Objective To explore the molecular mechanism of miR-125b promoting invasion,metastasis and epithelial-mesenchymal transition(EMT)of gastric cancer cells.Methods The expression of miR-125b in gastric cancer and its adjacent tissues was studied by qRT-PCR.After upregulating or downregulating miR-125b in gastric cancer cells,the protein expressions of DKK3 and SERPINA4 were detected by Western blotting.Dual luciferase reporting assay was used to verify whether miR-125b can target DKK3 and SERPINA4.MKN45 cells were co-transfected with miR-125b inhibitor and target gene siRNA.Migration and invasion experiments were conducted to explore whether miR-125b can regulate the biological function of MKN45 cells through DKK3 and SERPINA4.Then,the regulatory mechanism of SRF on miR-125b was investigated.Finally,by in vivo experiments,the expression of SRF in gastric cancer cells was upregulated or downregulated by lentivirus transfection;the number of lung metastases in nude mice was detected to explore the effect of SRF on gastric cancer cell metastasis.Results In this study,the expression of miR-125b increased in gastric cancer tissues,which was correlated with clinical stage and lymph node metastasis.Dual luciferase reporting experiments showed that DKK3 and SERPINA4 were the direct targets of miR-125b in gastric cancer cells,and could activate the Wnt/β-catenin signaling pathway,thereby promoting the transcription process of EMT-related transcription factors Twist1 and Slug,inducing the occurrence of EMT,and promoting the metastasis of gastric cancer.In vitro and in vivo experiments confirmed that SRF promoted the invasion and metastasis of gastric cancer cells by positively regulating the expression of miR-125b.Conclusion Taken together,SRF/miR-125b axis promotes the EMT and metastasis of gastric cancer cells,and these regulators represent new potential therapeutic targets or biomarkers for gastric cancer.

Objective To investigate the expression of circular RNA(Circ RNAS)Actinin α 4(ACTN4)and platelet thrombin protein 1(THBS1)in intrahepatic cholangiocarcinoma(ICC)and their relationship with clinicopathological characteristics and prognosis,and provide reference for clinical diagnosis and treatment.Method A retrospective analysis was conducted on 84 ICC patients diagnosed and treated in the Second Affiliated Hospital of Xian Jiaotong University from May 2017 to June 2020.The expressions of CircACTN4 mRNA,THBS1 mRNA and protein were detected by real-time fluorescent quantitative PCR and immunohistochemistry.Pearson correlation analysis was used to analyze the correlation between CircACTN4 mRNA and THBS1 mRNA in ICC cancer tissue.The prognostic differences of ICC patients with different CircACTN4 mRNA and THBS1 mRNA expressions were compared by the Kaplan-Meier method(log rank test).COX regression analysis was performed to identify prognostic factors in ICC patients.The prognostic value of CircACTN4 mRNA and THBS1 mRNA in evaluating the risk of death in ICC patients was assessed by receiver operating characteristic(ROC)curve analysis.Results The expression of CircACTN4 mRNA in ICC tissues(3.14±0.42)was higher than that in adjacent tissues(0.76±0.25),with significant difference(t=44.094,P＜0.001).The positive rates of THBS1 mRNA(2.82±0.36)and protein positive rate(92.86％)in ICC tissues were higher than those in adjacent tissues(0.81±0.24,7.14％),and the differences were statistically significant(t/x2=42.068,123.429,all P＜0.001).CircACTN4 mRNA was positively correlated with THBS1 mRNA in ICC cancer tissue(r=0.669,P＜0.001).The expressions of CircACTN4 mRNA,THBS1 mRNA in ICC cancer tissues with TNM stage Ⅲ,low differentiation,and lymph node metastasis were higher than those in TNM stage Ⅰ～Ⅱ,high differentiation,and non-lymph node metastasis cancer tissues,and the differences were statistically significant(x2=7.949,9.164,12.207;23.270,18.625,19.828,all P＜0.001).The 3-year cumulative survival rates of the high expression group of CircACTN4 mRNA and THBS1 mRNA were lower than those of the low expression group of CircACTN4 mRNA(25.00％vs 56.82％)and THBS1 mRNA(19.51％vs 62.79％),and the differences were statistically significant(Log rank x2=13.601,24.310,all P＜0.001).CircACTN4 mRNA(OR=1.839,95％CI:1.228～2.753),THBS1 mRNA(OR=1.744,95％CI:1.245～2.443),lymph node metastasis(OR=1.925,95％CI:1.316～2.816),TNM staging(OR=1.613,95％CI:1.223～2.126),and tumor differentiation(OR=1.510,95％CI:1.205～1.892)were independent factors affecting the prognosis of ICC.The area under the curve of the combination detection of circACTN4 and THBS1 mRNA on the prognosis of death in patients with ICC was 0.868,which was greater than that of the single index(0.812 and 0.784),with significant differences(Z=3.348,3.847,all P＜0.001).Conclusion The expressions of CircACTN4 mRNA and THBS1 mRNA were increased in ICC,and they were associated with TNM stage,differentiation,and lymph node metastasis.These markers may serve as novel indicators to evaluate the poor prognosis of ICC patients.

Objective To explore the relationship between serum Krüppel-like factor 5(KLF5)and actinin-4 with prognosis of hepatocellular carcinoma(HCC)treated with transcatheter arterial chemoembolization(TACE).Methods From March 2015 to March 2020,130 HCC patients admitted into the Second Affiliated Hospital were collected as study subjects and divided into HCC group and 86 patients with liver cirrhosis were collected as control group.After 3 years of postoperative follow-up,HCC patients were sub-divided into survival group(n=38)and mortality group(n=92)based on their survival outcomes.ELISA was applied to detect the level of serum KLF5 and actinin-4 in each group and analyzed the correlation between serum KLF5 and actinin-4 levels.Cox regression was applied to analyze the risk factors affecting the prognostic mortality of HCC patients;Receiver operating char-acteristic curve(ROC)was plotted to evaluate the predictive value of serum KLF5 and actinin-4 levels for the prognostic mortality of HCC patients.Results The serum level of KLF5 and actinin-4 in the mortality group was significantly higher than those in the survival group(P＜0.05);There was a positive correlation between serum level of KLF5 and actinin-4 in HCC patients(P＜0.001);Low differentiation,staging(CNLC)stage IIIa of China liver cancer staging(CNLC),vascular invasion and elevated level of KLF5 and actinin-4 were risk factors for prognostic mortality in HCC patients(P＜0.05);The area under the curve(AUC)of serum KLF5 and acti-nin-4 alone and in combination for predicting mortality in HCC patients was 0.835,0.866,and 0.936,respec-tively,showing a high level of combined predicting function(Zcombination-KLF5=2.792,P=0.005,Zcombi-nation-actinin-4=2.014,P=0.044).Conclusions Serum level of KLF5 and actinin-4 l found in HCC patients has a high predictive value for prognosis.

Objective:To investigate the effect of stilamin on mitochondrial injury of acute pancreatitis (AP)-related acinar cells and the possible mechanism.Methods:24 C57BL/6 mice were randomly divided into control group, AP group and AP+ stilamin treatment group (Stilamin group). AP model was prepared by intraperitoneal injection of l-arginine in AP group, 0.4 mg/kg stilamin was intraperitoneally injected at 2 h after modeling in stilamin group, and control group received intraperitoneal injection with saline in the same volume. Histopathological examination of pancreatic tissue was performed routinely. Serum levels of IL-6, IL-10 and TNF- α were detected by ELISA, serum levels of amylase and lipase, and serum levels of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione (GSH) reflecting oxidative stress were detected by biochemistry method, respectively. Mitotracker red fluorescent labeling was used to detect the number of mitochondria in pancreatic acinar cells, and western blot was used to detect the ND-3 protein expression reflecting the number of mitochondria. The expression levels of mitochondrial fusion protein (Mfn-2) and mitochondrial transcription factor A (TFAM) reflecting mitochondrial function were determined by immunohistochemical staining.Results:Compared with AP group, the pancreatic pathology scores of mice in Stilamin group were significantly decreased [(2.07±0.50) vs (3.93±0.64)], serum amylase and lipase levels were significantly decreased [(1 493±172)U/L vs (1 832±86)U/L, (225.4±83.2)U/L vs (671.0±164.5)U/L]; serum IL-6 and TNF-α levels were greatly decreased, while IL-10 levels were obviously increased [(99.09±39.65)ng/L vs (358.60±139.22)ng/L, (22.75±11.24)ng/L vs (40.83±1.62)ng/L, (15.12±5.03)ng/L vs (9.92±8.73)ng/L]. Mitotracker staining density and expression levels of ND-3, Mfn-2 and Tfam were increased [(71.67±17.62) vs (40.00±10.15), (0.45±0.16) vs (0.11±0.05), 78% vs 54%, 86% vs 47%], and MDA, SOD, and GSH levels were increased [(5.00±1.73)nmol/mg vs (7.33±2.08)nmol/mg protein, (17.33±3.21)U/mg vs (8.67±2.07)] U/mg protein. The ratio of (131.33±20.55)U/mg to (77.33±29.69)U/mg protein was statistically significant (all P<0.05). Conclusions:By protecting the mitochondria number and function of damaged pancreatic acinar cells and reducing the level of oxidative stress, stilamin could alleviate the level of pancreatic tissue damage and inflammatory response in mice with acute pancreatitis.

Objective To investigate the effect of time interval between diagnosis and surgical treatment on the prognosis of breast cancer.Methods A retrospective study that include a total of 252 female patients who underwent breast cancer diagnosis and treatment in the Second Affiliated Hospital of Xi'an Jiaotong University from April 2012 to August 2014 were included in the present study,the average age was (58.2 ± 10.8) years old,range from 31 to 67 years old.General demographic information and data of tumor were collected.Information on postoperative recurrence,metastasis,death,and disease-free survival status of breast cancer patients were followed up 5 years by outpatient follow-up or telephone follow-up.All participants were divided into four groups (＜2 weeks,2-4 weeks,4-8 weeks,≥8 weeks) by the time interval between diagnosis and surgical treatment,including 26,118,78 and 30 cases,respectively.In addition,according to the diameter of breast cancer tumors,all participants were divided into three groups (＜20 mm,20-40 mm,and ≥40 mm),including 99,124,and 29 cases,respectively.According to the results of pathological examination of the lymph nodes obtained during intraoperative dissection,the all participants were divided into three groups (lymph nodes without metastasis,1 to 3 metastasis,and ≥3 metastasis),including 66,124,and 62 cases,respectively.The Cox proportional regression risk models were used to assess the hazard ratio (HR) and its 95％ confidence interval (CI) of time interval between diagnosis and surgical treatment with the prognosis of breast cancer,with adjustment for age,education levels and body mass index.Further,stratified analysis by tumor characteristics,including pathological type,histological grade,tumor diameter,lymph node metastasis,and receptor expression were also conducted to evaluated the above association.Kaplan-Meier survival curve was used to evaluate the effects of time interval between diagnosis and surgical treatment on the prognosis of breast cancer.Results The Kaplan-Meier survival curves for the five-year follow-up of total survival time between 4 different time intervals groups showed significantly different (P ＜0.001),and patients with a pre-treatment interval of ＜2 weeks had the longest survival time,while those with ≥8 weeks had the lowest survival time.With a one-week interval before treatment,the overall risk of death in breast cancer patients increased by 6％ (HR =1.06,95％ CI:1.01-1.1 l),and the risk of breast cancer death increased by 8％ (HR =1.08,95％ CI:1.02-1.14),the risk of distant metastasis of breast cancer cells increased by 10％ (HR =1.10,95％ CI:1.08-1.13).With the increase in breast cancer tumor diameter (＜20 mm,20-40 mm,≥40 mm),the overall risk of death due to prolonged treatment interval increased gradually,with HR (95％CI) were 1.06 (1.03-1.09),1.08 (1.02-1.12) and 1.11 (1.05-1.17),respectively.With the increase of lymph node metastasis in breast cancer (no metastasis,metastasis at 1-3,≥ 3 metastasis),the total mortality risk caused by prolonged treatment time interval also showed an increasing trend,with HR (95％CI) were 1.04 (1.02-1.08),1.06 (1.04-1.08) and 1.08 (0.99-1.11),respectively.The same results were also shown in the effect of tumor diameter or distant lymph node metastasis on the association between treatment time interval and breast cancer survival and distant metastasis of breast cancer cells.Conclusion With the prolongation of the time interval between the diagnosis of the breast cancer and the surgical treatment of breast cancer patients,the risk of postoperative death is significantly increased,and the association is more pronounced in breast cancer patients with larger tumor volume or higher distant lymph node metastasis.

Objective To explore the application of Oxybuprocaine Hydrochloride Gel in colonoscopy examination. Methods 1000 patients received colonoscopy examination were randomly divided into experimental group and control group, 500 cases in each. Oxybuprocaine Hydrochloride Gel was applied on anal region in the experimental group before endoscopy for perianal anesthesia and lubrication. Paraffin oil was used in the control group to lubricate perianal and enteroscopy. The success rate of primary insertion, visual analogue pain score (VAS) score, examination time and postoperative complications were compared between the two groups. Results In the experimental group, the success rate of primary insertion (95%) was higher than that of the control group (76%), and the pain score was lower than that of the control group.The examination time was shortened, and the difference was statistically significant (P < 0.05). Conclusions Oxybuprocaine Hydrochloride Gel applied to colonoscopy can effectively reduce the patient's pain and discomfort, improve the success rate of primary insertion, shorten the examination time. And the method is worthy of clinical popularization and application.

Objective To investigate the mRNA expression of IKB kinase (IKK-α) and interferon-α (IFN-α) in the peripheral blood leukocytes of patients with systemic lupus erythematosus (SLE), and to explore the role of IKK-α in the production of IFN-α in SLE patients. Methods SYBR green dye I based real-time quantitative PCR was used to detect the mRNA expression levels of IKK-α and IFN-α in the peripheral blood leucocytes of SLE patients and healthy controls. Serum levels of IFN-α were measured with ELISA method. Results IKK-α mRNA expression levels in SLE patients were significantly higher than those of normal controls (P<0.05). IKK-α mRNA expression levels in SLE patients with active disease were significantly higher than patients with stable disease (P<0.01). IFN-α mRNA expression level in SLE patients was significantly lower than that of the normal controls (P<0.01). IFN-α mRNA expression levels in SLE patients with active disease were significantly higher than patients with stable disease (P<0.01). Serum levels of IFN-α in SLE patients with active disease was significantly higher than that of the normal controls and patients with stable disease (P<0.05). The anti-dsDNA antibody correlated positively, and complement C3 correlated negatively with serum concentration of IFN-α. IKK-α mRNA expression levels in SLE patients correlated positively with serum concentration of IFN-α. Conclusion IKK-α correlates positively with serum concentration of IFN-α. The IFN-α level is significantly correlated with disease activity, This suggests that IKK-α may play an important role in the pathogenesis of SLE.

Objective To evaluate the short-term efficacy and safety of etanercept treatment in Chinese patients with active ankylosing spondylitis ( AS ). Methods This was a 12-week multicenter,double-blind, placebo-controlled, randomized phase Ⅲ clinical study. The first part was a 6-week placebocontrolled period followed by a 6-week open-label period. The primary efficacy endpoint was the percentage of subjects achieving a 20% improvement in assessment in ankylosing spondylitis (ASAS) ( ASAS 20). The secondary efficacy endpoints were the percentage of patients achieving a 40% improvement in ASAS (ASAS 40), achieving a 50% improvement in ASAS( ASAS 50), achieving a 70% improvement in ASAS (ASAS 70), and ASAS 5/6 responses at all visits, and the improvement in subject global assessment,physician global assessment, nocturnal and total back pain, bath AS functional index ( BASFI ), bath AS disease activity index (BASDAI), spinal mobility, joint assessment and quality of life assessment. All subjects in the study were evaluated for safety. Results The primary endpoint, ASAS 20 at week 6, was achieved by 86. 5% (64/74) patients in the etanercept group compared to 29. 5% (23/78) patients in the placebo group(P ＜0. 001 ). As early as week 2, the percentages of patients achieving the ASAS 20 between the two groups were significantly different. Furthermore, the majority of secondary efficacy end points were also significantly improved. Most of adverse events (AE) were mild in nature, the commonest adverse events were elevated liver function levels, injection site reactions and nasopharyngitis. No death or serious AE were observed. Conclusion Etanercept can improve symptoms fastly,significantly and safely in Chinese patients with active AS.

Objective To compare the efficacy and safety ofetanercept injection 50 mg once weeklycombined with methotrexate (MTX) therapy for patients withactive rheumatoid arthritis.Methods This studyconsists of 2 parts：a 12-week double-blind treatmentperiod (part A) followed by a 12-week open-labelsafety study period (part B).The randomization oftreatments in double-blind treatment period was completedthrough the clinical operations randomization environment(CORE) system.During part A,the subjects wererandomly assigned to the etanercept 50 mg or placebo group. The dosage regimen for etanercept was 50 mgadministered subcutaneously once weekly while MTX wasadministered orally.All subjects who completed partA received 50 mg etanercept once weekly and MTX1 during theopen-label treatment.The primary endpointwas ACR 20 response at week 12.Secondary endpoint variablesincluded physician／patient global assessmentsof disease activities,duration of morning stiffness,painvisual analog scale (VAS),health assessment questi onnaire (HAQ),CRP level and tender and swollen joint counts .The results of safety between the two groupswere compared.The primary endpoint and other secondarybinary endpoints were analyzed using the Fisher’sexact test.For continuous endpoints.the change frombaseline was analyzed with analysis of covariance.Results One hundred and fifty six subjects satisfiedmodified intent-to-treat (mITT) population were enrolled during part A,of which 77 subjects were in theetanercept+MTX group,and 79 subjects were in theplacebo+MTX group respectively.A total of 149 subjectscompleted part A.As early as week 4.the ACR 20response achieved 39％ (30,77) in the etanerceptgroup,which was significantly higher than that of theplacebogroup [16％(13／79),P<0.001].At week 12,the ACR 20respouse achieved 62％(48,77)in the etanercept group and 23％(18／79) in the placebo group (P<0.01).Fromweek 4,other study endpoints including physician global assessment,patient globalassessment,duration of morning stiffness,painVAS,HAQ,CRPlevel,tender joint counts,swollen joint counts were alsocompared.The results showed that all above efficacyendpoints in the etanercept+MTX group were better than thoseof the placebo+MTX group(P<0.01).Butthere Was no significant difference in the total adverseeriects between the two groups.ConclusionEtanercept 50 mg once weekly + MTX treatment for 24 weeks iswell tolerated.During the first 12-weektreatment period,the etanercept group has shown a rapidefficacy onset and a significantly better therapeuticeffect compared to that of the placebo group.

Objective To evaluate the efficacy of medical supervised aerobic exercise on physical activity, quality of life and psychological status in patients with systemic lupus erythematosus (SLE). Methods SLE patients who fulfilled ACR criteria were recruited and divided into 2 groups: exercise group (n=24) and control group (n=25). The patients in the exercise group were supervised to have aerobic exercises. The intensiveness of exercise was determined by 20%-40% of maximum heart rate reservation. Visual analog scale (VAS), SLE disease activity index (SLEDAI) score, physical working capacity (PWC170), SF-36 and profile of mood states (POMS) of the two groups were used to evaluate the changes at the baseline, 1 month, 3 months and 6 months after this study. Results The 2 groups were homogeneous and comparable in disease activity at baseline. 1, 3 and 6 months after the study, the VAS, PWC170, POMS and SF -36 of SLE patients were improved in certain degrees in both groups, while the improvement of VAS (P<0.05), PWCITO (P< 0.01 ) and social function of SF-36 (P<0.05) of exercise group were significantly more evident than those of the control group in 6 months after study without any impact on disease condition. There was a high negative correlation between VAS and 5 categories of POMS (r=-6.26~-0.393, P<0.01 ) and a more relevant positive association between VAS and 2 categories of POMS (r=0.534~0.611, P<0.01). Conclusion The data demonstrate that the supervised aerobic exercise can ameliorate physical capacity, improve quality of life and psychological and emotional status in the state SLE patients without aggravating disease per se.