properties, including hydrodynamic size distribution, ultrastructure, iron concentration, protein content and activity of the CD163 monoclonal antibody, AS1517499 content, and selectivity for M2-type macrophages, and its biological applications. Results: The hydrodynamic size distribution was stable (average size = 95.37 nm). Regarding biological applications, the targeted nanocomplex selectively inhibited M2-type macrophages. Conclusions: The targeted nanocomplex AS1517499-USPION-CD163 showed high selectivity for M2-type macrophages. Therefore, iron oxide nanoparticles targeting TAMs may be an effective approach to TAM therapy.

Aberrant expression of LINC00346 regulates cell migration and proliferation via competitively binding to miRNA-148a-3p/Dnmt1 in Hirschsprung's disease.

BACKGROUND: Hirschsprung's disease (HSCR) is a common birth defect caused by dysplasia of neural crest cells in the gut. Long noncoding RNAs (lncRNAs) play an important role in cellular processes, including development and disease. Despite the known engagement of LINC00346 in several human diseases, its biological function in HSCR remains unknown. METHODS: The relative expression levels of LINC00346, miR-148a-3p and Dnmt1 in HSCR colon tissues were detected by quantitative real-time PCR. Western blot assays were conducted to investigate the Dnmt1 protein expression level. Knockdown of LINC00346 and overexpression of miR-148a-3p in SH-SY5Y and SK-N-BE(2) cell lines was conducted. Cell proliferation and migration were detected by cell counting Kit-8 assays, 5-ethynyl-2'-deoxyuridine assays and transwell assays. Cell apoptosis was verified by flow cytometric analysis. Furthermore, the competing endogenous RNA (ceRNA) activity of LINC00346 on miR-148a-5p was investigated via bioinformatics analysis and luciferase reporter assays. RESULT: Downregulation of LINC00346 and Dnmt1 was detected in HSCR tissues. Knockdown of LINC00346 and overexpression of miR-148a-3p in SK-N-BE(2) and SH-SY5Y cells inhibited cell migration and proliferation and promoted apoptosis. Moreover, the miR-148a-3p inhibitor rescued the downregulation of Dnmt1 in LINC00346 knockdown cell lines, which was evidence of the ceRNA regulatory mechanism of Dnmt1 by LINC00346. CONCLUSIONS: LINC00346 was downregulated in HSCR colon tissues and acted as a ceRNA to regulate the expression of Dnmt1 in vitro. Together, these findings indicate that LINC00346 could affect the occurrence of HSCR by participating in the development of enteric neural crest cells.

MMP-9 secreted by M2-type macrophages promotes Wilms' tumour metastasis through the PI3K/AKT pathway.

BACKGROUND: Wilms' tumour (WT) is a malignant tumour of childhood with the typical symptoms of an abdominal mass. Tumour-associated macrophages (TAMs) accumulate and imply a poor prognosis in WT, but the mechanism of how TAMs affect the prognosis has not been fully elucidated. In this study, we aimed to present the molecular mechanisms underlying the protumorigenic capacities of TAMs in WT. METHODS: TAMs were polarized into M1- and M2-type macrophages. The two types of macrophages were cocultured with SK-NEP-1 cells, and their cell viability and invasion ability were measured. Matrix metalloproteinase 9 (MMP9) expression was assessed in different types of macrophages, and the role of MMP9 in WT was explored. Then data from children diagnosed with WT in our department between February 2006 and July 2014 were retrospectively analysed, the tumour tissues were analysed to explore the distribution of MMP9. Kaplan-Meier analysis of the relationship between MMP9 expression and follow-up information was performed. RESULTS: The results showed that M2-type macrophages could improve the viability and invasive ability of SK-NEP-1 cells. MMP9 expression in M2-type macrophages was significantly higher than that in M1-type macrophages. MMP9 could activate the AKT/PI3K signalling pathway to initiate the epithelial-mesenchymal transition (EMT) process, and promote the proliferation and invasion of WT. In WT tissue, the MMP9 expression level was elevated and it was located in the tumour stroma, which was the same as M2-type macrophage location, and a high level of MMP9 predicted poor survival. CONCLUSION: M2-type macrophages facilitate tumour proliferation and metastasis by secreting MMP9 to enhance the EMT process via a PI3K/AKT dependent pathway in Wilms' tumour.

Self-Powered Cathodic Photoelectrochemical Aptasensor Comprising a Photocathode and a Photoanode in Microfluidic Analysis Systems.

An intriguing self-powered cathodic photoelectrochemical (PEC) microfluidic aptasensor with enhanced cathodic photocurrent response is proposed for sensitive detection of prostate-specific antigen (PSA). The self-powered system is constructed by a cadmium sulfide-sensitized zinc oxide nanorod array (CdS/ZnO NA) as a photoanode with an iodide-doped bismuth oxychloride flower-array (I0.2:BiOCI0.8) as a photocathode, which can generate the electrical output under visible light irradiation with no external power supply. In addition, the p-type semiconductor I0.2:BiOCI0.8 with a special internal electric field between the iodide ion layer and the [Bi2O2]2+ layer could increase the cathodic photocurrent response by facilitating the separation of electron/hole pairs under visible light excitation. It is worth noting that dissolved oxygen as an electron acceptor can be reduced by the photogenerated electron to form a superoxide radical (•O2-) in the self-powered cathodic PEC system. The further enhanced cathodic photocurrent response can be achieved by eliminating •O2- that reacts with the luminol anion radical (L•-) to produce chemiluminescence emission, which serves as an inner excitation light source. What is more exciting is that the integration of the photoanode and the photocathode into a microfluidic chip could realize automatic sample injection and detection. On this basis, the proposed aptasensor presents excellent reproducibility and high sensitivity for detecting PSA and exhibits a good linearity range (50 fg·mL-1 to 50 ng·mL-1) with a low detection limit (25.8 fg·mL-1), which opens up a new horizon of potential for sensitively detecting other kinds of disease markers.

Peadiatric transvesicoscopic dismembered ureteric reimplantation for ectopic upper ureter in duplication anomalies.

INTRODUCTION: Ectopic upper pole ureters in duplex kidneys can be managed surgically by ipsilateral distal ureteroureterostomy or reimplantation of both ipsilateral ureters when upper pole shows reasonable function. OBJECTIVE: This study aimed to evaluate the clinical outcomes of transvesicoscopic dismembered upper ureteric reimplantation for patients with ectopic upper pole ureters in duplication anomalies. PATIENTS AND METHODS: Between July 2015 and January 2019, laparoscopic transvesicoscopic ureteral reimplantation was performed in 20 patients with ectopic upper pole ureters. An incision was made in the bladder wall at 1.0 cm proximal to the lower ureteral orifice of affected side. The upper pole ureter was recognized, and the terminal portion of the upper ureter was ligated and cut. Then the proximal portion of the upper ureter was mobilized, a transverse submucosal tunnel was created and upper ureteric reimplantation was performed with 6/0 absorbable sutures. Patients were followed up with renal ultrasonography and voiding cystourethrogram for clinical outcomes and hydronephrosis trends. RESULTS: Median (range) age at surgery was 22.5 (10-53) months. All of the 20 operations were successful, and none required conversion to an extravesical approach or open surgery. Four patients presented with worsening upper pole hydroureteronephrosis but recovered three to six months postoperatively. Resolution of symptoms and improving hydroureteronephrosis were achieved in all patients and VUR of the upper and lower ureters was not detected at postoperative follow-up. DISCUSSION: To our knowledge, dismembered reimplantation of upper pole ureters has been reported only in a small series through extravesical technique. In our study, we performed intravesicoscopic upper ureter Cohen reimplantation for duplex system ureteral ectopia. Compared with the extravesical approach, the transvesicoscopic approach leave most of the pelvic structures intact and the creation of a submucosal tunnel for prevention of ureteral reflux is more reliable; in addition, this approach avoids any manipulation of the lower pole ureter compared to ipsilateral ureteroureterostomy. But this method does not seem applicable to children under 6 months of age because of the small bladder capacity. CONCLUSIONS: The laparoscopic intravesical technique of dismembered ureteral reimplantation was safe and feasible in our cases and may be an alternative surgical treatment for ectopic upper pole ureters in duplication anomalies.

Recognition of M2 type tumor-associated macrophages with ultrasensitive and biocompatible photoelectrochemical cytosensor based on Ce doped SnO2/SnS2 nano heterostructure.

Tumor-associated macrophages (TAMs) play central roles in the regulation of tumor growth. TAMs can be differentiated into M1 and M2 types, which are responsible for the inhibition and growth of tumor tissues, respectively. Recognition of M2-TAMs is significant for the diagnosis and therapy of cancer, which is however severely limited due to the deficiency of selective and sensitive photoelectrochemical sensors. In this work, using Ce doped SnO2/SnS2 nano heterostructure as the highly sensitive platform, a photoelectrochemical sensor enabling the recognition of M2-TAMs was fabricated for the first time. By the decoration of CD163 antibody on the platform, the ultrasensitive photoelectrochemical sensor can selectively detect the CD163 protein on the surface of M2-TAMs. To our best knowledge, this is the first demonstration for recognition of M2-TAMs using photoelectrochemical method. The fabricated cytosensor has ultra-sensitive photocurrent response, applicable biological compatibility, high selectivity and relatively wide linear sensing range (5 × 101 to 1 × 105 cells/ml) with a low detection limit (50 cells/ml) for the detection of M2-TAMS. This kind of PEC cytosensor would provide a novel analysis and detection strategy for M2-TAMs.

Downregulation of miR-140-5p affects the pathogenesis of HSCR by targeting EGR2.

BACKGROUND/AIMS: Hirschsprung's disease (HSCR) is the most common digestive disease caused by disorders of neural crest development. Despite the known involvement of miR-140-5p in many human diseases, its biological role in Hirschsprung's disease (HSCR) remains undefined. In this study, we sought to reveal the roles of miR-140-5p in the pathogenesis of HSCR. METHODS: Quantitative real-time PCR and western blotting were used to measure the relative expression levels of miRNAs, mRNAs, and proteins in stenotic and dilated sections of the colon of 32 HSCR patients. Targets and proteins were evaluated by western blotting, and Transwell, CCK-8, and flow cytometry assays were adopted to detect the functional effects of miR-140-5p on SH-SY5Y cells. RESULTS: miR-140-5p was significantly downregulated in HSCR tissue samples with increased expression of EGR2, and knockdown of miR-140-5p inhibited cell migration and proliferation and promoted apoptosis in SH-SY5Y cell lines. EGR2 expression was inversely correlated with that of miR-140-5p in cell lines. CONCLUSIONS: miR-140-5p may influence the pathogenesis of HSCR by targeting EGR2.

Relationship of tumour-associated macrophages with poor prognosis in Wilms' tumour.

BACKGROUND: Wilms' tumour (WT) is the most common childhood renal tumour. Tumour-associated macrophages (TAMs) are a critical component of tumour microenvironments and contain two main subtypes, classically (M1) or alternatively (M2) activated macrophages. Evidence has revealed TAMs in predicting poor prognosis in some malignant tumours. However, the role of TAMs in WT is still unclear, and the relationship of different types of TAMs with prognosis has not been elucidated. OBJECTIVE: The aim of the study was to explore the presence of two types of TAMs in WT and analyse the relationship of TAMs with prognosis. STUDY DESIGN: Overall, 61 paediatric patients with WT underwent nephrectomy before any chemotherapy from April 2006 to March 2014. The tumour tissues were analysed by Western blot, immunohistochemistry, and immunofluorescence to explore the distribution of M1 and M2 macrophages in different stages. Kaplan-Meier analysis with regard to the relationship between the presence of TAMs and follow-up information was performed. RESULTS: In the 61 patients (44 males and 17 females), there was a median age of 19 months (IQR 13-35.5); 47 patients are still alive, 11 died, 3 were lost to follow-up. According to the National Wilms Tumor Study (NWTS)-5 guidelines, the distribution of tumour stages was as follows: stage I, 27 patients; stage II, 18 patients; and stage III, 16 patients. The Western blot analysis showed that the density of M1 and M2 macrophages in tumour tissues were significantly greater than that in adjacent normal tissues. Immunohistochemistry showed the proportion of patients with positive M1-type macrophages across different stages: stage I, 66.7% (18/27); stage II, 44.4% (8/18); and stage III, 25% (4/16) (p = 0.027). The proportion of patients with positive M2-type macrophages across different stages: stage I, 25.9% (7/27); stage II, 55.6% (10/18); and stage III, 81.3% (13/16) (p = 0.002). Kaplan-Meier analysis suggested that patients with high densities of M2-type macrophages had shorter overall survival time than those with low densities (log-rank test, p = 0.011). DISCUSSION: TAMs play a pivotal comments in the tumour microenvironment and tumorigenesis. With the progression of clinical stage, M2 macrophage densities increased greatly, and M1 macrophage density decreased. M2 macrophages represent a poor prognosis and can be utilized as a new indicator in pathological examination. CONCLUSION: There is a high density of TAMs in WT, and M2-type macrophage density increases with tumour progression and implies a poor prognosis.

Association between different levels of lipid metabolism­related enzymes and fatty acid synthase in Wilms' tumor.

Wilms' tumor is one of the most common malignant tumors of the abdomen in children. However, there is currently no recognized specific biomarker for the clinical diagnosis and prognosis of this tumor. Lipid metabolism is involved in membrane synthesis and oxidation in tumor cells. This process plays an important role in the development of tumors, but it has not yet been investigated in Wilms' tumor. The aim of the present study was to characterize the changes in lipid metabolism and to contribute to the diagnosis and prognosis of Wilms' tumor. Proteomics analysis was performed to detect lipid­metabolizing enzymes in 9 tissue samples from Wilms' tumors and adjacent tissues, and proteomics revealed the presence of 19 differentially expressed lipid­metabolizing enzymes. Protein interaction analysis with the Search Tool for the Retrieval of Interacting Genes/Proteins was used to identify the interacting proteins. Immunohistochemistry (IHC), immunofluorescence and western blotting were used to further confirm whether the expression of fatty acid synthase (FASN) was significantly increased in the tumor tissues. Oncomine database and reverse transcription­PCR analyses further confirmed that the expression of FASN at the gene level was significantly increased in the tumors. Following collection of 65 pediatric cases of Wilms' tumor at the Shandong Provincial Hospital between 2007 and 2012, the association between the expression of FASN and the clinical characteristics was analyzed, and IHC analysis further demonstrated that FASN expression was significantly associated with tumor stage and size. The association between FASN and the prognosis of children with Wilms' tumor was analyzed using Kaplan­Meier survival curves. In addition, univariate survival analysis revealed that higher expression of FASN in Wilms' tumors was associated with poorer prognosis. Our findings revealed that FASN may be used as a prognostic biomarker in patients with Wilms' tumor. Furthermore, lipid metabolism may play an important role in the occurrence and development of Wilms' tumor.