Background and purpose:SEC14L1P1,a pseudogene of the SEC14 family,is closely associated with the development of various tumors,but its role in oral squamous cell carcinoma(OSCC)has not been clarified.This study aimed to gain insights into the expression characteristics and subcellular localization of SEC14L1P1 in OSCC cells,as well as its effects on OSCC cell proliferation and migration.Methods:The expression of SEC14L1P1 in head and neck squamous cell carcinoma(HNSCC)tissues was analyzed by the ENCORI database;The expression of SEC14L1P1 and its relationship with patient prognosis in HNSCC was further analyzed using the GDC and UCSC Xena databases.The expression of SEC14L1P1 in OSCC cell lines was detected by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR);RNA nucleoplasmic separation assay was performed to determine the localization of SEC14L1P1 in OSCC cells.SEC14L1P1 knockdown(SS-SEC14L1P1)group and knockdown control(SS-NC)group were established for CAL-27 cells,and SEC14L1P1 overexpression(SEC14L1P1)group and overexpression control(Vector)group were established for HN30 cells.The effects of SEC14L1P1 expression on the proliferation and migration abilities of cells in each group were assessed by cell counting kit-8(CCK-8)and transwell migration assays.RTFQ-PCR and Western blot experiments were used to detect the effects of altered SEC14L1P1 expression on the expression levels of epithelial-mesenchymal transition(EMT)-related genes.To investigate the effects of SEC14L1P1 on the proliferation of OSCC cells in vivo using a subcutaneous xenograft tumor model in nude mice,12 four-week-old BALB/c nude mice were randomly divided into two groups:the antisense oligonucleotide(ASO)-NC group and the ASO-SEC14L1P1 group,with 6 mice in each group.All mice were individually labeled.Further mechanistic studies were performed by analyzing molecules interacting with SEC14L1P1 through the RNAInter database,and the ENCORI database was queried for expression correlation between SEC14L1P1 and DHX9.The effect of altered SEC14L1P1 expression on the phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)pathway was detected by Western blot assay.Results:Database analysis showed that the expression of SEC14L1P1 was higher in HNSCC tissues than in normal tissues,and was strongly associated with poor patient prognosis.The RTFQ-PCR results showed that SEC14L1P1 was highly expressed in all six OSCC cell lines;RNA nucleoplasmic separation showed that SEC14L1P1 was mainly localized in the nucleus in CAL-27 and HN30 cells.Compared with SS-NC,the relative expression of SEC14L1P1 in the SS-SEC14L1P1 group was significantly lower and significantly inhibited cell proliferation and migration,while the relative expression of SEC14L1P1 in the SEC14L1P1 group was significantly higher compared with the Vector group,which also significantly increased cell proliferation and migration.The down-regulation of SEC14L1P1 was accompanied by increased mRNA and protein levels of E-cadherin,and decreased mRNA and protein levels of N-cadherin and vimentin,with the opposite result after SEC14L1P1 overexpression.In vivo experiments showed that the xenograft tumor weight and volume of the ASO-SEC14L1P1 group were significantly reduced.Further mechanistic studies revealed a positive correlation between SEC14L1P1 and DHX9 expressions,and DHX9 has been shown to activate the PI3K/AKT signaling pathway.Knockdown of SEC14L1P1 resulted in decreased protein expressions of phosphorylated-PI3K(p-PI3K)and phosphorylated-AKT(p-AKT),and overexpression of SEC14L1P1 increased protein expressions of p-PI3K and p-AKT.Conclusion:SEC14L1P1 showed high expression levels in OSCC cells and tissues and promoted the proliferation and migration of OSCC cells,a phenomenon that may be related to the regulation of the PI3K/AKT signaling pathway by SEC14L1P1,which in turn promotes EMT.

Background and purpose:SEC14L1P1,a pseudogene of the SEC14 family,is closely associated with the development of various tumors,but its role in oral squamous cell carcinoma(OSCC)has not been clarified.This study aimed to gain insights into the expression characteristics and subcellular localization of SEC14L1P1 in OSCC cells,as well as its effects on OSCC cell proliferation and migration.Methods:The expression of SEC14L1P1 in head and neck squamous cell carcinoma(HNSCC)tissues was analyzed by the ENCORI database;The expression of SEC14L1P1 and its relationship with patient prognosis in HNSCC was further analyzed using the GDC and UCSC Xena databases.The expression of SEC14L1P1 in OSCC cell lines was detected by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR);RNA nucleoplasmic separation assay was performed to determine the localization of SEC14L1P1 in OSCC cells.SEC14L1P1 knockdown(SS-SEC14L1P1)group and knockdown control(SS-NC)group were established for CAL-27 cells,and SEC14L1P1 overexpression(SEC14L1P1)group and overexpression control(Vector)group were established for HN30 cells.The effects of SEC14L1P1 expression on the proliferation and migration abilities of cells in each group were assessed by cell counting kit-8(CCK-8)and transwell migration assays.RTFQ-PCR and Western blot experiments were used to detect the effects of altered SEC14L1P1 expression on the expression levels of epithelial-mesenchymal transition(EMT)-related genes.To investigate the effects of SEC14L1P1 on the proliferation of OSCC cells in vivo using a subcutaneous xenograft tumor model in nude mice,12 four-week-old BALB/c nude mice were randomly divided into two groups:the antisense oligonucleotide(ASO)-NC group and the ASO-SEC14L1P1 group,with 6 mice in each group.All mice were individually labeled.Further mechanistic studies were performed by analyzing molecules interacting with SEC14L1P1 through the RNAInter database,and the ENCORI database was queried for expression correlation between SEC14L1P1 and DHX9.The effect of altered SEC14L1P1 expression on the phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)pathway was detected by Western blot assay.Results:Database analysis showed that the expression of SEC14L1P1 was higher in HNSCC tissues than in normal tissues,and was strongly associated with poor patient prognosis.The RTFQ-PCR results showed that SEC14L1P1 was highly expressed in all six OSCC cell lines;RNA nucleoplasmic separation showed that SEC14L1P1 was mainly localized in the nucleus in CAL-27 and HN30 cells.Compared with SS-NC,the relative expression of SEC14L1P1 in the SS-SEC14L1P1 group was significantly lower and significantly inhibited cell proliferation and migration,while the relative expression of SEC14L1P1 in the SEC14L1P1 group was significantly higher compared with the Vector group,which also significantly increased cell proliferation and migration.The down-regulation of SEC14L1P1 was accompanied by increased mRNA and protein levels of E-cadherin,and decreased mRNA and protein levels of N-cadherin and vimentin,with the opposite result after SEC14L1P1 overexpression.In vivo experiments showed that the xenograft tumor weight and volume of the ASO-SEC14L1P1 group were significantly reduced.Further mechanistic studies revealed a positive correlation between SEC14L1P1 and DHX9 expressions,and DHX9 has been shown to activate the PI3K/AKT signaling pathway.Knockdown of SEC14L1P1 resulted in decreased protein expressions of phosphorylated-PI3K(p-PI3K)and phosphorylated-AKT(p-AKT),and overexpression of SEC14L1P1 increased protein expressions of p-PI3K and p-AKT.Conclusion:SEC14L1P1 showed high expression levels in OSCC cells and tissues and promoted the proliferation and migration of OSCC cells,a phenomenon that may be related to the regulation of the PI3K/AKT signaling pathway by SEC14L1P1,which in turn promotes EMT.

Background and purpose:Long noncoding RNA(lncRNA)can regulate gene transcription,mRNA shear,stabilization and translation,and it is an important regulatory factor in a variety of biological processes.This study aimed to investigate the expression and clinical features of lncRNA FLJ30679 in oral squamous cell carcinoma(OSCC)and its effect on the malignant biological behavior of OSCC.Methods:The expression of FLJ30679 in head and neck squamous cell carcinoma(HNSCC)tissues and normal tissues was analyzed by the UCSC Xena database for expression and prognosis.The expression of FLJ30679 in OSCC cell lines was detected by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR).The subcellular localization of FLJ30679 in OSCC cells was detected by RNA nuclear-cytoplasmic fractionation assays.FLJ30679 Smart Silencer was used to establish the FLJ30679 knockdown group(SS-FLJ30679),and overexpression plasmid of FLJ30679 was used to establish FLJ30679 overexpression group(FLJ30679).The effects of altered FLJ30679 expression on the proliferative and migration capacity of OSCC cells were examined by cell counting kit-8(CCK-8)and transwell migration assays.RTFQ-PCR and Western blot were used to determine the effect of altered FLJ30679 expression on the expression of epithelial-mesenchymal transition(EMT)-related genes in OSCC cells.The effects of altered FLJ30679 expression on the phosphoinositide 3-kinase(PI3K)/protein kinase(AKT)pathway were detected by Western blot.Results:Online query of database showed that FLJ30679 expression was higher in HNSCC tissues compared to normal tissues(P＜0.01).HNSCC patients with higher FLJ30679 expression had lower overall survival(P＜0.01).The RTFQ-PCR results showed that FLJ30679 was expressed at a higher level in six OSCC cell lines compared with normal cells,and was predominantly localized in the nucleus.The ability of OSCC cells in the SS-FLJ30679 group to proliferate and migrate was significantly lower compared with the SS-NC group(P＜0.01).OSCC cells in the FLJ30679 overexpression group had significantly higher proliferative and migratory capacities than those in the vector group(P＜0.001).RTFQ-PCR and Western blot results showed that FLJ30679 knockdown resulted in upregulation of mRNA and protein expression levels of E-cadherin(P＜0.01)and downregulation of mRNA and protein expression levels of N-cadherin and vimentin(P＜0.01).FLJ30679 overexpression resulted in downregulation of protein expression levels of E-cadherin(P＜0.01)and upregulation of mRNA and protein expression levels of N-cadherin and vimentin(P＜0.05).Western blot results showed that knockdown of FLJ30679 resulted in decreased protein expression levels of phosphorylated-PI3K(p-PI3K)and phosphorylated-AKT(p-AKT)(P＜0.001),and overexpression of FLJ30679 resulted in increased protein expression levels of p-PI3K and p-AKT(P＜0.01).Conclusion:The expression of FLJ30679 was increased in OSCC tissues and cells.It promoted the proliferation and migration ability of OSCC cells,which may be caused by FLJ30679 promoting EMT via PI3K/AKT pathway.

AIM:This study aimed to investigate the expression and localization of ALOX12P2 in oral squa-mous cell carcinoma(OSCC),as well as its effects on cell viability,migration,and invasion.METHODS:The expres-sion of ALOX12P2 in head and neck squamous cell carcinoma(HNSCC)tissues and its correlation with clinicopathologi-cal features were analyzed using the UALCAN database(University of Alabama at Birmingham Cancer Data Analysis Por-tal).Additionally,the expression of ALOX12P2 in OSCC and its impact on survival prognosis were evaluated through the GDC and UCSC Xena databases.The expression levels of ALOX12P2 in OSCC cell lines were assessed via quantitative re-al-time PCR(RT-qPCR).The subcellular localization of ALOX12P2 was determined using nucleoplasmic RNA isola-tion.CAL-27 cells were used to establish an ALOX12P2 knockdown group(SS-ALOX12P2)and a control group(SS-NC).HN30 cells were employed to form an ALOX12P2 overexpression group(ALOX12P2)and a control group(vector).The effects of altered ALOX12P2 expression on the epithelial-mesenchymal transition(EMT)-related gene E-cadherin and the PI3K/AKT signaling pathway were assessed through Western blot analysis.RESULTS:ALOX12P2 expression was significantly higher in HNSCC and OSCC tissues compared to normal tissues,with its expression correlating with poor prog-nosis.RT-qPCR analysis indicated that the relative expression of ALOX12P2 in OSCC cells was comparable to that in nor-mal cells(P<0.05).RNA nucleoplasmic isolation confirmed that ALOX12P2 localized in the nucleus.In comparison to the SS-NC group,the SS-ALOX12P2 group exhibited a marked reduction in ALOX12P2 expression(P<0.01),alongside significant decreases in cell viability,migration,and invasion(P<0.01).Conversely,the ALOX12P2 group showed sub-stantially higher relative expression compared to the vector group(P<0.01),with enhanced cell viability,migration,and invasion abilities(P<0.01).Western blot analysis demonstrated that ALOX12P2 knockdown resulted in upregulation of E-cadherin and downregulation of N-cadherin and Vimentin(P<0.01),while overexpression of ALOX12P2 yielded the opposite effects(P<0.01).Knockdown of ALOX12P2 led to decreased protein expression of p-PI3K and p-AKT(P<0.01),whereas overexpression increased these protein levels(P<0.01).CONCLUSION:ALOX12P2 is highly ex-pressed in OSCC and promotes cell viability,migration,and invasion.This effect may be linked to the activation of the PI3K/AKT signaling pathway,which facilitates the epithelial-mesenchymal transition(EMT)process.

Objective:To investigated the expression and localization of the long non-coding RNA(lncRNA)STAG3L5P in oral squamous cell carcinoma(OSCC)cells and its effects on OSCC cell proliferation and migration.Methods:STAG3L5P expression in HNSC and OSCC was ana-lyzed online using gene expression profiling interactive analysis 2(GEPIA2)and the University of California Santa Cruz Xena(UCSC Xena)database,respectively.STAG3L5P expression in OSCC cell lines was detected using real-time fluorescence quantitative PCR(qPCR).Nuclear-cytoplasmic RNA fractionation assays were carried out to pinpoint the location of STAG3L5P.Cell counting kit-8(CCK-8)and Transwell migra-tion assays were used to assess OSCC cell proliferation and migration changes.The effect of STAG3L5P overexpression on epithelial-mesen-chymal transition(EMT)related gene expression was detected by qPCR and Western blot.The effect of STAG3L5P overexpression on PI3K/AKT pathway activity was also assessed by Western blot.Results:STAG3L5P was highly expressed in OSCC,and its expression correl-ated significantly with histological grade.STAG3L5P expression was significantly higher in OSCC cell lines than in normal cells.The level of cytoplasmic STAG3L5P in OSCC cells was significantly higher than that in the nucleus.The proliferation and migration capacity of OSCC cells overexpressing STAG3L5P were significantly enhanced compared to negative control OSCC cells.N-cadherin and vimentin mRNA and protein levels were significantly increased by STAG3L5P overexpression,while E-cadherin protein expression was decreased.Overexpression of STAG3L5P also increased activity of p-PI3K and p-AKT.Conclusions:STAG3L5P is up-regulated in OSCC,and STAG3L5P overexpression can promote OSCC cell proliferation and migration.This effect may be related to activation of the PI3K/AKT pathway,thus promoting EMT.

Objective To study the clinical value of double balloon endoscopy (DBE) in diagnosis and treatment of intestinal diseases. Methods Clinical data of 344 patients suspected of small bowel lesions from March 2006 to April 2014 was retrospectively analyzed. All the patients clinical manifestations including varying degrees of abdominal pain, bloating, nause and vomiting, diarrhea. The 344 patients underwent DBE for 397 times. Results The incidence of adverse reactions in checking was only 4.53%, and there was no complications occurred after checking. Positive results shown in 214 cases by DBE, the other 130 patients shown negative results. While 28 of the other 130 patients were found small bowel lesions by capsule endoscopy and CT scans, the remaining 102 cases had no obvious abnormalities. The misdiagnosis rate of DBE was 8.13%, and mainly focuses on polyps and vascular malformation. 242 of the 344 patients were diagnosed with intestinal diseases, and mostly of them were tumor, ulcers and polyps. The detect rate of DBE examination for tumor and ulcer diseases was greater than that of small intestinal polyposis, the difference was statistically significant (P < 0.05). Conclusion DBE in diagnosis and treatment of intestinal diseases is a relatively safe and effective way. It is worthy being popularized and applied.

Objective To explore the difference in short-term efficacy between peroral endoscopic myotomy (POEM) and pneumatic dilatation (PD) in achalasia patients.Methods A retrospective analysis was applied.From September 2010 to March 2015,patients with POEM or PD were enrolled and divided into POEM group (n=26) and PD group (n=40).High-resolution manometry (HRM) before and one month after treatment were compared between POEM group and PD group.Before and three months after treatment,Eckardt score and gastroesophageal reflux symptom was compared between groups (Eckardt score ≤ 3 as the standard for successful treatment).Wilcoxon signed rank test was performed for before and after treatment comparison in the same group.Mann-Whitney U test was used to compare between two groups and Fisher's exact test was used for rate comparison.Results The successful treatment rates at one and three month after POEM group were 92.3 ％ (24/26) and 96.2％ (25/26),respectively.The successful treatment rates at one and three month after PD were 87.5％ (35/40) and 75.0％ (30/40).At three month after treatment,the successful treatment rate of POEM group was higher than that of PD group (Fisher's exact test,P =0.02).At three months after treatment,the Eckardt score of POEM group was lower than that of PD group (1.35,0 to 4.00,vs2.73,0 to 6.00;U=-3.921,P＞0.01).By the end of three months after treatment,the rate of gastroesophageal reflux symptom of POEM group was higher than that of PD group (7/26,26.9 ％ vs 2/40,5.0％;Fisher's exact test,P=0.01).The postoperative 4 second integrated relaxation pressure (4s-IRP) and lower esophageal sphincter pressure (LESP) of POEM group were both lower than those of PD group (7.01 mmHg,3.48 to 10.40 mmHg vs 10.11 mmHg,5.75 to 12.91 mmHg,U=-4.541,P＜0.01;11.61 mmHg,4.21 to 14.64 mmHg vs 17.85 mmHg,8.39 to 24.57 mmHg,U=-6.142,P＜0.01).The analysis of achalasia subtypes indicated that the efficacy of POEM was better than that of PD both in type Ⅰ and type Ⅱ.Conclusion During short-term follow-up,the efficacy of POEM was better than that of PD in achalasia patients,however there was a higher incidence of post-operative gastroesophageal reflux after POEM.

Objective To investigate the characteristics,treatment and prognosis of the gastrointes-tinal iatrogenic perforation resulting from endoscopic operations.Methods The clinical data of the 107 pa-tients with gastrointestinal iatrogenic perforations caused by endoscopic operations from October 2003 to Octo-ber 2013 were retrospectively studied,and the characteristics,treatment and prognosis of these patients were analysed.Results The incidence of the gastrointestinal iatrogenic perforation resulting from endoscopic oper-ations was 0.041%(107 /263 549,among which 0.006% was diagnostic(13 /232 011),and 0.298% was therapeutic (94 /31 538).A total of 107 patients with gastrointestinal iatrogenic chose conservative medical management,endoscopic clipping or surgery according to different conditions after perforations.All patients had good prognosis.Conclusion Endoscopic therapeutic operations are more likely to lead to the occurrence of iatrogenic gastrointestinal perforations compared with diagnostic operations.And most patients could get good prognosis as long as the appropriate treatment is performed when the perforation occurs.

Objective To investigate the effects of peroral endoscopic myotomy (POEM) on esophageal dynamics in patients with achalasia (AC) and appraise the role of high resolution manometry (HRM)in assessment of POEM therapy.Methods From July 2011 to September 2012,20 patients with achalasia underwent POEM in the department of gastroenterology,Tianjin Medical University General Hospital.Preoperation esophageal dynamics of all the patients were evaluated by high resolution manometry (HRM) system and one month after POEM operation the test was repeated.Data of lower esophageal sphincter (LES)and esophageal body were analyzed,with 15 healthy volunteers as a contrast study.Results Mter POEM,the LES pressure (LESP) and 4-second integrated relaxation pressure (4sIRP) significantly decreased:LESP [pre-operation (24.5 ± 13.1) mm Hg vs.post-operation (8.5 ± 3.1) mm Hg,P ＜ 0.05] ; 4sIRP [pre-operation (20.7 ± 6.8) mm Hg vs.post-operation (5.0 ± 3.4) mm Hg,P ＜ 0.05].The LES relax rate (LESRR) significantly elevated [pre-operation (12.7 ± 9.8) ％ vs.post-operation (39.6 ± 18.1) ％,P ＜ 0.05].However,the esophageal aperistalsis remained after POEM.The total of 20 patiems were all type Ⅰ achalasia and responded well to POEM therapy.Post-operation data on the symptom scores markedly decreased and a significant correlation was found between the decreasing level of symptom scores and LESP,as well as the scores 4sIRP in the 20 patients (LESP:r =0.751,P ＜ 0.05 ; 4sIRP:r =0.500,P ＜ 0.05).Conclusion POEM can significantly improve esophageal dynamics in patients with achalasia and the treatment outcome is definite.HRM plays an important role in evaluation of POEM therapy on achalasia.

ObjectiveTo review the clinical characteristics of patients with gastric mucosa-associated lymphoid tissue(MALT) lymphoma and to investigate the effects of Helicobacter pylori (Hp) eradication therapy in these patients.MethodsClinical data of ninety-five patients with gastic MALT lymphoma were reviewed with respect to their clinical manifestations,endoscopic features,histopathological features and Hp infection.The follow-up data of patients treated by Hp eradication therapy were analyzed.The survival curve was calculated with Kaplan-Meier,while the predictive factors for resistance to Hp eradiation were then analyzed by using Cox proportional hazards model.ResultsThe clinical manifestations and endoscopic features of 85 patients with gastric MALT lymphoma were non-specific.Hp positive was found in 97.9％ (93/95) patients at presentation,and 36 patients were treated by Hp eradication with a median follow-up duration of ( 58.1 ± 29.9) months.Total remission was achieved in 94.4％ ( 34/36),including 24 complete remission 10 partial remission.Therapy failure occurred in 2 patients.3-year survival rate of patients treated by Hp eradication were 86.2％ (25/29). Cox multivariate analysis showed that age ( ≥60 years),multiple lesions and non-superficial lesions were independent predictors of resistance to Hp eradication therapy.ConclusionGastric MALT lymphoma is associated with Hp infection.Eradication of Hp can effectively induce remission in these patients.