Circulating Exosomal Transfer RNA-Related Fragments as Diagnostic Biomarkers for Non-small Cell Lung Cancer.

PURPOSE: Exosomal transfer RNA-derived fragments [exo-tRF] possess the capacity to be employed as biomarkers for several types of cancer. We aim to ascertain the diagnostic significance of exosomal 5'tRF-TyrGTA and 5'tRF-ValTAC in non-small cell lung cancer [NSCLC]. METHODS: Ultracentrifugation was deployed to obtain serum exosomes from NSCLC patients and healthy donors. The acquired exosomes were then confirmed by transmission electron microscopy [TEM], qNano, and western blot [WB] techniques. The level of exo- tRF expression was validated by the use of microarrays and RT-qPCR. The diagnostic performance of exo-tRFs for NSCLC was determined through the receiver operating characteristic curve [ROC]. RESULTS: Exosomal 5'tRF-TyrGTA and 5'tRF-ValTAC were significantly downregulated in both early- and late-stage NSCLC patients compared to healthy donors, representing favorable diagnostic efficiency for NSCLC. In addition, the exosomal 5'tRF-TyrGTA level was correlated with tumor stage and lymph node metastasis. CONCLUSION: Exosomal 5'tRF-TyrGTA and 5'tRF-ValTAC can serve as potential biomarkers for NSCLC.

Multi-modal molecular determinants of clinically relevant osteoporosis subtypes.

Due to a rapidly aging global population, osteoporosis and the associated risk of bone fractures have become a wide-spread public health problem. However, osteoporosis is very heterogeneous, and the existing standard diagnostic measure is not sufficient to accurately identify all patients at risk of osteoporotic fractures and to guide therapy. Here, we constructed the first prospective multi-omics atlas of the largest osteoporosis cohort to date (longitudinal data from 366 participants at three time points), and also implemented an explainable data-intensive analysis framework (DLSF: Deep Latent Space Fusion) for an omnigenic model based on a multi-modal approach that can capture the multi-modal molecular signatures (M3S) as explicit functional representations of hidden genotypes. Accordingly, through DLSF, we identified two subtypes of the osteoporosis population in Chinese individuals with corresponding molecular phenotypes, i.e., clinical intervention relevant subtypes (CISs), in which bone mineral density benefits response to calcium supplements in 2-year follow-up samples. Many snpGenes associated with these molecular phenotypes reveal diverse candidate biological mechanisms underlying osteoporosis, with xQTL preferences of osteoporosis and its subtypes indicating an omnigenic effect on different biological domains. Finally, these two subtypes were found to have different relevance to prior fracture and different fracture risk according to 4-year follow-up data. Thus, in clinical application, M3S could help us further develop improved diagnostic and treatment strategies for osteoporosis and identify a new composite index for fracture prediction, which were remarkably validated in an independent cohort (166 participants).

Rhodium-catalyzed C-H carboxymethylation of anilines with vinylene carbonate.

A rhodium-catalyzed synthesis of phenylacetate has been realized by direct C-H carboxymethylation of anilines bearing removable directing groups. The reaction occurred most efficiently in air, without any external base or oxidant. This methodology is expected to provide a facile and general access to various bioactive 2-amino aromatic acetic acid derivatives.

FOXO3a­modulated DEPDC1 promotes malignant progression of nephroblastoma via the Wnt/ß­catenin signaling pathway.

DEP domain containing 1 (DEPDC1) and forkhead box transcription factor 3a (FOXO3a) serve a role in tumor cells. To the best of our knowledge, however, the expression of DEPDC1 and FOXO3a in nephroblastoma and their role and potential mechanisms in nephroblastoma cells have not been reported. The aim of the present study was to characterize the expression of DEPDC1 and FOXO3a in nephroblastoma, as well as the underlying mechanisms. The expression levels of DEPDC1 and FOXO3a were detected using reverse transcription­quantitative PCR and western blotting. Cell viability, proliferation, invasion and migration were detected using Cell Counting Kit­8, colony formation, Transwell and wound healing assays, respectively. The activity of DEPDC1 promoter was detected by dual­luciferase reporter assay and the association between FOXO3a and DEPDC1 was detected using immunoprecipitation. DEPDC1 expression was significantly increased in nephroblastoma cells, particularly WiT49 cells. Compared with the negative control, DEPDC1 knockdown significantly inhibited proliferation, invasion and migration of WiT49 cells, while DEPDC1 overexpression (Ov) reversed these effects. By contrast, expression of FOXO3a was decreased in WiT49 cells and immunoprecipitation showed that FOXO3a bound to the DEPDC1 promoter. Ov­FOXO3a inhibited WiT49 cell proliferation, invasion and migration, as well as protein expression levels of phosphorylated­glycogen synthase kinase­3ß, Wnt3a and ß­catenin, while DEPDC1 Ov reversed the inhibitory effects of FOXO3a Ov on WiT49 cells. In conclusion, DEPDC1 promoted malignant progression of nephroblastoma via the Wnt/ß­catenin signaling pathway; this may be regulated by FOXO3a.

Carboplatin-based Nanomedicine to Enhance the Anticancer Effect in SK-NEP-1 WilmsꞌTumor Cells.

Wilms tumor (WT) is the most common pediatric malignant primary renal tumor. Carboplatin (CRB), a platinum compound is widely used in the treatment of multiple cancers including ovarian, lung, head and neck, and wilm's tumor. However, lower uptake of CRB in cancer cells and toxicity concerns in healthy cells often limited its clinical outcome. The aim of this study was to investigate the antitumor effect of CRB on SK-NEP-1 wilm's cancer cells. Earlier, CRB was formulated in nanoparticulate formulations and characterized its biophysical parameters. SK-NEP-1 cell growth in vitro was assessed by MTT. Then, apoptosis potential was investigated by TUNEL, Hoechst, and colony formation assay. CRB treatment resulted in inhibition of cell proliferation of SK-NEP-1cells in a dose-dependent manner. TUNEL, Hoechst, and colony formation assay demonstrated that CRB was more effective in killing wilm's cancer cell when encapsulated in nanoparticle formulations. Overall, the present study demonstrates that CRB treatment resulted in marked inhibition of cell proliferation and cell apoptosis. These results may pave way for the effective treatment of wilm's tumor in clinical models.

MicroRNA-602 regulating tumor suppressive gene RASSF1A is overexpressed in hepatitis B virus-infected liver and hepatocellular carcinoma.

AIMS: It is important to understand the role of microRNA in the transformation from chronic HBV hepatitis to hepatocellular carcinoma in hepatocarcinogenesis. Relationship of microRNA-602 with chronic HBV hepatitis, liver cirrhosis and HCC was investigated in this article. RESULTS: (1) 14 MicroRNAs were aberrantly expressed in HCC and CL compared with NL. Among these, microRNA-602 expression in CH, LC, NT and HCC was 2.939, 3.234, 2.439 and 4.134 times of that in NL respectively, which was significantly different (p < 0.01 for all vs. NL); RASSF1A expression in LC and HCC was lower than that in NL, while P73 protein expression in CL was higher than that in NL and HCC. (2) MicroRNA-602 expression in HepG2 2.2.15 and HepG2-HBX was 2.643 and 3.48 times of that in HepG2 (p < 0.05 for both). (3) MicroRNA-602 inhibition in HepG2 cells was associated with RASSF1A mRNA and protein expression increased to 4.37, 3.01 times respectively of those not, with cell apoptosis increased and cell proliferation rate decreased significantly, changes were similar in HepG2-HBX cells. METHODS: (1) MicroRNA expression was investigated in normal (NL), chronic HBV hepatitis (CH), HBV-positive cirrhotic (CL), HBV-positive HCC and corresponding normal para-tumorous livers (NT) and hepatoma cells was evaluated with microRNA microarray and verified by real-time PCR, and microRNA-602 was selected for further study. Expression of miR602-target genes RASSF1A and P73 were detected with RT-PCR and western blot. (2) MicroRNA-602 expression in HepG2 and HepG2-HBX was inhibited by miR-602 inhibitor transfection; RASSF1A and P73 expression was detected and cell apoptosis and proliferation were detected. CONCLUSIONS: MicroRNA-602 plays a pro-carcinogenic role in HBV-related hepatocarcinogenesis by inhibiting RASSF1A. MicroRNA-602 might be an early diagnostic marker for HBV-mediated HCC.

Expression of Runx2 and type X collagen in vertebral growth plate of patients with adolescent idiopathic scoliosis.

The different expression of type X collagen and Runx2 between the convex and concave side of vertebral growth plate in scoliosis may help to improve our understanding of the role that growth plate tissue play in the development or progression of idiopathic scoliosis. In this investigation, there were significant differences of the total expression of type X collagen, Runx2 protein, and Runx2 mRNA between convex side and concave side growth plates of the apex vertebrae (p < 0.05). The total expression of type X collagen in the concave side growth plates of the lower end vertebrae was higher than that in the same side growth plates of apex (p < 0.05). The total expression of Runx2 in the concave side growth plates in the upper and lower end vertebrae were higher than that in the concave side growth plates of apex (p < 0.05). The expression of type X collagen, Runx2, and Runx2 mRNA, the cell density of type X collagen and Runx2 positive chondrocytes, and histological changes between convex side and concave side of the vertebral growth plate indicated that the vertebral growth plate was affected by mechanical forces, which was a secondary change and could contribute to progression of adolescent idiopathic scoliosis.

[Digital skeletal age and histological evaluation for residual spine growth potential in idiopathic scoliosis].

OBJECTIVE: To ascertain the correlation between histological grades (HGs) of vertebral growth plates and Risser grades in idiopathic scoliosis (IS) patients; to identify whether digital skeletal age (DSA) is a reliable indicator for accurate evaluation of the spinal residual growth potential. METHODS: Twenty eight Chinese female patients were available for this study. Superior and inferior growth plates were obtained at each level when anterior approach surgeries were performed. Histological examinations were conducted after the specimens were processed. The patients were evaluated by DSA stages in this study. Correlations between histological grades, menarchal status, and chronological age were analyzed. RESULTS: There was a negative correlation between the following: HGs and DSA stages in 28 cases (r = -0.541, P = 0.003), and HGs and menarchal status in patients in DSA stage III (r = -0.591, P = 0.006). Statistical significance of growth activity of growth plates was found between patients in DSA-stage II and those in DSA-stage III (P = 0.014). CONCLUSIONS: DSA may be a reliable indicator for predicting the spinal residual growth potential in IS patients, but it should be correlated with menarchal status and chronological ages.

[Assessment of the residual spine growth potential in idiopathic scoliosis by risser sign and histological grading].

OBJECTIVE: To investigate the correlation between histological grade (HG) of vertebral growth plates and Risser grade in the female idiopathic scoliosis (IS) patients; and to identify whether Risser grading is a reliable indicator for accurate evaluation of the spinal residual growth potential. METHODS: Thirty-nine samples of vertebral growth plates obtained during operation from 15 females IS patients, all female, aged 15.1 (12.4 - 18.0), underwent HE staining and light microscopy to determine the values of HG. Xray photography of pelvis was conducted before operation to identify the Risser sign. The correlation of Risser grade with pubertal status was analyzed. RESULTS: All the vertebral growth plates of the IS patients with the Risser grade of 0 showed growth activity. The vertebral growth plates showed HG II activity in 6 of the 16 IS patients with the Risser grade of 4. All the vertebral growth plates showed no growth activity in the 3 IS patients with the Risser grade of 5. There was a negative correlation between the HG and Risser grade in all 39 patients (r = -0.645, P =0.000). The HG of the patients with the Risser grade of 4 was negatively correlated with the menarchal status (time between menarche and operation) (r = -0.710, P = 0.002). The residual growth potential of spinal growth plates of the patients with the Risser grades of 2 - 5 was significantly lower than of the patients with the Risser grades of 0 - 1 (P = 0.020). CONCLUSION: Risser sign may be a reliable indicator for predicting the spinal residual growth potential in IS patients, but it should be correlated with menarchal status and chronological ages.

Histomorphological study of the spinal growth plates from the convex side and the concave side in adolescent idiopathic scoliosis.

Asymmetrical growth of the vertebrae has been implicated as one possible etiologic factor in the pathogenesis of adolescent idiopathic scoliosis. The longitudinal vertebral growth derives from the endochondral ossification of the vertebral growth plate. In the present study, the growth plates from the convex and concave side of the vertebrae were characterized by the method of histology and immunohistochemistry to evaluate the growth activity, cell proliferation, and apoptosis. Normal zoned architectures were observed in the convex side of the growth plate and disorganized architectures in the concave side. The histological grades were significantly different between the convex and the concave side of the growth plate in the apex vertebrae (P < 0.05). The histological difference was also found significant statistically between end vertebrae and apex vertebrae in the concave side of vertebral growth plates (P < 0.05). The proliferative potential indexes and apoptosis indexes of chondrocytes in the proliferative and hypertrophic zone in the convex side were significantly higher than that in the concave side in the apex vertebral growth plate (P < 0.05). There was a significant difference of the proliferative potential index (proliferating cell nuclear antigen, PCNA index) between convex side and concave side at the upper end vertebra (P < 0.05). The difference of the proliferative potential index and apoptosis index were found significant statistically in the concave side of the vertebral growth plate between end vertebrae and apex vertebrae (P < 0.05). The same result was also found for the apoptosis index (terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate biotin nick end labeling assay, TUNEL index) in the convex side of vertebral growth plate between end vertebrae and apex vertebrae (P < 0.05). Some correlation were found between radiographic measurements and proliferation and apoptosis indexes. The difference in histological grades and cellular activity between the convex and concave side indicated that the bilateral growth plate of the vertebrae in AIS patients have different growth kinetics which may affect the curve progression.

Histologic, risser sign, and digital skeletal age evaluation for residual spine growth potential in Chinese female idiopathic scoliosis.

STUDY DESIGN: A prospective study. OBJECTIVE: To ascertain the correlation between histologic grades (HGs) of vertebral growth plates and Risser grades as well as DSA stages in the Chinese female idiopathic scoliosis (IS) patients; to identify whether digital skeletal age (DSA) is a reliable indicator for accurate evaluation of the spinal residual growth potential. SUMMARY OF BACKGROUND DATA: DSA is considered one of the more important indicators for representing the peak height velocity (PHV) typically and predicting spinal growth potential. The correlation between HGs of growth plates and DSA stages in IS patients is unclear. METHODS: Thirty-nine Chinese female patients were available for this study. Superior and inferior growth plates were obtained at each level when anterior approach surgeries were performed. Histologic examinations were conducted after the specimens were processed. Of these patients, 28 cases were evaluated by DSA stages in this study. Correlations between histologic grades, Risser grades, menarchal status, and chronologic age were analyzed in 39 patients. Correlations between histologic grades, DSA, menarchal status, and chronologic age were analyzed in 28 patients. RESULTS: There was a negative correlation between the following: HGs and Risser grades in 39 patients (r = -0.645, P = 0.000-0.05), HGs and menarchal status in patients in Risser 4 (r = -0.710, P = 0.002-0.05), HGs and DSA stages in 28 cases (r = -0.541, P = 0.003-0.05), and HGs and menarchal status in patients in DSA Stage III (r = -0.591, P = 0.006-0.05). Statistical significance of growth activity of growth plates was found between patients in Risser Grades 0 to 1 and those in Risser Grades 2 to 5 (P = 0.020-0.05) and patients in DSA Stage II and those in DSA Stage III (P = 0.014-0.05). CONCLUSION: DSA may be a reliable indicator for predicting the spinal residual growth potential in IS patients, but it should be correlated with menarchal status and chronologic ages.