In silico analysis of intestinal microbial instability and symptomatic markers in mice during the acute phase of severe burns.

BACKGROUND: Severe burns may alter the stability of the intestinal flora and affect the patient's recovery process. Understanding the characteristics of the gut microbiota in the acute phase of burns and their association with phenotype can help to accurately assess the progression of the disease and identify potential microbiota markers. METHODS: We established mouse models of partial thickness deep III degree burns and collected faecal samples for 16 S rRNA amplification and high throughput sequencing at two time points in the acute phase for independent bioinformatic analysis. RESULTS: We analysed the sequencing results using alpha diversity, beta diversity and machine learning methods. At both time points, 4 and 6 h after burning, the Firmicutes phylum content decreased and the content of the Bacteroidetes phylum content increased, showing a significant decrease in the Firmicutes/Bacteroidetes ratio compared to the control group. Nine bacterial genera changed significantly during the acute phase and occupied the top six positions in the Random Forest significance ranking. Clustering results also clearly showed that there was a clear boundary between the communities of burned and control mice. Functional analyses showed that during the acute phase of burn, gut bacteria increased lipoic acid metabolism, seleno-compound metabolism, TCA cycling, and carbon fixation, while decreasing galactose metabolism and triglyceride metabolism. Based on the abundance characteristics of the six significantly different bacterial genera, both the XGboost and Random Forest models were able to discriminate between the burn and control groups with 100% accuracy, while both the Random Forest and Support Vector Machine models were able to classify samples from the 4-hour and 6-hour burn groups with 86.7% accuracy. CONCLUSIONS: Our study shows an increase in gut microbiota diversity in the acute phase of deep burn injury, rather than a decrease as is commonly believed. Severe burns result in a severe imbalance of the gut flora, with a decrease in probiotics and an increase in microorganisms that trigger inflammation and cognitive deficits, and multiple pathways of metabolism and substance synthesis are affected. Simple machine learning model testing suggests several bacterial genera as potential biomarkers of severe burn phenotypes.

Accurate and rapid quantification of PD-L1 positive exosomes by a triple-helix molecular probe.

Programmed death ligand-1 (PD-L1) positive exosomes (P-Exo) have been widely used for tumor diagnosis. However, accurate and rapid quantification of P-Exo remains challenging due to the heterogeneity of clinical individuals and isolation techniques. In this study, the triple-helix molecular probe (THMP) coupled with high-affinity silica-based TiO2 magnetic beads was used to isolate exosomes and to analyze the relative abundance of P-Exo in total exosomes (T-Exo). By employing this strategy, the entire analysis was completed within 70 min and the detection limit for P-Exo was 880 particles µL-1. Additionally, the relative abundance of P-Exo in T-Exo (RAP-Exo/T-Exo) was calculated from their fluorescence ratio, which could avoid errors due to differences in samples and separation methods, and identify 1.5 × 103 P-Exo from 5 × 106 T-Exo per microliter. RAP-Exo/T-Exo values were not only effective in distinguishing healthy volunteers from breast cancer patients, but also highly positively correlated with the stage of breast carcinoma. Overall, this strategy opens a new avenue for rapid and quantitative analysis of P-Exo, providing an opportunity for precise diagnosis and prediction of treatment efficacy in cancer.

[Clinicopathological Research and Expression of PTEN/PI3K/Akt Signaling Pathway in Non-small Cell Lung Cancer.].

BACKGROUND: It has been known that abnormality of PTEN/PI3K/Akt signal pathway played an important role in initiation of some malignant tumors. The aim of this study is to examine the expression and clinicopathological significance of PTEN, PI3K and Akt in non-small cell lung cancer (NSCLC). METHODS: Expression levels of PTEN, PI3K and Akt protein were determined using immunohistochemistry S-P in 61 specimens of NSCLC with follow-up. RESULTS: (1)The levels of PTEN protein was higher than that of control group, and levels of PI3K and Akt protein were lower than that of control group; (2)Expression of PTEN and PI3K were related to histotype, clinical stage, lymphonode metastasis and survival rate; Expression of Akt was related to clinical stage, lymphonode metastasis and survival rate; (3)The Cox Monovariable Analyses revealed that both smoking and negative expression of PTEN were the risking factors on the death of the NSCLC patients after surgery; (4)The expression of PTEN protein was negatively correlated to that of PI3K and Akt respectively, while the expression of PI3K was positively correlated to that of Akt. CONCLUSIONS: In NSCLC, the lack of PTEN induced up-regulation of PI3K and Akt, which demonstrated that PTEN/PI3K/Akt signaling pathway contributed to the tumorigenesis and development of NSCLC. They could be used as the indicators of prognosis and targets of therapy.

[Expression of PTEN, p53 and P-glycoprotein in Non-small cell Lung Cancer and Their Predictive Values.].

BACKGROUND: Multiple drug resisting (MDR) phenotype is the sign of intrinsic or acquired resistance and is the key factor which leads to chemotherapy failure. It has been proven that PTEN, p53, P-gp expressions were related to drug resistance and prognosis of the patients with lung cancer. The aim of this study is to analyze the association of the expression of PTEN, p53, P-gp with postoperative survival in patients with non-small cell lung cancer (NSCLC), and to explore the relationship between the characteristics and drug resistance of NSCLC patients. METHODS: A total of 61 patients with NSCLC were followed up. Immunohistochemical staining using polyclonal PTEN, p53 and P-gp antibody were performed on paraffinimbedded specimens from 61 patients with NSCLC and 20 specimens of tumor-surrounding normal lung tissue were used as control. RESULTS: The PTEN expression was significantly lower in NSCLC than in tumor-surrounding normal lung tissue and the expression of p53 and P-gp were in the opposite aspects (P<0.05). Expression of PTEN was positively related with histology, clinical stage, lymphatic metastasis, p53 expression was positively related with gender, lymphatic metastasis and the three kinds of protein expression were related with prognosis (P<0.05). The expression of PTEN was related with p53 expression (r=-0.282, P<0.05), but the expression of PTEN and p53 were not related with P-gp in NSCLC (P>0.05). CONCLUSIONS: To some extent, the expression level of PTEN, p53 and P-gp may predict the effect of radiotherapy, chemotherapy and prognosis of NSCLC.