Mechanism of Reversal of Precancerous Lesions of Gastric Cancer by Celastrus orbiculatus Extract via Regulating Lgr5/Wnt/β-catenin Signaling Pathway / 中国实验方剂学杂志

ObjectiveThis study aims to investigate the mechanism in which Celastrus orbiculatus extract （COE） affects the proliferation and differentiation of gastric organoids and the expression of Lgr5 and thus reverses the precancerous lesions of gastric cancer （PLGC） by regulating the leucine-rich repeat containing G protein-coupled receptor 5 （Lgr5）/Wingless （Wnt）/β-catenin signaling pathway based on a gastric organoid injury model. MethodGastric organoids were established based on stem cells of the mouse gastric gland. Gastric organoid injury models were constructed by treating gastric organoids with N-methyl-N'-nitro-N-nitrosoguanidine （MNNG， 0.02 mg·L-1）. Gastric organoid injury models were randomly divided into normal group， model group （0.02 mg·L-1 MNNG）， low， medium， and high dose （5， 10， 20 mg·L-1） groups of COE， and Wnt inhibitor Dickkopf-related protein 1 （DKK1） （0.5 mg·L-1） group， and they were treated with respective agents for 24 h. The number and volume of gastric organoids under different drug concentrations were observed under a microscope. The viability of the gastric organoid injury models was detected by Methyl thiazolyl tetrazolium （MTT） assay. The morphology and pathology of gastric organoids were observed using Hematoxylin and Eosin （HE） staining. The expression levels of Lgr5， Mucin2 （MUC2）， Mucin5AC （MUC5AC）， Mucin6 （MUC6）， Wnt， and β-catenin in gastric organoids under different drug concentrations were detected by Western blot （WB）. ResultCompared with the normal group， the number， volume， and activity of gastric organoids in the model group were decreased （P<0.01）， while the expressions of Lgr5， MUC2， Wnt， and β-catenin were significantly increased （P<0.01）. The expressions of MUC5AC and MUC6 were significantly decreased （P<0.01）. Compared with the model group， the number and volume of gastric organoids in the low， medium， and high dose groups of COE were all improved （P<0.01）， and the vitality of gastric organoids was significantly enhanced （P<0.01）. The effect was the most significant at a COE concentration of 20 mg·L-1 （P<0.01）. The expressions of Lgr5 and MUC2 in the medium and high dose groups of COE were significantly reduced （P<0.01）， while the expression of MUC5AC and MUC6 were significantly increased in the low， medium， and high dose groups of COE （P<0.05， P<0.01）. Compared with the model group， Wnt inhibitors could promote the expression of MUC5AC and MUC6 in gastric organoids （P<0.05， P<0.01） and reduce the expression of MUC2， Wnt， and β-catenin. In addition， the combined use of COE at high concentrations and Wnt inhibitors could further promote this trend （P<0.01）. ConclusionCOE inhibits the Wnt/β-catenin pathway by inhibiting the expression of Lgr5， MUC2， Wnt， and β-catenin and promoting the expression of MUC5AC and MUC6， thus promoting the proliferation and differentiation of gastric organoids and reversing the PLGC process.

Dilemma and Reflection on Children Exercising the Right of Medical Decision-making / 中国医学伦理学

Whether children can exercise their medical decision-making power has always been a controversial topic in law and ethics, and it is also the focus of attention of people from all walks of life. In this regard, combined with the problems existing in the exercise of children’s medical decision-making power, such as conflict with the right to life and health, insufficient guarantee of the right to informed consent system, and the legal guardian’s exercise of children’s medical decision-making power may not be in the best interests of children. This paper discussed the dilemma and feasibility of children’s exercise of medical decision-making power from three aspects: children’s right to life and health, the evaluation of informed consent and medical decision-making ability, and the thinking of children’s informed consent and medical decision-making ability, and pointed out that children who are able to make self-determination should be fully endowed with legal medical decision-making power, so as to ensure their best interests in medical clinic.

TJP3 promotes T cell immunity escape and chemoresistance in breast cancer: a comprehensive analysis of anoikis-based prognosis prediction and drug sensitivity stratification.

BACKGROUND: Overcoming anoikis is a necessity during the metastasis and invasion of tumors. Recently, anoikis has been reported to be involved in tumor immunity and has been used to construct prognosis prediction models. However, the roles of anoikis in regulating tumor immunity and drug sensitivity in breast cancer are still not clear and therefore worth uncovering. METHODS: TCGA and GEO data are the source of gene expression profiles, which are used to identify anoikis-related-gene (ARG)-based subtypes. R4.2 is used for data analysis. RESULTS: Breast cancer is divided into three subgroups, amongst which shows prognosis differences in pan-cancer cohort, ACC, BLCA, BRCA, LUAD, MESO, PAAD, and SKCM. In breast cancer, it shows significant differences in clinical features, immune cell infiltration and drug sensitivity. Machine learning constructs prognosis prediction model, which is useful to perform chemotherapy sensitivity stratification. Following, TJP3 is identified and verified as the key ARG, up-regulation of which increases tolerance of paclitaxel-induced cell toxicity, accompanied with increased expression of caspas3 and cleaved-caspase3. In addition, Down-regulation of TJP3 weakens the cell migration, which accompanied with increased expression of E-cad and decreased expression of vimentin, twist1, zeb1, and MMP7. Furthermore, the expression level of PD-L1 is negative correlated with TJP3. CONCLUSION: ARGs-based subgroup stratification is useful to recognize chemotherapy sensitive cohort, and also is useful to predict clinical outcome. TJP3 promotes chemoresistance, tumor metastasis and potential immunotherapy escape in breast cancer.

Effects of Celastrus orbiculatus extract on gastric organoids and expression of E －caherin in mice / 安徽医科大学学报

Objective@#To investigate the effect of Celastrus orbiculatus extract ( COE) on the growth of gastric organoids and the expression of E-cadherin in gastric epithelial cells of mice．@*Methods@#The gastric pylorus of 8-week-old C57 mice was isolated and cultured into gastric organoids．The dynamic changes of gastric organoid formation were observed under light microscope ; the intercellular structure of gastric epithelium was observed by HE staining ; the expression of epithelial-cadherin E-cadherin in gastric epithelial cells was observed by immunofluorescence staining．After passage to the third-generation ，the organoids were treated with different concentrations of COE (0，5 ，10，20 μg/ ml) ，the organoids were collected ，their numbers were counted ，their diameters were measured，their cellular activities were measured by methyl thiazolyl tetrazolium( MTT) colorimetry，and Western blot was used to detect the expression of E-cadherin in organoids after COE treatment. @*Results @#At 24 to 48 h， cystlike structures were formed and three-dimensional cell clusters with cystic gland-like central structure appeared， and gradually budding and forming gastric organoids after 72 h，suggesting that the organoids were successfully constructed．The epithelial cell marker E-cadherin was expressed in the organoid，which further confirmed the formation of organoid．Compared with the control group，the number and diameter of gastric organoids in the COE group significantly increased，cell activity was significantly enhanced (P＜0. 05) ，and the expression of E-cadherin increased with the increase of COE dose (P＜0. 01) ．@*Conclution @#Low dose COE can promote the expression of E-cadherin and the growth and formation of organoids，which may affect the repair of gastric mucosa injury.

Dilemma and Reflection on Children Exercising the Right of Medical Decision-making / 中国医学伦理学

Whether children can exercise their medical decision-making power has always been a controversial topic in law and ethics, and it is also the focus of attention of people from all walks of life. In this regard, combined with the problems existing in the exercise of children’s medical decision-making power, such as conflict with the right to life and health, insufficient guarantee of the right to informed consent system, and the legal guardian’s exercise of children’s medical decision-making power may not be in the best interests of children. This paper discussed the dilemma and feasibility of children’s exercise of medical decision-making power from three aspects: children’s right to life and health, the evaluation of informed consent and medical decision-making ability, and the thinking of children’s informed consent and medical decision-making ability, and pointed out that children who are able to make self-determination should be fully endowed with legal medical decision-making power, so as to ensure their best interests in medical clinic.

Identification of the Immune-Related Genes in Tumor Microenvironment That Associated With the Recurrence of Head and Neck Squamous Cell Carcinoma.

Head and neck squamous cell carcinomas (HNSCC) are still one of the most common malignant tumors in China, with a high metastasis rate and poor prognosis. The tumor immune microenvironment can affect the occurrence, development and prognosis of tumors, but the underlying mechanism is still unclear. In this study, we tried to describe the correlation between the recurrence of HNSCC and the tumor microenvironment (TME). The expression data [estimate the level of tumor stromal and immune infiltration, expression data (ESTIMATE)] algorithm was used to identify and estimate highly correlated stromal cells, immune cells, and prognostic scores in 116 samples of head and neck cancer patients from The Cancer Genome Atlas (TCGA) dataset. The functional enrichment analysis and protein-protein interaction (PPI) networks of differential expressed genes (DEGs) were constructed. Subsequently, the abundance of various infiltrating immune cells was estimated with the tumor immune estimation resource (TIMER) and the infiltration pattern of immune cells were explored in HNSCC. A total of 407 immune-related genes were identified to involve in the TME. We found that CCR5, CD3E, CD4, and HLA -DRB1 were the most obvious DEGs and the dendritic cells (DCs) showed the highest abundance in the TME of HNSCC. In addition, the unsupervised cluster analysis determined 10 clusters of immune infiltration patterns, and indicated that immune infiltrated CD4 + T and B cells may be related to the prognosis of HNSCC. In conclusion, our research determined the list of immune genes and immune infiltrating cells related to the prognosis of HNSCC, and provided a perspective for HNSCC evolution, anti-tumor drugs selection, and drug resistance research.