Vertebral fracture severity assessment on anteroposterior radiographs with a new semi-quantitative technique.

We developed a new tool to assess the severity of osteoporotic vertebral fracture using radiographs of the spine. Our technique can be used in patient care by helping to stratify patients with osteoporotic vertebral fractures into appropriate treatment pathways. It can also be used for research purposes. PURPOSE: The aim of our study was to propose a semi-quantitative (SQ) grading scheme for osteoporotic vertebral fracture (OVF) on anteroposterior (AP) radiographs. METHODS: On AP radiographs, the vertebrae are divided into right and left halves, which are graded (A) vertical rectangle, (B) square, (C) traverse rectangle, and (D) trapezoid; whole vertebrae are graded (E) transverse band or (F) bow-tie. Type A and B were compared with normal and Genant SQ grade 1 OVF, Type C and D with grade 2 OVF, and Type E and F with grade 3 OVF. Spine AP radiographs and lateral radiographs of 50 females were assessed by AP radiographs SQ grading. After training, an experienced board-certified radiologist and a radiology trainee assessed the 50 AP radiographs. RESULTS: The height-to-width ratio of the half vertebrae varied 1.32-1.48. On lateral radiographs, 84 vertebrae of the 50 patients had OVFs (38 grade 1, 24 grade 2, and 22 grade 3). On AP radiographs, the radiologist correctly assigned 84.2%, 91.7%, and 77.2% and the trainee correctly assigned 68.4%, 79.2%, and 81.8% of grade 1, 2, and 3 OVFs, respectively. Compared with lateral radiographs, the radiologist had a weighted Kappa of 0.944 including normal vertebrae and 0.883 not including normal vertebrae, while the corresponding Kappa values for the trainee were 0.891 and 0.830, respectively. CONCLUSION: We propose a new semi-quantitative grading system for vertebral fracture severity assessment on AP spine radiographs.

The effect of small molecule inhibitor NSC348884 on nucleophosmin 1-mutated acute myeloid leukemia cells.

OBJECTIVE: Nucleophosmin 1 (NPM1) is a common shuttling protein. Mutation in the NPM1 gene is the most frequent gene alteration in acute myeloid leukemia (AML). This study aims to explore the inhibitory effects of small molecule NSC348884 on wild-type and NPM1-mutated AML cells. MATERIALS AND METHODS: Immunofluorescence was used to determine the intracellular localization of NPM1 protein in wild-type (OCI-AML2) and NPM1-mutated (OCI-AML3) AML cell lines. The oligomerization state of NPM1 was assessed by Western blot analysis, and the inhibitory effect of NSC348884 on the proliferation of AML cells was evaluated by Cell-counting kit-8 (CCK-8). Flow cytometry was used to detect the proapoptotic effect of NSC348884 on AML cells. RESULTS: Western blot results showed a significant reduction in the levels of the oligomeric NPM1 protein after the treatment with NSC348884. NSC348884 had an inhibitory effect on the proliferation of both wild-type and NPM1-mutant AML cells. The inhibitory effect on OCI-AML3 cells was stronger, compared to OCI-AML2 cells. Flow cytometry showed that NSC348884 could significantly induce AML cell apoptosis and had a stronger proapoptotic effect on OCI-AML3 cells. CONCLUSIONS: NSC348884 had inhibitory and proapoptotic effects on both wild-type and NPM1-mutated AML cells. The effect of NSC348884 on AML cells, carrying NPM1 mutation was significantly stronger.

Functional identification of ANR genes in apple (Malus halliana) that reduce saline-alkali stress tolerance.

As one of the major abiotic stresses restricting the development of global agriculture, saline-alkali stress causes osmotic stress, ion poisoning, ROS damage and high pH damage, which seriously restrict sustainable development of fruit industry. Therefore, it is essential to develop and cultivate saline-alkali-resistant apple rootstocks to improve the yield and quality of apples in China. Based on transcriptome data, MhANR (LOC114827797), which is significantly induced by saline-alkali stress, was cloned from Malus halliana. The physicochemical properties, evolutionary relationships and cis-acting elements were analysed. Subsequently, the tolerance of MhANR overexpression in Arabidopsis thaliana, tobacco, and apple calli to saline-alkali stress was verified through genetic transformation. Transgenic plants contained less Chl a, Chl b and proline, SOD, POD and CAT activity, and higher relative electrical conductivity (REC) compared to WT plants under saline-alkali stress. In addition, expression of saline-alkali stress-related genes in overexpressed apple calli were also lower than in WT calli, including the antioxidant genes (MhSOD and MhCAT^), the Na+ transporter genes (MhCAX5, MhCAX5, MhSOS1, MhALT1), and the H+ -ATPase genes (MhAHA2 and MhAHA8), while expression of the K+ transporter genes (MhSKOR and MhNHX4) were higher. Expression of MhANR reduced tolerance of A. thaliana, tobacco, and apple calli to saline-alkali stress by regulating osmoregulatory substances, chlorophyll content, antioxidant enzyme activity, and expression of saline-alkali stress-related genes. This research provides a theoretical basis for cultivating apple rootstocks with effective saline-alkali stress tolerance.

Analysis of clinical characteristics of thyroid phenotype in Pendred syndrome based on multiple databases.

OBJECTIVE: This study aimed to provide statistical data support for the development of thyroid phenotype-related follow-up and reference for follow-up duration and project selection by analyzing the clinical characteristics of thyroid phenotype in Pendred syndrome (PDS) based on multiple databases. MATERIALS AND METHODS: PDS-related pathogenic or possibly/pathogenic mutations were searched by Deafness Variation Database (DVD), ClinVar, and PubMed databases, the mutation sites were counted and the characteristics and thyroid phenotypes were analyzed. RESULTS: The median age of hearing phenotype onset in PDS cases reported in multiple databases was 1.0 (1.0, 2.0) years, the median age of thyroid phenotype onset was 14.5 (5.8, 21.0) years, and the median age that thyroid phenotype was more delayed than hearing phenotype was 10.0 (4.0, 17.0) years. There were significant differences in the distribution of onset time between the two phenotypes (Z=-4.560, p<0.01). In these patients, the positive rates of goiter, thyroid nodules, abnormal thyroid function, and perchlorate discharge test (PDT) were 78%, 78%, 69%, and 78%, respectively. Moreover, the number of thyroid phenotype-positive items in the genotype group with frameshift mutation was not significantly higher than that in the group without frameshift mutation (Z=-1.452, p=0.147). CONCLUSIONS: The early missed diagnosis of PDS may be due to the late onset of thyroid phenotype and the non-100% positive rate of examination items. Therefore, multi-item follow-up of the thyroid gland into adulthood will benefit patients. At present, the relationship between genotype and phenotype is still unclear, and prognosis cannot be determined according to genotype.

The impact of instant neutrophil gelatinase-associated lipocalin level on the severity of septic acute kidney injury.

OBJECTIVE: The clinical value of increased levels of neutrophil gelatinase-associated lipocalin (NGAL) in patients with septic acute kidney injury (AKI) is still unclear. This study aimed to assess the link between illness severity and NGAL in patients with septic AKI. PATIENTS AND METHODS: This is a retrospective observational study that took place at the Fourth Hospital of Hebei Medical University, Shijiazhuang, China. The cohort included 365 patients who were admitted to the ICU during the 21-month period. Of them, 18 patients were diagnosed with sepsis (septic group). The average age of patients in the septic group was over 65, and 60.00% of them eventually progressed to septic AKI. Plasma NGAL (pNGAL) and urine NGAL (uNGAL) levels at defined time points were measured. AKI staging was done based on the Kidney Disease Improving Global Outcomes (KDIGO) classification. The Sequential Organ Failure Assessment (SOFA) and Acute Physiology and Chronic Health Evaluation (APACHE) II scores were determined. Patterns and associations between NGAL levels with SOFA scores and different stages of septic AKI were investigated. RESULTS: Both pNGAL and uNGAL showed a positive correlation with SOFA and proved to be reliable predictors of the same. Furthermore, the accuracy of severe sepsis (SOFA ≥ 8) was 0.67 for pNGAL and 0.66 for uNGAL. Real-time detection of pNGAL and uNGAL indicated that they were good biomarkers of severe septic AKI. Area under the receiver operating characteristic (AUROC) for pNGAL and uNGAL were 0.72 (0.69-0.85), and 0.83 (0.71-0.95), respectively. However, only patients with KDIGO 3 AKI presented significantly elevated levels of pNGAL (p < 0.05). Furthermore, the uNGAL level at each stage of septic AKI was higher than that of the non-AKI period (p < 0.01). CONCLUSIONS: In patients with septic AKI, levels of NGAL correlated with SOFA. Levels of pNGAL were good predictors of severe kidney injury and uNGAL levels could detect mild stages of AKI.

Predicting the risk of non-specific low back pain in the young population: development and assessment of a new predictive nomogram.

OBJECTIVE: Non-specific low back pain is a common disorder that affects more than 80% of the world's population. But the potential risk factors remain unclear. The aim of this study is to develop a nomogram for the risk prediction of low back pain in young population. PATIENTS AND METHODS: A total of 264 young participants (18-45 years old) were recruited and randomly divided into a training set (n=188) and a validation set (n=76) by a ratio of 7:3. The nomogram was developed based on the training set. The independent predictors of low back pain were identified by LASSO and logistic regression analysis. A nomogram was developed according to the predictors. To assess the reliability of the nomogram, the area under the curve (AUC), calibration curve, and decision curve analysis (DCA) were applied. The validation set was used to validate the results. RESULTS: Sixteen factors were included in the characteristics of the eligible subjects. LASSO showed that five independent predictors including working posture, exercising hours per week, Tuffier's line, six lumbar vertebrae anomaly, and lumbar lordosis angle were the independent risk factors of low back pain in young population, which were identified by multivariate logistic regression analysis and were used to establish the nomogram. The AUC values of the nomogram were 0.867 (95% CI: 0.809-0.924) and 0.868 (95% CI: 0.775-0.961) in the training and validation set, respectively. The calibration curve revealed that the prediction model of the nomogram was greatly consistent with the actual observation. In addition, the DCA indicated that the nomogram was clinically useful. CONCLUSIONS: Working posture, exercising hours per week, Tuffier's line, six lumbar vertebrae anomaly, and lumbar lordosis angle are identified as independent predictors of non-specific low back pain in young population. And the nomogram based on the above five predictors can accurately predict the risk of low back pain in young people.

Polarization due to emergent polarity in elemental semiconductor thinfilms under bending.

Polarization via strain engineering provides a facial way to functionalize materials. We investigate the origin of electronic polarization in the bent elemental semiconductor thinfilms by combining analytical modeling with quantum mechanical simulation. A bond orbital model reveals a polarity of covalent bonds induced by strain gradient such that polarization along the strain gradient dimension can be induced, giving rise to the flexoelectric effect. At strain gradient1/R=0.01 nm-1, the net charge differences between the two sides are5×10-4e,2.5×10-3eand7.2×10-3efor C, Si and Ge films respectively. On the other hand, due to the emergent bond polarity, the polarization can be effectively tuned by normal strain applied to the bent film, mimicking the piezoelectric effect. Simulations using the generalized Bloch theorem strongly support this revelation. Findings have important implications for delineating the formation of polarization and related phenomena in semiconductors.

Retraction Note: MicroRNA-132 stimulates the growth and invasiveness of trophoblasts by targeting DAPK-1.

The article "MicroRNA-132 stimulates the growth and invasiveness of trophoblasts by targeting DAPK-1, by Y.-P. Wang, P. Zhao, J.-Y. Liu, S.-M. Liu, Y.-X. Wang, published in Eur Rev Med Pharmacol Sci 2020; 24 (19): 9837-9843-DOI: 10.26355/eurrev_202010_23193-PMID: 33090386" has been retracted by the authors due to some inaccuracies in the miRNA-132 primer sequence. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/23193.

Identification of Malus halliana R2R3-MYB gene family under iron deficiency stress and functional characteristics of MhR2R3-MYB4 in Arabidopsis thaliana.

Iron (Fe) is an essential element for plant growth and development. Fe deficiency can trigger leaf chlorosis and reduce fruit yield. Therefore, it is necessary to explore transcription factors in response to Fe deficiency stress. A total of 29 MhR2R3-MYB transcription factors were identified based on the transcriptome of Malus halliana under Fe deficiency stress. A comprehensive analysis of physical and chemical properties, gene structures, conserved motif composition, evolutionary relationship and chromosome distribution was performed. Subsequently, based on the transcriptome, 14 genes with the most significant expression under Fe deficiency stress were screened for qRT-PCR verification. Among them,the functional characteristics of MhR2R3-MYB4 (MD05G1089600) were further studied in Arabidopsis thaliana. Expression of 13 out of these 14 genes was upregulated, only one was downregulated. Maximum upregulation of MhR2R3-MYB4 under Fe deficiency was 36.39-fold and 58.21-fold compared with day 0 in leaves and roots, respectively. Overexpression of MhR2R3-MYB4 enhanced tolerance to Fe deficiency in A. thaliana and led to multiple biochemical changes: transgenic lines have higher chl a, chl b and Fe2+ content, higher enzyme activity (SOD, POD, CAT and FCR) and lower chlorosis than the wild type in Fe deficiency conditions. We suggest that MhR2R3-MYB4 plays an important part in Fe deficiency stress, which may contribute to improve Fe deficiency tolerance of apple in future.

Hsa_circ_0000337 promotes proliferation, migration and invasion in glioma by competitively binding miRNA-942-5p and thus upregulates MAT2A.

OBJECTIVE: CircRNAs are vital factors involved in the pathological processes. This study aims to elucidate the biological functions of hsa_circ_0000337 in affecting the malignant progress of glioma. PATIENTS AND METHODS: Relative levels of hsa_circ_0000337 in 45 cases of glioma and 24 cases of normal tissues were tested. The correlation between hsa_circ_0000337 and clinical features of glioma was assessed. Proliferative and metastatic abilities of U87 and U251 cells regulated by hsa_circ_0000337 were examined by 5-Ethynyl-2'-deoxyuridine (EdU) and transwell assay, respectively. Potential molecular mechanism of hsa_circ_0000337 on regulating glioma cell functions was clarified by bioinformatic analysis, which was further verified through rescue experiments. RESULTS: Hsa_circ_0000337 was highly expressed in glioma cases. Its level was correlated to poor prognosis of glioma. In vitro experiments obtained the conclusion that hsa_circ_0000337 accelerated proliferative and metastatic abilities of glioma cells. Serving as a ceRNA, hsa_circ_0000337 sponged miRNA-942-5p to upregulate MAT2A, thus inducing the malignant phenotypes of glioma. CONCLUSIONS: Hsa_circ_0000337/miRNA-942-5p / MAT2A axis is responsible for the deterioration of glioma. Hsa_circ_0000337 may be a potential therapeutic target for glioma.

LncRNA-TMPO-AS1 promotes apoptosis of osteosarcoma cells by targeting miR-329 and regulating E2F1.

OBJECTIVE: This study aims to explore the clinical value and mechanism of lncRNA-TMPO-AS1 in osteosarcoma. PATIENTS AND METHODS: We collected 51 samples of cancer tissues and 51 samples of matched adjacent tissues from 51 patients with osteosarcoma undergoing surgery in our hospital from February 2018 to February 2019. The expression of TMPO-AS1 in tissue samples was tested to analyze its value in the diagnosis and prognosis prediction of osteosarcoma. We transfected osteosarcoma cells with stable and transient vectors containing overexpressed or inhibited genes. Then, we measured cell proliferation by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay, cell invasion by transwell assay, and cell apoptosis by flow cytometry (FCM). The relationship between TMPO-AS1 and miR-329 or between miR-329 and E2F1 was determined by the Dual-Luciferase reporter (DLR) assay. RESULTS: TMPO-AS1 was up-regulated in osteosarcoma cells. Serum TMPO-AS1 could work as a diagnostic marker for osteosarcoma, with an area under the receiver operating characteristic (ROC) curve (AUC) of more than 0.8. Osteosarcoma cells were transfected with siRNA-TMPO-AS1, pcDNA3.1-TMPO-AS1, miR-329-mimics, and miR-329-inhibitor. The results revealed that inhibited TMPO-AS1/overexpressed miR-329/inhibited E2F1 suppressed the proliferation and invasion of osteosarcoma cells and enhanced cell apoptosis. The DLR assay demonstrated that TMPO-AS1 could target miR-329 and miR-329 could target E2F1. In vitro experiments revealed that TMPO-AS1 could regulate the progression of epithelial-mesenchymal transition (EMT) in osteosarcoma through the miR-329/E2F1 axis. CONCLUSIONS: TMPO-AS1 can function as a diagnostic and prognostic marker for osteosarcoma. LncRNA-TMPO-AS1 can promote apoptosis of osteosarcoma cells by targeting miR-329 and regulating E2F1, which is a potent treatment option for osteosarcoma.

MicroRNA-132 stimulates the growth and invasiveness of trophoblasts by targeting DAPK-1.

OBJECTIVE: The purpose of this study was to elucidate the regulatory effects of microRNA-132 on the growth and invasiveness of trophoblasts, thus influencing the development of preeclampsia (PE). PATIENTS AND METHODS: Placenta tissues from 24 PE pregnancies and 24 healthy pregnancies were collected. Expression levels of microRNA-132 and DAPK-1 in collected placenta tissues were detected. Then, the regulatory effects of microRNA-132 and DAPK-1 on expression levels of apoptosis-associated genes, viability, and invasiveness in trophoblasts were assessed. Finally, through Dual-Luciferase reporter assay, the binding relationship between microRNA-132 and DAPK-1 was determined. RESULTS: The results showed that microRNA-132 was downregulated in placenta of PE pregnancies, while DAPK-1 was upregulated. Overexpression of microRNA-132 stimulated viability and invasiveness, but inhibited apoptosis in trophoblasts. Besides, it was found that DAPK-1 was the target gene binding microRNA-132, and a negative correlation was identified between their expression levels. Notably, the overexpression of DAPK-1 inhibited viability and invasiveness, but stimulated apoptosis in trophoblasts. CONCLUSIONS: MicroRNA-132 stimulates proliferative and invasive capacities and inhibits apoptosis in trophoblasts by targeting DAPK-1.

Carbachol alleviates myocardial injury in septic rats through PI3K/AKT signaling pathway.

OBJECTIVE: To explore the effect of carbachol on myocardial injury in septic rats, and to further study its influence on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway. MATERIALS AND METHODS: A total of 48 healthy male Sprague-Dawley rats were randomly divided into sham group (n=16), model group (n=16), and carbachol group (n=16). The rat model of sepsis was established via cecal ligation and puncture. Carbachol was intraperitoneally injected (10 µg/kg) immediately after operation in carbachol group, and no cecal ligation was performed in sham group. At 48 h after operation, the survival rate of rats in each group was recorded, the activity of plasma creatine kinase-MB (CK-MB) was detected, and the cardiac function in each group was determined. Moreover, the heart was isolated, and the myocardial tissues were taken to detect the apoptosis level using the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) apoptosis kit. The content of inflammatory factors in myocardial tissues was determined using enzyme-linked immunosorbent assay (ELISA) kits, and the expression levels of apoptosis-related proteins and the PI3K/AKT signaling pathway-related proteins were detected via Western blotting. RESULTS: Carbachol could significantly raise the survival rate of septic rats (p<0.01), remarkably decrease the activity of CK-MB (p<0.01), markedly reduce the left ventricular internal diameter at end-systole (LVIDs), and markedly increase the left ventricular ejection fraction (LVEF, %) and left ventricular fractional shortening (LVFS, %). Besides, carbachol could evidently lower the apoptosis level of myocardial cells of septic rats (p<0.01), reduce the content of inflammatory factors including tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and IL-6 (p<0.01), notably decrease the expression of Caspase-3 in myocardial tissues (p<0.01), remarkably increase the expression of Bcl-2/Bax (p<0.01), and distinctly inhibit the expressions of phosphorylated (p-)PI3K, p-AKT, Nod-like receptor protein 3 (NLRP3), and Caspase-1 (p<0.01). CONCLUSIONS: Carbachol can reduce the release of inflammatory factors in myocardial cells, the expression of apoptotic proteins and the apoptosis of myocardial cells, and improve the cardiac function and survival rate of septic rats by inhibiting the PI3K/AKT signaling pathway.

Utilizing integrating network pharmacological approaches to investigate the potential mechanism of Ma Xing Shi Gan Decoction in treating COVID-19.

Beginning in December 2019, coronavirus disease 2019 (COVID-19), due to 2019-nCoV infection, emerged in Wuhan and spread rapidly throughout China and even worldwide. Employing combined therapy of modern medicine and traditional Chinese medicine has been proposed, in which Ma Xing Shi Gan Decoction (MXSGD) was recommended as a basic prescription and applied widely in the clinical treatment of COVID-19. We investigated the underlying mechanism of MXSGD in treating COVID-19 utilizing the approaches of integrating network pharmacology. A total of 97 active ingredients of MXSGD were screened out, and 169 targets were predicted. The protein-protein interaction network exhibited hub targets of MXSGD, such as Heat shock protein 90, RAC-alpha serine/threonine-protein kinase, Transcription factor AP-1, Mitogen-activated protein kinase 1, Cellular tumor antigen p53, Vascular endothelial growth factor A, and Tumour necrosis factor. Gene Ontology functional enrichment analysis demonstrated that the biological processes altered within the body after taking MXSGD were closely related to the regulation of such processes as the acute inflammatory response, chemokine production, vascular permeability, response to oxygen radicals, oxidative stress-induced apoptosis, T cell differentiation involved in the immune response, immunoglobulin secretion, and extracellular matrix disassembly. KEGG enrichment analysis indicated that the targets of MXSGD were significantly enriched in inflammation-related pathways, immunomodulation-related pathways, and viral infection-related pathways. The therapeutic mechanisms of MXSGD on COVID-19 may primarily involve the following effects: reducing inflammation, suppressing cytokine storm, protecting the pulmonary alveolar-capillary barrier, alleviating pulmonary edema, regulating the immune response, and decreasing fever.

Circular RNA circDENND4C facilitates proliferation, migration and glycolysis of colorectal cancer cells through miR-760/GLUT1 axis.

OBJECTIVE: Colorectal cancer is a common malignant tumor of the digestive tract, and its incidence is closely related to lifestyle inheritance and colorectal adenoma. Circular RNA (circRNA) has been proved to participate in the progression of colorectal cancer cells. Our study aimed to investigate the function and the underlying mechanism of circRNA circDENND4C in colorectal cancer cells. PATIENTS AND METHODS: The expression of circDENND4C, glucose transporter 1 (GLUT1), and miR-760 was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Western blot was used to measure the protein levels of GLUT1, the proliferation-related protein (Cyclin D1) and matrix metallopeptidase 9 (MMP-9). Cell Counting Kit-8 (CCK-8) assay and transwell assay were performed to evaluate cell proliferation and migration. The glucose uptake and lactate production were detected by the corresponding kits. The targets between circDENND4C and miR-760 and miR-760 and GLUT1 were predicted by starBase 3.0 and TargetScan, and then confirmed by Dual-Luciferase reporter assay. Animal experiment revealed the effect of circDENND4C on colorectal cancer cells in vivo. RESULTS: The expression of circDENND4C and GLUT1 was upregulated in colorectal cancer tissues and cells. Functionally, the knockdown of circDENND4C suppressed proliferation, migration, and glycolysis of colorectal cancer cells. Similarly, silence of GLUT1 also inhibited cell proliferation, migration, and glycolysis. Notably, the overexpression of GLUT1 reversed the functional effects of circDENND4C knockdown on colorectal cancer cells. More importantly, miR-760 acted as a direct target of circDENND4C, and miR-760 could bind to GLUT1, and circDENND4C regulated GLUT1 by sponging miR-760. Finally, circDENND4C knockdown decreased the growth of colorectal cancer cells in vivo. CONCLUSIONS: CircRNA circDENND4C accelerated proliferation, migration, and glycolysis of colorectal cancer cells through regulating GLUT1 by sponging miR-760.

Circular RNA_LARP4 inhibits the progression of non-small-cell lung cancer by regulating the expression of SMAD7.

OBJECTIVE: Researchers have uncovered the importance of circular RNAs (circ) in malignant tumors. Circ LARP4 has been found to serve as a tumor suppressor gene in gastric cancer. However, the exact function of circ LARP4 in non-small-cell lung cancer (NSCLC) has not been fully elucidated. The aim of this study was to uncover the role of circ LARP4 in the tumorigenesis of NSCLC. PATIENTS AND METHODS: Expression level of circ LARP4 in NSCLC tissues was detected through Real Time-quantitative Polymerase Chain Reaction (RT-qPCR). Subsequently, the association between expression and patients' prognosis was analyzed. Circ LARP4 lentivirus was constructed and transfected into NSCLC cells. The effect of circ LARP4 on NSCLC cell migration and invasion was detected by function assays. Furthermore, Western blot was performed to analyze the expression of predicted protein of circ LARP4. RESULTS: Compared with adjacent tissues, circ LARP4 was lowly expressed in NSCLC tissues. Meanwhile, expression of circ LARP4 was associated with the prognosis of NSCLC patients. Downregulated circ LARP4 was found in NSCLC cell lines as well. The migration and invasion abilities of NSCLC cells were significantly inhibited via overexpression of circ LARP4. SMAD7, the predicted protein of circ LARP4, increased remarkably via overexpression of circ LARP4. CONCLUSIONS: Circ LARP4 could suppress the metastasis of NSCLC by up-regulating SMAD7.

MiRNA-92a promotes cell proliferation and invasion through binding to KLF4 in glioma.

OBJECTIVE: Glioma is one of the most frequent brain tumors in adults, and it has a low 5-year survival rate. MicroRNA-92a (miR-92a) has been reported to be upregulated and acted as an oncogene in many cancers. The purpose of this study was to explore the molecular mechanisms of miR-92a and kruppel-like factor 4 (KLF4) in glioma. PATIENTS AND METHODS: Western blotting assay and quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) were applied to calculate the relative expression of interest proteins and mRNAs. Luciferase ability assay was conducted to evaluate whether miR-92a was targeting to KLF4. RESULTS: A higher expression of miR-92a was observed in glioma tissues compared with the corresponding adjacent non-tumor tissues. The upregulation of miR-92a predicted poor prognostic characteristics of glioma. The overexpression miR-92a significantly promoted cell proliferation an invasion, while the knockdown of miR-92a presented the opposite results. MiR-92a bound to KLF4 and mediated the expression of KLF4 in glioma cells. The knockdown of miR-92a inhibited cell invasion-mediated EMT. Furthermore, the knockdown of miR-92a suppressed cell proliferation through the KLF4/AKT/mTOR signal pathway. CONCLUSIONS: MiR-92a promoted the proliferation through the KLF4/AKT/mTOR signal pathway in glioma. The newly identified miR-92a/KLF4/AKT/mTOR axis provides novel insight into the pathogenesis of glioma.

LncRNA MEG3 promotes proliferation and differentiation of osteoblasts through Wnt/ß-catenin signaling pathway.

OBJECTIVE: We aimed at detecting the expression of long non-coding ribonucleic acid (lncRNA) maternally expressed 3 (MEG3) in the serum of fracture patients, and at investigating its impacts on the proliferation and differentiation of osteoblasts and the specific molecular mechanism of action. PATIENTS AND METHODS: The serum samples of 48 fracture patients diagnosed in our hospital (Fracture group) and 30 healthy people receiving physical examination (Health group) were collected. The expression level of serum lncRNA MEG3 in Fracture group and Health group was measured via reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, a small interfering RNA (siRNA) was applied to construct mouse osteoblast cell line MC3T3-E1 with a stable knockout of MEG3. The growth status of the cell was observed, and the impacts of MEG3 knockout on the osteoblast proliferation were determined using cell counting kit-8 (CCK-8), a proliferation activity detection kit. Meanwhile, 5-ethynyl-2'-deoxyuridine (EdU) staining was applied to detect the proportion of EdU positive cells in the osteoblasts in Control group and MEG3 knockout group (MEG3 siRNA group). In addition, RT-PCR was performed to measure the messenger RNA (mRNA) levels of differentiation-related genes. Finally, RT-PCR and Western blotting assay were adopted to analyze the expression of the Wnt/ß-catenin signaling pathway. RESULTS: The expression of serum lncRNA MEG3 in fracture patients was increased markedly (p<0.05). Results of in-vitro cell experiment indicated that intervention with MEG3 siRNA could obviously promote the proliferation and differentiation of osteoblast cell line MC3T3-E1. The results of RT-PCR and Western blotting assay revealed that the role of MEG3 in promoting differentiation and proliferation might be mediated by the activation of the Wnt/ß-catenin signaling pathway in osteoblasts. CONCLUSIONS: LncRNA MEG3 can promote the proliferation and differentiation of osteoblasts by activating the Wnt/ß-catenin signaling pathway, so it is expected to become a new target for accelerating the fracture healing.

MiR-124 inhibits spinal neuronal apoptosis through binding to GCH1.

OBJECTIVE: To explore the effect of micro ribonucleic acid (miR)-124 on the spinal neuronal apoptosis and to explore its related mechanism. MATERIALS AND METHODS: The rat model of spinal cord injury (SCI) was established, agomir-124 was injected intrathecally and the effect of agomir-124 on motor function recovery of rats was evaluated using the Basso-Beattie-Bresnahan (BBB) score. The gene expression levels of miR-124 and GTP-cyclohydrolase 1 (GCH1) in spinal cord tissues were detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR), and the correlation between them was detected using the Pearson correlation coefficient. Then, the direct interaction between miR-124 and GCH1 mRNA was detected using the TargetScan software and luciferase reporter assay. The changes in apoptosis in each group were examined via flow cytometry and Western blotting. Moreover, the changes in the tetrahydrobiopterin (BH4) content in each group were detected via high-performance liquid chromatography, and the changes in the nitrite level in the supernatant in each group were detected using the Griess reagent. Finally, the changes in the activity of the inducible nitric oxide synthase (iNOS) protein were detected using the iNOS kit. RESULTS: Compared with that in the model group, the BBB score was significantly increased in agomir-124 group at 21, 28, 35 and 42 d. In the agomir-124 group, the relative expression level of miR 124 in spinal cord tissues was significantly increased at 7-28 d and reached the peak at 21 d, while the mRNA level of GCH1 in spinal cord tissues declined and touched the bottom at 21 d. According to the Pearson correlation coefficient, there was a significant negative correlation between the expression of miR-124 and mRNA expression of GCH1 (r =- 0.87, p = 1.5e-6). It was found in the prediction using TargetScan software that GCH1 might be a potential target for miR-124, which was further verified by the luciferase reporter assay. The results of flow cytometry and Western blotting showed that miR-124 significantly reduced the LPS-induced primary spinal neuronal apoptosis, while the miR-124 inhibitor remarkably increased the primary spinal neuronal apoptosis. Moreover, it was also found that the knockout of GCH1 reduced the LPS-induced spinal neuronal apoptosis. In addition, the GCH1 overexpression assay revealed that miR-124 inhibited spinal neuronal apoptosis by suppressing the GCH1 expression. LPS + miR-124 remarkably decreased the BH4 content, nitrite level, and iNOS activity while LPS + miR-124 + GCH1 remarkably increased the BH4 content, nitrite level, and iNOS activity. CONCLUSIONS: MiR-124 inhibits neuronal apoptosis in SCI by binding to GCH1. The results in the present study may provide a new mechanism for the therapeutic effect of miR-124, and miR-124 may have a potential therapeutic value in the treatment of SCI in the future.

Zyxin: a mechanotransductor to regulate gene expression.

OBJECTIVE: Cells answer to biochemical, electrical and mechanical signals in the environment, which regulate their behavior. Mechanical signals can propagate through mechanically stiff structures like focal adhesions (FAs). Zyxin, a LIM domain protein, is localized primarily at focal adhesion plaques. Growing evidence suggests that zyxin is a vital mechanotransductor to regulate the gene expression. In this review, we summarize the features of zyxin and the molecular mechanism of how zyxin participate in the cellular activity. MATERIALS AND METHODS: An English-language literature search is based on a keyword-based query of multiple databases (MEDLINE, Embase) and bibliographies from identified publications. The references in the selected paper are also considered as an additional source of data. The search was last updated in April 2018; no limitations are applied. RESULTS: Zyxin enhances actin polymerization with the aid of Enabled (Ena)/vasodilator-stimulated phosphoprotein (VASP) proteins in response to mechanical tension, to perform its role in stress fibers (SFs) remodeling and repair. Zyxin can translocate from focal adhesions (FAs) to the nucleus responds to stretch, and regulate gene transcription by interaction with transcription factors like nuclear matrix protein 4 (NMP4). Misregulation of nuclear functions of zyxin appears to be associated with pathogenic effects and diseases, such as prostate cancer and non-small-cell lung cancer. CONCLUSIONS: Zyxin is a crucial ingredient of the cellular mechanotransducing system and can modulate the gene expression. Given its clinical relevance, zyxin is also a promising target for the diagnosis and treatment of certain diseases. Understanding the role of zyxin in force sensing and gene expression regulating provides a compelling challenge for future biomechanics studies, and offers attractive evidence for zyxin as a potential diagnostic marker and therapeutic target for clinical diseases.