[Establishment of a CFTR-based detection method for the second messenger cAMP in the cytoplasm].

Objective: To establish a detection method based on Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) that can sensitively detect the second messenger cyclic AMP (cAMP) in the cytoplasm. Methods: The eukaryotic expression vectors of CFTR and YFP-H148Q / I152L were constructed respectively. FRT cells co-expressing CFTR and YFP-H148Q / I152L were obtained by liposome transfection. The expression of CFTR and YFP-H148Q / I152L in FRT cells was observed by an inverted fluorescence microscopy, and flow cytometry was used to detect the purity of cells; The cell model was identified by the fluorescence quenching kinetics test. The validation of the cell model which could screen CFTR modulators was verified by the fluorescence quenching kinetics experiments. The radioimmunoassay was used to detect the cAMP concentration in cytoplasm after adding CFTR activator. Results: The results of the inverted fluorescence microscope showed that CFTR was expressed in the cell membrane and YFP-H148Q / I152L was expressed in the cytoplasm of FRT cells. The FRT cell model stably co-expressing ANO1 and YFP-H148Q / I152L was successfully constructed. The model could screen CFTR modulators, and the slope of fluorescence change and the concentration of CFTR modulators were in a dose-dependent manner. The slope of the fluorescence could reflect the cAMP concentration in the cytoplasm. The cell model could sensitively detect the intracellular cAMP concentration. Conclusion: The cell model could efficiently and sensitively detect the second messenger cAMP concentration in the cytoplasm, and it provided a simple and efficient method for the study of other targets associated cAMP signal.

miR-139-5p Loss-Mediated WTAP Activation Contributes to Hepatocellular Carcinoma Progression by Promoting the Epithelial to Mesenchymal Transition.

Background: Hepatocellular carcinoma (HCC) is a primary aggressive gastrointestinal neoplasm that affects patients worldwide. It has been shown that Wilms' tumor 1-associating protein (WTAP) is frequently upregulated in various cancers. However, the potential role of WTAP in HCC remains largely unknown. Methods: The expression levels of WTAP in human HCC tissues were determined by the western blotting and immunohistochemical (IHC) staining. A correlation between the WTAP expression, clinicopathological features, and the HCC prognosis was analyzed. The WTAP expression was silenced by short hairpin RNA (shRNA), and effects of the knockdown of WTAP on the proliferation and invasion of HCC cells were assessed. The microRNAs (miRNAs) involved in the regulation of the WTAP expression were identified by a bioinformatics analysis and further confirmed by in vitro assays. Results: The expression levels of WTAP in liver cancer tissues were significantly elevated and compared with those in the adjacent normal tissues and significantly correlated with the clinical stage and prognosis in patients with HCC. Further investigation revealed that the knockdown of WTAP drastically suppressed HCC cell proliferation and invasion abilities. Luciferase reporter assay and validation experiments confirmed that WTAP was a direct target of miR-139-5p. Moreover, the overexpression of WTAP could partly abolish the inhibitory effects of miR-139-5p on the HCC cell growth and invasion. Mechanistically, we revealed that the miR-139-5p/WTAP axis regulated the HCC progression by controlling the epithelial to mesenchymal transition (EMT). Conclusions: In summary, the results indicate that WTAP is a potential oncogene in HCC and miR-139-5p negatively regulates the WTAP expression. MiR-139-5p/WTAP can be utilized as a potential therapeutic target for HCC.

[Vesselplasty for the treatment of spinal metastases complicated by posterior wall destruction of vertebral body].

OBJECTIVE: To evaluate the early clinical efficacy and safety of vesselplasty for the treatment of spinal metastases complicated by posterior wall destruction of vertebral body. METHODS: The clinical data of 19 patients(21 segments) with spinal metastases complicated by posterior wall destruction of vertebral body treated from January 2016 to January 2017 were retrospectively analyzed. There were 15 males and 4 females, aged 40 to 85 years old with a mean of (66.00±10.25) years . All patients had severe low back pain before the operation, which were diagnosed by CT as damage-type metastatic tumor of the vertebral posterior wall. All patients were treated by vesselplasty technique. Nineteen vertebrae received percutaneous unilateral pedicle puncture and two vertebrae received percutaneous bilateral pedicle puncture. VAS, ODI were recorded before operation, 1 d and 3 d after operation respectively. X-ray and CT scan were used to observe bone cement leakage and complications. RESULTS: All the operations were successful and postoperative pain was significantly relieved. Postoperative VAS score and ODI of the two groups were significantly improved (P<0.05). A small amount of bone cement leakage occurred in one vertebral body, which was a vertebral venous plexus leakage, but no clinical symptoms after operation. CONCLUSION: Vesselplasty for the treatment of spinal metastases complicated by posterior wall destruction of vertebral body can significantly reduce the symptoms of thoracolumbar back pain, improve the quality of life, reduce the incidence of bone cement leakage, and has high clinical efficacy and safety.

Cardioprotective role of metformin against sodium arsenite-induced oxidative stress, inflammation, and apoptosis.

Arsenic is a universal component and a notable natural poison. Its exposure to people occurs primarily through natural, therapeutic, and occupational sources. This investigation intended to discover the defensive impact of metformin (100 and 150 mg/kg body weight) against sodium arsenite (SA)-induced cardiotoxicity in experimental animals. The study was conducted on Sprague Dawley rats (n = 50), which were separated into five different groups as follows: control, met-150, SA, SA + met-100, and SA + met-150. The results demonstrated that SA caused a significant increase in the level of malondialdehyde and also reduced activities of antioxidative enzyme. SA similarly increased inflammatory reactions by increasing the level of interleukin-6, tumor necrosis factor-α, and interleukin-1ß. In addition, SA provoked the apoptosis by expanding the p53 and Bax levels, terminal deoxynucleotidyl transferase dUTP nick end labeling, and expression of caspase-3. SA altered the histological integrity of cardiac tissue and 8-hydroxy-2'-deoxyguanosine expression. In conclusion, metformin significantly reduced oxidative stress, inflammatory reaction, and apoptotic pathway. The present investigation showed that metformin has a cardioprotective impact because of its protective role against oxidation, inflammation, and apoptosis.

Novel near-infrared spectrum analysis tool: Synergy adaptive moving window model based on immune clone algorithm.

This paper presents a novel spectrum analysis tool named synergy adaptive moving window modeling based on immune clone algorithm (SA-MWM-ICA) considering the tedious and inconvenient labor involved in the selection of pre-processing methods and spectral variables by prior experience. In this work, immune clone algorithm is first introduced into the spectrum analysis field as a new optimization strategy, covering the shortage of the relative traditional methods. Based on the working principle of the human immune system, the performance of the quantitative model is regarded as antigen, and a special vector corresponding to the above mentioned antigen is regarded as antibody. The antibody contains a pre-processing method optimization region which is created by 11 decimal digits, and a spectrum variable optimization region which is formed by some moving windows with changeable width and position. A set of original antibodies are created by modeling with this algorithm. After calculating the affinity of these antibodies, those with high affinity will be selected to clone. The regulation for cloning is that the higher the affinity, the more copies will be. In the next step, another import operation named hyper-mutation is applied to the antibodies after cloning. Moreover, the regulation for hyper-mutation is that the lower the affinity, the more possibility will be. Several antibodies with high affinity will be created on the basis of these steps. Groups of simulated dataset, gasoline near-infrared spectra dataset, and soil near-infrared spectra dataset are employed to verify and illustrate the performance of SA-MWM-ICA. Analysis results show that the performance of the quantitative models adopted by SA-MWM-ICA are better especially for structures with relatively complex spectra than traditional models such as partial least squares (PLS), moving window PLS (MWPLS), genetic algorithm PLS (GAPLS), and pretreatment method classification and adjustable parameter changeable size moving window PLS (CA-CSMWPLS). The selected pre-processing methods and spectrum variables are easily explained. The proposed method will converge in few generations and can be used not only for near-infrared spectroscopy analysis but also for other similar spectral analysis, such as infrared spectroscopy.