Prognostic immunogenic characteristics of iron pendant disease modifiers in colon cancer.

Background: We explored the prognostic and immunogenic characteristics of iron pendant disease regulators in colon cancer to provide a scientific basis for the prediction of tumor prognosis-related markers and potential immunotherapeutic drug targets. Methods: RNA sequencing and matched complete clinical information of colon cancer (COAD) were retrieved from the UCSC Xena database, and genomic and transcriptomic data of colon cancer from the TCGA database were downloaded. Then univariate and multifactorial Cox regression were used to process these data. The prognostic factors were analyzed by single-factor and multi-factor Cox regression, followed by Kaplan-Meier survival curves with the aid of R software "survival" package. Then we use FireBrowse online analysis tool to analyze the expression variation of all cancer genes, and draw a histogram according to the influencing factors to predict the 1, 3, and 5 year survival rates of patients. Results: The results show that age, tumor stage and iron death score were significantly correlated with prognosis (p<0.05). Further multivariate cox regression analysis confirmed that age, tumor stage and iron death score were still significantly correlated with prognosis (p<0.05); The calibration curve results show that the deviation between the predicted values of 1 year, 3 years and 5 years and the diagonal of the figure is very small; the ROC curve results show that the AUC values of the 1-year and 5-year ROC curves of the bar graph are high; the DCA curve results show that the net yield of the bar graph is the largest; The scores of T cells and B cells in the high iron death score group were significantly lower than those in the low iron death score group, and the activities of immune related pathways were significantly reduced. There was a significant difference in the iron death score between the iron death molecular subtype and the gene cluster subtype. Conclusions: The model showed a superior response to immunotherapy in the high-risk group, revealing a potential relationship between iron death and tumor immunotherapy, which will provide new ideas for the treatment and prognostic assessment of colon cancer patients.

[Fusion expression of ß-mannanase Man5A and xylanase Tlxyn11B in Pichia pastoris].

Mannanase and xylanase, the main hemicellulolytic enzymes, are widely used in food, feed, textile and papermaking industries, and usually they are used in combination. Mannanase Man5A from Talaromyces leycettanus JCM12802 consist of the carbohydrate binding module (CBM), linker region and catalytic domain. The CBM coding region of Man5A was removed and fused to C-terminal of the xylanase gene Tlxyn11B. The fusion gene Tlxyn11B-linker-man5A was successfully expressed in Pichia pastoris and the fusion protein Tlxyn11B-Man5A was purified and characterized. The theoretical molecular weights of Tlxyn11B, Man5A without CBM region, and Tlxyn11B-Man5A are 21.6 kDa, 41.0 kDa, and 62.6 kDa, respectively. The fusion protein had high xylanase and mannanase activities. The optimal temperature of the fused xylanase is 70 °C, which is 5 °C higher than Tlxyn11B-w (xylanase before fusion). The fused mannanase exhibited maximal activity at 90 °C, which is similar to Man5A-w (mannanase before fusion). More than 48% of xylanase activity of Tlxyn11B-Man5A was residual after the condition of 60 °C with 1 h, which is significantly higher than Tlxyn11B-w (only 20% of activity was left at 60 °C for 20 minutes). The optimal pHs of Tlxyn11B-Man5A for xylanase and mannanase activity are 4.0 and 5.0, respectively, which are 0.5 and 1.0 units higher than those of Tlxyn11B-w and Man5A-w. The pH range of fused enzymes got wider and the pH stability is improved. The specific activities of xylanase and mannanase of Tlxyn11B-Man5A are 1 784.3 U/mg and 1 639.6 U/mg, respectively, which is lower than those of Tlxyn11B and Man5A (8 300.0 U/mg and 1 979.0 U/mg). It may be due to of the high molecular weight of fusion enzyme. The Km and Vmax of the fused xylanase and mannanase are 1.2 mg/mL and 1.7 mg/mL, 2 000.0 µmol/(min·mg) and 2 831.6 µmol/(min·mg), respectively. Tlxyn11B and Man5A were successfully fusion expressed in P. pastoris, and the good properties of fusion of xylanase and mannanase make it has great application potential in animal feed, food and other industrial production, and it provided new ideas for the improvement of enzyme performance.

Synergistic effect of acetyl xylan esterase from Talaromyces leycettanus JCM12802 and xylanase from Neocallimastix patriciarum achieved by introducing carbohydrate-binding module-1.

Wheat bran is an effective raw material for preparation xylooligosaccharides; however, current research mainly focuses on alkali extraction and enzymatic hydrolysis methods. Since ester bonds are destroyed during the alkali extraction process, xylanase and arabinofuranosidase are mainly used to hydrolyze xylooligosaccharides. However, alkali extraction costs are very high, and the method also causes pollution. Therefore, this study focuses on elucidating a method to efficiently and directly degrade destarched wheat bran. First, an acidic acetyl xylan esterase (AXE) containing a carbohydrate-binding module-1 (CBM1) domain was cloned from Talaromyces leycettanus JCM12802 and successfully expressed in Pichia pastoris. Characterization showed that the full-length acetyl xylan esterase AXE + CBM1 was similar toe uncovered AXE with an optimum temperature and pH of 55 °C and 6.5, respectively. Testing the acetyl xylan esterase and xylanase derived from Neocallimastix patriciarum in a starch-free wheat bran cooperative experiment revealed that AXE + CBM1 and AXE produced 29% and 16% reducing sugars respectively, compared to when only NPXYN11 was used. In addition, introduced the CBM1 domain into NPXYN11, and the results indicated that the CBM1 domain showed little effect on NPXYN11 properties. Finally, the systematically synergistic effects between acetyl xylan esterase and xylanase with/without the CBM1 domain demonstrated that the combined ratio of AXE + CBM1 coming in first and NPXYN11 + CBM1 s increased reducing sugars by almost 35% with AXE and NPXYN11. Furthermore, each component's proportion remained the same with respect to xylooligosaccharides, with the largest proportion (86%) containing of 49% xylobiose and 37% xylotriose.

[Automatic Method for Selecting Characteristic Lines Based on Genetic Algorithm to Quantify Laser-Induced Breakdown Spectroscopy].

Selecting proper characteristic lines from enormous spectral intensities is crucially important to implement quantitative analysis of Laser-Induced Breakdown Spectroscopy using internal standard method. Manual selecting of characteristic lines by researchers is time consuming and energy consuming, which cannot guarantee the best result. An automatic method to select analytical and reference lines for internal standard method from the original spectra based on Genetic Algorithm was proposed in this paper. This method was utilized to select analytical and reference lines for internal standard methods from LIBS of Mn, Ni, Cr, Si and Fe of low alloy steels. The optimal characteristic lines optimized by this method were the analytical line 403.306 8 nm of Mn and the corresponding reference line 368.745 7 nm of Fe, the analytical line 288.157 7 nm of Si and the corresponding reference line 427.176 1 nm of Fe, the analytical line 286.510 0 nm of Cr and the corresponding reference line 272.753 9 nm of Fe and the analytical line 352.453 6 nm of Ni and the corresponding reference line 358.698 5 nm of Fe, respectively. Then these elements were quantified by the internal standard method using these selected lines. The results showed that this proposed method for selecting characteristic lines can automatically select the optimal analytical and reference lines and could guarantee the best quantitative result obtained by internal standard method.