[Effects and mechanism of mitochondrial transcription factor A and cytochrome c oxidase pathway in the energy production of hypoxic cardiomyocytes of rats regulated by tumor necrosis factor receptor associated protein 1].

Objective: To investigate the effects and mechanism of mitochondrial transcription factor A (TFAM) and cytochrome c oxidase (COX) pathway in the energy production of hypoxic cardiomyocytes of rats regulated by tumor necrosis factor receptor associated protein 1 (TRAP1). Methods: The cardiomyocytes were isolated from 135 neonatal Sprague-Dawley rats (aged 1-3 d) and cultured for the following experiments. (1) Cells were collected and divided into normoxia blank control (NBC) group, hypoxia blank control (HBC) group, hypoxia+ TRAP1 over-expression control (HTOC) group, and hypoxia+ TRAP1 over-expression (HTO) group according to the random number table (the same grouping method below), with 1 bottle in each group. Cells in NBC group were cultured routinely, cells in HBC group were cultured in hypoxic condition for 6 hours after routine culture, cells in HTOC and HTO groups were respectively added with TRAP1 over-expression empty virus vector and TRAP1 over-expression adenovirus vector virus suspension for transfection for 48 hours after routine culture and then cultured in hypoxic condition for 6 hours. The protein expression of TFAM of cells in each group was detected by Western blotting. (2) Cells were collected and divided into NBC, HBC, HTOC, HTO, HTO+ TFAM interference control (HTOTIC), and HTO+ TFAM interference (HTOTI) groups, with 1 well in each group. Cells in the former 4 groups were dealt with the same methods as the corresponding groups in experiment (1). Cells in HTOTIC and HTOTI groups were respectively added with TFAM interference empty virus vector and TFAM interference adenovirus vector virus suspension for transfection for 48 hours, and the other processing methods were the same as those in HTO group. The content of ATP of cells in each group was determined by ATP determination kit and microplate reader, and the COX activity of cells in each group was determined by COX activity assay kit and microplate reader. (3) Cells were collected and divided into NBC group, normoxia+ sodium azide (NSA) group, HBC group, and hypoxia+ sodium azide (HSA) group, with 1 well in each group. Cells in NBC and HBC groups were respectively dealt with the same methods as the corresponding groups in experiment (1). Cells in NSA and HSA groups were respectively added with 32 nmol sodium azide at 30 min before experiment or hypoxia, and then cells in HSA group were cultured in hypoxic condition for 6 hours. The content of ATP was determined by the same method as above. The above three experiments were repeated for three times. Data were statistically analyzed with one-way analysis of variance and least significant difference test. Results: (1) Compared with that in NBC group, the protein expression of TFAM of cells in HBC group was significantly decreased (P<0.01). Compared with that in HBC group or HTOC group, the protein expression of TFAM of cells in HTO group was significantly increased (P<0.01). (2) Compared with 0.552±0.041 and 1.99±0.15 in NBC group, the COX activity (0.270±0.044) and ATP content (1.09±0.11) of cells in HBC group were significantly decreased (P<0.01). Compared with 0.269±0.042 and 1.17±0.12 in HBC group and those in HTOC group, the COX activity (0.412±0.032 and 0.404±0.016) and ATP content (1.75±0.06 and 1.69±0.07) of cells in HTO and HTOTIC groups were significantly increased (P<0.01). Compared with those in HTO and HTOTIC groups, the COX activity (0.261±0.036) and ATP content (1.23±0.07) of cells in HTOTI group were significantly decreased (P<0.01). (3) Compared with that in NBC group, the ATP content of cells in NSA and NBC groups was significantly decreased (P<0.01). Compared with that in HBC group, the ATP content of cells in HSA group was significantly decreased (P<0.01). Conclusions: TRAP1 can increase the COX activity of cardiomyocytes by raising the expression of TFAM, and finally alleviate the impairment in energy production of cardiomyocytes caused by hypoxia.

CircRNA_009934 induces osteoclast bone resorption via silencing miR-5107.

OBJECTIVE: We aimed to explore the expression of circRNA_009934 in osteoclast, as well as its potential roles in regulating osteoclastogenesis and bone resorption via regulating miR-5107. MATERIALS AND METHODS: We performed qRT-PCR analysis to examine the expression of circRNA_009934 in osteoclast in distinctive stages. We used CCK-8 assay to detect the cell proliferation ability. Correlation analysis between the expression levels of circRNA_009934 and miR-5107 was performed using statistical analysis. Bioinformatics prediction was performed to predict the binding site of circRNA_009934 and miR-5107, subsequently followed by Luciferase assay for validation. The mice TRAF6 3'-UTR were cloned into the Luciferase reporter vector and miR-5107 binding mutants were constructed to validate the inhibited regulation of miR-5107 to the expression of TRAF6. RESULTS: Our results showed that expression of circRNA_009934 was increased during osteoclast differentiation. CircRNA_009934 expression was closely correlated with osteoclastogenesis and bone resorption activity. Bioinformatics prediction and Luciferase assay demonstrated that circRNA_009934 served as a ceRNA of miR-5107 and regulated its downstream TRAF6 expression. CONCLUSIONS: We first demonstrated that circRNA_009934 expression was increased in osteoclasts, which promoted osteoclastogenesis by serving as a ceRNA of miR-5107 and regulated the expression of TRAF6.

[The (18)F-FDG positron emission tomography integrated computed tomography associate with intravoxel incoherent motion in prediction of EGFR expression in lung cancer].

Objective: To discuss the diagnostic value potential of (18)F-FDG PET/CT and intravoxel incoherent motion (IVIM) in genotype EGFR of lung cancer. Methods: A total of 116 cases proved pathologically pulmonary space occupying lesions (72 males, 44 females; age range was 31-85 years; 37 cases of high differentiation; 53 cases of middle differentiation, 26 cases of low differentiation) were prospectively collected from August 2017 to March 2019 in Henan Provincial People's Hospital. EGFR gene detection was performed in 50 patients (wild type 20 cases, mutant 30 cases). Whole body PET/CT and lung MR-imaging were performed before treatment in all patients. Comparison of PET/CT, IVIM semi-quantitative parameters between positive and negative of EGFR by Student-t test or Mann-Whitney U test. Evaluation diagnostic efficacy of each parameter to EGFR by drawing ROC curves. The optimum diagnostic threshold, specificity, sensitivity, positive predictive value, negative predictive value and the diagnostic accuracy were calculated. The diagnostic accuracy of (18)F-FDG PET/CT combined with IVIM was calculated. Results: The quantitative parameters standard intake(SUV), apparent diffusion coefficient (ADC), true diffusion coefficient (D), pseudo-diffusion coefficient (D(*)), perfusion fraction (f) were 8.7±3.5, (1.59±0.26) ×10(-3) mm(2)/s, (1.04±0.12)×10(-3) mm(2)/s, (3.9±3.0)×10(-2) mm(2)/s, 0.43±0.16 and the above parameters of the wild group were 14.3±3.3, (1.34±0.26)×10(-3) mm(2)/s, (0.89±0.12)×10(-3) mm(2)/s, (3.5±2.5)×10(-2) mm(2)/s, 0.40±0.11, respectively. The difference of quantitative parameters SUV (t=4.196, P=0.0001), ADC (t=2.502, P=0.0018), D (t=3.158, P=0.006) between the EGFR mutant group and the wild group was statistically significant. The difference of quantitative parameters D(*) (t=0.361, P=0.721), f (t=0.627, P=0.536) was not statistically significant. ROC curve was drawn and the area under the curves for diagnosis of EGFR mutations by SUV, ADC, D, D(*), f was 0.880, 0.755, 0.820, 0.575, 0.550. The diagnostic accuracy of these parameters above mentioned was 80.0%, 76.7%, 73.3%, 66.7%, 53.3% respectively. The diagnostic accuracy of SUV combined with ADC, D values was 83.3% and 80.0%. Conclusion: PET/CT and IVIM have certain diagnostic value for EGFR typing of lung cancer gene.

[Advances in the research of diagnosis techniques of burn depth].

The accurate diagnosis of burn depth is one of the important problems in the field of burn surgery. The diagnosis accuracy rate is directly related to the treatment plan and effect. The existed clinical diagnosis methods mainly depend on the experience of burn surgeon, making the accuracy rate from 50% to 65%. In order to improve the accuracy rate of clinical burn depth diagnosis, a large number of diagnosis methods based on imaging are proposed, however, all of the methods are still in the stage of experimental research. In this paper, the research advances on the diagnosis techniques of burn depth are summarized, both the advantages and the shortcomings are pointed, and the development trend of diagnosis techniques of burn depth is expected.