Anoikis regulator GLI2 promotes NC cell immunity escape by TGF-ß-mediated non-classic hedgehog signaling in colorectal cancer: based on artificial intelligence and big data analysis.

BACKGROUND: Anoikis is a speed-limited procedure to inhibit tumor metastasis during epithelial-mesenchymal transition (EMT). Previous studies have explored anoikis-related genes (ARG) in predicting prognosis and distinguishing tumoral immunity in many types of cancer. However, the role of ARGs in regulating NK cell exhaustion (NKE) and in predicting chemotherapy sensitivity is not clear. Therefore, it is necessary to work on it. METHODS: Gene expression profiles and clinical features are collected from TCGA and GEO, and data analysis is performed in R4.2.0. RESULTS: The ARGs-based no-supervised learning algorithm identifies three ARG subgroups, amongst which the prognosis is different. WCGNA and Artificial intelligence (AI) are applied to construct an NKE-related drug sensitivity stratification and prognosis identification model in digestive system cancer. Pathways association analysis screens out GLI2 is a key gene in regulating NKE by non-classic Hedgehog signaling (GLI2/TGF-ß/IL6). In vitro experiments show that down-regulation of GLI2 enhances the CAPE-mediated cell toxicity and accompanies with down-regulation of PD-L1, tumor-derive IL6, and snial1 whereas the expression of cleaved caspas3, cleaved caspase4, cleaved PARP, and E-cadherin are up-regulated in colorectal cancer. Co-culture experiments show that GLI2- decreased colorectal tumor cells lead to down-regulation of TIM-3 and PD1 in NK cells, which are restored by TGF-bate active protein powder. Besides, the Elisa assay shows that GLI2-decreased colorectal tumor cells lead to up-regulation of IFN-gamma in NK cells.

Effect of the rolB gene on phenotypic development and tropane alkaloids biosynthesis in Atropa belladonna / 药学学报

The rol genes on pRiA4 plasmid of Agrobacterium rhizogenes are potent genes that promote secondary metabolism. Molecular breeding of Atropa belladonna can be conducted by introducing rol genes to increase tropane alkaloids (TAs) content in A. belladonna. In this study, the rolB gene was overexpressed in A. belladonna plants to study the effect of rolB gene on the biosynthesis of TAs. The phenotype, TAs content and expression levels of key enzyme genes in the pathway of TAs biosynthesis of transgenic A. belladonna were analyzed. The results showed that transgenic A. belladonna had developed root system, enlarged leaves, increased leaf fresh weight, deepened leaf color, enlarged flowers, changed flower shape, reduced pistil height and decreased pollen vitality. The content of TAs in the stems of transgenic A. belladonna was significantly higher than that of the control, and the contents of scopolamine, anisodamine, hyoscyamine can reach 2.11-2.91, 1.23-2.37 and 4.88-5.20 times of the control, respectively. Compared with the control group, the expressions of key enzymes putrescine N-methyltransferase (PMT), type III polyketide synthase (PYKS), tropinone reductase I (TRI), aromatic amino acid aminotransferase 4 (ArAT4), UDP-glycosyltransferase 1 (UGT1) and hyoscyamine 6-β-hydroxylase (H6H) in the TAs biosynthesis pathway were up-regulated, and the expression of tropinone reductase II (TRII) as a metabolic shunting gene was down-regulated. The results indicated that rolB gene enhanced TAs synthesis ability in roots and accumulation in stems of A. belladonna by enhancing metabolic flow of TAs synthesis pathway and weakening the metabolic shunt of competing pathway. This study laid a foundation for molecular breeding of A. belladonna with high-yield TAs content using rolB gene.

In vitro study of genistein inducing apoptosis in MDA-MB-231 breast cancer cells via NF-κB and MAPK signaling pathways / 中国药理学通报

Aim To study the effect of genistein on apoptosis in human breast cancer MDA-MB-231 cells and the underlying mechanisms. Methods MTT as-say was used to observe the inhibitory rate on human breast cancer MDA-MB-231 cells treated with genistein. Colony assay was used to determine the cell colony formation rate on human breast cancer MDA-MB-231 cells treated with genistein. Western blot was used to detect the expression of Bcl-2, Bax, caspase-3,NF-κB, ERK, p-ERK, JNK and p-JNK in human breast cancer MDA-MB-231 cells treated with genistein. Results The results of MTT assay showed that genistein inhibited the viability of breast cancer MDA-MB-231 cells in a time- and concentration-de-pendent manner. Colony assay suggested that genistein had an antiproliferative effect on MDA-MB-231 cells. The expression levels of Bcl-2, NF-κB and p-ERK were significantly down-regulated compared with con-trol(P < 0.01). However, the expression of Bax, caspase-3 and p-JNK was significantly up-regulated(P<0.01). Conclusions Genistein could inhibit the growth of human breast cancer MDA-MB-231 cells and induce apoptosis,and the mechanism may be related to the inhibition of NF-κB, ERK/MAPK signaling path-ways and the activation of JNK/MAPK signaling path-way.

Rosmarinic acid derivative RAD-9 induces apoptosis in gastric cancer MGC-803 cells via PI3K/Akt and p38 MAPK signaling pathway / 中国药理学通报

Aim To study the apoptosis-inducing effect of rosmarinic acid derivative RAD-9 on gastric cancer MGC-803 cells and the underlying mechanisms.Methods MTT assay was taken to detect the survival of gastric cancer MGC-803 cells effected by RAD-9.Cell apoptosis was detected by flow cytometry.The apoptotic morphology of MGC-803 cells was observed by Hoechst 33258 staining.The protein expression levels of Bcl-2,Bax,caspase-3,Akt,p-Akt,p38 MAPK and p-p38 MAPK were measured by Western blot.Results The results of MTT assay showed that RAD-9 inhibited the viability of gastric cancer MGC-803 cells in a time and concentration-dependent manner.Flow cytometry showed that RAD-9 significantly promoted apoptotic cell percentage in gastric cancer MGC-803 cells (P ＜ 0.01).Hoechst 33258 staining showed that the nucleus of MGC-803 cells could be observed with typical apoptotic morphological changes after RAD-9 administration.Compared with the control group,the protein expression levels of Bcl-2,Akt,p-Akt were significantly down-regulated (P ＜ 0.01),while those of Bax,caspase-3,p38 MAPK,p-p38 MAPK were significantly up-regulated (P ＜ 0.01).Conclusion RAD-9 can inhibit the growth and further induce apoptosis in gastric cancer MGC-803 cells,which may involve inhibiting PI3K/Akt and activating p38 MAPK signaling pathway.

Assessment of renal cortex blood flow with arterial spin labeling MRI in patients with type 2 diabetes / 中国医学影像技术

Objective To investigate the value of arterial spin labeling (ASL) MRI in evaluation of renal cortex perfusion in patients with type 2 diabetes.Methods Fifty patients with type 2 diabetes were enrolled and divided into simple diabetes (SD) group (n=25) and diabetes kidney disease (DKD) group (n=25) according to suffering from DKD or not.Based on estimated glomerular filtration rate (eGFR),DKD group were further divided into mild disease subgroup (n=11,eGFR≥ 60 ml/[min · 1.73m2]) and moderate-severe disease subgroup (n=14,eGFR＜60 ml/[min · 1.73m2]).Twenty-five healthy volunteers were recruited as control group at the same time.ASL MRI were performed on all participants.The cortical renal blood flow (RBF) of bilateral kidneys were measured by 2 radiologists.The consistency between 2 radiologists was analyzed.Statistical analysis were conducted to analysis the differences in cortical RBF among different groups.Correlation analysis were performed to evaluate the relationship between RBF and eGFR in type 2 diabetes patients.Results Cortical RBF values measured by two radiologists showed high consistency (all ICC＞0.90).There was significant difference in cortical RBF among control group ([269.71±33.28]ml/[100 g · min]),SD group ([258.52±42.30]ml/[100 g · min]),mild disease group ([242.86±56.86]ml/[100 g · min]) and moderate-severe disease group ([173.39±27.16]ml/ [100 g· min];F=20.66,P＜0.01).Moreover,the RBF in moderate-severe disease group was significantly lower than those in other groups (all P＜0.01).And no significant differences of RBF was found among the remainder groups (P=0.064,0.320).RBF in type 2 diabetes patients was positively correlated to eGFR (r=0.646,P＜0.001).Conclusion ASL MRI is a valuable tool to quantitatively assess the renal perfusion in patients with type 2 diabetes mellitus,which can provide potential imaging indicator as RBF for the functional evaluation of kidney.

Analysis of early complications after surgery for portal hypertension / 中国普通外科杂志

Objective To investigate the relevant factors for development of early postoperative complications in patients with portal hypertension due to advanced schistosomiasis, and to offer a theoretical basis for preventing the occurrence of postoperative complications. Methods The cases of advanced schistosomiasis with portal (hypertension) operated upon over the past 40 years were retrospectively reviewed. Results In 8 240 cases, the complication rate was 19.66% at one month after operation, and the mortality rate was 15.31%. The main causes of death were MDOS, massive hemorrhage into abdominal cavity, hepatorenal syndrome and (recurrence) of bleeding of upper alimentary tract. Conclusions The relevant factors for development of early complications after operation in patients with portal hypertension is related to the cause of portal hypertension, whether or not there is a history of ascites, the timing of operation and the stage of liver function(Child′s classification).