Sirtuin 1 alleviates endoplasmic reticulum stress-mediated apoptosis of intestinal epithelial cells in ulcerative colitis

BACKGROUND Sirtuin 1(SIRT1)is a nicotinamide adenine dinucleotide(NAD+)-dependent protein deacetylase that is involved in various diseases,including cancers,metabolic diseases,and inflammation-associated diseases.However,the role of SIRT1 in ulcerative colitis(UC)is still confusing.AIM To investigate the role of SIRT1 in intestinal epithelial cells(IECs)in UC and further explore the underlying mechanisms.METHODS We developed a coculture model using macrophages and Caco-2 cells.After treatment with the SIRT1 activator SRT1720 or inhibitor nicotinamide(NAM),the expression of occludin and zona occludens 1(ZO-1)was assessed by Western blot analysis.Annexin V-APC/7-AAD assays were performed to evaluate Caco-2 apoptosis.Dextran sodium sulfate(DSS)-induced colitis mice were exposed to SRT1720 or NAM for 7 d.Transferase-mediated dUTP nick-end labeling(TUNEL)assays were conducted to assess apoptosis in colon tissues.The expression levels of glucose-regulated protein 78(GRP78),CCAAT/enhancerbinding protein homologous protein(CHOP),caspase-12,caspase-9,and caspase-3 in Caco-2 cells and the colon tissues of treated mice were examined by quantitative real-time PCR and Western blot.RESULTS SRT1720 treatment increased the protein levels of occludin and ZO-1 and inhibited Caco-2 apoptosis,whereas NAM administration caused the opposite effects.DSS-induced colitis mice treated with SRT1720 had a lower disease activity index(P<0.01),histological score(P<0.001),inflammatory cytokine levels(P<0.01),and apoptotic cell rate(P<0.01),while exposure to NAM caused the opposite effects.Moreover,SIRT1 activation reduced the expression levels of GRP78,CHOP,cleaved caspase-12,cleaved caspase-9,and cleaved caspase-3 in Caco-2 cells and the colon tissues of treated mice.CONCLUSION SIRT1 activation reduces apoptosis of IECs via the suppression of endoplasmic reticulum stress-mediated apoptosis-associated molecules CHOP and caspase-12.SIRT1 activation may be a potential therapeutic strategy for UC.

Resveratrol alleviates intestinal mucosal barrier dysfunction in dextran sulfate sodium-induced colitis mice by enhancing autophagy

BACKGROUND Intestinal mucosal barrier dysfunction plays an important role in the pathogenesis of ulcerative colitis(UC).Recent studies have revealed that impaired autophagy is associated with intestinal mucosal dysfunction in the mucosa of colitis mice.Resveratrol exerts anti-inflammatory functions by regulating autophagy.AIM To investigate the effect and mechanism of resveratrol on protecting the integrity of the intestinal mucosal barrier and anti-inflammation in dextran sulfate sodium(DSS)-induced ulcerative colitis mice.METHODS Male C57BL/6 mice were divided into four groups:negative control group,DSS model group,DSS+resveratrol group,and DSS+5-aminosalicylic acid group.The severity of colitis was assessed by the disease activity index,serum inflammatory cytokines were detected by enzyme-linked immunosorbent assay.Colon tissues were stained with haematoxylin and eosin,and mucosal damage was evaluated by mean histological score.The expression of occludin and ZO-1 in colon tissue was evaluated using immunohistochemical analysis.In addition,the expression of autophagy-related genes was determined using reverse transcription-polymerase chain reaction and Western-blot,and morphology of autophagy was observed by transmission electron microscopy.RESULTS The resveratrol treatment group showed a 1.72-fold decrease in disease activity index scores and 1.42,3.81,and 1.65-fold decrease in the production of the inflammatory cytokine tumor necrosis factor-α,interleukin-6 and interleukin-1β,respectively,in DSS-induced colitis mice compared with DSS group(P<0.05).The expressions of the tight junction proteins occludin and ZO-1 in DSS model group were decreased,and were increased in resveratrol-treated colitis group.Resveratrol also increased the levels of LC3B(by 1.39-fold compared with DSS group)and Beclin-1(by 1.49-fold compared with DSS group)(P<0.05),as well as the number of autophagosomes,which implies that the resveratrol may alleviate intestinal mucosal barrier dysfunction in DSS-induced UC mice by enhancing autophagy.CONCLUSION Resveratrol treatment decreased the expression of inflammatory factors,increased the expression of tight junction proteins and alleviated UC intestinal mucosal barrier dysfunction;this effect may be achieved by enhancing autophagy in intestinal epithelial cells.

Diagnostic efficacy of the Japan Narrow-band-imaging Expert Team and Pit pattern classifications for colorectal lesions: A meta-analysis

BACKGROUND Pit pattern classification using magnifying chromoendoscopy is the established method for diagnosing colorectal lesions. The Japan Narrow-band-imaging(NBI) Expert Team(JNET) classification is a novel NBI magnifying endoscopic classification that focuses on the vessel, and surface patterns.AIM To determine the diagnostic efficacy of each category of the JNET and Pit pattern classifications for colorectal lesions.METHODS A systematic literature search was performed using PubMed, Embase, the Cochrane Library, and Web of Science databases. The pooled sensitivity, specificity, diagnostic odds ratio, and area under the summary receiver operating characteristic curve of each category of the JNET and Pit pattern classifications were calculated.RESULTS A total of 19227 colorectal lesions in 31 studies were included. The diagnostic performance of the JNET classification was equivalent to the Pit pattern classification in each corresponding category. The pooled sensitivity, specificity,and area under the curve(AUC) for each category of the JNET classification were as follows: 0.73(95%CI: 0.55-0.85), 0.99(95%CI: 0.97-1.00), and 0.97(95%CI: 0.95-0.98), respectively, for Type 1;0.88(95%CI: 0.78-0.94), 0.72(95%CI: 0.64-0.79), and 0.84(95%CI: 0.81-0.87), respectively, for Type 2 A;0.56(95%CI: 0.47-0.64), 0.91(95%CI: 0.79-0.96), and 0.72(95%CI: 0.68-0.76), respectively, for Type 2 B;0.51(95%CI: 0.42-0.61), 1.00(95%CI: 1.00-1.00), and 0.90(95%CI: 0.87-0.93), respectively, for Type 3.CONCLUSION This meta-analysis suggests that the diagnostic efficacy of the JNET classification may be equivalent to that of the Pit pattern classification. However, due to its simpler and clearer clinical application, the JNET classification should be promoted for the classification of colorectal lesions, and to guide the treatment strategy.

EUS for choosing best endoscopic treatment of mesenchymal tumors of upper gastrointestinal tract

AIM: To evaluate the value of endoscopic ultrasonography (EUS) in the choice of endoscopic therapy strategies for mesenchymal tumors of the upper gastrointestinal tract. METHODS: From July 2004 to September 2010, 1050 patients with upper gastrointestinal mesenchymal tumors (GIMTs) were diagnosed using EUS. Among them, 201 patients underwent different endoscopic therapies based on the deriving layers, growth patterns and lesion sizes. RESULTS: Using EUS, we found 543 leiomyomas and 507 stromal tumors. One hundred and thirty-three leiomyomas and 24 stromal tumors were treated by snare electrosection, 6 leiomyomas and 20 stromal tumorswere treated by endoloop, 10 stromal tumors were treated by endoscopic mucosal resection and 8 stromal tumors were treated by endoscopic submucosal dissection. Complete resection of the lesion was achieved in all cases. Of the mesenchymal tumors, 90.38% diagnosed by EUS were also identified by pathohistology. All wounds were closed up nicely and no recurrence was found in the follow-up after 2 mo. CONCLUSION: EUS is an effective means of diagnosis for upper GIMTs and is an important tool in choosing the endoscopic therapy for GIMTs, by which the lesions can be treated safely and effectively.

Endoscopic ultrasound-guided deep and large biopsy for diagnosis of gastric infiltrating tumors with negative malignant endoscopy biopsies

AIM: To assess the diagnostic yield and safety of adeep and large biopsy technique under the guidance of endoscopic ultrasound(EUS) for diagnosis of gastric infiltrating tumors with negative malignant endoscopy biopsies.METHODS: From January 2009 to March 2014, 36 patients in whom gastric infiltrating tumors had been diagnosed by EUS received negative results for malignancy after endoscopic biopsies. The deep and large biopsy technique combined bite-on-bite technique with or without endoscopic mucosal resection(EMR) to obtain submucosal tissue from lesions. EUS was used to select the appropriate biopsy sites. If the lesion protruded into the cavity, EMR was performed for removal of the overlying mucosa and then bite-onbite technique was conducted in the resected area to obtain submucosal tissue. If the lesion appeared to be flat or was difficult to lift by injection, the bite-on-bite technique was directly used.RESULTS: Twenty-eight of the 36 patients were treated by EMR followed by bite-on-bite technique, while 8 patients only underwent bite-on-bite technique. Histological results showed 23 of the 36 lesions were poorly differentiated adenocarcinomas, 2 diffuse large B cell lymphomas, 4 mucosa-associated lymphoid tissue-type lymphomas, and 7 undiagnosed. The deep and large biopsy technique provided a definitive and conclusive diagnosis in 29(80.6%) of the 36 patients. The 12 gastric linitis plastica and 6 lymphoma patients received chemotherapy and avoided surgery. Minor oozing of blood in 2 mucosal resection wounds was managed by argon plasma coagulation and in 5 cases after deep biopsies by epinephrine(0.001%). Neither severe hemorrhage nor perforation occurred in any patient.CONCLUSION: The deep and large biopsy technique is superior to ordinary endoscopic biopsy for achieving an accurate diagnosis of gastric infiltrating tumors.This procedure guided by EUS is an effective and safe diagnostic method for gastric infiltrating tumors in which endoscopic biopsy results were negative for malignancy.

Blockage of ETS homologous factor inhibits the proliferation and invasion of gastric cancer cells through the c-Met pathway

BACKGROUND Gastric cancer(GC)is one of the most common and deadliest types of cancer worldwide due to its delayed diagnosis and high metastatic frequency,but its exact pathogenesis has not been fully elucidated.ETS homologous factor(EHF)is an important member of the ETS family and contributes to the pathogenesis of multiple malignant tumors.To date,whether EHF participates in the development of GC via the c-Met signaling pathway remains unclear.AIM To investigate the role and mechanism of EHF in the occurrence and development of GC.METHODS The expression of EHF mRNA in GC tissues and cell lines was measured by quantitative PCR.Western blotting was performed to determine the protein expression of EHF,c-Met,and its downstream signal molecules.The EHF expression in GC tissues was further detected by immunohistochemical staining.To investigate the role of EHF in GC oncogenesis,small interfering RNA(siRNA)against EHF was transfected into GC cells.The cell proliferation of GC cells was determined by Cell Counting Kit-8 and colony formation assays.Flow cytometry was performed following Annexin V/propidium iodide(PI)to identify apoptotic cells and PI staining to analyze the cell cycle.Cell migration and invasion were assessed by transwell assays.RESULTS The data showed that EHF was upregulated in GC tissues and cell lines in which increased expression of c-Met was also observed.Silencing of EHF by siRNA reduced the proliferation of GC cells.Inhibition of EHF induced significant apoptosis and cell cycle arrest in GC cells.Cell migration and invasion were significantly inhibited.EHF silencing led to c-Met downregulation and further blocked the Ras/c-Raf/extracellular signal-related kinase 1/2(Erk1/2)pathway.Additionally,phosphatase and tensin homolog was upregulated and glycogen synthase kinase 3 beta was deactivated.Moreover,inactivation of signal transducer and activator of transcription 3 was detected following EHF inhibition,leading to inhibition of the epithelial-to-mesenchymal transition(EMT).CONCLUSION These results suggest that EHF plays a key role in cell proliferation,invasion,apoptosis,the cell cycle and EMT via the c-Met pathway.Therefore,EHF may serve as an antineoplastic target for the diagnosis and treatment of GC.

Proteomic analysis of liver mitochondria from rats with nonalcoholic steatohepatitis

AIM: To explore mitochondrial dysfunction in nonalcoholic steatohepatitis (NASH) by analyzing the proteome of liver mitochondria from a NASH model.

Mesh migration into the sigmoid colon after inguinal hernia repair presenting as a colonic polyp:A case report and review of literature

Mesh migration and penetration into abdominal visce-ra rarely occur after laparoscopic inguinal hernia repair. We present the first case of mesh migration into the sigmoid colon identified as a colonic polyp at initial co-lonoscopic examination. The patient complained of mild abdominal distention in the lower abdomen over the previous year without changes in bowel habits or stool appearance and without weight loss. By complement-ary endoscopic ultrasonography, a cavity--like structure beneath the suspected polyp was further confirmed. Enhanced abdominal computed tomography merely re-vealed local bowel wall thickening and inflammation of the colosigmoid junction. The migrating mesh, which was lodged in the sigmoid colon and caused intra--abdomi-nal adhesion in the lower abdominal cavity, was finally identified via exploratory surgery. The components of inflammatory granulation tissue around the mesh mate-rial were diagnosed based on histological examination of the surgical specimen after sigmoidectomy. In this patient, nonspecific endoscopic and imaging outcomes during clinical work--up led to the diagnostic dilemma of mesh migration. Therefore, the clinical, radiological and endoscopic challenges specific to this case as well as the underlying reasons for mesh migration are discussed in detail.

Diagnostic value of liquid-based cytology and smear cytology in pancreatic endoscopic ultrasound-guided fine needle aspiration: A meta-analysis

BACKGROUND Smear cytology(SC)using endoscopic ultrasound-guided fine needle aspiration(EUS-FNA)is the established and traditional choice for diagnosing pancreatic lesions.Liquid-based cytology(LBC)is a novel alternative cytological method,however,the comparative diagnostic efficacy of LBC remains inconclusive.AIM To examine the diagnostic efficacy of LBC and SC for pancreatic specimens obtained through EUS-FNA via a systematic review and meta-analysis.METHODS A systematic literature search was performed using PubMed,EMBASE,the Cochrane Library,and Web of Science.The numbers of true positives,false positives,true negatives,and false negatives for each cytological test(LBC and CS)were extracted from the included studies.The pooled sensitivity and specificity and the area under the summary receiver operating characteristic curve(AUC)were calculated,and the AUC was compared by Tukey's multiple comparisons test.The quality of the included studies was assessed using the Quality Assessment of Diagnostic Accuracy Studies II tool.RESULTS A total of 1656 patients in eight studies were included.The pooled sensitivity and specificity and the AUC for LBC were 0.76(95%CI:0.72-0.79),1.00(95%CI:0.98-1.00),and 0.9174,respectively,for diagnosing pancreatic lesions.The pooled estimates for SC were as follows:Sensitivity,0.68(95%CI:0.64-0.71);specificity,0.99(95%CI:0.96-100.00);and AUC,0.9714.Similarly,the corresponding values for LBC combined with SC were 0.87(95%CI:0.84-0.90),0.99(95%CI:0.96-1.00),and 0.9894.Tukey’s multiple comparisons test was used to compare the sensitivities and AUCs of the three diagnostic methods;statistically significant differences were found between the three methods,and LBC combined with SC was superior to both LBC(P<0.05)and SC(P<0.05).The pooled sensitivity and AUC did not change significantly in the sensitivity analysis.CONCLUSION LBC may be sensitive than SC in the cytological diagnosis of pancreatic lesions,however,the superior diagnostic performance of their combination emphasizes their integrated usage in the clinical evaluation of pancreatic lesions.

Adipose-derived mesenchymal stem cells alleviate TNBS-induced colitis in rats by influencing intestinal epithelial cell regeneration, Wnt signaling, and T cell immunity

BACKGROUND Conventional Crohn’s disease(CD)treatments are supportive rather than curative and have serious side effects.Adipose-derived mesenchymal stem cells(ADSCs)have been gradually applied to treat various diseases.The therapeutic effect and underlying mechanism of ADSCs on CD are still not clear.AIM To investigate the effect of ADSC administration on CD and explore the potential mechanisms.METHODS Wistar rats were administered with 2,4,6-trinitrobenzene sulfonic acid(TNBS)to establish a rat model of CD,followed by tail injections of green fluorescent protein(GFP)-modified ADSCs.Flow cytometry,qRT-PCR,and Western blot were used to detect changes in the Wnt signaling pathway,T cell subtypes,and their related cytokines.RESULTS The isolated cells showed the characteristics of ADSCs,including spindle-shaped morphology,high expression of CD29,CD44,and CD90,low expression of CD34 and CD45,and osteogenic/adipogenic ability.ADSC therapy markedly reduced disease activity index and ameliorated colitis severity in the TNBS-induced rat model of CD.Furthermore,serum anti-sacchromyces cerevisiae antibody and panti-neutrophil cytoplasmic antibody levels were significantly reduced in ADSCtreated rats.Mechanistically,the GFP-ADSCs were colocalized with intestinal epithelial cells(IECs)in the CD rat model.GFP-ADSC delivery significantly antagonized TNBS-induced increased canonical Wnt pathway expression,decreased noncanonical Wnt signaling pathway expression,and increased apoptosis rates and protein level of cleaved caspase-3 in rats.In addition,ADSCs attenuated TNBS-induced abnormal inflammatory cytokine production,disturbed T cell subtypes,and their related markers in rats.CONCLUSION Successfully isolated ADSCs show therapeutic effects in CD by regulating IEC proliferation,the Wnt signaling pathway,and T cell immunity.

Effect of Biostimble Therapy on Gastrointestinal Function Recovery after Gynecological Surgery

Objective:To clarify the effectiveness and safety of the Biostimble therapeutic instrument in postoperative gastrointestinal dysfunction in gynecology.Methods:Eighty cases of gynecological surgery patients from Changle County People's Hospital in Shandong Province were selected and divided into the experimental group and the control group,with 40 patients in each group.Low-frequency pulse electrical stimulation on the Zu San Li acupoint with Biostimble therapy instrument was carried in the experimental group.Electrode pads were arranged at the location of the bilateral Zu San Li acupoints,and the Biostimble treatment was applied twice a day for 30 minutes each time.The patients in the control group received stand postoperative nursing.The time of first anal venting,the time of recovery of bowel sounds,the duration of abdominal distension,the duration of abdominal pain,and the postoperative symptom score were observed in the two groups.Adverse events were recorded.Results:Compared to the control group,the experimental group demonstrated significantly shorter bowel sound recovery time,first anal exhaust time(all P<0.001).The duration of abdominal distension was 23.79±5.12 hours,and the duration of abdominal pain was 30.15±4.24 hours in the experimental group.While the duration of abdominal distension was 29.07±5.06 hours,and the duration of abdominal pain was 37.17±5.24 hours in the control group.There was a significant difference between the two groups(All P<0.001).The mean postoperative symptoms scores in the experimental group(1.17±1.03)were lower than those in the control group(2.08±1.71)at the 72-hour after the operation(P=0.005).The complete recovery rate in the experimental group was higher than that in the control group.The difference in treatment outcome between experimental and control was statistically significant(P=0.013).Conclusion:Biostimble therapy instrument electrostimulation on Zu San Li acupoints can promote rapid recovery of gastrointestinal function in patients after general gynecological surgery.

Quality standard study of Li folk medicine callicarpa nudiflora powder

Objective:To establish the quality standard of Li folk medicine callicarpa nudiflora powder by modern analytical method.Methods:TLC was used to identify the powder.Five heavy metals and harmful elements(Pb,Cd,As,Hg and Cu)in the powder were determined by flame atomic absorption spectrometry and hydride atomic fluorescence spectrometry.The content of luteolin and Mullein glycoside in the powder was determined by HPLC.Results:the fluorescent spots of TLC identification of powder were clear,and the separation of the two main components was good.It has good specificity,reproducibility and durability,which can be used for the identification of the powder.The inspection items of heavy metals for the identification of powder were established.It was stipulated that:according to the determination method,the lead should not exceed 5 mg/kg;the cadmium should not exceed 0.3 mg/kg;the arsenic should not exceed 2 mg/kg;the mercury should not exceed 0.2 mg/kg;the copper should not exceed 20 mg/kg.The content of luteolin and Mullein glycoside in the powder were determined by HPLC and the content inspection item of the powder was established.It was stipulated that the content of luteolin and Mullein glycoside in the powder should not be less than 0.48%and 5.83%,which can be used for the content limit inspection of the powder.Conclusion:the quality standard method established by our institute is advanced and operable,and can be used for the quality control of the powder.