OBJECTIVES: To investigate the role of SF3B3 in gastric cancer (GC) progression and prognosis and its possible mechanisms. METHODS: SF3B3 expression levels in pan-cancer and GC were analyzed using TIMER2.0, GEPIA, and UALCAN databases and validated using immunohistochemistry in GC tissues. Survival curves of GC patients were established using Kaplan-Meier Plotter and the data of a patient cohort our hospital. The independent risk factors for 5-year postoperative survival were identified using Cox regression, and their predictive values were evaluated using ROC analysis. SF3B3-associated biological processes were predicted by bioinformatics enrichment analyses. In GC HGC-27 cells, the effects of lentivirus-mediated SF3B3 knockdown and overexpression on cell proliferation and migration were investigated, and the changes in the key glycolytic proteins and extracellular acidification rate (ECAR) were detected. The influence of SF3B3 expression level on tumorigenesis and glycolytic protein expression in vivo were evaluated in a nude mouse xenograft model. RESULTS: High expression of SF3B3 in GC was associated with poor patient prognosis (P<0.05). The factors affecting 5-year survival outcomes following gastric oncological resection included high SF3B3 expression, a CEA level ≥5μg/L, a CA19-9 level ≥37 kU/L, tumor stage T3-4, and lymph node metastasis stage N2-3 (P<0.05). Bioinformatics analysis showed significant enrichment of SF3B3 in glycolysis. In HGC-27 cells, SF3B3 knockdown significantly inhibited while SF3B3 overexpression enhanced cell proliferation, migration, and invasion. SF3B3 knockdown obviously decreased the expressions of HK2, PKM2 and LDHA proteins and ECAR in HGC-27 cells, whereas SF3B3 overexpression produced the opposite effect. In nude mouse xenograft models, SF3B3 knockdown significantly reduced tumor mass and downregulated expression of HK2, PKM2 and LDHA proteins, and SF3B3 overexpression induced the opposite changes. CONCLUSIONS SF3B3 overexpression is associated with poor prognosis of GC patients and promotes GC cell proliferation, migration and invasion possibly by enhancing glycolysis.
Stomach Neoplasms/diagnosis* ; Humans ; Cell Proliferation ; Prognosis ; Animals ; Mice, Nude ; Cell Line, Tumor ; Mice ; Cell Movement ; Male ; Female

OBJECTIVES: To investigate YEATS2 expression in gastric cancer (GC), its prognostic value, and its regulatory role in epithelial-mesenchymal transition (EMT) of GC cells. METHODS: YEATS2 expression in GC was analyzed using publicly available databases. Paired GC and adjacent tissues were collected from 100 patients undergoing radical surgery for immunohistochemical detection of YEATS2 expression, and its correlations with the patients' clinicopathological parameters and Ki67 expression were analyzed. The prognostic value of YEATS2 was assessed using Kaplan-Meier analysis, Cox regression and ROC curves, and its regulatory mechanisms were analyzed using KEGG enrichment analysis. In cultured GC cell lines (HGC-27 and AGS), the effect of YEATS2 knockdown and overexpression on migration, invasion and EMT of the cells were examined with scratching assay, Transwell assay and Western blotting. RESULTS: YEATS2 was significantly overexpressed in GC tissues with a positive correlation with Ki67 (P<0.05). High YEATS2 expression was associated with elevated CEA (≥5 μg/L), CA19-9 (≥37 kU/L), T3-4 stage, and N2-3 stage (all P<0.05). Patients with high YEATS2 expression had significantly reduced 5-year survival (P<0.001); ROC analysis showed that YEATS2 expression levels had a sensitivity of 80.00% and a specificity of 66.67% for predicting patient survival (P<0.05). Cox regression identified high YEATS2 as an independent risk factor for poor postoperative 5-year survival outcome of GC patients (HR: 1.675, 95%CI: 1.013-2.771; P=0.045). KEGG enrichment analysis suggested involvement of YEATS2 in EMT in GC and Wnt/β-catenin signaling. In cultured GC cells, YEATS2 overexpression significantly promoted cell migration and invasion, upregulated the expressions of vimentin, N-cadherin, Wnt and active β-catenin, and downregulated E-cadherin expression, and these changes were obviously suppressed by treatment with XAV-939 (a Wnt/β-catenin inhibitor). CONCLUSIONS High YEATS2 expression activates Wnt/β-catenin signaling to promote EMT in GC and is correlated with poor prognosis of GC patients.
Humans ; Stomach Neoplasms/pathology* ; Epithelial-Mesenchymal Transition ; Wnt Signaling Pathway ; Cell Line, Tumor ; Prognosis ; Cell Movement ; Male ; Female ; beta Catenin/metabolism*

Objective To investigate the effect and molecular mechanism of isolongifolene(ISO)on the apoptosis of intestinal epithelial cells and 2,4,6-trinitrobenzenesulfonic acid(TNBS)-induced Crohn's disease(CD)-like colitis in mice.Methods In the animal experiments,mice were randomly assigned to the wild type(WT)group,TNBS group and TNBS+ISO group,with 8 mice in each group.Colitis models of mice were established in the TNBS group and the TNBS+ISO group by rectal injection of TNBS.After modeling,the mice in the TNBS+ISO group were given ISO intervention via intragastric gavage(10 mg/kg),and the other two groups were given the same amount of normal saline via intragastric gavage.The mice were sacrificed on the 7th day.The changes in body mass,disease activity scores(DAI),and the colon length of mice were measured,and transepithelial electrical resistance(TEER)of the colon tissues was determined.The score of colon inflammation was calculated according to HE staining.The levels of intestinal mucosal inflammatory factors,including tumor necrosis factor alpha(TNF-α),interferon(IFN)-γ,interleukin(IL)-1 β,and IL-6,were measured by RT-PCR and ELISA.The apoptosis of colon tissue cells was determined by TUNEL assay.The expressions of apoptotic proteins(cleaved caspase-3/caspase-3 and Bax),an anti-apoptotic protein(Bcl-2),and tight junction proteins(ZO-1 and claudin-1)were detected by Western blot and immunofluorescence.In the cell experiment,TNF-α was used to induce intestinal epithelial cell Caco-2 apoptosis model,which was treated with ISO.Then,intervention with the AMPK inhibitor Compound C was given.TUNEL assay,Western blot assay,and immunofluorescence assay were performed to measure apoptosis and the expression of apoptosis proteins in the Caco-2 cells.Gene Ontology(GO)enrichment analysis was performed to predict the biological function of ISO.Then,the mechanism involved was verified by examination of the mice and Caco-2 cells.Western blot was performed to determine the expression levels of p-AMPK/AMPK and p-PGC1α in the colon tissues from the mice of different groups and Caco-2 cells.The apoptosis of the cells was determined by TUNEL assay.Results According to the results of the animal experiment,ISO could alleviate experimental colitis and intestinal barrier dysfunction,leading to improvements in body mass loss,colon length shortening,DAI score,inflammatory rating,and TEER values(all P＜0.05)in mice.Furthermore,ISO decreased the expression of pro-inflammatory factors TNF-α,IFN-γ,IL-1β,and IL-6 and increased the expression of the tight junction proteins ZO-1 and claudin-1(all P＜0.05).In the cell experiment,in a TNF-α-induced intestinal epithelial cell model,ISO was also found to protect intestinal barrier against damage.ISO reduced the proportion of apoptotic intestinal epithelial cells,reduced the expression of cleaved-caspase-3/caspase-3 and Bax,and upregulated the level of Bcl-2(all P＜0.05).GO enrichment predictive analysis showed that the role of ISO in improving CD-like enteritis might be associated with the negative regulation of apoptosis.Verification of the mechanism showed that the expression of p-AMPK and p-PGC1α in the mice colon tissue was significantly upregulated after ISO intervention(P＜0.05).In contrast,the AMPK inhibitor Compound C increased the apoptosis rate of ISO-treated Caco-2 cells and decreased the relative expression levels of ZO-1 and claudin-1(P＜0.05).Conclusion ISO reduces intestinal epithelial cell apoptosis at least in part by activating AMPK/PGC1α signaling pathway,thereby alleviating TNBS-induced intestinal barrier dysfunction and CD-like colitis in mice.

Objective:To compare influence of valsartan (Val) combined amlodipine (Am) or hydrochlorothiazide (Hyd) on blood pressure variability and quality of life in aged patients with hypertension .Methods:A total of 127 aged patients with hypertension stage 2 and 3 were randomly divided into Val + Am group (n=64) and Val + Hyd group (n=63) .Circadian rhythm and variability of blood pressure and quality of life were observed in both groups before and after treatment .Results:(1) After treatment ,the 24hSBP ,DBP ,SBPV , DBPV and daytime SBP , DBP ,SBPV ,DBPV ,and nighttime ,morning surge of SBP and DBP all significantly reduced in both groups;(2) Compared with Val+ Hyd group ,there were significant reductions in 24hSBP [(120.6 ± 10.2) mmHg vs . (110.9 ± 11.3) mmHg] ,daytime SBP [(120.6 ± 11.3) mmHg vs .(111.6 ± 11.37) mmHg] ,nighttime SBP [(118.5 ± 11.6) mmHg vs .(108.6 ± 11.9) mmHg] ,morning surge of SBP [ (26.2 ± 13.7) mmHg vs .(23.0 ± 10.4) mmHg] (P<0.05 or <0.01);24hSBPV [ (10.7 ± 2.2) mmHg vs .(8.2 ± 2.0) mmHg] ,daytime SBPV [ (10.4 ± 1.9) mmHg vs .(8.1 ± 2.1) mmHg] (P<0.01 all);significant rise in improved percentages of physical health ,mental health and total function in quality of life (P<0.05 or <0.01) in Val + Am group .Conclusion:Valsartan combined am-lodipine or hydrochlorothiazide can both effectively improve blood pressure variability and quality of life in aged pa-tients with hypertension ,and the effect of the former is even better .

Objective To better understand the role of CREB signaling pathway in chronic mild stress (CMS), we investigated the alteration of CREB and p-CREB in CMS rats with and without fluoxetine hydrochloride. Methods Fifty adult male Sprague-Dawley rats were randomly divided into three groups:CMS group (26), fluoxtine group (12) and con-trol group (12). The rats in CMS group and fluoxtine group received 8 weeks of chronic mild stress. Rats in fluoxtine group were administered daily injections of fluoxetine 10mg/kg I.P. Sucrose preference tests and open-field test were car-ried out after the 8th week. Based on endpoint sucrose-intake, animals were further divided into 4 groups:CMS sensitive group, CMS resilient group, fluoxtine group and control group. Western blot was used to detect the expression levels of CREB and p-CREB in the hippocampus and prefrontal cortex. Results The sucrose consumption was significantly de-creased in CMS resilience group compared to sensitive group, control group and fluoxetine-intervention group (all P<0.05). Similarly, the numbers in total arm entries, percentage of entries into open arms and time spent in open arms was significantly lower in CMS resilience group compared to control group(all P<0.05), but not different compared to CMS sensitive group(all P<0.05). The p-CREB in the hippocampus was significantly lower in CMS sensitive rat compared to CMS resilience group, control group and fluoxetine-intervention group(all P<0.05), but CREB was not dfferent among the four groups(all P<0.05). Conclusions The elevated phosphorylation of CREB in the hippocampus and prefrontal cortex of resilience CMS rats may contribute to the mood alteration induced by stress.

Objective: To explore the diagnostic value of head-up tilt test with sublingual isosorbide dinitrate （HUTSI）in patients with vasovagal syncope (VVS). Methods: A total of fifty-two consecutive patients with clinically vasovagal syncope (VVS group) and thirty-eight control subjects without prior experience of syncope (normal control group) were evaluated by baseline head-up tilt table test (BHUT) , then the BHUT negative subjects underwent HUTSI test. . Results: (1) Positive BHUT test rate of VVS group and normal control group were 25% (13/52) and 0 respectively; from supine to HUT positive, there were significant decrease in heart rate [(73.1±8.5) times/min vs. (56.2±11.2) times/min] and mean arterial pressure [MAP, (81.2± 10.8) mmHg vs. (50.2±10.4) mmHg，P<0.05 both] in BHUT positive subjects. There were 21 HUTSI positive cases（53.9%）among the other 39 VVS patients, and two cases（5.3%）among the 38 subjects of normal control group. From supine to HUTSI positive, there were significant decrease in heart rate [(65.2±7.5) times/min vs. (52.9±10.5) times/min] and MAP [(78.3±10.7) mmHg vs. (48.8±11.2) mmHg, P<0.05 both] in HUTSI positive subjects. Duration from tilt started to positive reaction occurred in HUTSI positive group was significantly shorter than that of BHUT positive group [(10.8±9.3) min vs. (21.1±11.5) min，P<0.05]. All subjects can tolerate the test and only two cases in VVS group and one case in normal control group occurred headache and face red. Conclusion: Head-up tilt test with sublingual isosorbide dinitrate is a practical and easy-to- perform method with high sensitivity, specificity and few side effects for diagnosis of vasovagal syncope.