Increased serum extrachromosomal circular DNA SORBS1circle level is associated with insulin resistance in patients with newly diagnosed type 2 diabetes mellitus.

BACKGROUND: Extrachromosomal circular DNAs (eccDNAs) exist in human blood and somatic cells, and are essential for oncogene plasticity and drug resistance. However, the presence and impact of eccDNAs in type 2 diabetes mellitus (T2DM) remains inadequately understood. METHODS: We purified and sequenced the serum eccDNAs obtained from newly diagnosed T2DM patients and normal control (NC) subjects using Circle-sequencing. We validated the level of a novel circulating eccDNA named sorbin and SH3-domain- containing-1circle97206791-97208025 (SORBS1circle) in 106 newly diagnosed T2DM patients. The relationship between eccDNA SORBS1circle and clinical data was analyzed. Furthermore, we explored the source and expression level of eccDNA SORBS1circle in the high glucose and palmitate (HG/PA)-induced hepatocyte (HepG2 cell) insulin resistance model. RESULTS: A total of 22,543 and 19,195 eccDNAs were found in serum samples obtained from newly diagnosed T2DM patients and NC subjects, respectively. The T2DM patients had a greater distribution of eccDNA on chromosomes 1, 14, 16, 17, 18, 19, 20 and X. Additionally, 598 serum eccDNAs were found to be upregulated, while 856 eccDNAs were downregulated in T2DM patients compared with NC subjects. KEGG analysis demonstrated that the genes carried by eccDNAs were mainly associated with insulin resistance. Moreover, it was validated that the eccDNA SORBS1circle was significantly increased in serum of newly diagnosed T2DM patients (106 T2DM patients vs. 40 NC subjects). The serum eccDNA SORBS1circle content was positively correlated with the levels of glycosylated hemoglobin A1C (HbA1C) and homeostasis model assessment of insulin resistance (HOMA-IR) in T2DM patients. Intracellular eccDNA SORBS1circle expression was significantly enhanced in the high glucose and palmitate (HG/PA)-induced hepatocyte (HepG2 cell) insulin resistance model. Moreover, the upregulation of eccDNA SORBS1circle in the HG/PA-treated HepG2 cells was dependent on generation of apoptotic DNA fragmentation. CONCLUSIONS: These results provide a preliminary understanding of the circulating eccDNA patterns at the early stage of T2DM and suggest that eccDNA SORBS1circle may be involved in the development of insulin resistance.

Identifying and Validating a Novel miRNA-mRNA Regulatory Network Associated with Prognosis in Lung Adenocarcinoma.

Objective Many studies have revealed the crucial roles of miRNA in multiple human cancers, including lung adenocarcinoma (LUAD). In this study, we sought to explore new miRNA-mRNA pairs that are associated with LUAD prognosis. Methods A novel miRNA-mRNA regulatory network associated with prognosis in LUAD was identified and validated using the bioinformatic tools including OncomiR database, StarBase, miRnet, GEPIA2, UALCAN. Results Twenty key miRNAs were compiled after the analysis of the expression and prognostic value in OncomiR and StarBase. Targeted mRNAs of these key miRNAs were predicted in miRnet, and the resulting mRNAs were also analyzed for their prognostic values and expression patterns in GEPIA2 and UALCAN, respectively. Further expression correlation analysis was performed in StarBase. Subsequently, a new miRNA-mRNA network was built, of which each RNA pair showed negative expression correlation, opposite expression pattern, and prognostic value. Protein-protein interaction network was under construction for the mRNAs, and 19 hub genes were determined. Enrichment analysis showed that "Cell Cycle, Mitotic" was the most significantly enriched term. Then, a miRNA-hub gene sub-network was built. We selected and validated the regulatory relationship of some miRNA-hub pairs, including hsa-miR-1976/RFC2, hsa-let-7c-5p/RFC2, hsa-let-7c-5p/ESPL1, hsa-let-7c-5p/CDC25A, and hsa-miR-101-3p/KIF2C. Moreover, over-expression of hsa-miR-1976 and hsa-let-7c-5p resulted in significant cell cycle arrest. Conclusions Our results determined new prognosis-associated miRNA-mRNA pairs and might shed further light on the mechanism via which miRNA-mRNA network influences prognosis in LUAD.

[Study on the Enhancement Intensity of Cd in Rice with Microwave-Assisted Laser-Induced Breakdown Spectroscopy].

As food safety problem has become the focus of attention all over the world, green detection methods of the contaminants in food is in accordance with the sustainable development of environment. Heavy metal pollutant Cd element in rice was used as the object of study in this work, laser induced breakdown spectroscopy (LIBS) and microwave assisted laser induced breakdown spectroscopy (MA-LIBS) were utilized to detect the blank and laboratory polluted rice samples respectively. The characteristic line of Cd Ⅰ 228.802 nm was employed as analytical line to discuss the enhancement effect of plasmas emission intensity for the analytical line of target element. Meanwhile, the actual concentration of Cd in rice was measured by anodic stripping voltammetry. The result displayed that LIBS can just detect the plasmas signals of the sample which contained 13.69 µg·g-1 cadmium for the laboratory polluted rice samples which concentration range from 2.16 to 13.69 µg·g-1, however, in the same experimental conditions, MA-LIBS can detect the plasmas signals of Cd in all of the contaminated rice samples successfully, and compared with LIBS, the plasmas emission intensity of Cd element was enhanced from 9 to 27 times. The results showed that the plasmas emission intensity of Cd element in rice can be enhanced effectively by MA-LIBS, and the detection sensitivity can be effectively improved.

[Influence of Spectral Pre-Processing on PLS Quantitative Model of Detecting Cu in Navel Orange by LIBS].

Cu in navel orange was detected rapidly by laser-induced breakdown spectroscopy (LIBS) combined with partial least squares (PLS) for quantitative analysis, then the effect on the detection accuracy of the model with different spectral data ptetreatment methods was explored. Spectral data for the 52 Gannan navel orange samples were pretreated by different data smoothing, mean centralized and standard normal variable transform. Then 319~338 nm wavelength section containing characteristic spectral lines of Cu was selected to build PLS models, the main evaluation indexes of models such as regression coefficient (r), root mean square error of cross validation (RMSECV) and the root mean square error of prediction (RMSEP) were compared and analyzed. Three indicators of PLS model after 13 points smoothing and processing of the mean center were found reaching 0. 992 8, 3. 43 and 3. 4 respectively, the average relative error of prediction model is only 5. 55%, and in one word, the quality of calibration and prediction of this model are the best results. The results show that selecting the appropriate data pre-processing method, the prediction accuracy of PLS quantitative model of fruits and vegetables detected by LIBS can be improved effectively, providing a new method for fast and accurate detection of fruits and vegetables by LIBS.

[Effect of Characteristic Variable Extraction on Accuracy of Cu in Navel Orange Peel by LIBS].

Heavy metals pollution in foodstuffs is more and more serious. It is impossible to satisfy the modern agricultural development by conventional chemical analysis. Laser induced breakdown spectroscopy (LIBS) is an emerging technology with the characteristic of rapid and nondestructive detection. But LIBS' s repeatability, sensitivity and accuracy has much room to improve. In this work, heavy metal Cu in Gannan Navel Orange which is the Jiangxi specialty fruit will be predicted by LIBS. Firstly, the navel orange samples were contaminated in our lab. The spectra of samples were collected by irradiating the peel by optimized LIBS parameters. The laser energy was set as 20 mJ, delay time of Spectral Data Gathering was set as 1.2 micros, the integration time of Spectral data gathering was set as 2 ms. The real concentration in samples was obtained by AAS (atom absorption spectroscopy). The characteristic variables Cu I 324.7 and Cu I 327.4 were extracted. And the calibration model was constructed between LIBS spectra and real concentration about Cu. The results show that relative error of the predicted concentrations of three relational model were 7.01% or less, reached a minimum of 0.02%, 0.01% and 0.02% respectively. The average relative errors were 2.33%, 3.10% and 26.3%. Tests showed that different characteristic variables decided different accuracy. It is very important to choose suitable characteristic variable. At the same time, this work is helpful to explore the distribution of heavy metals between pulp and peel.

[Determination of Cu in Shell of Preserved Egg by LIBS Coupled with PLS].

In this work, the content of copper in the shell of preserved eggs were determined directly by Laser induced breakdown spectroscopy (LIBS), and the characteristics lines of Cu was obtained. The samples of eggshell were pretreated by acid wet digestion, and the real content of Cu was obtained by atomic absorption spectrophotometer (AAS). Due to the test precision and accuracy of LIBS was influenced by a serious of factors, for example, the complex matrix effect of sample, the enviro nment noise, the system noise of the instrument, the stability of laser energy and so on. And the conventional unvariate linear calibration curve between LIBS intensity and content of element of sample, such as by use of Schiebe G-Lomakin equation, can not meet the requirement of quantitative analysis. In account of that, a kind of multivariate calibration method is needed. In this work, the data of LIBS spectra were processed by partial least squares (PLS), the precision and accuracy of PLS model were compared by different smoothing treatment and five pretreatment methods. The result showed that the correlation coefficient and the accuracy of the PLS model were improved, and the root mean square error and the average relative error were reduced effectively by 11 point smoothing with Multiplicative scatter correction (MSC) pretreatment. The results of the study show that, heavy metal Cu in preserved egg shells can be direct detected accurately by laser induced breakdown spectroscopy, and the next step batch tests will been conducted to find out the relationship of heavy metal Cu content in the preserved egg between the eggshell, egg white and egg yolk. And the goal of the contents of heavy metals in the egg white, egg yolk can be knew through determinate the eggshell by the LIBS can be achieved, to provide new method for rapid non-destructive testing technology for quality and satety of agricultural products.

[Quantitative analysis of Cu in water by collinear DP-LIBS].

The purpose of this research is to study the influence of double pulse laser induced breakdown spectroscopy (DP-LIBS) on the sensitivity of Cu in water. The water solution of Cu was tested by collinear DP-LIBS in this article. The results show that spectral intensity of Cu can be enhanced obviously by DP-LIBS, compared with single pulse laser induced breakdown spectroscopy (SP-LIBS). Besides, the experimental results were significantly impacted by delay time between laser pulse and spectrometer acquisition, delay time of double laser pulse and energy of laser pulse and so on. The paper determined the best conditions for DP-LIBS detecting Cu in water. The optimal acquisition delay time was 1 380 ns. The best laser pulse delay time was 25 ns. The most appropriate energy of double laser pulse was 100 mJ. Characteristic analysis of spectra of Cu at 324.7 and 327.4 nm was done for quantitative analysis. The detection limit was 3.5 microg x mL(-1) at 324.7 nm, and the detection limit was 4.84 microg x mL(-1) at 327.4 nm. The relative standard deviation of the two characteristic spectral lines was within 10%. The calibration curve of characteristic spectral line, established by 327.4 nm, was verified with 500 microg x mL(-1) sample. Concentration of the sample was 446 microg x mL(-1) calculated by the calibration curve. This research shows that the detection sensitivity of Cu in water can be improved by DP-LIBS. At the same time, it had high stability.

[Analysis of Cr in soil by LIBS based on conical spatial confinement of plasma].

The present study is to improve the sensitivity of detection and reduce the limit of detection in detecting heavy metal of soil by laser induced breakdown spectroscopy (LIBS). The Cr element of national standard soil was regarded as the research object. In the experiment, a conical cavity with small diameter end of 20 mm and large diameter end of 45 mm respectively was installed below the focusing lens near the experiment sample to mainly confine the signal transmitted by plasma and to some extent to confine the plasma itself in the LIBS setup. In detecting Cr I 425.44 nm, the beast delay time gained from experiment is 1.3 micros, and the relative standard deviation is below 10%. Compared with the setup of non-spatial confinement, the spectral intensity of Cr in the soil sample was enhanced more than 7%. Calibration curve was established in the Cr concentration range from 60 to 400 microg x g(-1). Under the condition of spatial confinement, the liner regression coefficient and the limit of detection were 0.997 71 and 18.85 microg x g(-1) respectively, however, the regression coefficient and the limit of detection were 0.991 22 and 36.99 microg x g(-1) without spatial confinement. So, this shows that conical spatial confinement can/improve the sensitivity of detection and enhance the spectral intensity. And it is a good auxiliary function in detecting Cr in the soil by laser induced breakdown spectroscopy.

[Experimental study on chromium in gannan navel orange by laser-induced breakdown spectroscopy].

It is a relatively new task to apply the laser induced breakdown spectroscopy (LIBS) to fruit samples. To apply LIBS technique in the field related to analysis of trace heavy metal element in fruits samples, we have done primary experiments using Gannan navel orange samples. The authors put the samples into different concentration gradient K2Cr2O7 solution and left it for 30 hours, and then we did the LIBS experiment, discriminated characteristic spectra of chromium element and recorded the peak intensity information. Weighing three grams of sample and determined chromium concentration in the samples by atomic absorption spectrophotometer using wet digestion. The calibration curve of the line intensities versus the concentrations of the Cr element was acquired by the Origin software. The authors found that the linear correlation coefficient is 0. 981 66. The calibration curve can be used for the quantitative analysis of chromium element with an unknown concentration in Gannan navel orange. The LIBS detection limit of Cr in the solution was 11.64 mg x g(-1) from the measured calibration curve. Experiment results showed that LIBS technique is a valid means for measuring and quantitatively analyzing the content of heavy metal elements in fruit samples.

[Quantitative analysis of Ba and Sr in soil using laser-induced breakdown spectroscopy].

Laser induced breakdown spectroscopy (LIBS) was applied to measure barium and strontium elements in soil. The emission spectrum was emitted as the plasma was cooling off, which was collected and analyzed by fiber spectrometer. Spectral lines of barium (BaII line at 455.41 nm) and strontium (SrI line at 460.73 nm) were separately used for the determining. In order to reduce the error, fitting of the spectra by using Lorenizian function was used, the datum was preprocessed and the net intensity value of the spectra was collected. A signal intensity in the proper range of the characteristic spectrum was chosen as the interior label. A calibration curve was drawn according to the ratio between the value of the analytical line and the interior scaling line. The correlation coefficient between the content and the intensity ratio of Ba and Sr were up to 0.990 0 and 0.990 6. The content of Ba and Sr was inversed by means of the intensity scaling method. As a result, the relative deviations between the content values and the standard values of Ba and Sr were 5.7% and 5.1% respectively.

[Determination of parameter range in detecting Cu of Gannan navel orange by LIBS setup].

In order to improve the precision and sensitivity of laser-induced breakdown spectroscopy device on detecting heavy metal, parameter optimization should be done. After selecting the initial delay time parameter in detecting navel orange experiment of laser-induced breakdown spectroscopy device, a cell range from 1.10 to 1.30 micros of delay time containing the initial parameter was extracted and experiment on it was conducted. There are more than one peak in the curve of element line intensity and signal-to-background ratio vs the delay time in the cell range, and the peak values are higher than the initial one. Confidence interval of the delay time calculated indirectly by interval estimation in Statistics is (1.13, 1.25). The relative standard deviation decreases from 0. 103 269 548 to 0. 025 322 279 5 when the delay time adjusted from 1.20 to 1.14 micros in the confidence interval. It is suggested that parameters range of LIBS experimental device can be adjusted freely in the range while detecting measured elements so as to improve the precision and sensitivity.