Methylation-mediated silencing of miR-125a-5p facilitates breast cancer progression by inducing autophagy.

BACKGROUND: microRNA-125a-5p (miR-125a) is a tumor suppressor gene whose role in autophagy remains poorly understood. In the current study, we aimed to investigate the methylation status of miR-125a, its transfection into SK-BR3 cells, and its effects on autophagy. METHODS: Sixty samples of tumor and non-tumor adjacent tissue were collected and the methylation status of miR-125a was evaluated by methylation-specific PCR (MSP). The effect of 5-Aza-dC on miR-125a expression was investigated in the SK-BR3 cells. Cells were also transfected with miR-125a mimic/antimiR. The expression of miR-125a and its target genes was evaluated by Real-Time PCR. Protein levels of ATG5 and LC3 were assessed by Western blotting. HER2 expression was investigated by immunocytochemistry (ICC). RESULTS: The data showed that the miR-125a promoter CpG Island was significantly hypermethylated in breast cancer tissues (p < 0.01) and in SK-BR3 cells. The 5-Aza-dC could significantly increase miR-125a expression by decreasing its methylation (p < 0.05). In addition, Western blot analysis indicated the expression of ATG5 and LC3 II/ LC3I, as autophagy biomarkers, was significantly reduced in SK-BR3 cells transfected with miR-125a (p < 0.05). CONCLUSIONS: Our data showed miR-125a expression was significantly decreased in tumor tissues due to its promoter hypermethylation. Overexpression of miR-125a was associated with a reduction in autophagy, which could provide a new therapeutic avenue for advanced-stage breast cancer treatment.

The utility of PAX8 in comparison with PAX5 immunohistochemical staining in the diagnosis of Hodgkin lymphoma.

BACKGROUND: Hodgkin's lymphoma (HL) is among the most prevalent lymphomas worldwide. PAX5 has a great value in separating this entity from other T cell lymphoma; however, it is weakly expressed in neoplastic cells. Polyclonal PAX8 was positive in a variety of lymphoid neoplasms in several previous studies and the staining paralleled that of PAX5 but monoclonal PAX8 was negative in the same neoplasms. The aim of this study was to compare immunohistochemical patterns of monoclonal PAX8 with PAX5 in Classical and NLPHL samples. MATERIAL AND METHODS: This retrospective study was conducted on 89 formalin-fixed paraffin embedded blocks from HL patients (69 Classical and 20 NLPHL) admitted to Imam Khomeini and Dr. Shariati hospitals, Tehran, Iran during 2016-2020. Diagnoses were confirmed by reviewing previous immunohistochemistry (IHC) studies, including PAX5. All samples were stained for PAX8 (clone MRQ-50). Expression intensity scoring was made for both antibodies in neoplastic and background cells based on nuclear staining percentage. RESULTS: PAX8 was positive in neoplastic and background B lymphocytes of all classical and NLPHL samples. PAX8 Expression intensity was significantly higher in neoplastic and background cells compared to PAX5 in classical HL samples (P = 0.001). PAX5 expression intensity in neoplastic cells was significantly higher in NLPHL samples compared to classical HL (P = 0.040); however, no significant difference in PAX8 expression between neoplastic cells of NLPHL and HL was seen. PAX8 expression intensity was not significantly correlated with gender, histologic subtype, tumor location, and relapse. CONCLUSIONS: PAX8 monoclonal antibody (clone MRQ-50) showed strong nuclear reactivity in neoplastic and background cells of classical HL and NLPHL samples. Therefore, this marker can be utilized as a valuable alternative for PAX5 in differentiating HL from other T cell lymphoma in challenging cases.

Breast Cytology and Pathology in Pregnancy and Lactation.

Breast tissue reveals some physiologic changes during pregnancy and lactation due to hormonal alterations. Whole range of breast diseases including inflammatory, benign and malignant neoplasms can be seen in pregnancy but due to concurrent physiologic changes, may lead to diagnostic challenges. This chapter reviews sampling methods and histologic features of common benign breast lesions in pregnancy and lactation periods.

Histology of Pregnancy-Associated Breast Cancer.

Breasts are one of the most common sites of neoplastic lesions in women during pregnancy and lactation. This chapter reviews carcinomas of the breast during pregnancy and lactation while focusing on histologic features, biomarker profiles and some involved molecular pathways. Also, a brief review of previous studies on this field is performed.

Safety and toxicity of silymarin, the major constituent of milk thistle extract: An updated review.

Milk thistle (Silybum marianum) is a medicinal plant from the Asteraceae family. Silymarin is the major constituent of milk thistle extract and is a mixture of some flavonolignans such as silybin, which is the most active component of silymarin. It is most commonly known for its hepatoprotective effect. Also, studies have shown other therapeutic effects such as anticancer, anti-Alzheimer, anti-Parkinson, and anti-diabetic, so its safety is very important. It has no major toxicity in animals. Silymarin was mutagen in Salmonella typhimurium strains in the presence of metabolic enzymes. Silybin, silydianin, and silychristin were not cytotoxic and genotoxic at concentration of 100 µM. Silymarin is safe in humans at therapeutic doses and is well tolerated even at a high dose of 700 mg three times a day for 24 weeks. Some gastrointestinal discomforts occurred like nausea and diarrhea. One clinical trial showed silymarin is safe in pregnancy, and there were no anomalies. Consequently, caution should be exercised during pregnancy, and more studies are needed especially in humans. Silymarin has low-drug interactions, and it does not have major effects on cytochromes P-450. Some studies demonstrated that the use of silymarin must be with caution when co-administered with narrow therapeutic window drugs.

Turmeric (Curcuma longa) and its major constituent (curcumin) as nontoxic and safe substances: Review.

Curcumin is the major constituent of turmeric (Curcuma longa). Turmeric has been widely used as a spice in foods and for therapeutic applications such as anti-inflammatory, antihyperlipidemic, and antimicrobial activities. Turmeric and curcumin are nonmutagenic and nongenotoxic. Oral use of turmeric and curcumin did not have reproductive toxicity in animals at certain doses. Studies on human did not show toxic effects, and curcumin was safe at the dose of 6 g/day orally for 4-7 weeks. However, some adverse effects such as gastrointestinal upsets may occur. Moreover, oral bioavailable formulations of curcumin were safe for human at the dose of 500 mg two times in a day for 30 days, but there are still few trials and more studies are needed specially on nanoformulations and it should be discussed in a separate article. In addition, curcumin is known as a generally recognized as safe substance. This review discusses the safety and toxicity of turmeric and curcumin in medicine. Turmeric and curcumin are nontoxic for human especially in oral administration. Turmeric and curcumin are also safe in animals. They are nonmutagenic and are safe in pregnancy in animals but more studies in human are needed.

Expression and Clinical Significance of IRE1-XBP1s, p62, and Caspase-3 in Colorectal Cancer Patients.

Background: Three main cell signaling pathways including the endoplasmic reticulum stress (ERS) response, autophagy, and apoptosis play critical roles in both cell survival and death. They were found to crosstalk with one another during tumorigenesis and cancer progression. This study aimed to investigate the expression of the spliced form of X-box binding protein 1 (XBP1s), p62, and caspase-3, as the essential biomarkers of ERS, autophagy, and apoptosis in patients with colorectal cancer (CRC), as well as the correlation between their expression and clinicopathological data. Methods: This retrospective study was conducted on formalin-fixed paraffin-embedded (FFPE) blocks, which were collected from patients and their tumor margins, from the tumor bank of Imam Khomeini Hospital (Tehran, Iran) from 2017 to 2019. Tissue microarray (TMA) was used to measure the XBP1s, p62, and caspase-3 biomarkers. Data were analyzed using SPSS software version 20, and P≤0.05 was considered statistically significant. Results: Evaluating the total of 91 patients, a significant relationship was found between XBP1s expression and TNM stage (P=0.003), primary tumor (pT) (P=0.054), and the degree of differentiation (P=0.006); and between caspase-3 with pT (P=0.004), and lymphovascular invasion (P=0.02). However, no significant correlation was found between p62 and clinicopathological data. Furthermore, a positive relationship between XBP1s and p62 was confirmed (correlation coefficient: 22.2% and P=0.05). Conclusion: Our findings indicated that XBP1s could be considered as a target for therapy in personalized medicine.

Evaluation of PD-L1 antigen expression using immunohistochemistry technique in medullary thyroid carcinoma samples.

BACKGROUND: Markers related to the mechanism of tumoral cell escape from the immune system have received more attention. The PD-L1 molecule encoded by the "CD274" gene binds to T lymphocytes and can inhibit these cells. Therefore, increasing the expression of this marker on inflammatory or tumor cells can indicate tumor progression invasiveness and long-term consequences. The present study aimed to determine the expression of the PD-L1 marker in thyroid medullary tumors and to evaluate its role in predicting long-term outcomes after cancer. METHODS: This retrospective longitudinal study was performed on pathology samples of patients with medullary thyroid carcinoma referred to the Cancer Institute of Imam Khomeini Hospital from 2015 to 2020. Slides related to medullary thyroid tumors were examined. A tissue microarray was used to evaluate the immunohistochemistry of PD-L1. Patients were followed up to assess the occurrence of recurrence. Out of 207 patients evaluated in the present study, histopathological information of 144 patients was available. RESULTS: The expression rate of PD-L1 in our community was 14.6% in lymphocyte cells, 35.4% in tumor cells, and 12.5% in both cells. The presence of metastasis at the time of diagnosis was reported in 35 cases (72.9%), and the occurrence of tumor recurrence was reported in 38 cases (79.2%). There was no relationship between the expression of this marker and the sex and age of patients. In addition, PD-L1 expression was unrelated to the two main characteristics of this cancer, namely tumor size and its focality. The presentation of tumor PD (L1) (but not lymphocytic) was a prognostic marker for synchronous metastasis at cancer diagnosis but could not predict tumor recurrence. CONCLUSION: PD-L1 tumor marker expression is predictable in 14.6% of lymphocyte cells, 35.4% of tumor cells, and 12.5% in the selected Iranian population with medullary thyroid cancer. The expression of this marker is not related to the morphological characteristics of the tumor, such as tumor size or focality.

A review on oxidant and antioxidant effects of antibacterial agents: impacts on bacterial cell death and division and therapeutic effects or adverse reactions in humans.

Reactive oxygen species (ROS) are produced in the mitochondrial respiratory pathway and cellular metabolism. They are responsible for creating oxidative stress and lipid peroxidation. In living organisms, there is a balance between oxidative stress and the antioxidant system, but some factors such as medicines disturb the balance and cause many problems. These effects can impact bacterial death and division and also in humans can induce therapeutic or adverse reactions. Web of Science and Pubmed databases were used for searching. This review focuses on the oxidant and antioxidant effects of different classes of antibacterial agents and the mechanisms of oxidative stress. Some of these agents have beneficial effects on killing bacteria due to their antioxidant or oxidant effects. However, some of their side effects may be due to their oxidative effects. Based on the results of this review, minocycline is an antioxidant, but aminoglycosides, chloramphenicol, glycopeptides, antituberculosis drugs, fluoroquinolones, and sulfamethoxazole agents have oxidant effects. Furthermore, cephalosporins, penicillins, metronidazole, and macrolides have both oxidant and antioxidant effects in different studies. It is concluded that some antibacterial agents have oxidant and other antioxidant effects. These activities may affect their therapeutic effects or side effects. Some antioxidants can prevent the adverse effects of antibacterial agents. Clarifying the exact oxidant and antioxidant effects of some antimicrobial agents needs more research projects.

MCM6 versus Ki-67 in diagnosis of luminal molecular subtypes of breast cancers.

BACKGROUND: Currently, breast cancers are divided into four major molecular subtypes. The distinction between the luminal A and luminal B subtypes is mainly based on the cellular proliferation indices and is assessed by the Ki-67 scoring. Due to the limitations in the assessment and expression of Ki-67, we hypothesized that minichromosome maintenance protein 6 (MCM6) might be taken as a surrogate marker to differentiate molecular subtypes and aid in more precise grading of tumors. METHODS: We performed a retrospective, cross-sectional study on 124 samples of breast cancer and 40 samples of normal breast tissue. Relevant clinical information was retrieved from the Cancer Institute database. RESULTS: MCM6 could discriminate between various categories of histologic grades, tubule formation, mitotic indices, and nuclear pleomorphism (P = 0.002 for tubule formation and P < 0.001 for other). Moreover, the MCM6 score exhibited a significant correlation with the mitotic count (P < 0.001). However, the Ki-67 score could not discriminate subgroups of the mitotic index and nuclear pleomorphism. Compared to the luminal A subtype, luminal B exhibited a higher MCM6 score (P = 0.01). Besides, MCM6 scores were higher for certain subtypes with more aggressive behaviors, such as hormone receptor (HR)-negative disease, and human epidermal growth factor receptor 2 (HER2)-enriched and triple-negative breast cancers, as there was a significantly higher MCM6 mean score in the HR-negative in comparison to the luminal breast cancers (P < 0.001). Similarly, higher MCM6 scores were observed among samples with more advanced nuclear grades, tubule formation, and overall grades. CONCLUSION: MCM6 can differentiate luminal A and luminal B subtypes and is correlated with mitotic counts. However, this study was unable to prove the superiority of MCM6 in differentiating between molecular subtypes compared to the Ki-67 score. Nevertheless, in our study, MCM6 was superior to Ki-67 in exhibiting correlations with the mitotic grade, tubule formation, and nuclear grades. More studies are needed to standardize its assessment methods, determine more robust cut-off values, and evaluate its associations with prognostic features of breast cancer.

Burned-out Post Pubertal Teratoma Presenting as a Liver Metastases in a 34-Year-Old Male.

Germ cell teratomas belong to non-seminomatous germ cell tumors and account for 95% of malignant testicular tumors. Regarding the current World Health Organization (WHO) criteria, testicular teratomas are divided into prepubertal and post-pubertal subtypes based on patients' age. The term "burned-out testicular tumor" is a very rare condition referring to a regressed testicular tumor which presents with its metastases without any clinical finding in the testicle. Metastasis can be the presentation of post-pubertal teratoma in 22-37% of cases. In scar associated teratoma (burn-out component), the metastasis rate is 66%. We reported a rare case of post-pubertal teratoma in a 34-year-old male who presented with multiple liver masses initially. Liver biopsy revealed poorly differentiated adenocarcinoma probably of gastrointestinal (GI) tract origin. The upper and lower GI endoscopy were normal. Scrotal ultrasonography showed a hypoechoic cystic intratesticular lesion in the left testis. The patient underwent radical orchiectomy and the histopathologic examination revealed post-pubertal teratoma with burned out component. The patient underwent proper treatment and is still under follow up. As a result, in a young male patient who presented with a retroperitoneal mass or poorly differentiated carcinomas of an unknown primary site, using light microscopy and immunohistochemical profiling alone may be inadequate. Therefore, scrotal screening and physical examination of the scrotum and bilateral testis should be considered to exclude possibility of a metastatic progression from a testicular germ cell neoplasia.

Erdheim-Chester disease with long-standing diabetes insipidus and generalized edema.

Erdheim-Chester disease (ECD) is a rare non-Langerhans histiocytosis. ECD is detected more frequently due to increased awareness of healthcare providers and improved diagnostic tools. This report describes a 51-year-old woman with a history of weakness, bone pain, xanthelasma palpebrarum, and diabetes insipidus. ECD is a multisystemic condition with a poor prognosis. This disease should be considered in patients with diabetes insipidus, bone pain, and multiorgan involvements.

Optical anisotropy measurement in normal and cancerous tissues: backscattering technique.

Investigating the deformation of tissue architecture is one of the most important clinical methods for cancer diagnosis. Optical methods are now widely developed for rapid, precise, and real-time assessment of these alterations at the microscopic scale. One of the proposed methods is enhanced backscattering (EBS) technique that allows in-vivo measurement of the optical scattering characteristics. Here, EBS technique is employed to evaluate the optical anisotropy of human epithelial tissues as a measure to distinguish between normal and cancerous one. Orientation dependence of the mean scattering length is assessed in healthy and cancerous tissues of five different human organs i. e. uterus, bladder, colon, kidney, and liver. Helicity preserving channel and rotating ground glass diffuser are utilized to eliminate the polarization induced anisotropy and the background speckle noises respectively. Analysis of the backscattering cones recorded by a high-resolution CCD camera reveals the modification of the strength and degree of optical anisotropy in different tissues during cancer progression. Pathology data affirm the correlation between the experimental results and the morphological alteration of the epithelial cells in each carcinoma type. In general, tissues with fibrous constructional cells are subject to a decrease in anisotropy due to cancer, whereas those with cuboidal cells experience an increase in anisotropy. This complementary information enhances the potency of the EBS technique as a fast, non-destructive, and easily accessible tool for real-time tissue diagnosis.

LINK-A long non-coding RNA and VEGF RNA expression in epithelial ovarian cancer patients.

LINK-A (long intergenic non-coding RNA for kinase activation) is a newly identified long non-coding RNA with oncogenic function, which leads to the hyperactivation of AKT and HIF1α. thereby, fosters cell proliferation, mobility and metastasis. VEGF (vascular endothelial growth factor), a well-known cytokine has an important role in angiogenesis. In this study, we quantified RNA expression of LINK-A and VEGF on 45 tumor specimens obtained from Iranian patients diagnosed with Epithelial Ovarian Cancer (EOC). Our goal was to evaluate expression of LINK-A lncRNA and VEGF mRNA in ovarian cancer tissues and find the probable correlation of LINK-A with VEGF as a major transcription target for HIF1α. LINK-A and VEGF were remarkably overexpressed in EOC tissues compared to normal tissues (P value: 0.004, 0.0001, respectively), but we did not find correlation between LINK-A and VEGF RNA expressions in this study. LINK-A was significantly overexpressed in higher stages of cancer and tumor grades. VEGF was only significantly elevated in higher stages. This study confirms importance of novel lncRNA of LINK-A in Iranian EOC patients.

Depth-Based Whole Body Photoplethysmography in Remote Pulmonary Function Testing.

OBJECTIVE: We propose a novel depth-based photoplethysmography (dPPG) approach to reduce motion artifacts in respiratory volume-time data and improve the accuracy of remote pulmonary function testing (PFT) measures. METHOD: Following spatial and temporal calibration of two opposing RGB-D sensors, a dynamic three-dimensional model of the subject performing PFT is reconstructed and used to decouple trunk movements from respiratory motions. Depth-based volume-time data is then retrieved, calibrated, and used to compute 11 clinical PFT measures for forced vital capacity and slow vital capacity spirometry tests. RESULTS: A dataset of 35 subjects (298 sequences) was collected and used to evaluate the proposed dPPG method by comparing depth-based PFT measures to the measures provided by a spirometer. Other comparative experiments between the dPPG and the single Kinect approach, such as Bland-Altman analysis, similarity measures performance, intra-subject error analysis, and statistical analysis of tidal volume and main effort scaling factors, all show the superior accuracy of the dPPG approach. CONCLUSION: We introduce a depth-based whole body photoplethysmography approach, which reduces motion artifacts in depth-based volume-time data and highly improves the accuracy of depth-based computed measures. SIGNIFICANCE: The proposed dPPG method remarkably drops the error mean and standard deviation of FEF , FEF , FEF, IC , and ERV measures by half, compared to the single Kinect approach. These significant improvements establish the potential for unconstrained remote respiratory monitoring and diagnosis.

Dicrocoelium Dentriticum in Explanted Liver: Report of an Unusual Finding.

Dicrocoelium dentriticum, a member of trematode type helminths, is a liver parasite of ruminants. Humans are infected accidentally by ingestion of intermediate host, through infected ants via eating of raw vegetables or drinking of contaminated water. Infection is often asymptomatic or results in subtle symptoms; therefore, infections are usually unrecognized. However, it can produce chronic cholangitis and swelling or adenomatous proliferation in the bile ducts and lead to abdominal pain, diarrhea, fatigue, jaundice, and other symptoms. We report a 49-year-old female patient with end-stage hepatic cirrhosis from viral hepatitis B and D coinfection who underwent liver transplant. Shortly after transplant, she developed symptoms suggesting an obstructed biliary duct. Liver needle biopsy was done 24 hours after transplant to rule out rejection. Biopsy of her explanted liver was also examined pathologically. Microscopic examination of the liver needle biopsy ruled out rejection. Prepared sections of explanted liver revealed a helminth in the common bile duct. Morphologic reconstruction of helminth by microscopic findings and consultation with an expert parasitologist supported the diagnosis of Dicrocoelium dentriticum.

Remote, Depth-Based Lung Function Assessment.

OBJECTIVE: We propose a remote, noninvasive approach to develop pulmonary function testing (PFT) using a depth sensor. METHOD: After generating a point cloud from scene depth values, we construct a three-dimensional model of the subject's chest. Then, by estimating the chest volume variation throughout a sequence, we generate volume-time and flow-time data for two prevalent spirometry tests: forced vital capacity (FVC) and slow vital capacity (SVC). Tidal volume and main effort sections of volume-time data are analyzed and calibrated separately to remove the effects of a subject's torso motion. After automatic extraction of keypoints from the volume-time and flow-time curves, seven FVC ( FVC, FEV1, PEF, FEF 25%, FEF 50%, FEF 75%, and FEF [Formula: see text]) and four SVC measures ( VC, IC, TV, and ERV) are computed and then validated against measures from a spirometer. A dataset of 85 patients (529 sequences in total), attending respiratory outpatient service for spirometry, was collected and used to evaluate the proposed method. RESULTS: High correlation for FVC and SVC measures on intra-test and intra-subject measures between the proposed method and the spirometer. CONCLUSION: Our proposed depth-based approach is able to remotely compute eleven clinical PFT measures, which gives highly accurate results when evaluated against a spirometer on a dataset comprising 85 patients. SIGNIFICANCE: Experimental results computed over an unprecedented number of clinical patients confirm that chest surface motion is linearly related to the changes in volume of lungs, which establishes the potential toward an accurate, low-cost, and remote alternative to traditional cumbersome methods, such as spirometry.

Toward Respiratory Assessment Using Depth Measurements from a Time-of-Flight Sensor.

Introduction: There is increasing interest in technologies that may enable remote monitoring of respiratory disease. Traditional methods for assessing respiratory function such as spirometry can be expensive and require specialist training to perform and interpret. Remote, non-contact tracking of chest wall movement has been explored in the past using structured light, accelerometers and impedance pneumography, but these have often been costly and clinical utility remains to be defined. We present data from a 3-Dimensional time-of-flight camera (found in gaming consoles) used to estimate chest volume during routine spirometry maneuvres. Methods: Patients were recruited from a general respiratory physiology laboratory. Spirometry was performed according to international standards using an unmodified spirometer. A Microsoft Kinect V2 time-of-flight depth sensor was used to reconstruct 3-dimensional models of the subject's thorax to estimate volume-time and flow-time curves following the introduction of a scaling factor to transform measurements to volume estimates. The Bland-Altman method was used to assess agreement of model estimation with simultaneous recordings from the spirometer. Patient characteristics were used to assess predictors of error using regression analysis and to further explore the scaling factors. Results: The chest volume change estimated by the Kinect camera during spirometry tracked respiratory rate accurately and estimated forced vital capacity (FVC) and vital capacity to within ± <1%. Forced expiratory volume estimation did not demonstrate acceptable limits of agreement, with 61.9% of readings showing >150 ml difference. Linear regression including age, gender, height, weight, and pack years of smoking explained 37.0% of the variance in the scaling factor for volume estimation. This technique had a positive predictive value of 0.833 to detect obstructive spirometry. Conclusion: These data illustrate the potential of 3D time-of-flight cameras to remotely monitor respiratory rate. This is not a replacement for conventional spirometry and needs further refinement. Further algorithms are being developed to allow its independence from spirometry. Benefits include simplicity of set-up, no specialist training, and cost. This technique warrants further refinement and validation in larger cohorts.

Expression and prognostic significance of Oct4 and Nanog in neuroblastoma.

Neuroblastoma is the most common extracranial solid tumor of children, accounting for an estimated 15% cancer-related deaths in this period. It has been hypothesized that drug resistance of cancer stem cells may be responsible for chemotherapy failure, sustained tumor growth, and recurrence in many solid tumors. In this study, we investigated the expression of Octamer-binding transcription factor 4 (Oct4) and Nanog, two stem cell markers, in 47 neuroblastic tumors by immunohistochemistry and correlated their expression by other prognostic factors especially with NMYC amplification using both fluorescent and chromogenic in situ hybridization methods. Twenty three cases (48.9%) showed Oct4 signals and eight cases (17%) showed Nanog expression. All Nanog positive tumors showed Oct4 expression. Seven cases (14.1%) had NMYC amplification. There was also no association between positive Oct4 and Nanog reactivity and tumor morphology, age, mitosis-karyorrhexis index, NMYC amplification, favorable or unfavorable histology, and risk groups (p > 0.05). Cancer stem cells hypothesis is a challenging issue and controversies exist about their significance. Although our study did not show strong association between prognostic factors and expression of stem cell markers, performing of further large-scale studies of various neuroblastic tumors with various stages is suggested.