The transition of cancer cells from epithelial state to mesenchymal state awarded hepatocellular carcinoma (HCC) stem cell properties and induced tumorigenicity, drug resistance, and high recurrence rate. Reversing the mesenchymal state to epithelial state by inducing mesenchymal-epithelial remodeling could inhibit the progression of HCC. Using high-throughput screening, chrysin was selected from natural products to reverse epithelial-mesenchymal transition (EMT) by selectively increasing CDH1 expression. The target identification suggested chrysin exerted its anti-HCC effect through covalently and specifically binding threonine 205 (Thr205) of alpha-enolase (ENO1). For the first time, we revealed that ENO1 bound β-catenin mRNA, and recruited YTHDF2 to identify the m6A modified β-catenin in the 3'-UTR region to degrade β-catenin mRNA. Eventually, the CDH1 gene expression was improved through the regulation of β-catenin mRNA. ENO1/β-catenin mRNA interaction might be a promising target for cellular plasticity reprogramming. Moreover, chrysin could mediate mesenchymal‒epithelial remodeling through increasing degradation of β-catenin mRNA by promoting the binding of ENO1 and β-catenin mRNA. To the best of our knowledge, chrysin is the first reported small molecule inducing β-catenin mRNA degradation through binding to ENO1. The water-soluble derivative of chrysin may be a natural product-derived lead compound for circumventing metastasis, recurrence, and drug resistance of HCC by mediating mesenchymal‒epithelial remodeling.

AIM:To investigate the effects and mechanism of long noncoding RNA PCED1B antisense strand 1(lncRNA PCED1B-AS1)on the proliferation,migration,invasion and apoptosis of papillary thyroid carcinoma(PTC)cells.METHODS:Human PTC cells were cultured in vitro.The expression of PCED1B-AS1 and fused in sarcoma(FUS)was measured by RT-qPCR.The effects of knockdown/overexpression of PCED1B-AS1/FUS on the migration and invasion of PTC cells were detected via Transwell assay.The effects of knockdown/overexpression of PCED1B-AS1/FUS on PTC cell proliferation were analysed via CCK-8 and plate colony assay.The effect of knockdown PCED1B-AS1 on PTC cell apoptosis was determined by flow cytometry.The target binding of PCED1B-AS1 and FUS was determined with bioin-formatics and RNA immunoprecipitation(RIP)experiments.Fluorescence in situ hybridization experiment was performed to verify whether PCED1B-AS1 colocalises with FUS.The mitogen-activated protein kinase(MAPK)signaling pathway-re-lated proteins were detected via Western blot.RESULTS:(1)PCED1B-AS1 expression was significantly higher and FUS expression was significantly lower in PTC cells compared with normal thyroid Nthy-ori3-1 cell(P<0.05).(2)Knockdown of PCED1B-AS1 and overexpression of FUS inhibited PTC cell migration,invasion and proliferation,and promoted apopto-sis(P<0.05).(3)Bioinformatics analysis and RIP assay verified the existence of targeted binding of PCED1B-AS1 to FUS(P<0.05).(4)PCED1B-AS1 and FUS colocalised in the cytoplasm.(5)Inhibition of PCED1B-AS1 decreased the expression of MAPK signaling pathway-related proteins p-ERK 1/2,p-JNK and p-P38(P<0.05).CONCLUSION:ln-cRNA PCED1B-AS1 inhibits the proliferation,migration and invasion,and promotes the apoptosis of PTC cells,and its mechanism may be related to the expression of FUS and the MAPK signaling pathway.

Objective To investigate the effect of long intergenic non-coding RNA COX2 (lincRNA-COX2) on apoptosis and polarization of Listeria monocytogenes (Lm)-infected RAW264.7 cells. Methods RAW264.7 cells were cultured and divided into control group (uninfected cells), Lm infection group, negative control of small interfering RNA (si-NC) group, si-NC and Lm infection group, small interfering RNA of lincRNA-COX2 (si-lincRNA-COX2) group, si-lincRNA-COX2 and Lm infection group. RAW264.7 cells were infected with MOI=10 Lm for 6 hours, and then the inhibition efficiency of siRNA transfection was detected by fluorescence microscope and quantitative real-time PCR (qRT-PCR). The expression levels of cleaved-caspase-3(c-caspase-3), caspase-3, B-cell lymphoma-2 (Bcl2), Bcl2 associated X protein (BAX), arginase 1 (Arg1), inducible nitric oxide synthase (iNOS) were detected by Western blot analysis. Results c-caspase-3/caspase-3, BAX/Bcl2 and iNOS were significantly up-regulated, while the level of Arg1 was down-regulated in Lm-infected RAW264.7 cells compared with control group. LincRNA-COX2 knockdown inhibited the increase of protein levels for BAX/Bcl2, c-caspase-3/caspase-3 and iNOS in Lm-infected RAW264.7 cells, while the level of Arg1 in Lm-infected RAW264.7 cells was up-regulated. Conclusion Knockdown of lincRNA-COX2 can inhibit cell apoptosis and suppress the macrophage polarization into M1 type in Lm-infected RAW264.7 cells.
Apoptosis/genetics* ; bcl-2-Associated X Protein/metabolism* ; Caspase 3/metabolism* ; Cyclooxygenase 2/metabolism* ; Listeria monocytogenes/pathogenicity* ; Macrophages/microbiology* ; RNA, Long Noncoding/metabolism* ; RNA, Small Interfering/genetics* ; Animals ; Mice

Objective To investigate the effect of Trichomonas vaginalis macrophage migration inhibitory factor (TvMIF) on THP-1 macrophages.. Methods Recombinant TvMIF protein was prokaryotic expressed and purified, and endotoxin was removed after identification. Following exposure to TvMIF at concentrations of 0, 1, 5, 10, 50 and 100 ng/mL, the cytotoxicity of the recombinant TvMIF protein to THP-1 macrophages was tested using cell counting kit (CCK)-8 assay, and the apoptosis of THP-1 macrophages and reactive oxygen species (ROS) were detected using flow cytometry. The relative expression of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), caspase-1, interleukin-1β (IL-1β) and IL-18 genes was quantified using real-time fluorescent quantitative PCR (qPCR) assay, and the expression of caspase-1, NLRP3, gasdermin D (GSDMD), gasdermin D N-terminal (GSDMD-NT) and pro-IL-1β proteins were determined using Western blotting assay. Results Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) displayed successful expression and purification of the recombinant TvMIF protein with a molecular weight of 15.5 kDa, and the endotoxin activity assay showed the successful removal of endotoxin in the recombinant TvMIF protein (endotoxin concentration < 0.1 EU/mL), which was feasible for the subsequent studies on protein functions. Flow cytometry revealed that the recombinant TvMIF protein at a concentration of 10 ng/mL and less promoted the apoptosis of THP-1 macrophages, and the highest apoptotic rate of THP-1 macrophages was seen following exposure to the recombinant TvMIF protein at a concentration of 5 ng/mL, while the recombinant TvMIF protein at concentrations of 50 and100 ng/mL inhibited the apoptosis of THP-1 macrophages. Exposure to the recombinant TvMIF protein at a concentration 1 ng/mL resulted in increased ROS levels in THP-1 macrophages. qPCR assay quantified significantly elevated caspase-1, NLRP3, IL-18 and IL-1β expression in THP-1 macrophages 8 hours post-treatment with the recombinant TvMIF protein at a concentration 1 ng/mL, and Western blotting determined increased caspase-1, NLRP3, pro-IL-1β, GSDMD and GSDMD-NT protein expression in THP-1 macrophages following exposure to the recombinant TvMIF protein at a concentration 1 ng/mL. Pretreatment with MCC950 significantly reduced GSDMD and GSDMD-NT protein expression. Conclusions High-concentration recombinant TvMIF protein inhibits macrophage apoptosis, while low-concentration recombinant TvMIF protein activates NLRP3 inflammasome and promotes macrophage pyroptosis.

Objective To investigate the effect of tetrahydrobiopterin (BH₄), superoxide dismutase-polyethylene glycol (PEG-SOD) and N(G)-nitro-L-arginine (L-NNA) on impaired endothelial progenitor cell (EPC) functions in DOCA-salt hypertensive mice. Methods DOCA-salt hypertension was created and systolic blood pressure was measured by tail-cuff methods. EPC was isolated from bone marrow of mice and characterized by flow cytometry and fluorescence microscopy. EPC of DOCA-salt mice was incubated with BH₄, PEG-SOD, and L-NNA for 24 h, then in vitro EPC function assays were performed. Results Compared with control group, systolic blood pressure was significantly increased in DOCA-salt mice. Both EPC adhesion and angiogenesis functions were impaired in DOCA-salt mice compared to control animals, which were reversed by incubation with BH₄, PEG-SOD and L-NNA. Conclusion BH₄, PEG-SOD and L-NNA rescued the impairments from EPC functions in DOCA-salt hypertensive mice.

PURPOSE: To identify the potential therapeutic role of postoperative radiotherapy (RT) in patients with locally advanced (stage II and stage III) gastric signet ring cell carcinoma (SRC).MATERIALS AND METHODS: Patients with locally advanced gastric SRC from the Surveillance, Epidemiology, and End Results program database between 2004 and 2012 were included in our study. Univariate and multivariate Cox proportional models were performed, and survival curves were generated to evaluate the prognostic effect of postoperative RT and surgery alone on SRC patients. Propensity score matching (PSM) was used to avoid selection bias among the study cohorts.RESULTS: We found that patients with postoperative RT had better probability of survival compared with those who did not receive RT (overall survival [OS], P<0.001; cancer-specific survival [CSS], P<0.001). After PSM, analysis of both overall and CSS showed that patients who underwent postoperative RT had better prognosis than those receiving surgery alone in the matched cohort (OS, P=0.00079; CSS, P=0.0036). Multivariate Cox proportional model indicated that postoperative RT had better effect on prognosis compared with surgery alone with respect to both overall (hazard ratio [HR], 0.716; 95% confidence interval [95% CI], 0.590–0.87; P=0.001) and CSS (HR, 0.713; 95% CI, 0.570–0.890; P=0.003).CONCLUSIONS: Postoperative RT had better prognosis compared with surgery alone for both overall and CSS for patients with locally advanced gastric SRC.
Carcinoma, Signet Ring Cell ; Cohort Studies ; Humans ; Nomograms ; Prognosis ; Propensity Score ; Radiotherapy ; SEER Program ; Selection Bias ; Stomach Neoplasms

Objective To investigate the effect of polyphosphazene(PAGP) microspheres controled release of growth factors on the adhesion and proliferation of bone marrow mesenchymal stem cells (BMSCs).Methods Two kinds of functional poly(alanine ethyl ester-co-glycine ethyl ester) phosphazene microspheres with different ratios of side-substituent groups were synthesized using the emulsion technique.The rate of degradation/hydrolysis of the polymers was carefully tuned to suit the desired application for controlled release.The enzyme-linked immune sorbent assay was utilized to investigate the characterization of the temporal controlled release strategy of microspheres loaded with transforming growth factor-beta 1(TGF-β1) and insulin-like growth factor-l(IGF-1) respectively.The cell adhesion and proliferation stimulated by different growth factors were evaluated by acridine orange staining.Resuits The morphological difference between two PAGP microspheres was identified according to SEM images.The average diameter of two microspheres was 54.22 ± 19.19 μm and 34.11 ± 18.82 μm respectively.The release assay showed that two kinds of microspheres had different release characteristics,with earlier outburst of TGF-β1 and IGF-1 for them to cooperate and later sustained release of TGF-β1 to stimulate the differentiation of stem cells.The result of the acridine orange staining demonstrated that PAGP microspheres supported cell adhesion and growth without obvious cytotoxicity.Meanwhile,the growth factors release strategy significantly improved the proliferation of BMSCs.Conclusion The two polyphosphazene microspheres have a great release-control effect and their controlled release system will have a promising prospect in the future tissue engineering field.

Objective To survey the awareness and knowledge of Helicobacter pylori (Hp) infection among medical staff in Shanghai.Methods A questionnaire survey was conducted among 316 medical staff in Shanghai,including 74 gastroenterologists (GI),158 general practitioners (GP),and 68 gastroenterology nurses(GN),from October 2014 to September 2015.The questionnaire was designed according to the Fourth Helicobacter Pylori Infection Treatment Consensus Report of China (the Consensus).There were 4 parts and 29 questions in the questionnaire,including the knowledge and performance of the Consensus (8 questions),the indications of Hp eradication (8 questions),detection methods of Hp infection (7 questions)and the therapy of Hp eradication (6 questions).Results Total 300 valid questionnaires were received with a response rate of 94.9％ (300/316).The awareness rate of the Consensus in GI,GP and GN groups was 81.1％ (60/74),57.6％ (91/158) and 26.4％ (18/68),respectively (χ2 =43.67,P=0.001).GI had higher awareness rate than GP and GN in indications of Hp eradications (for peptic ulcer,mucosa-associated lymphoid malignancies,post-resection patients of early gastric cancer,and family history of gastric cancer,the χ2 values were 16.68,35.60,33.46 and 39.22,respectively;all P ＜0.05).In part of Hp infection detection methods,the responses of GI,GP and GN groups in C14 or C13 urea breathing test were 97.3％ (72/74),47.5％ (75/158) and 82.1％ (55/68),respectively (χ2 =72.38,P =0.001);in gastric mucosa tissue rapid urease test were 70.3％ (52/74),13.9％ (22/158) and 25.4％ (17/68),respectively (χ2 =78.22,P =0.001);in serological test were 58.1％ (43/74),20.9％ (33/158)and 44.8％ (30/68),respectively (χ2 =40.30,P =0.001);in gastric mucosa tissue section staining were 56.8％ (42/74),13.3％ (21/158) and 22.4％ (15/68),respectively (χ2 =50.35,P =0.00).In part of Hp eradication therapy the responses of GI,GP and GN groups in recommended bismuth quadruple therapy were 71.6％ (53/74),47.5％ (75/158) and 40.3％ (25/62),respectively (χ2 =15.93,P =0.001);in triple therapy were 27.0％ (20/74),51.6％ (81/158) and 42.0％ (26/62),respectively (χ2 =12.42,P =0.002);in 10 or 14 d for treatment duration were 78.4％ (58/74),78.5％ (124/158)and 67.6％ (46/68),respectively (χ2 =3.36,P =0.186).Conclusion Gastroenterologists are more likely to adhere with the Consensus than general practitioners and gastroenterological nurses in the management of Hp infection.The survey suggests that more attention should be paid for popularization and implementation of Hp infection guidelines and consensus among Shanghai medical staff,especially for GP and nurses.

Objective:To explore the effect of interleukin-1α (IL-1 α) on the senescence of human umbilical vein endothelial cells (HUVECs) and the function of high mobility group protein 1 (HMGB 1).Methods:HUVECs were randomly divided into a control group,a IL-1α group (10 ng/mL IL-1α),a HMGB 1 group (100 ng/mL HMGB 1),and a HMGB 1 +IL-1α group (100 ng/mL of HMGB 1 plus 10 ng/mL of IL-lα).Senescence associated β-galactosidase (SA β-gal) staining was used to assess the number of senescent cells,Western blot were performed to detect the protein levels of silent information regulator 1(SIRT1),and quantitative real-time PCR (qRT-PCR) was used to detect the mRNA levels of p53,p21 and p 16.Restlts:Compared with the control group,the number of SA β-gal positive cells were significantly increased in the IL-1α group (P＜0.05),while the expression of SIRT1 protein significantly decreased (P＜0.01).Compared with the IL-1 α group,the expression of SA β-gal positive cells in the HMGB 1+IL-1α group was decreased and the mRNA levels of p21 and p53 were down-regulated (all P＜0.05),however,there was no statistical significance in the mRNA expression ofp16 (P＞0.05).Conclusion:IL-1α can induce the senescence of HUVECs,and HMGB1 may inhibit IL-1α-induced endothelial cell senescence via p53-p21 pathway.

OBJECTIVETo investigate the factors which may influence the imatinib plasma concentration in Chinese patients with gastrointestinal stromal tumor(GIST), and to illuminate the significance of monitoring imatinib plasma concentration in adjuvant therapy for patients with GIST.

METHODSA cross-sectional study with 60 GIST patients who accepted the imatinib therapy after surgery was conducted. They were respectively administrated in 10 domestic hospitals from December 2014 to April 2016, including The First Affiliated Hospital of Nanjing Medical University(n=28), The Affiliated Hospital of Nantong University(n=9), The Affiliated Hospital of Xuzhou Medical College(n=6), Nanjing Drum Tower Hospital(n=5), The Second Affiliated Hospital of Nanjing Medical University (n=2), Jingling Hospital (n=2), The Second People's Hospital of Lianyungang(n=2), Shandong Provincial Hospital(n=2), Jiangsu Province Tumor Hospital(n=2), and The First Affiliated Hospital of Zhejiang University(n=2). Some specific time points for collecting blood sample before and after taking imatinib were determined, then liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was used for monitoring imatinib plasma concentration in patients with GIST. Linear regression analysis was used for the correlation analysis of imatinib plasma concentration with dosage, clinicopathologic feature and side effect.

RESULTSPatients who could not tolerate 400 mg imatinib per day(n=3) received 300 mg per day. There was no significant difference in imatinib plasma concentration between patients with 300 mg and those with 400 mg imatinib(n=53)(P=0.527). However, the imatinib plasma concentration in patients with 600 mg imatinib per day (n=4) was significantly higher as compared to those with 400 mg(P=0.000). Linear regression analysis indicated a negative correlation between the imatinib plasma concentration in patients with 400mg imatinib per day for 90 days continuously and body surface area(R=0.074, P=0.035), but no significant correlations of with age, creatinine clearance and serum albumin concentration were observed (all P>0.05). The differences in imatinib plasma concentration were not statistically significant between patients of different gender and those taking proton-pump inhibitor (PPI) or not (both P>0.05). Difference in imatinib plasma concentration between patients with different surgery was significant (P=0.026). Compared to patients who underwent wedge resection, enterectomy and other surgeries, the imatinib plasma concentration of patients with subtotal gastrectomy or total gastrectomy decreased significantly (all P<0.05). After 90 days of taking imatinib continuously, linear regression analysis revealed a negative correlation between imatinib plasma concentration in patients with 400 mg imatinib per day and white blood cell count (R=0.103, P=0.013), and a positive correlation with serum alanine aminotransferase (ALT) concentration (R=0.076, P=0.033).

CONCLUSIONSThe imatinib plasma concentration in patients with larger body surface area, subtotal gastrectomy or total gastrectomy may be lower. For these patients, dosage of imatinib should be considered to increase in order to achieve effective plasma concentration. Excessive imatinib plasma concentration can result in some side effects, such as decrease of white blood cells and liver damage. Therefore, it is significant for receiving optimal clinical therapeutic efficacy to monitor imatinib plasma concentration, adjust imatinib dosage timely and keep imatinib plasma concentration in effective and safe range.

Adult ; Antineoplastic Agents ; administration & dosage ; pharmacokinetics ; Benzamides ; Combined Modality Therapy ; Cross-Sectional Studies ; Female ; Gastrectomy ; Gastrointestinal Stromal Tumors ; drug therapy ; surgery ; Humans ; Imatinib Mesylate ; administration & dosage ; pharmacokinetics ; Male ; Middle Aged ; Piperazines ; Pyrimidines ; Tandem Mass Spectrometry