IL-23p19 plays important roles in intestinal antimicrobial immunity, while its over-expression can lead to intestinal inflammation. However, the bacterial compounds and the type of pattern recognition receptor involved in the inducible expression of IL-23p19 in Paneth cells remain unclear. Here we show that the mRNA expression of IL-23p19 was increased in Paneth cell (PC)-like cells stimulated by Toll-like receptor 2 (TLR2) ligands, peptidoglycan (PGN) and Pam3CSK4, and was further increased in the presence of nucleotide-binding oligomerization domain 2 (NOD2)-ligand muramyl dipeptide (MDP). However, its mRNA expression was decreased in NOD2-knockdown PC-like cells. Additionally, the c-Rel activation was increased in Pam3CSK4- or PGN-stimulated PC-like cells, but the PGN-induced c-Rel activation was decreased in NOD2-knockdown PC-like cells and had no significant difference compared with Pam3CSK4-induced c-Rel activation. Our results suggest that NOD2 up-regulates TLR2-mediated IL-23p19 expression via increasing c-Rel activation in PC-like cells. This finding might provide us with a novel therapeutic target for inflammatory bowel disease to inhibit IL-23p19 over-expression via the NOD2-c-Rel pathway.
Objective To study the influence of the small interfering RNA (siRNA) interference TMPRSS4 expression on human pancreatic cancer SW1990 cell's proliferation and invasion. Methods The four eukaryotic expression vector of TMPRSS4 gene were synthesized in vitro and were transfected transiently into human pancreatic cancer SW1990 cells. TMPRSS4 mRNA expression of transfected cells was detected by real-time RT-PCR. The most efficient eukaryotic expression vector was used to be transfected into SW1990 cells. By using G418, cell strain that can silence TMPRSS4 gene stably was screened. The TMPRSS4 mRNA expression of the stable cell strain was detected by real time PCR TMPRSS4 protein expression was detected by western blot. The proliferation ability of transfected SW1990 cells was detected by CCK-8 method. By Transwell, the invasion change of SW1990 cell was detected. Results A stable cell strain, SW1990/psi TMPRSS4, was successfully constructed, in which the expression level of TMPRSS4 could be reduced stably by RNA interference. Cell transfection efficiency was 82.9%. Compared with the control group, the TMPRSS4 mRNA and protein levels were reduced by 80.1% and 60% ,and number of penetrating cells was 118.6 ±13.4 in SW1990/psi TMPRSS4 group, which was significantly lower than those in the negative control group (157.4 ± 12.9) and control group (157.0±9.5, P <0.01). Cells invasion inhibitory rate was 24.5% in SW1990/psi TMPRSS4 group. The cell proliferation was not significantly different among all the groups. Conclusions A stable cell strain is screened successfully in which the expression level of TMPRSS4 can be reduced stably. The down-regulation of TMPRSS4 gene expression level can inhibit the invasion of SW1990 cells, but has no effect on cell proliferation.
Key words:
Pancreatic neoplasms; Small interfering RN A; Gene silencing; TMPRSS4; SW1990 cell
Background: Gut microbiota play a crucial role in the development and progression of nonalcoholic steatohepatitis (NASH) and associated hepatocellular carcinoma (HCC). Akkermansia muciniphila as a next-generation probiotic was reported to inhibit inflammation-associated cancer in the intestine. The anti-NASH ability of A. muciniphila has recently been found. Thus, we were to investigate whether supplementation of A. muciniphila could prevent NASH-associated HCC.Methods: In a model we called STAM, male C57BL/6J mice were administered a single subcutaneous injection of 200 µg streptozotocin at 2 days after birth followed by high-fat diet to induce NASH (10 weeks of age) and HCC (20 weeks of age). Faeces from mice and patients with NASH-related HCC were collected for 16S rRNA sequencing. STAM mice were orally administered either saline or A. muciniphila twice a day starting at 4 or 10 weeks of age. The effects of A. muciniphila on the immune responses were also evaluated.Findings: The abundance of faecal A. muciniphila was significantly decreased in patients and mice with NASH-related HCC. Administration of A. muciniphila could improve NASH severity. Interestingly, A. muciniphila treatment suppressed the progression of NASH to HCC, with an increased hepatic natural killer T (NKT) cell and decreased macrophage infiltration. The antitumor ability of A. muciniphila was not evident in NKT cell-deficient mice (CD1d knockout). In vitro, A. muciniphila promoted the killing of hepG2 cells by NKT cells.Interpretation: Our study will provide theoretical basis for the application of A. muciniphila in the treatment of NASH and prevention for its progression to HCC.Funding Information: This work was supported by the National Natural Science Foundation of China (No. 82102684), China Postdoctoral Science Foundation (No. 2021M691472), Guangdong Science and Technology Project (No. 2017B020209003), Innovation Leader Team Program of Guangzhou (No. 201809010014) and R & D Plan for Key Areas in Guangdong Province (No. 2019B020204003).Declaration of Interests: The authors declare no conflict of interest.Ethics Approval Statement: All animal procedures were performed in accordance with the ‘‘Guide for the Care and Use of Laboratory Animals’’ prepared by the National Academy of Sciences and published by the National Institutes of Health (revised 1985). The study conformed to the ethical guidelines of the Declaration of Helsinki and approval by the Ethics Committee.
AIMTo evaluate the cytological diagnostic capacity and sample quality of the slow-pull technique and compare them with different suction techniques. METHODSFrom July 2010 to December 2015, 102 patients with pancreatic solid lesions who underwent endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) with 22-gauge needles were retrospectively evaluated.EUS-FNA diagnosis was based on a cytological examination, and final diagnosis was based on a comprehensive standard of cytological diagnosis, surgical pathology and clinical or imaging follow-up.Cytological specimens were characterized for cellularity and blood contamination.The cytological diagnostic capacity and sample quality of the slow-pull technique and suction techniques with 5-mL/10-mL/20-mL syringes were analyzed.
Objective The topical antispasmodic agent l ‐menthol is commonly used for gastric peristalsis suppression during diagnostic upper gastrointestinal (GI) endoscopy. We evaluated the efficacy and safety of a single dose l ‐menthol solution in suppressing gastric peristalsis during upper GI endoscopy in Chinese patients. Methods In this phase III, multicenter, randomized, double‐blind, placebo‐controlled study (ClinicalTrials.gov: NCT03263910), 220 patients scheduled to undergo upper GI endoscopy at five Chinese referral centers received a single dose of either 160 mg of l ‐menthol ( n = 109) or placebo ( n = 111). Both treatments were sprayed endoscopically on the gastric mucosa. An independent committee evaluated the degree of gastric peristalsis (peristaltic score: grade 1–5). Results At baseline, the proportion of patients with grade 1 peristalsis (no peristalsis) did not differ between the groups. The proportion of patients with grade 1 peristalsis post‐treatment was significantly higher in the l ‐menthol group (40.37%, 44/109) versus the placebo group (16.22%, 18/111; P < 0.001); the difference between the groups was 24.15% (95% confidence interval: 12.67%–35.63%; P < 0.001). In the l ‐menthol group, 61.47% of patients had grade 1 peristalsis after endoscopy versus 24.55% in the placebo group ( P < 0.001). The ease of intragastric examination correlated significantly with the grade of peristalsis. The incidence of adverse events was comparable between the groups ( P = 0.340). Conclusions During upper GI endoscopy, a single dose of l ‐menthol solution (160 mg) sprayed on the gastric mucosa significantly attenuated gastric peristalsis versus placebo, thereby improving the visual stability without any safety concerns.
Abstract Although the abnormal expression of miRNAs in cancer cells is a widely accepted phenomenon, the molecular mechanisms underlying miR-3648 progression and metastasis in gastric cancer (GC) remain unclear. miR-3648 expression is downregulated and its ectopic expression in GC cells significantly suppressed cell proliferation and metastasis. Mechanistic analyses indicated that miR-3648 directly targets FRAT1 or FRAT2 and inhibits FRAT1- or FRAT2-mediated invasion and motility in vitro and in vivo. Moreover, FRAT1 physically interacted with FRAT2. Furthermore, FRAT1 overexpression promoted GC cell invasion, whereas siRNA-mediated repression of FRAT2 in FRAT1-overexpressing GC cells reversed its invasive potential. Besides, miR-3648 inactivated the Wnt/β-catenin signalling pathway by downregulating FRAT1 and FRAT2 in GC. Interestingly, c-Myc, a downstream effector of Wnt/β-catenin signalling, was also downregulated by miR-3648 overexpression. In turn, c-Myc negatively regulated miR-3648 expression by binding to the miR-3648 promoter. In addition, miR-3648 expression levels were negatively correlated with c-Myc, FRAT1, and FRAT2 expression in fresh gastric samples. Our studies suggest that miR-3648 acts as a tumour-suppressive miRNA and that the miR-3648/FRAT1-FRAT2/c-Myc negative feedback loop could be a critical regulator of GC progression.
Transcription factors (TFs) and long noncoding RNAs (lncRNAs) contribute to gastric cancer (GC). However, the roles of TFs and lncRNAs in the invasion and metastasis of GC remain largely unknown. Here, we observed that the transcription factor VAX2 is significantly upregulated in GC cells and tissues and acts as an oncogene. Moreover, high VAX2 expression is associated with the advancement of tumors in GC. In terms of functionality, the enforced expression of VAX2 promotes the proliferation and metastasis of GC cells. Mechanistically, VAX2 specifically interacts with the LINC01189 promoter and represses LINC01189 transcription. Furthermore, LINC01189 exhibits significant downregulation in GC and functions as a suppressor gene. Functionally, it inhibits migratory and invasive abilities in GC cells. In the context of GC metastasis, VAX2 plays a role in modulating it by trans-repressing the expression of LINC01189. Additionally, LINC01189 binds to hnRNPF to enhance hnRNPF degradation through ubiquitination. The cooperation between LINC01189 and hnRNPF regulates GC cell invasion and migration. In addition, both VAX2 and hnRNPF are highly expressed, while LINC01189 is expressed in at low levels in GC tissues compared to normal gastric tissues. Our study suggests that VAX2 expression facilitates, while LINC01189 expression suppresses, metastasis and that the VAX2-LINC01189-hnRNPF axis plays a contributory role in GC development.
Corrigendum on: Li T, Lin X, Shen B, Zhang W, Liu Y, Liu H, Wang Y, Zheng L and Zhi F (2022) Akkermansia muciniphila suppressing nonalcoholic steatohepatitis associated tumorigenesis through CXCR6 + natural killer T cells. Front. Immunol. 13:1047570. doi: 10.3389/fimmu.2022.1047570In the published article, there was an error in [Figure 1F] as published. [According to the experimental design of this paper, the original purpose of Figure 1F was to confirm the decreased abundance of A. muciniphila in the colon of STAM mice (20 weeks old) compared to control mice. However, in Figure 1F, we included FISH results of STAM mice at 4, 10, 16 weeks of age by mistake, which was not consistent with the description in the Results part. According to the description in the Results part, the comparision of the FISH results of STAM mice (20 weeks old) with control mice should be include in Figure 1F, which makes the results more readable and rigorous.]. The corrected [Figure 1F] and its caption appear below.
To evaluate the value of deep small-bowel endoscopy (DSBE) in the diagnosis of Crohns disease (CD).The endoscopic and clinical data of 54 patients with CD receiving capsule endoscopy (CE) and double-balloon enteroscopy (DBE) between January, 2004 and December, 2008 were summarized and analyzed retrospectively.The main indications for DSBE in our series were suspected CD (42.6%) and obscure gastrointestinal bleeding (25.9%). DSBE was obviously superior to barium imaging. The detection rate of CD was significantly higher with DSBE (92.6%) than with ileocolonoscopy (75.9%, P=0.017), and DSBE provides much more detailed descriptions of specific endoscopic features such as segmental distribution and lumen changes. DSBE significantly improve the diagnostic efficiency, giving priority to offer a guide and raise suspected diagnosis for CD.DSBE is a valuable modality for detecting CD lesions in the jejunum and ileum and for evaluating lesion involvement and severity. The combination with a comprehensive analysis of routine imaging findings, gastro endoscopy, and clinical data can further enhance the diagnostic efficiency of DSBE.
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