Background The aryl hydrocarbon receptor (AhR) plays a pivotal role in modulating immune responses and influencing tumor development by detecting metabolites derived from tryptophan breakdown. In patients suffering from pancreatic ductal adenocarcinoma (PDAC), elevated levels of AhR are strongly correlated with poor clinical outcomes. Despite this, the cell-autonomous functions of AhR in pancreatic tumor cells, particularly its role in modulating anti-tumor immunity within the tumor microenvironment, remain poorly characterized and require further investigation. Methods CRISPR-Cas9 technology was used to generate Ahr-/-/AHR-/- Pan02, B16-F10 and Panc-1 cells. Subcutaneous tumor models in nude and C57BL/6J mice were used to evaluate the effect of tumor cell intrinsic Ahr on the anti-tumor immune response. Transcriptome sequencing, ATAC sequencing, CUT&Tag sequencing, ChIP and IP-MS were used to elucidate how AhR controls the expression and accessibility of major histocompatibility complex class I (MHC-I). Results Elevated AhR levels in PDAC patients correlate with worse outcomes. AhR protein is located in both cytoplasm and nucleus of tumor cells. Deletion of Ahr in Pan02 and B16-F10 leads to a significant upregulation of MHC-I and related gene expression. AhR drives tumor growth by suppressing T cell-mediated immunity. Mechanistically, AhR specifically suppressed MHC-I expression in pancreatic tumor cells by interacting with protein arginine methyltransferase 5 (PRMT5) to decrease chromatin accessibility, which led to impaired antigen presentation. We demonstrated that AhR inhibition improved the therapeutic efficacy of immune checkpoint blockade, chemotherapy, and PRMT5-targeted therapy in pancreatic cancer models. Conclusions Tumor cell intrinsic AhR inhibits MHC-I expression in PDAC cells by epigenetic regulation mediated by PRMT5, resulting in reduced immunogenicity of pancreatic tumor cells. The discovery of the AhR-PRMT5-H4R3me2s-MHC-I axis provides critical mechanistic insights into how tumor-intrinsic epigenetic regulation of antigen presentation promotes the progression of PDAC.

BACKGROUND The incidence rate of acute pancreatitis（AP）, which is a pathophysiological process with complex etiology, is increasing globally. miR-125b-5p, a bidirectional regulatory miRNA, is speculated to exhibit anti-tumor activity. However,exosome-derived miR-125b-5p in AP has not been reported.AIM To elucidate the molecular mechanism of exosome-derived miR-125b-5p promoting AP exacerbation from the perspective of the interaction between immune cells and acinar cells.METHODS Exosomes derived from AR42J cells were isolated and extracted in active and inactive states by an exosome extraction kit, and were verified via transmission electron microscopy, nanoparticle tracking analysis, and western blotting. RNA sequencing assay technology was used to screen differentially expressed miRNAs in active and inactive AR42J cell lines, and bioinformatics analysis was used to predict downstream target genes of miR-125b-5p. The expression level of miR-125b-5p and insulin-like growth factor 2（IGF2） in the activated AR42J cell line and AP pancreatic tissue were detected by quantitative real-time polymerase chain reaction and western blots. The changes in the pancreatic inflammatory response in a rat AP model were detected by histopathological methods. Western Blot was used to detect the expression of IGF2, PI3K/AKT signaling pathway proteins, and apoptosis and necrosis related proteins.RESULTS miR-125b-5p expression was upregulated in the activated AR42J cell line and AP pancreatic tissue,while that of IGF2 was downregulated. In vitro experiments confirmed that miR-125b-5p could promote the death of activated AR42J cells by inducing cell cycle arrest and apoptosis. In addition,miR-125b-5p was found to act on macrophages to promote M1 type polarization and inhibit M2type polarization, resulting in a massive release of inflammatory factors and reactive oxygen species accumulation. Further research found that miR-125b-5p could inhibit the expression of IGF2 in the PI3K/AKT signaling pathway. Additionally, in vivo experiments revealed that miR-125b-5p can promote the progression of AP in a rat model.CONCLUSION miR-125b-5p acts on IGF2 in the PI3K/AKT signaling pathway and promotes M1 type polarization and inhibits M2 type polarization of macrophage by inhibiting IGF2 expression, resulting in a large release of pro-inflammatory factors and an inflammatory cascade amplification effect, thus aggravating AP.

This study aimed to clarify the role of glutamine in atherosclerosis and its participating mechanism. Forty C57BL/6J mice were divided into wild control (wild Con), ApoE − / − control (ApoE − / − Con), glutamine + ApoE − / − control (Glut + ApoE − / − Con), ApoE − / − high fat diet (ApoE − / − HFD), and glutamine + ApoE − / − HFD (Glut + ApoE − / − HFD) groups. The degree of atherosclerosis, western blotting, and multiomics were detected at 18 weeks. An in vitro study was also performed. Glutamine treatment significantly decreased the degree of aortic atherosclerosis ( p = 0.03). O ‐GlcNAcylation ( O ‐GlcNAc), IL‐1β, IL‐1α, and pyruvate kinase M2 (PKM2) in the ApoE − / − HFD group were significantly higher than those in the ApoE − / − Con group ( p < 0.05). These differences were attenuated by glutamine treatment ( p < 0.05), and aggravated by O ‐GlcNA transferase (OGT) overexpression in the in vitro study ( p < 0.05). Multiomics showed that the ApoE − / − HFD group had higher levels of oxidative stress regulatory molecules (guanine deaminase [GUAD], xanthine dehydrogenase [XDH]), proinflammatory regulatory molecules (myristic acid and myristoleic acid), and stress granules regulatory molecules (caprin‐1 and deoxyribose‐phosphate aldolase [DERA]) ( p < 0.05). These differences were attenuated by glutamine treatment ( p < 0.05). We conclude that glutamine supplementation might alleviate atherosclerosis through downregulation of O ‐GlcNAc, glycolysis, oxidative stress, and proinflammatory pathway.

Background Cancer cell viability is strongly modulated by the Hippo pathway, which includes mammalian STE20-like protein kinase 1 (Mst1) and yes-associated protein (Yap). Although the roles of Mst1 and Yap in thyroid carcinoma cell death have been fully addressed, no study has determined whether differential modification of Mst1 and Yap could further suppress thyroid carcinoma progression. The aim of our study was to explore the antiapoptotic effects exerted by combined Mst1 overexpression and Yap knockdown in thyroid carcinoma MDA-T32 cells in vitro. Methods Mst1 adenovirus and Yap shRNA were transfected into MDA-T32 cells to overexpress Mst1 and inhibit Yap, respectively. Cell viability and death were determined via an MTT assay, a TUNEL assay and western blotting. Mitochondrial function, mitochondrial fission and pathway studies were performed via western blotting and immunofluorescence. Results The results of our study showed that combined Mst1 overexpression and Yap knockdown further augmented MDA-T32 cell death by mediating mitochondrial damage. In addition, cancer cell migration and proliferation were suppressed by combined Mst1 overexpression and Yap knockdown. At the molecular level, mitochondrial membrane potential, ATP production, respiratory function, and caspase-9-related apoptosis were activated by combined Mst1 overexpression and Yap knockdown. Further, we found that fatal mitochondrial fission was augmented by combined Mst1 overexpression and Yap knockdown in a manner dependent on the JNK-MIEF1 pathway. Inhibition of JNK-MIEF1 pathway activity abolished the proapoptotic effects exerted by Mst1/Yap on MDA-T32 cells. Conclusions Taken together, our data suggest that Mst1 activation and Yap inhibition coordinate to augment thyroid cancer cell death by controlling the JNK-MIEF1-mitochondria pathway, suggesting that differential regulation of the core Hippo pathway components is potentially a novel therapeutic tool for the treatment of thyroid cancer.

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the authors as "some data in this study are not solid enough and need to be further explored". The authors stated that they "found abnormalities in the re-identification of CAF-EVs, the extracted extracellular vesicles may be polluted and the elliptical structure under electron microscope is not owned by CAF-EVs. The identification of CAF-EVs by Western Blots did not refer to the definition of international society." The authors informed the journal that, after the re-experiment, they found that "there is no vesicle specific protein expression, whether the results of subsequent experiments are generated by CAF-EVs needs to be re-tested".

Malignant tumors usually have no obvious clinical symptoms in the early stage. Most patients are already in the advanced stage when they are diagnosed. Some patients have lost the opportunity for operation, resulting in poor prognosis. Therefore, how to find the best therapeutic target for such patients and improve the prognosis of patients has gradually become the focus of scholar′s attention. Recently, Kruppel-like factor (KLF) is a transcriptional regulator that can bind to the target DNA, and its family plays an important role in the occurrence and development of malignant tumors. It has also been confirmed that the KLF family affects the proliferation, differentiation and migration of tumor cells, but the specific mechanism is still not fully elucidate. Consequently, in order to further explored the effect of the KLF family on tumors, this study intends to briefly review the roles and regulatory mechanisms of the KLF family in the cell proliferation, differentiation and migration of malignant tumors, hoping to provide new target for the biological treatment of tumors.

Our purpose was to study the effect of hyperglycemia on macrophage TBK-HIF-1 alpha-mediated IL-17/IL-10 signaling and its correlation with coronary atherosclerosis. A total of 135 patients with coronary heart disease (CHD) were divided into a stable CHD (SCHD) group (n = 30) and an acute myocardial infarction (AMI) group (n = 105) [nondiabetes mellitus (non-DM)-AMI, n = 60; DM-AMI, n = 45] from January to September 2020. The SYNTAX score and metabolic and inflammatory markers were quantified and compared. THP-1 cell studies and an animal study of coronary intimal hyperplasia were also carried out. We found that the DM-AMI group showed a higher SYNTAX score than the non-DM-AMI group (P<.05). The DM-AMI group showed the highest expression levels of TANK-binding kinase 1 (TBK1), hypoxia-inducible factor 1 alpha (HIF-1 alpha), and interleukin (IL)-17 and the lowest expression level of IL-10, followed by the non-DM-AMI group and the SCHD group (P<.05). THP-1 cell studies showed that BAY87-2243 (a HIF-1 alpha inhibitor) reversed the increase in IL-17 and decrease in IL-10 expression induced by hyperglycemia. Amlexanox (a TBK1 inhibitor) reversed the increase in HIF-1 alpha expression induced by hyperglycemia. Amlexanox treatment resulted in lower coronary artery intimal hyperplasia and a larger lumen area in a diabetic swine model. We conclude that hyperglycemia might aggravate the complexity of coronary atherosclerosis through activation of TBK1-HIF-1 alpha-mediated IL-17/IL-10 signaling. Thus, TBK1 may be a novel drug therapy target for CHD complicated with DM.

Objective:To observe the effects of engulfment and cell motility protein 2 (ELMO2) on the chemotactic motility, migration motility, cell adhesion ability and invasion ability of pancreatic cancer cell line pancreatic cancer cell line-1 (PANC-1) [American type culture collection (ATCC)], and investigate the underlying mechanism.Methods:The small interfering RNA technique was used to knock down the ELMO2 protein in the pancreatic cancer cell line PANC-1. The experimental group was designed and PANC-1 cells were transfected with small interfering RNA knockdown strands and negative control strands, respectively, and composed of ELMO2 protein knockdown group, negative control group and normal group. We established concentration gradients of CXC chemokine ligand 12 (CXCL12) with different concentrations, and the chemotaxis assay, adhesion assay and cell invasion assay were used to detect the role of ELMO2 on chemotaxis and metastasis of the pancreatic cancer cells. Exogenous co-immunoprecipitation was performed to show the ELMO2 interaction with the Gαi2 subunit. Two-way ANOVAwas used for statistical analysis.Results:RNAi interfering strands were transfected into PANC-1 cell line, and the expression of ELMO2 protein in the cells was significantly reduced. After the expression of ELMO2 protein was reduced in PANC-1 cells, when the chemokine concentration was 10 μg/L, the number of mobile cells capable of directing transmembrane in the knockdown group was significantly reduced as compared with that in the negative control group [(36.67±1.21) vs. (75.67±1.76), t=18.270, P<0.01]. Under the condition of chemokine concentration of 10 μg/L, the adhesion rate of PANC-1 cells in the ELMO2 knockdown group was significantly lower than that in the negative control group [(0.47±0.02) vs. 0.79±0.01), t=19.850, P<0.01]. When the concentration of chemokine CXCL12 was 10 μg/L, the invasion ability of PANC-1 cells in ELMO2 knockdown group was significantly weakened as compared with that in the negative control group [(84.33±2.03) vs. (132.00±0.58), t=22.610, P<0.01]. A GV362-ELMO2 overexpression plasmid vector was constructed. The band of Gαi2 protein was found in the immunoprecipitation of anti-flag antibody. Conclusion:ELMO2 knockdown inhibited chemotaxis, adhesion and invasion of pancreatic cancer cells. Our results confirmed the physical association between ELMO2 and Gαi2 in pancreatic cancer cells through exogenous co-immunoprecipitation.

Objective:To investigate the operation timing of laparoscopic cholecystectomy for mildly biliary pancreatitis.Methods:Pubmed, MEDLINE, Embase, Cochrane Library, CBMdisc, Wanfang database were searched to collect published English and Chinese literature on randomized controlled trials (RCT) of early and delayed cholecystectomy for mildly biliary pancreatitis. The primary outcome was the readmission rate due to recurrence of biliary complication. Secondary outcomes included intraoperative and postoperative complications, conversion rate, biliary colic, degree of surgical difficulty, mortality, operation time and hospital stay. The above indicators were analyzed by RevMan 5.2 software.Results:Five RCTs were identified, which included 629 patients, 318 in the early cholecystectomy (EC) group and 311 in the delayed cholecystectomy (DC) group. Compared with patients in DC group, the readmission rate due to recurrence of biliary complications in EC group was significantly reduced (OR=0.15, 95% CI: 0.08-0.28, P<0.001). However, there was no statistically significant difference between the two groups in the incidence of intraoperative complications (OR=0.57, 95% CI: 0.17-1.84, P=0.34), postoperative complications (OR= 0.77, 95% CI: 0.38-1.55, P=0.46), and conversion rate (OR=1.57, 95% CI: 0.69-3.54, P=0.28).Conclusion:Early cholecystectomy for mildly biliary pancreatitis is a safe and effective treatment, which can reduce the readmission rate due to recurrence of biliary complications without increasing the incidence of intraoperative and postoperative complications.

•Patients with DM-STEMI had a higher GRACE score compared with non-DM STEMI.•DM-STEMI had higher levels of PKM2 and NLRP3 inflammasomes compared with non-DM STEMI.•DM-STEMI had lower level of G3BP1 compared with non-DM STEMI.•PKM2 might be a new target for the treatment of patients with CHD and DM.

Objective This study aimed to explore whether treatment with the glucagon-like peptide-1 (GLP-1) analog liraglutide reduces intimal hyperplasia after coronary stent implantation via regulation of glycemic variability, the NLRP3 inflammasome, and IL-10 in diabetic swine. Methods Fifteen pigs were divided into a diabetes mellitus (DM) group (n = 6), a DM + liraglutide treatment group (L group) (n = 6) and a sham group (n = 3). A total of 24 everolimus-eluting stents were implanted in the left anterior descending and right coronary arteries at 3 weeks. A novel continuous glucose monitoring system (GMS) was used for 2 weeks. The means and standard deviations (SDs) were measured and calculated by the GMS. At 22 weeks, the lumen area (LA), neointimal thickness (NIT), neointimal area (NIA), and percent area stenosis (%AS) were analyzed by optical coherence tomography. Plasma tumor necrosis factor-alpha, interleukin-6, and interleukin-10 were assayed by ELISA. The intima protein expression levels of NLRP3, interleukin-1 beta, interleukin-18 and interleukin-10 were examined using Western blot analysis. Histology was used to evaluate the healing response. In an in vitro study, THP-1 cells were divided into control, high glucose (HG), HG + liraglutide, and HG + liraglutide + Exe(9-39) (a GLP-1 receptor inhibitor) groups. Results The L group had a lower SD, NIT, NIA, and %AS; a larger LA; reduced inflammation and injury scores; lower expression levels of tumor necrosis factor-alpha, interleukin-6, NLRP3, interleukin-1 beta, and interleukin-18; and higher expression of interleukin-10 compared with those of the DM group (p < 0.05). In the in vitro study, similar results were obtained in the HG + liraglutide group, and Exe(9-39) abolished the effect of liraglutide (p < 0.05). Conclusions Liraglutide treatment reduces intimal hyperplasia after stent implantation via regulation of glycemic variability, the NLRP3 inflammasome, and IL-10 in diabetic pigs in a GLP-1 receptor-dependent manner. Reducing the inflammation induced by glycemic variability may be one of the cardioprotective mechanisms of liraglutide.

BACKGROUND:Pancreatic cancer is the 7th leading cause of cancer-related death worldwide. Aberrant expressions of transmembrane 176A (TMEM176A) were found in multiple cancer types. However, the expression and function of TMEM176A remain unclear in pancreatic cancer.MATERIALS AND METHODS:Immunohistochemistry, flow cytometry, western blot, and transwell were applied on investigating samples from pancreatic cancer patients and pancreatic cancer cell lines.RESULTS:Analysis based on the TCGA database showed that a high level of TMEM176A was associated with a better relapse-free survival rate (P = 0.012). Further, the results of tissue microarray showed patients with a high level of TMEM176A were associated with lymph node metastasis (P = 0.045) and a better overall survival rate (P = 0.032). Overexpression of TMEM176A in Capan-1 and PANC-1 cells suppressed cell proliferation, cell invasion, and migration, and induced apoptosis in pancreatic cancer cells. TMEM176A suppressed ERK signaling in pancreatic cancer.CONCLUSION:TMEM176A suppresses the growth and migration of pancreatic cancer cells by inhibiting ERK signaling.

Abstract. Background. Pancreatic stellate cells (PSCs) activation plays a critical role in the development of chronic pancreatitis. Previous studies confirmed that thromboxane A2 receptor (TxA2r) was overexpressed in activated PSCs in rats. The purpose of this study was to investigate the role of TxA2r in the activation of PSCs induced by 8-epi-prostaglandin F2α (8-epi-PGF2α). Methods. TxA2r expression in both quiescent and activated PSCs was detected by immunocytochemistry and immunoblot assay. Isolated PSCs were treated with 8-epi-PGF2α (10−6, 10−7, 10−8 mol/L) for 48 h, and SQ29548 (10−4, 10−6, and 10−7 mol/L), a TxA2r-specific antagonist for 48 h, respectively, to identify the drug concentration with the best biological effect and the least cytotoxicity. Then isolated PSCs were treated with SQ29548 (10−4 mol/L) for 2 h, followed by 10−7 mol/L 8-epi-PGF2α for 48 h. Real-time polymerase chain reaction was performed to detect the messenger RNA (mRNA) levels of α-smooth muscle actin (α-SMA) and collagen I. Comparisons between the groups were performed using Student's t test. Results. TxA2r was up-regulated in activated PSCs in vitro compared with quiescent PSCs (all P < 0.001). Compared with the control group, different concentrations of 8-epi-PGF2α significantly increased mRNA levels of α-SMA (10−6 mol/L: 2.23 ± 0.18 vs. 1.00 ± 0.07, t = 10.70, P < 0.001; 10−7 mol/L: 2.91 ± 0.29 vs. 1.01 ± 0.08, t = 10.83, P < 0.001; 10−8 mol/L, 1.67 ± 0.07 vs. 1.00 ± 0.08, t = 11.40, P < 0.001) and collagen I (10−6 mol/L: 2.68 ± 0.09 vs. 1.00 ± 0.07, t = 24.94, P < 0.001; 10−7 mol/L: 2.12 ± 0.29 vs. 1.01 ± 0.12, t = 6.08, P < 0.001; 10−8 mol/L: 1.46 ± 0.15 vs. 1.00 ± 0.05, t = 4.93, P = 0.008). However, different concentrations of SQ29548 all significantly reduced the expression of collagen I (10−4 mol/L: 0.55 ± 0.07 vs. 1.00 ± 0.07, t = 10.47, P < 0.001; 10−6 mol/L: 0.56 ± 0.10 vs. 1.00 ± 0.07, t = 6.185, P < 0.001; 10−7 mol/L: 0.27 ± 0.04 vs. 1.00 ± 0.07, t = 15.41, P < 0.001) and α-SMA (10−4 mol/L: 0.06 ± 0.01 vs. 1.00 ± 0.11, t = 15.17, P < 0.001; 10−6 mol/L: 0.28 ± 0.03 vs. 1.00 ± 0.11, t = 11.29, P < 0.001; 10−7 mol/L: 0.14 ± 0.04 vs. 1.00 ± 0.11, t = 12.86, P < 0.001). After being treated with SQ29548 (10−4 mol/L) and then 8-epi-PGF2α (10−7 mol/L), the mRNA levels of α-SMA (0.20 ± 0.08 vs. 1.00 ± 0.00, t = 17.46, P < 0.001) and collagen I (0.69 ± 0.13 vs. 1.00 ± 0.00, t = 4.20, P = 0.014) in PSCs were significantly lower than those of the control group. Conclusions. The results show that 8-epi-PGF2α promoted PSCs activation, while SQ29548 inhibited PSCs activation induced by 8-epi-PGF2α. The result indicated that TxA2r plays an important role during PSC activation and collagen synthesis induced by 8-epi-PGF2αin vitro. This receptor may provide a potential target for more effective antioxidant therapy for pancreatic fibrosis.

The present study identified whether glycemic variability (GV) was associated with vascular calcification and explored the underlying mechanisms. Eighty-four consecutive type 2 diabetic patients with unstable angina (UA) were included from January 2018 to June 2018 to calculate calcification scores using computerized tomographic angiography (CTA), and the patients were divided into 2 groups: high calcification score group (HCS group) and low calcification score group (LCS group). Intergroup differences in GV were determined via comparisons of the standard deviation (SD) of blood glucose. Calcification staining, content measurement, apoptosis evaluation and Western blot analysis of endoplasmic reticulum (ER) stress-related apoptosis, Wnt1, galectin-3 and bone morphogenetic protein-2 (BMP-2) were compared in cell cultures from rat vascular smooth muscle cells in the different degrees of GV. The SD increased significantly with the increases in calcification scores from human studies (HCS group 2.37 ± 0.82 vs. LCS group 1.87 ± 0.78, p = 0.007). Multivariate logistic regression analysis suggested that increased SD and serum creatinine were independent predictors of calcification. The high GV group had a higher apoptotic rate, higher calcification content and higher expressions of glucose-regulated protein, caspase-3, Wnt1, galectin-3 and BMP-2 markers compared to the low GV group in the in vitro studies (p < 0.001). We report the novel finding that GV is associated with vascular calcification, and ER stress-related apoptosis, Wnt1, galectin-3 and BMP-2 may be involved in this regulation.

Background. Laparoscopy has been widely applied in gastrointestinal surgery, with benefits such as less intraoperative blood loss, faster recovery, and shorter length of hospital stay. However, it remains controversial if laparoscopic major gastrointestinal surgery could be conducted for patients with chronic obstructive pulmonary disease (COPD) which was traditionally considered as an important risk factor for postoperative pulmonary complications. The present study was conducted to review and assess the safety and feasibility of laparoscopic major abdominal surgery for patient with COPD. Materials and Methods. Databases including PubMed, EmBase, Cochrane Library, and Wan-fang were searched for all years up to Jul 1, 2018. Studies comparing perioperative results for COPD patients undergoing major gastrointestinal surgery between laparoscopic and open approaches were enrolled. Results. Laparoscopic approach was associated with less intraoperative blood loss (MD = -174.03; 95% CI: −232.16 to -115.91, P < 0.00001; P < 0.00001, I2=93% for heterogeneity) and shorter length of hospital stay (MD = -3.30; 95% CI: −3.75 to -2.86, P < 0.00001; P = 0.99, I2=0% for heterogeneity). As for pulmonary complications, laparoscopic approach was associated with lower overall pulmonary complications rate (OR = 0.58; 95% CI: 0.48 to 0.71, P < 0.00001; P = 0.42, I2=0% for heterogeneity) and lower postoperative pneumonia rate (OR = 0.53; 95% CI: 0.41 to 0.67, P < 0.00001; P = 0.57, I2=0% for heterogeneity). Moreover, laparoscopic approach was associated with lower wound infection (OR = 0.51; 95% CI: 0.42 to 0.63, P < 0.00001; P = 0.99, I2=0% for heterogeneity) and abdominal abscess rates (OR = 0.59; 95% CI: 0.44 to 0.79, P < 0.0004; P = 0.24, I2=30% for heterogeneity). Conclusions. Laparoscopic major gastrointestinal surgery for properly selected COPD patient was safe and feasible, with shorter term benefits.

Objective To observe the effects of proximal gastric electrical stimulation (GES) on body weight and gastrointestinal motility in SD rats,and investigate the regulation of gastric nerve stimulation and serum gastrointestinal hormones by neuro-humoral regulation.Methods 12 SD rats were divided into experimental group (n =6) and control group (n =6),with gastric electrical stimulator implanted,and in experimental group dual-channel GES was activated.General status was observed for 4 weeks after GES activation,including body weight,feeding and water intake,urine and stool volumes,the resting gastric volume and gastric emptying were monitored via the establishment of intestinal fistula,and serum gastrointestinal hormones change was detected.Results During 4-week GES process,one rat had gastric retention and died at 1 week after GES activated.Compared with the control group,body weight,food intake,urine and stool volumes levels of the rats in experimental group decreased significantly (t =4.005,2.530,3.350,all P＜0.05).Resting gastric volume was significantly lower than that in the control group [(2.93 ± 0.50) ml vs.(5.10 ± 0.53) ml,Z =2.460,P =0.014],and the intestinal juice drainage was lower than the control group [(0.18 ±0.15)ml vs.(0.44 ±0.05)ml,Z =2.513,P =0.012],while serum GLP-1 levels were similar between the two groups [(0.44 ± 0.05) ml vs.(0.18 ± 0.15) ml,Z =1.026,P =0.305],but Ghrelin was significantly higher than that in the control group [(1.65 ± 0.58) vs.(0.65 ±0.36),Z =2.380,P =0.017].Conclusion The proximal GES may lead to the change of the body weight,food intake,gastrointestinal function and motility,possibly by stimulating nerve reflex inducing gastrointestinal hormones secretion and affect gastrointestinal function.

Background Breast cancer stem cells (BCSCs) are associated with the invasion of breast cancer. In recent years, studies have demonstrated different phenotypes among BCSCs. Furthermore, BCSCs of diverse phenotypes are present at different tumour sites and different histological stages. Fibroblasts are involved in the phenotypic transformation of BCSCs. Cancer-associated fibroblasts (CAFs) participate in the induction of epithelial–mesenchymal transition, thereby promoting the acquisition of stem cell characteristics, but little is known about the role of normal fibroblasts (NFs) in the phenotypic transformation of BCSCs or about the effect of CAFs and NFs on BCSC phenotypes. Methods A total of six pairs of primary CAFs and NFs were isolated from surgical samples of breast cancer patients and subjected to morphological, immunohistochemical, cell invasion and proteomics analyses. After establishing a cell culture system with conditioned medium from CAFs and NFs, we used the mammosphere formation assay to explore the effect of CAFs and NFs on the self-renewal ability of BCSCs. The effect of CAFs and NFs on the phenotypic differentiation of BCSCs was further analysed by flow cytometry and immunofluorescence. Results The isolated CAFs and NFs did not show significant differences in cell morphology or alpha-smooth muscle actin (α-SMA) expression, but cell invasion and proteomics analyses demonstrated heterogeneity among these fibroblasts. Both CAFs and NFs could promote the generation of BCSCs, but CAFs displayed a greater ability than NFs in promoting mammosphere formation. Conditioned medium from CAFs increased the proportion of aldehyde dehydrogenase-1 positive (ALDH1+) BCSCs, but conditioned medium from NFs was more likely to promote the generation of CD44+CD24− BCSCs from MCF-7 cells. Discussion This study validated the heterogeneity among CAFs and NFs and expanded on the conclusion that fibroblasts promote the generation of cancer stem cells. Our results particularly emphasized the effect of NFs on the phenotypic transformation of BCSCs. In addition, this study further highlighted the roles of CAFs and NFs in the induction of different phenotypes in BCSCs.

Objective To explore the potential mechanism of epithelial-mesenchymal transition (EMT) of pancreatic cancer induced by pancreatic stellate cell (PSC).Methods The effect of PSC on EMT of pancreatic cancer was determined by direct co-culture.We also added signal pathway inhibitors and Notch-3 small interfere RNA (siRNA) to the co-culture system.Morphological change observation,two-chamber migration assays to assess the cell migration,fluorescent immunohistochemistry,Reverse transcriptase PCR (RT-PCR) and Western blotting to assess the expression level of Notch-3,E-cadherin,Vimentin was performed to determine whether the EMT happened.Results After co-culture with PSC,EMT phenomenon of PANC-1 happened with increased motility [mono-culture vs.co-culture,(16.4 ±2.6)/HP vs.(51.6 ±4.5)/HP,P=0.001].L1790 (5 μmol/L,Notch signaling pathway inhibitor) can significantly inhibit the increased motility of PANC-1 cells,and suppress the occurrence of EMT.Notch-3 siRNA successfully inhibited Notch-3 gene expression,blocked the increased migration ability and EMT phenomenon of PANC-1 induced by PSC.Conclusion Notch-3 plays critical role in EMT of PANC-1 induced by PSC.