Euxanthone Impairs the Metastatic Potential of Osteosarcoma by Reducing COX-2 Expression

Osteosarcoma (OS) is one of the most common malignancies of bone. This study was aimed to explore the anti-metastatic effect of euxanthone on OS. Adhesion assay and Transwell assay were used to examine the effect of euxanthone on adhesion, migration and invasion of OS cells. COX-2-over-expressing plasmid was applied to transfect OS cells to assess whether COX-2 affects the anti-metastatic function of euxanthone. PDCD4 knockdown and miR-21 mimic were applied to assess whether euxanthone suppresses the transactivation of c-jun via modulating miR-21-PDCD4 signaling. The effect of euxanthone in vivo was also examined by lung metastasis assay. Euxanthone, a xanthone derivative extracted from Polygala caudata, has been found to exhibit anti-neoplastic activities. In present study, our results showed that euxanthone suppressed cell adhesion, migration, and invasion in OS cells. Our experimental data also showed that repression of COX-2 by euxanthone mediated its anti-metastatic activities. Moreover, our findings revealed that euxanthone modulated the COX-2 expression through the miR-21/PDCD4/c-jun signaling pathway. The anti-metastatic activities of euxanthone were also validated in a pulmonary metastasis model. Taken together, our results highlighted the potential of euxanthone to be used in the treatment of OS. Anat Rec, 302:1399–1408, 2019. © 2018 Wiley Periodicals, Inc.     

Biallelic loss of function variants in PPP1R21 cause a neurodevelopmental syndrome with impaired endocytic function

Next‐generation sequencing (NGS) has been instrumental in solving the genetic basis of rare inherited diseases, especially neurodevelopmental syndromes. However, functional workup is essential for precise phenotype definition and to understand the underlying disease mechanisms. Using whole exome (WES) and whole genome sequencing (WGS) in four independent families with hypotonia, neurodevelopmental delay, facial dysmorphism, loss of white matter, and thinning of the corpus callosum, we identified four previously unreported homozygous truncating PPP1R21 alleles: c.347delT p.(Ile116Lysfs*25), c.2170_2171insGGTA p.(Ile724Argfs*8), c.1607dupT p.(Leu536Phefs*7), c.2063delA p.(Lys688Serfs*26) and found that PPP1R21 was absent in fibroblasts of an affected individual, supporting the allele's loss of function effect. PPP1R21 function had not been studied except that a large scale affinity proteomics approach suggested an interaction with PIBF1 defective in Joubert syndrome. Our co‐immunoprecipitation studies did not confirm this but in contrast defined the localization of PPP1R21 to the early endosome. Consistent with the subcellular expression pattern and the clinical phenotype exhibiting features of storage diseases, we found patient fibroblasts exhibited a delay in clearance of transferrin‐488 while uptake was normal. In summary, we delineate a novel neurodevelopmental syndrome caused by biallelic PPP1R21 loss of function variants, and suggest a role of PPP1R21 within the endosomal sorting process or endosome maturation pathway.     

Morphological difference between pleural mesothelioma cells in effusion smears with either BAP1 loss or 9p21 homozygous deletion and reactive mesothelial cells without the gene alterations

We previously characterized the morphological characteristics of malignant pleural mesothelioma (MPM) cells with 9p21 homozygous deletion (HD) using a combination of the virtual microscopy and fluorescence in situ hybridization (FISH). In this study, we investigated whether MPM cells with BRCA1‐associated protein 1 (BAP1) loss show the same morphological characteristics identified in MPM cells with 9p21 HD. MPM cells with either BAP1 loss detected by immunocytochemistry (ICC) or 9p21 HD detected by FISH were identified via virtual microscopy prior to ICC or FISH, followed by analysis and quantification of their morphological characteristics. MPM cells with BAP1 loss or 9p21 HD exhibited significantly more frequent cell‐in‐cell engulfment, multinucleation, and larger multicellular clusters composed of more than 10 cells than reactive mesothelial cells. In conclusion, MPM cells with BAP1 loss or 9p21 HD share similar cytological features, indicating that the same morphological criteria can be used to detect MPM cells harboring such genetic aberrations.     

Referencing the trochlear groove based on three-dimensional computed tomography imaging improves the reliability of the measurement of the tibial tuberosity–trochlear groove distance in patients with higher grades of trochlea dysplasia

BackgroundTo determine whether 3D-CT imaging technique is valid and reproducible compared to conventional CT measurement technique (CCT) for the detection of a femoropatellar instability.MethodsPatients who had undergone surgery for femoropatellar instability (patellar instability group) between 2010 and 2016 (n = 37 knees of 35 patients) were retrospectively enrolled. For the matched control group, patients who had acute anterior cruciate ligament injury (< 4 weeks previously; n = 30) were recruited. Preoperative CT data had been obtained in all patients. Inter-rater reliability was calculated for both measurement protocols, and inter-method reliability was calculated between the two imaging modalities. The results are reported using intraclass correlation coefficients (ICCs) and Bland–Altman 95% limits of agreement.ResultsAll patients in the patellar instability group had femoral trochlear dysplasia (Dejour types A: four, B: 19, C: seven, and D: six), but no dysplasia was noted in the control group. In the patellar instability group, the CCT technique showed a poor inter-rater agreement (ICC = 0.74), and the 3D-CT technique still showed excellent inter-rater agreement (ICCs = 0.91). In the sub-analysis of the patellar instability group according to the trochlear dysplasia grade, ICCs were markedly decreased with severe trochlear dysplasia when using CCT technique; however, the 3D-CT technique could provide excellent reliability even with severe trochlear dysplasia.ConclusionThe 3D-CT imaging technique for the measurement of the TT–TG distance can be suggested as a better measurement technique for patellar instability patients with bone abnormality.     

血清TK-1、SE-CAD表达及联合HE4、CEA、CYFRA21-1检测在非小细胞肺癌诊断中的意义

目的探讨可溶性E-钙黏蛋白(soluble E-cadherin,SE-CAD)、胸苷激酶-1(thymidine kinase 1,TK-1)在非小细胞肺癌(NSCLC)患者血清中的水平,并分析其与附睾蛋白4(HE4)、癌胚抗原(CEA)、细胞角蛋白19片段(CYFRA21-1)联合检测在NSCLC诊断中的临床应用价值。方法收集本院2016年1月至2018年1月期间初诊NSCLC患者86例、小细胞肺癌患者45例、同期良性疾病患者42例、同期初诊其他恶性肿瘤(除肺癌外)患者30例、健康受试者的血清标本40例,应用液态芯片技术对受试者血清标本进行CEA、CYFRA21-1水平检测,ELISA方法检测SE-CAD、TK-1、HE4。结果①非小细胞肺癌患者血清中SE-CAD水平明显高于良性对照组、正常对照组( P <0.05),差异具有统计学意义,但与小细胞肺癌组及其他恶性肿瘤组比较差异无统计学意义( P >0.05);②非小细胞肺癌组患者血清 TK1 与CYFRA21-1水平明显高于良性对照组、小细胞肺癌组、其他恶性肿瘤组和健康对照组,差异均有统计学意义(均 P <0.05)。③单项检测时,CYFRA21-1对非小细胞肺癌的诊断灵敏度最高,为87.2%,TK-1的诊断特异性最高。5种肿瘤标志物联合检测,灵敏度高达93.2%,特异性有所降低为85.7%。结论 SE-CAD、TK1 与CYFRA21-1均可作为鉴别良恶性肿瘤的检验指标,联合HE4、CEA、CYFRA21-1等指标对非小细胞肺癌诊断的灵敏度远高于单项检测。     

联合检测血清FGF-21、RBP4、SF水平在2型糖尿病评估中的价值研究

目的探究并分析联合检测血清人成纤维细胞生长因21 (FGF-21)、重组人视黄醇结合蛋白-4(recombinant human retinol binding protein-4,RBP4)、血清铁蛋白(serum ferritin,SF)水平在2型糖尿病(type 2 diabetes,T2DM)评估中的价值。方法选取2016年6月至2017年6月我院收治的2型糖尿病患者48例,作为T2DM组;同时选取34例糖耐量异常(impaired glucose tolerance,IGT)患者,作为IGT组,另选取我院进行体检的健康志愿者30例,作为对照组。观察并记录各组受试者FGF-21、RBP4、SF水平,比较其在不同组之间的表达情况。分析并比较单独FGF-21、RBP4、SF以及联合FGF-21、RBP4、SF检测对2型糖尿病的评估效果。采用ROC曲线分析各方法的评估效率。结果 T2DM组血清FGF-21、RBP4、SF水平显著高于IGT组和健康人群(P <0.05),而IGT组患者FGF-21、RBP4、SF水平显著高于健康人群(P<0. 05);用单独的FGF-21、RBP4、SF检测进行评估,特异性差异具有统计学意义(P <0.05),其中,RBP4评估的特异性显著高于FGF-21、SF的评估特异性(P<0.05);对于平行联合检测,FGF-21+RBP4+SF评估的灵敏度、阴性预测值均显著高于FGF-21+SF、RBP4+SF(P<0.05);对于系列联合检测,FGF-21+RBP4+SF评估的特异性显著高于FGF-21+SF(P<0.05);采用FGF-21+RBP4+SF评估时,系列联合检测的灵敏度显著低于平行联合检测(P <0.05),特异性显著高于平行联合检测(P<0.05);FGF-21+SF、RBP4+SF及FGF-21+RBP4+SF评估T2DM,平行联合检测ROC曲线下面积分别为0.761、0.763和0.915,系列联合检测ROC曲线下面积为0.750、0.721和0.904,FGF-21+RBP4+SF评估的ROC曲线下面积显著高于FGF-21+SF、RBP4+SF(P<0.05),具有统计学意义。结论 FGF-21+RBP4+SF联合检测评估效率显著高于各单项检查,能有效提高糖尿病评估灵敏度、特异性及准确率,值得临床推广。     

Enamelin maps to human chromosome 4q21 within the autosomal dominant amelogenesis imperfecta locus

Amelogenesis imperfecta is a group of hereditary enamel defects. Of the autosomal dominant forms, only the local hypoplastic type has been mapped to human chromosome 4q 13-4q21. Enamelin is a large enamel matrix protein secreted by ameloblasts. The purpose of this study was to determine the human chromosomal localization of enamelin to establish an association with various forms of amelogenesis imperfecta. Chromosomal mapping was performed by polymerase chain reaction (PCR) amplification using somatic hybrid and deletion/derivation cell line panels with an enamelin primer set based on 100% conserved regions between pig and mouse cDNAs. Sequence-tagged site content mapping using eight markers within the critical local hypoplastic amelogenesis imperfecta region was then performed using an isolated human enamelin genomic BAC clone. The human enamelin amplicon was confirmed by DNA sequence analysis, revealing 81% and 73% identity to pig and mouse cDNAs, respectively. PCR amplification using a somatic cell hybrid panel placed enamelin on chromosome 4 with analysis of a regional chromosome 4 mapping panel refining the localization to 4q 13.1-q21.23. An identified human enamelin BAC genomic clone was shown to contain markers D4S2604 and D4S2670, as well as the first exon of the human ameloblastin gene, placing enamelin in the critical amelogenesis imperfecta locus between markers HIS1 and D4S2604 at 4q21. Our results suggest that enamelin is a strong candidate gene for this disease. Furthermore, human 4q21 may contain a second cluster of enamel matrix genes located proximally to the identified cluster of dentin and bone genes.     

Apoptosis and cell cycle arrest in oral lichen planus Hypothesis on their possible influence on its malignant transformation

The quantitative importance of cell cycle arrest and apoptosis mechanisms in oral lichen planus (OLP) was analysed in order to assess the cell response to T lymphocyte aggression and establish a hypothesis on the influence of these phenomena in the malignant transformation process. The TUNEL assay and immunohistochemical methods were used to detect caspase-3, bax, and p21 in 32 tissue samples of oral mucosa with OLP and in 20 samples of normal oral mucosa. Positivity for TUNEL, caspase-3 and p21 was significantly more frequent in cases than in controls (p<0.001). Both TUNEL and caspase-3 positivity was significantly greater in the basal versus suprabasal layer (p=0.004 and 0.052, respectively). The basal and suprabasal expression of p21 was significantly higher in cases with a more intense liquefaction degeneration (p<0.01). There was no significant difference in basal expression of bax between cases and controls. The quantitative importance of apoptosis was small in OLP. Epithelial cells attacked in OLP have a very low response to apoptosis and cell cycle arrest mechanisms, which may produce an epithelial substrate that favours malignant transformation.     

HPV16 E6/E7对永生化口腔上皮细胞p53、p21表达的影响

目的：为了探讨人乳头状瘤病毒(HPV)转化口腔上皮细胞的作用机制，研究HPV16 E6、E7对永生化口腔上皮细胞细胞周期调节因子p53、p21表达的影响。方法：免疫细胞化学法检测HPV16 E6、E7诱导的永生化口腔上皮细胞HIOEC中野生型和突变型p53的表达；Westem印迹检测HIOEC HPV16 E6、E7、p53、p21蛋白表达，并以正常口腔上皮细胞和转染空白载体的口腔上皮细胞为对照。结果：HIOEC细胞表达HPV16 E6、E7蛋白．野生型p53和p21表达水平高于正常口腔上皮细胞和转染空白载体的口腔上皮细胞。结论：p53的失活可能不是HPV16 E6、E7诱导口腔上皮细胞永生化的主要原因。