Fibronectin-1 modulated by the long noncoding RNA OIP5-AS1/miR-200b-3p axis contributes to doxorubicin resistance of osteosarcoma cells.

Chemoresistance has been an obstacle in the further improvement of 5-year survival rates of osteosarcoma (OS) patients, but the underlying mechanism of chemo-resistance remains unclear. A comprehensive analysis of mRNAs and noncoding RNAs related to OS chemo-resistance could help solve this problem. In the current study, we first identified that fibronectin-1 (FN1), screened by microarray analysis in three paired chemo-resistant and chemo-sensitive OS cell lines, was significantly upregulated in the chemo-resistant OS cell lines and tissues and was related to unfavourable prognosis. Further functional assays revealed that FN1 inhibition greatly increased the sensitivity of OS cells to doxorubicin in vitro and in vivo, whereas FN1 overexpression had the opposite effect. Moreover, mechanistic investigation demonstrated, by a series of assays that included luciferase reporter gene, RNA immunoprecipitation, RNA pull-down and rescue assays, that FN1 expression was regulated by the oncogenic long noncoding RNA (lncRNA) OIP5-AS1 through sponging miR-200b-3p. Thus, these results indicated the role and potential application of the lncRNA OIP5-AS1/miR-200b-3p/FN1 regulatory pathway as a promising target in treatment of OS chemo-resistance.

Screening circular RNA related to chemotherapeutic resistance in osteosarcoma by RNA sequencing.

AIM: To identify circular RNAs (circRNAs) related to osteosarcoma (OS) chemoresistance. MATERIALS & METHODS: CircRNA expression profile was performed in three paired human chemoresistant and chemosensitive OS cell lines by next-generation sequencing. Quantitative real-time-PCR (qRT-PCR) was used to confirm next-generation sequencing data. Bioinformatics analysis was conducted to predict their functions. RESULTS: Eighty circRNAs were dysregulated in the chemoresistant OS cells compared with the control, after validated by qRT-PCR. Bioinformatics analysis showed that some pathways related to drug metabolism were significantly enriched. Additionally, hsa_circ_0004674 was distinctly increased in OS chemoresistant cells and tissues, related to poor prognosis. CircRNA-miRNA-mRNA pathways related to hsa_circ_0004674 were constructed by TargetScan and miRanda. CONCLUSION: CircRNAs may play a role in OS chemoresistance and hsa_circ_0004674 might be a candidate target.

Circular RNA circ_HIPK3 is down-regulated and suppresses cell proliferation, migration and invasion in osteosarcoma.

Circular RNA (circRNA) is associated with human cancers, however, few studies have reported its value in the diagnosis and prognosis prediction of osteosarcoma (OS). In this study, we investigated the expression level of eight selected cancer-related circRNAs including circ-Cdr1as, circ_HIPK3 and circ-ITCH in OS cell lines, tissues and plasmas by quantitative real-time polymerase chain reaction (qRT-PCR) and found that only circ_HIPK3 could stably down-regulate in the OS cell lines, tissues and plasmas than the corresponding controlled. One-way analysis of variance was further conducted to analyze the relationship between circ_HIPK3 expression level and clinic pathological factors of OS patients. Receiver operating characteristic (ROC) curve was built to evaluate the diagnostic values of circ_HIPK3. Circ_HIPK3 expression was significantly correlated with Enneking stage (P=0.042) and lung metastasis (P=0.036). The area under the ROC curve was 0.783 and the sensitivity and specificity were 0.56 and 0.84, respectively. Kaplan-Maier analysis also showed that lower expression of circ_HIPK3 correlated with shorter overall survival time and poor prognosis of OS patients. Besides, function analysis demonstrated that circHIPK3 overexpression significantly suppressed OS cell proliferation, migration and invasion in vitro. Overall, our data suggest that circ_HIPK3 may become a novel potential biomarker for diagnosis and treatment target of OS.

Overexpressed circPVT1, a potential new circular RNA biomarker, contributes to doxorubicin and cisplatin resistance of osteosarcoma cells by regulating ABCB1.

Circular RNAs (circRNAs) represent a widespread class of non-coding RNAs generated from back-splicing, with a circular loop structure. Many circRNAs have been reported to play essential roles in cancer development and have the potential to serve as a novel class of biomarkers for clinical diagnosis. However, the role of circRNA in osteosarcoma (OS) remains largely unknown. In the current study, we examined the expression level of circular RNA PVT1 (circPVT1), previously screened and identified the oncogenic role in gastric cancer, in OS and found that circPVT1 was significantly up-regulated in the OS tissues, serums and chemoresistant cell lines, correlated with poor prognosis of OS patients. Besides, ROC curve demonstrated that circPVT1 may be a better diagnostic biomarker than alkaline phosphatase (ALP) in OS with more sensitivity and specificity. In addition, functional assays revealed that circPVT1 knockdown by siRNA could weaken the resistance to doxorubicin and cisplatin of OS cells through decreasing the expression of classical drug resistance-related gene ABCB1. These findings may provide a new insight into the role of circPVT1 as a biomarker for the diagnosis and treatment target of OS.

Preparation and performance detection of small-diameter tissue-engineered blood vessels / 中国组织工程研究

BACKGROUND:Due to limited sources,poor hemocompatibility and poor anticoagulation performance,small-diameter tissue-engineered blood vessels cannot be applied in clinical practice.OBJECTIVE:To explore the physicochemical and mechanical properties of sheep carotid arteries after the decellularization in order to find appropriate materials for the preparation of tissue-engineered blood vessels.METHODS:Fresh carotid arteries from sheep were randomly divided into two groups:control group,in which,the sheep carotid arteries were cryopreserved for use after trimming and cleaning;experimental group,in which,after trimming and cleaning,the carotid arteries were decallularized by Triton X-100.sodium deoxycholate and EDTA for 24 hours,rinsed for 72 hours,digested with RNA/DNA enzymes for 24 hours,rinsed for 24 hours and reserved for later use.In both groups,blood samples were subjected to hematoxylin-eosin staining,collagen fiber staining,elastic fiber dyeing,and electron microscopy observation.The physical and chemical properties of the blood vessels are tested by tensile strength,wall tension and thickness.RESULTS AND CONCLUSION:(1) The collagen fibers in both two groups were neat and compact in alignment,with no obvious fracture.(2) Hematoxylin-eosin staining showed that:in the control group,the nuclei were distributed in the inner membrane,middle lamella and outer membrane of the vessels,and the fibers ran regularly;in the experimental group,the fibers ran in order but loosely,and there were no nuclei in the inner membrane,middle lamella and outer membrane of the vessels.(3) Elastic fibers in the control group were regular in alignment and mainly distributed in the middle lamella and outer membrane of the vessels,while in the experimental group,the elastic fibers ran regularly but loosely,and mainly distributed in the middle lamella and outer membrane of the vessels.(4) Under the scanning electron microscope,the originally formed vessels were observed in the experimental group,with no cell residues,and the collagen fibers ran orderly with no fracture and with uniform pore structure.(5) The vessel thickness was lower in the experimental group than the control group (P ＜ 0.01),but the tensile strength showed no difference between the two groups,which was 46.55 kPa in the two groups.To conclude,the decelluarized sheep carotid artery can retain the necessary mechanical properties of the blood vessels after achieving the maximum removal of antigenicity.

Antisense lncRNA FOXF1-AS1 Promotes Migration and Invasion of Osteosarcoma Cells Through the FOXF1/MMP-2/-9 Pathway.

Osteosarcoma (OS) is the most common primary malignant bone cancer in children and adolescents. Long non-coding RNAs (lncRNAs) have been shown to play significant role in various cancers, including OS. In a previous study, we have reported that a novel antisense lncRNA FOXF1-AS1, also known as FENDRR, could sensitize doxorubicin-resistance of OS cells through down-regulating ABCB1 and ABCC1. Here in, the critical role of FOXF1-AS1 in regulating OS progression was further investigated. Firstly, we found that FOXF1-AS1 and its antisense transcript FOXF1 expression were positively up-regulated in OS tissues and cell lines and correlated with poor prognosis of OS patients. Besides, FOXF1-AS1 as well as FOXF1 silencing significantly inhibited cell proliferation, migration, invasion of OS cells and tumor growth both in vitro and vivo through decreasing the expression of MMP2 and MMP9, whereas enhanced expression of FOXF1-AS1 had the opposite effects. In addition, mechanistically, both of FOXF1-AS1 and FOXF1 could regulate the expression of MMP2 and MMP9 at mRNA and protein levels, whereas FOXF1-AS1 could influence the FOXF1expression but FOXF1 did not have the same effect on FOXF1-AS1. Rescue assay further showed that FOXF1-AS1 overexpression efficiently reversed the knockdown of MMP2 and MMP9 expression induced by si-FOXF1. Thus, we concluded that FOXF1-AS1 may promote migration and invasion of OS cells through the FOXF1/MMP-2/-9 pathway. Taken together, these findings demonstrated the underlying mechanism of FOXF1-AS1 in the regulation of OS progression and provide a novel potential target in the OS therapy.

LncRNA FENDRR sensitizes doxorubicin-resistance of osteosarcoma cells through down-regulating ABCB1 and ABCC1.

Long noncoding RNAs (LncRNAs) act as crucial regulators in various cancers including osteosarcoma (OS), yet their potential roles and molecular mechanisms in OS chemoresistance remain unclear. In the present study, we investigated the role and potential regulatory mechanism of the most down-regulated expressed lncRNA, FENDRR screened by our previous lncRNA microarray analysis between the paired doxorubicin-resistant and sensitive human osteosarcoma cell lines (MG63/DXR vs MG63). FENDRR expression was down-regulated in the doxorubicin-resistant OS cell lines and tissues and negatively correlated to the poor prognosis of OS patients. Overexpression of FENDRR suppressed doxorubicin-resistance, G2/M phase of cell cycle, and promoted cell apoptosis of osteosarcoma cells in vitro and tumor growth in vivo whereas FENDRR knockdown had the opposite effects. In addition, we found that FENDRR was mainly located in the cytoplasm and could regulate the drug resistance of osteosarcoma cells by negatively affecting posttranscriptional expression of ABCB1 and ABCC1. Together, our study demonstrated that lncRNA FENDRR may act as an inhibitory molecule of doxorubicin-resistance through down-regulating the expression of ABCB1 and ABCC1 genes in osteosarcoma cells. These findings may extend the function of FENDRR in tumor progression and provide a novel target for reversing OS chemoresistance.

Correlation between estrogen receptor status and clinicopathologic parameters in endometrial cancer: a comparative study by immunohistochemistry using different scoring systems / 中华病理学杂志

<p><b>OBJECTIVE</b>To compare the efficiency of three different estrogen receptor (ER) immunostaining scoring systems by analyzing the correlation between ER status and clinicopathologic features for prediction of prognosis of patients with endometrial carcinoma (EC).</p><p><b>METHODS</b>ER immunostaining (EnVision method) was performed in 160 type I EC and 39 type II EC paraffin samples and was scored by ASCO/CAP criterion, H-Score and Allred scoring system. Correlation between ER status and clinicopathologic features was statistically analyzed.</p><p><b>RESULTS</b>ASCO/CAP criterion, H-Score and Allred (cutoff point: 4-8) scoring system showed high concordance in the following aspects. In EC patients, ER status was significantly associated with presurgical CA125 levels (P = 0.015, P = 0.007, P = 0.023), histologic grades (all P < 0.01) and PR status (all P < 0.01). In type I EC cohort, ER status was significantly correlated with PR status (P = 0.008, P < 0.01, P < 0.01) and p53 status (P = 0.042, P = 0.001, P < 0.01). As of the predictive value of ER status for type I EC patient age, ASCO/CAP (P = 0.027) and H-Score criteria (P = 0.035) were both superior to Allred score system (P = 0.064). Among well-known predictive clinicopathologic parameters, including FIGO stage, lympho-vascular involvement, lymph node metastasis, depth of myometrial invasion and omental involvement, ASCO/CAP scoring offered a better correlation (P = 0.005, P = 0.002, P = 0.021, P = 0.067, and P = 0.067, respectively) than H-Score (P > 0.05) and Allred scoring system (P > 0.05).</p><p><b>CONCLUSIONS</b>Compared with H-Score and Allred scoring system, ASCO/CAP criterion is more closely correlated with predictive clinicopathologic parameters. Therefore it may be used as a simple, highly efficient prognostic indicator for EC patients in routine practice.</p>

Application of ultrasmall superparamagnetic iron oxide in MRI diagnosis of central nervous system: Recent progress / 第二军医大学学报

Ultrasmall superparamagnetic iron oxide (USPIO), as the contrast agent of MRI, possesses two major properties: long half time in the plasma and specific binding with macrophages. Compared with gadolinium, widely-used in clinic presently, USPIO has its unique advantages in diagnosis of central nervous system diseases, though which still need further clinical verification. This article reviews the application of USPIO in MRI diagnosis of some central nervous system diseases.

Applications of the hospital statistics management system / 中国医疗器械杂志

The Hospital Statistics Management System is built on an Office Automation Platform of Shandong provincial hospital system. Its workflow, role and popedom technologies are used to standardize and optimize the management program of statistics in the total quality control of hospital statistics. The system's applications have combined the office automation platform with the statistics management in a hospital and this provides a practical example of a modern hospital statistics management model.

Application of fluorescence in-situ hybridization and reverse transcription-polymerase chain reaction in molecular diagnosis of Ewing's sarcoma and primitive neuroectodermal tumor / 中华病理学杂志

<p><b>OBJECTIVE</b>To detect tumor specific chromosome translocations and associated fusion transcripts in paraffin-embedded tissue by interphase fluorescence in-situ hybridization (FISH) and reverse transcription polymerase chain reaction (RT-PCR), respectively, and to evaluate their diagnostic values for Ewing's sarcoma/primitive neuroectodermal tumor (ES/PNET).</p><p><b>METHODS</b>Nuclei of the tumor cells and total RNA were extracted from 10 cases of ES/PNET. Interphase FISH was utilized to analyze the EWS gene translocation with a dual color, break apart probe (Vysis company). RT-PCR was used to detect t (11; 22) (q24; q12) and t (21; 22) (q22; q12) fusion transcripts.</p><p><b>RESULTS</b>Among 10 cases of ES/PNET, the EWS-FLI1 fusion transcript was detected in 8 by RT-PCR. EWS-ERG fusion transcript was not detected in any of the cases. EWS gene translocation was found in 9 of 10 cases by FISH.</p><p><b>CONCLUSIONS</b>Interphase FISH and RT-PCR can be reliably applied to paraffin-embedded tissues for molecular diagnosis of ES/PNET. Between the two approaches, interphase FISH provides a more sensitive and stable result.</p>