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The interaction between intestinal epithelial cells (IECs) and lymphocytes is known to regulate mucosal barrier function, immune responses, and by extension, to play an etiologic role in the development of chronic inflammation in Ulcerative colitis (UC). However, the biologic function of lymphocytes and their relationship with IECs is insufficiently understood. In this study, we report, for the first time, the establishment of 3-dimensional (3D) co-culture systems of lymphocytes and mouse primary colonic organoids.C57BL/6 mice were treated with dextran sulfate sodium to induce acute colonic inflammation. Primary colonic organoids were established from cells isolated from mouse colons. Lymphocytes isolated from spleens by Ficoll density gradient assays. Colonoids and lymphocytes were co-cultured. The effects of the tested drugs on the co-culture system were assayed by Enzyme-linked Immune Sorbent Assay (ELISA) for inflammatory molecules. Inflammatory stimulation resulted in a significant increase in inflammatory cytokines in the supernatant of the colonoid-lymphocyte organoid system. Treatment with drugs known to be effective in patients with UC demonstrated a reduction in inflammatory cytokines. These findings argue that the colonoid-lymphocyte system can be utilized to model inflammation in vitro as well as for drug discovery. To validate these findings, we tested three other drugs. We produced experimental evidence that, indeed, these three drugs also induced a decrease in inflammatory cytokines of varying degrees. We report, for the first time, the successful establishment of a 3D colonoid-lymphocyte system and we validate its ability to predict the anti-inflammatory properties of several small molecules.

Certain conflicting conclusions have been drawn that gastric xanthelasma is related to H. pylori, atrophic gastritis, intestinal metaplasia, and early gastric cancer. The aim of this study was to examine the relationship between gastric xanthelasma and upper gastrointestinal (GI) endoscopic or pathological features. A cross‑sectional study was completed. A total of 8,634 patients who underwent stomach biopsy and who had no gastrectomy history were enrolled in the study. The patients were divided into two groups according to the presence or absence of gastric xanthelasma. The relationship between gastric xanthelasma and demographic characteristics (including age and sex), endoscopic features (including peptic ulcer, bile reflux, and gastric poly), or pathological features (including atrophy, intestinal metaplasia, H. pylori, dysplasia, and gastric cancer) was analyzed. Age/sex matched analysis was also performed to exclude the influence of age and sex. The results revealed that out of the 8,634 patients, 3.54% patients had xanthelasma. Gastric xanthelasma was significantly associated with age (55.76 vs. 49.17 years, P0.0001), duodenal ulcer (OR 0.860, 95% CI 0.369‑0.923), atrophy (OR 1.839, 95% CI 1.432‑2.362), and intestinal metaplasia (OR 3.296, 95% CI 2.612‑4.159). Binary logistic analysis revealed that age (OR 1.027, 95% CI 1.017‑1.037) and intestinal metaplasia (OR 2.700, 95% CI 2.090‑3.487) were independently related to gastric xanthelasma. Age/sex matched control binary logistic analysis revealed that gastric xanthelasma was significantly associated with presence of intestinal metaplasia (OR 2.338, 95% CI 1.659‑3.297). There was no difference in the number (P=0.427) and location (P0.05) of gastric xanthelasma for intestinal metaplasia. In conclusion, gastric xanthelasma may be an independent endoscopic warning sign of intestinal metaplasia.

Patient screening is important for early diagnosis of colorectal cancer (CRC). The present study aimed to compare the multitarget stool DNA (mt-sDNA) test with the fecal occult blood test (FOBT) for CRC screening. A total of 151 individuals were screened using colonoscopy, mt-sDNA and FOBT for the detection of CRC and adenoma. The results of the mt-sDNA test and FOBT were compared with colonoscopy to examine their sensitivity and specificity. Subsequently, the sensitivity and specificity of the mt-sDNA test were compared with those of FOBT in CRC and large adenoma. Stool samples were collected from patients with CRC (n=50) or large adenoma (n=51), as well as from normal controls (n=50). The mt-sDNA test outperformed FOBT in detecting CRC with a sensitivity of 90.0% (45/50) vs. 42.0% (21/50), advanced adenoma with a sensitivity of 70.6% (36/51) vs. 19.6% (10/51), stage I-III CRC with a sensitivity of 91.9% (34/37) vs. 29.7% (11/37), and stage IV CRC with a sensitivity of 84.6% (11/13) vs. 76.9% (10/13). In addition, the mt-sDNA test exhibited a specificity of 94.0% (47/50) in detecting CRC, which was superior to FOBT with a specificity of 90.0% (45/50). Therefore, the mt-sDNA test may have higher sensitivity and specificity compared with FOBT in diagnosing both CRC and advanced adenoma.

Peripheral neurotoxicity injury caused by local anesthetics is a common complication of clinical anesthesia. The study of its mechanism is helpful to prevent and treat the neurotoxic injury of local anesthetics. Previous studies on peripheral neurotoxicity injury caused by local anesthetics have mainly focused on in vitro cell experiments. Due to the lack of an animal model of peripheral neurotoxicity damage caused by local anesthetics, there are few in vivo experimental studies regarding this topic. Herein, 1% ropivacaine hydrochloride was injected into the sciatic nerve by direct incision and exposure of the sciatic nerve to create a local anesthetic neurotoxic injury model. The results showed that 1% ropivacaine hydrochloride could reduce the lower limb motor score and mechanical paw withdrawal threshold in mice 48 hours after injection. Pathological sections showed that 48 hours after treatment with 1% ropivacaine hydrochloride, the sciatic nerve showed increased axonal edema and degeneration, edema between nerve fiber bundles, increased degeneration of axon and myelin sheath vacuoles, edema of nerve bundle membrane and local degeneration and necrosis, and a large number of inflammatory cells around the nerve adventitia were soaked. The above results show that under open vision, 1% ropivacaine hydrochloride can cause injury to the sciatic nerve after 48 h of treatment, which can simulate the neurotoxic damage of local anesthetics. This animal model provides a research tool for studying the mechanism of neurotoxic injury caused by local anesthetics.

Background and Aim: A proper colonoscopy referral criterion is essential for flexible sigmoidoscopy‐based colorectal cancer screening. We aimed to compare the predictive capability of four existing criteria to detect proximal neoplasia (PN) and advanced proximal neoplasia (APN) in a Chinese population. Methods: Asymptomatic Chinese participants aged 50–75 years, who received screening colonoscopy, were consecutively recruited. The four criteria included (i) UK flexible sigmoidoscopy; (ii) Italian Screening for COlon REctum; (iii) NORwegian Colorectal Cancer Prevention trial; and (iv) US clinical index. The sensitivity, specificity, positive/negative predictive value, and the number of subjects needed to screen (NNS)/refer (NNR) to detect one APN/PN were examined. The area under receiver operating characteristic curve was evaluated. Results: Among 5833 subjects, 749 (12.8%) and 151 (2.6%) cases were found to have PN and APN, respectively. US criteria achieved the highest sensitivity for PN (49%) and APN (66%), while UK criteria attained the highest specificity (93%) for PN/APN. The lowest NNS was required by US criteria for PN (16 vs 19–38) and APN (58 vs 69–86), while the lowest NNR was required by UK criteria for PN (3.2 vs 4.0–4.8) and APN (7 vs 10–16). The receiver operating characteristic of all four criteria was 0.57–0.61 for PN and 0.68–0.70 for APN. Conclusions: Among all the four criteria, US criteria had the highest sensitivity and lowest NNS, while UK criteria achieved the highest specificity and lowest NNR. Their limited discriminatory capability highlighted the need for a new score to predict PN/APN in Chinese populations.

Following the publication of the above article, the authors have drawn to our attention that, in Fig. 6 on p. 83, the 'Ad5/F‑35 vector' and 'Control' experiment data panels for the MKN45 cell line were inadvertently derived from the same original source. The Ad5/F35‑vector and Control groups were both negative controls; Ad5/F35‑vector was the empty vector condition, whereas 'Control' represented the 'no vector' experiment. A representative image belonging to the Control MKN45 experiment was inadvertently used to show the 'Ad5‑F35‑vector' experiment. An examination of the original data files confirmed that the data shown in the histogram for Fig. 6 were correct; therefore, this error in terms of figure placement did not influence the statistical analysis shown for the invasive ability of the cells. A corrected version of Fig. 6, containing correct representative data for the 'Ad5‑F35‑vector' experiment, is shown opposite. This error did not affect the results or the conclusions reported in this study. The authors sincerely apologize for this mistake, and regret any inconvenience this mistake has caused. [the original article was published in Oncology Reports 34: 77‑86, 2015; DOI: 10.3892/or.2015.3943].

Background The Asia-Pacific Colorectal Screening (APCS) score and a modified version that incorporates body mass index (BMI) has been widely recognised. This study aimed to compare the external validity of the APCS and the modified APCS to predict advanced colorectal neoplasia (ACN) in an asymptomatic Chinese population. Methods A prospective, multi-centre colonoscopy study was conducted in 7 endoscopic units across North Shanghai, China from 2016 to 2017. Average-risk subjects were recruited during their attendance for screening colonoscopy. Information on the predictors, including age, gender, family history of colorectal cancer and smoking status were collected by self-reported questionnaires. Body weight and height were measured by trained professionals. All screening colonoscopy procedures were performed by senior colonoscopists. The histology of all resected lesions was reported using the same diagnostic criteria by pathologists who were blinded to the study design. Cumulative scores were derived for each subject and categorised as average, moderate and high risk. The risk of ACN was assessed for each risk tier and the excess risk in moderate and high-risk tiers was compared with subjects with average risk. The C-statistics was employed to evaluate the discriminatory capability of both APCS scores. Results Among a total of 2813 enrolled subjects (mean age 58.7±8.4 years), 679 cases (24.1%) were found to have colorectal neoplasia, of which 142 (5.0%) had ACN and 29 patients (1.0%) had invasive cancers (table 1). In both APCS scoring systems, the risk of ACN was significantly different in low, moderate and high-risk tier (p Conclusions Both the APCS and modified APCS scores were similarly efficient to identify those subjects with high risk of can. Their feasibility and acceptability in different population groups should be further explored.

Previous studies have shown the colonoscopy quality affected by the endoscopist's fatigue. This study was aimed to evaluate this potential factor in a colorectal cancer-screening cohort of Chinese patients.The attendances at department of gastroenterology for colorectal cancer screening between 2013 and 2015 were retrospectively analyzed. The procedure time-of-day and hours elapse were recorded. The primary outcome was defined as adenoma detection rate (ADR).A total of 1342 screening colonoscopies were performed by 19 gastroenterologists in the study. Detection rates were 7.7% for all polyps and 20.0% for adenomas. Time-of-day was not significantly associated with ADR. With time elapsing, the first climax for ADR was presented at 09:00-10:00, and persistently rose again after the lunch break. Significant inclined trend in ADR was noted for each hour blocks of a full day (p = .0021).The procedure time-of-day, morning versus afternoon, did not affect the diagnostic efficacy of screening endoscopy in Chinese patients.

We tested our hypothesis that Myc-interacting zinc finger protein 1 (MIZ1), a cell cycle regulator, suppressed inflammation, and therefore, represented a useful prognostic marker in patients with acute necrotizing pancreatitis (ANP) complicated by acute lung injury.Sprague-Dawley rats were randomly divided into control and ANP groups at different time points. The MIZ1 protein expression was measured by Western blot and ELISA, and confirmed using immunohistochemistry. The severity of pancreatic and lung injury was evaluated by the injury score and wet/dry weight ratio. The severity of disease was evaluated by serum C-reactive protein (CRP). The MPO activity of lung tissue amylase levels and the degree of inflammation were evaluated by serum tumor necrosis factor (TNF)-α and interleukin (IL)-6 expression. The risk due to multiple factors was investigated by relationship analysis.The serum levels of CRP, amylase, TNF-α, and IL-6 were gradually increased at 6, 24, and 48 h in ANP when compared with the control rats. The MIZ1 expressions were greatly decreased in ANP rats, especially at 24 h. Statistical analysis showed that there were time-dependent differences in ANP rats when compared with control rats (6 vs. 24 or 48 h, P0.01). MIZ1 showed close negative correlation with the degree of pancreatic and lung injury, serum amylase, CRP, TNF-α, and IL-6 (P0.01, respectively).The decreasing MIZ1 expression was closely correlated with inflammatory response, and development of ANP. Decreasing MIZ1 levels indicate a risk for ANP.

Accumulating evidence indicates that resistin-like molecule-α (RELM-α) is involved in angiogenesis, while the clinical significance and the exact role of RELM-α in gastric cancer remain obscure. The aim of the present study was to evaluate the clinical significance of RELM-α in gastric cancer, and to investigate its effective mechanisms in order to identify a potential therapeutic target. The expression levels of RELM-α in 92 gastric cancer and adjacent normal tissues were investigated and the relationship between RELM-α expression and the clinicopathological characteristics was explored. To investigate the potential role of RELM-α in gastric cancer cell biological behavior, the cell proliferation, migration and invasion assays were conducted using two gastric cancer cell lines (SGC7901 and MKN45). We also assessed whether RELM-α gene silencing modulates angiogenesis using small interference RNA in cancer cell lines, and investigated its effect on nuclear factor (NF)-κB activation and vascular endothelial growth factor (VEGF) and MMP-9 expression. Contrasting sharply with the strong RELM-α-positive tumors, adjacent normal tissues and cell lines exhibited negative or weakly positive expression (P

Protein inhibitor of activated signal transducer and activator of transcription-1 (PIAS1) is an important regulator of the inflammatory signaling network, the expression of which was decreased in gastric cancer and implicated in the development of cancer. However, its mechanism has not been elucidated. The aim of the present study was to investigate the effect of PIAS1 on epithelial-mesenchymal transition (EMT) of gastric cancer cells within the inflammatory microenvironment. Recombinant adenovirus Ad5/F35-PIASl and Ad5/F35-null plasmids were constructed to transfect SGC7901 cells. Subsequently, these plasmids were confirmed by reverse transcription polymerase chain reaction and western blotting. The cells were treated with IL-6 or Ad5/F35-PIASl+IL-6, and the control cells were treated with Ad5/F35-null+IL-6. The morphological changes to the cells were observed using inverted microscopy. The effect of PIAS1 on cell migration and invasion was evaluated by scratch wound healing and Transwell chamber assays, and the protein expression of EMT markers and phosphatidylinositol 3-kinase (PI3K)/serine/threonine-protein kinase (Akt)/matrix metalloproteinase (MMP)-9 signaling pathway was examined by western blotting. Transfection with Ad5/F35-PIASl markedly increased the PIAS1 expression in SGC7901 cells. The cells acquired the more typical spindle-shape phenotype of mesenchymal cells following co-culture with IL-6; the cells co-cultured with IL-6 and Ad5/F35-PIASl acquired changes concordant with an epithelial phenotype. The overexpression of PIAS1 significantly decreased the migratory and invasive capacities of the SGC7901 cells (P

Vascular endothelial growth factor-C (VEGF-C) is considered as a prime mediator of lymphangiogenesis and has been implicated in carcinogenesis and metastasis. Various studies examined the relationship between VEGF-C overexpression and the clinical outcome in patients with gastric cancer, but yielded conflicting results. Electronic databases updated to July 2013 were searched to find relevant studies. A meta-analysis was conducted with eligible studies which quantitatively evaluated the relationship between VEGF-C overexpression and survival of patients with gastric cancer. Survival data were aggregated and quantitatively analyzed. We performed a meta-analysis of 13 studies that evaluated the correlation between VEGF-C overexpression and survival in patients with gastric cancer. Combined hazard ratios suggested that VEGF-C overexpression had an unfavorable impact on overall survival (OS) (hazard ratio (HR) = 1.38, 95 % confidence interval (CI) = 1.08–1.68), but not disease-free survival (DFS) (HR = 1.25, 95 % CI = 0.89–1.62) in patients with gastric cancer. No significant heterogeneity (P = 0.132) was observed among 11 studies for OS and among 5 studies for DFS (P = 0.105). VEGF-C overexpression indicates a poor prognosis for overall survival, but not disease-free survival in patients with gastric cancer.

The purposes of this study were to determine the expression profiles of microRNA-34a (miR-34a) in human gastric cancer cell line (SGC-7901) and cisplatin-resistant cell lines (SGC-7901/DDP), and to establish the correlation between miR-34a expression profile and the sensitivity of human gastric cancer cell to cisplatin-based pattern, thereby providing new methods and strategies for treating gastric cancer. Gastric cancer cell line (SGC-7901) and cisplatin-resistant cell line (SGC-7901/DDP) were cultivated in vitro, respectively. Quantitative real-time PCR (qRT-PCR) and Western blot were utilized to determine the expression profiles of miR-34a and survivin in both gastric cancer cell lines. With miR-34a mimic and miR-34a inhibitor transfected into SGC-7901 and SGC-7901/DDP for 48 h, post-transfection changes of miR-34a expression was determined; the effects of miR-34a ectopic expression on the viability of cisplatin-induce gastric cancer cell were assayed by the MTT method. The effects of miR-34a ectopic expression on apoptosis of cisplatin-induce gastric cancer cell were determined by Annexin V/propidium iodide (PI) double staining method and flow cytometry. The effects of miR-34a ectopic expression on the AKT and p-AKT expression of cisplatin-induce gastric cancer cells were determined by Western blot and flow cytometry with the PI3K pathway inhibitor Wortmannin. As shown by qRT-PCR and Western blot analyses, the expression of miR-34a in cisplatin-resistant cell lines decreased significantly in comparison to that of SGC-7901 cell line (p < 0.05), while significant up-regulation of survivin expression was also observed (p < 0.05). Compared with the control group, the expression of miR-34a increased significantly in SGC-7901 cells transfected with miR-34a mimic for 48 h (p < 0.01). After miR-34a inhibitor transfection, the expression of miR-34a decreased significantly (p < 0.05). The viability of cisplatin-induce gastric cancer cells increased significantly (p < 0.05) with significant decrease of apoptosis after miR-34a expression inhibition, as demonstrated by MTT and flow cytometry with miR-34a over-expression, the viability of cisplatin-induce gastric cancer cells decreased significantly (p < 0.05), with significant apoptosis increase (p < 0.05). As shown by Western blot and flow cytometry, in comparison to the control group, Wortmannin could inhibit miR-34a inhibitor and DDP induced up-regulation of p-AKT significantly (p < 0.05) and stimulated apoptosis. In conclusion, miR-34a expression was down-regulated in cisplatin-resistant cell lines. miR-34a over-expression could improve the sensitivity of gastric cancer cells against cisplatin-based chemotherapies, with PI3K/AKT/survivin signaling pathway possibly involved in the mechanism.

Whether screening participants with distal hyperplastic polyps (HPs) detected by flexible sigmoidoscopy (FS) should be followed by subsequent colonoscopy is controversial. We evaluated the association between distal HPs and proximal neoplasia (PN)/advanced proximal neoplasia (APN) in asymptomatic, average-risk patients.We searched Ovid Medline, EMBASE, and the Cochrane Library from inception to 30 June 2016 and included all screening studies that examined the relationship between different distal findings and PN/APN. Data were independently extracted by two reviewers with disagreements resolved by a third reviewer. We pooled absolute risks and odds ratios (ORs) with a random effects meta-analysis. Seven subgroup analyses were performed according to study characteristics. Heterogeneity was characterized with the IWe analyzed 28 studies (104,961 subjects). When compared with normal distal findings, distal HP was not associated with PN (OR=1.16, 95% confidence interval (CI)=0.89-1.51, P=0.14, IDistal HP is not associated with PN/APN in asymptomatic screening population when compared with normal distal findings. Hence, the presence of distal HP alone detected by FS does not automatically indicate colonoscopy referral for all screening participants, as other risk factors of PN/APN should be considered.

We aimed to investigate the expression of microRNA-34a (miR-34a) in human gastric cancer cells and to evaluate the effects of miR-34a, acting via its gene survivin, on gastric cancer cell HGC-27 to provide potential new strategies for treating gastric cancer. In vitro cultures of the human gastric cancer cell lines MGC80-3, HGC-27, NCI-N87, and SGC-7901 and the normal human gastric epithelial cell line GES-1 were established. The expression of miR-34a in each gastric cancer cell line and GES-1 normal human gastric epithelial cell line was detected using quantitative real-time polymerase chain reaction (qRT-PCR). After the HGC-27 cells were transfected with a miR-34a mimic for 48 h, the changes in the expression levels of miR-34a were detected using qRT-PCR. The effect of miR-34a on HGC-27 cell viability was measured using a tetrazolium-based colorimetric [−(4,5)-dimethylthiahiazo-(−z-y1)-3,5-di-phenytetrazoliumromide (MTT)] assay. Flow cytometry was used to analyze the effects of miR-34a on HGC-27 cell proliferation. Annexin V/propidium iodide double staining and flow cytometry were used to analyze the effects of miR-34a on HGC-27 cell apoptosis. A Transwell invasion chamber was used to detect the effects of miR-34a on HGC-27 cell invasion. Finally, western blotting was used to analyze the effects of miR-34a on survivin protein expression. The qRT-PCR test determined that miR-34a expression in gastric cancer cells was significantly reduced compared to the normal gastric epithelial cell line GES-1 (p < 0.01). Compared to the control group, cellular miR-34a expression levels were significantly increased in HGC-27 human gastric carcinoma cells after transfection with a miR-34a mimic for 48 h (p < 0.01). The MTT assay demonstrated that after overexpressing miR-34a in HGC-27 cells, cellular viability was significantly reduced (p < 0.05). Flow cytometry analysis determined that upon miR-34a overexpression, the proliferation index decreased significantly (p < 0.05), and cellular apoptosis was significantly increased (p < 0.01). The Transwell invasion chamber assay illustrated that after increasing the expression of miR-34a, the number of cells passing through the Transwell chamber was significantly reduced (p < 0.01). Based on western blotting, compared with the control group, survivin protein expression levels were significantly decreased in the HGC-27 cells transfected with the miR-34a mimic for 48 h (p < 0.01). In conclusion, the expression level of miR-34a was downregulated in human gastric cancer cell lines. miR-34a can negatively regulate survivin protein expression and inhibit gastric cancer cell proliferation and invasion. Therapeutically enhancing miR-34a expression or silencing the survivin gene may benefit patients with gastric cancer.

The expression of N-myc downstream-regulated gene 2 (NDRG2) is present in normal tissues but low or undetectable in various cancers and thus poses a potential tumor suppressor gene. However, the expression of NDRG2 in colorectal tissues remains unknown. Here, our results showed that NDRG2 was down-regulated in colorectal cancer compared to benign colorectal tissues by using immunohistochemical staining and semi-quantitative RT-PCR analyses. Bisulfite sequencing analysis showed that the reduced NDRG2 expression was accompanied by de novo DNA methylation at the NDRG2 promoter. We also found that microRNA-650 (miR-650) targets a homologous DNA region in the promoter region of the NDRG2 gene and represses its expression at the transcriptional level. Reporter assay with 3'untranslated region of NDRG2 cloned downstream of the luciferase gene showed reduced luciferase activity in the presence of miR-650, providing strong evidence that miR-650 is a direct regulator of NDRG2. In conclusion, these results suggest that NDRG2 expression is regulated by promoter methylation and miR-650 in human colorectal cancer cells, and endogenous small noncoding RNA induced control of transcription may be a potential system for expressional regulation in human colorectal cancer cells.

MicroRNAs (miRNAs) have emerged as post-transcriptional regulators that are critically involved in the pathogenesis of a number of human cancers. Recently, cyclin-dependent kinase 6 (CDK6) is found to be up-regulated in several types of human tumors and has been implicated in cancer initiation and progression. We have identified miR-107 as a potential regulator of CDK6 expression. A bioinformatics search revealed a putative target site for miR-107 within the CDK6 3' untranslated region. Expression of miR-107 in gastric cancer cell lines was found inversely correlated with CDK6 expression. miR-107 could significantly suppress CDK6 3' UTR luciferase reporter activity, and this effect was not detectable when the putative 3' UTR target site was mutated. Consistent with the results of the reporter assay, ectopic expression of miR-107 reduced both mRNA and protein expression levels of CDK6, inhibited proliferation, induced G1 cell cycle arrest, and blocked invasion of the gastric cancer cells. Our results suggest that miR-107 may have a tumor suppressor function by directly targeting CDK6 to inhibit the proliferation and invasion activities of gastric cancer cells.

International audience; Human telomeres are protected from DNA damage by a nucleoprotein complex that includes the repeat-binding factor TRF2. Here, we report that TRF2 regulates the 5' exonuclease activity of its binding partner, Apollo, a member of the metallo-beta-lactamase family that is required for telomere integrity during S phase. TRF2 and Apollo also suppress damage to engineered interstitial telomere repeat tracts that were inserted far away from chromosome ends. Genetic data indicate that DNA topoisomerase 2alpha acts in the same pathway of telomere protection as TRF2 and Apollo. Moreover, TRF2, which binds preferentially to positively supercoiled DNA substrates, together with Apollo, negatively regulates the amount of TOP1, TOP2alpha, and TOP2beta at telomeres. Our data are consistent with a model in which TRF2 and Apollo relieve topological stress during telomere replication. Our work also suggests that cellular senescence may be caused by topological problems that occur during the replication of the inner portion of telomeres.

The telomere-binding protein TRF2 regulates both telomere protection and telomere length. The fact that TRF2 is up-regulated in some tumors indicates that TRF2 plays a role in cancer. However, the role of TRF2 in gastric cancer has not been fully understood. The aim of this study is to evaluate the expression pattern of TRF2 in gastric cancer and examine the potential mechanism of TRF2 regulation. Our study revealed that the expression of TRF2 analyzed by immunohistochemistry was significantly higher in gastric cancers compared to noncancerous tissues. Moreover, TRF2 methylation was detected in six of 30 (20%) primary gastric cancers and 18 of 30 (60%) paired normal tissues, and the downregulation of TRF2 was strongly correlated with the methylation status (P < 0.001). Our results suggest that hypomethylation might contribute to the upregulation of TRF2 in gastric cancers and this overexpression may play a role in the pathogenesis of gastric cancers.

An approach to fingerprinting and the quantitative analysis of volatiles in Shexiang Baoxin Pill (SBP) was proposed by using gas chromatography (GC) with a flame ionization detector and a mass spectrometric detector (MS). Using the proposed method, the chemical fingerprint of SBP extract was established, in which the separation of more than 30 volatiles was realized in about 30 min, and 26 peaks were identified by GC/MS analysis. Seven major volatiles, including borneol, isoborneol, isopropyl methylphenol, mascone, cinnamaldehyde, cinnamic acid, and benzyl benzoate, were further quantified. The linearity, precision, stability, repeatability, and accuracy were acceptable. The proposed method was successfully applied to the determination of volatiles in 11 batches of SBP samples. To evaluate their quality, principal components analysis (PCA) was performed on the basis of the data of chemical fingerprints, and the score plot clearly revealed the variations of samples produced in different years. Moreover, the results of the quantitative analysis of various samples were also comparatively studied. The results indicated that, to ensure the quality of SBP, more efficient storage and package techniques are needed. The proposed method enables chemical fingerprinting and the simultaneous determination of multicomponents to be performed in one run, and can be applied as a comprehensive quality control technique for traditional Chinese medicine containing volatile constituents.

Double-balloon enteroscopy (DBE) is a novel endoscopic technique developed to investigate small bowel disease. There are limited available data on its impact in the diagnosis and management of obscure gastrointestinal bleeding (OGIB). The aim of this study was to evaluate the diagnostic yield and therapeutic impact of DBE in the management of patients with OGIB.This study is a retrospective analysis of patients referred to our hospital from December 2003 to January 2005 for the investigation of overt or occult OGIB who underwent DBE after negative upper endoscopy and colonoscopy. Demographic, clinical, procedural, and outcome data were collected for analysis.One hundred fifty-two patients (73 women and 79 men) were studied, with a mean age of 48.2 yr. Seventeen patients presented with occult OGIB while 135 patients had overt OGIB. A total of 191 DBEs was performed. Antegrade and retrograde approaches were performed in 60 and 53 patients, respectively, and 39 patients had a combination of both routes. DBE demonstrated a potential bleeding site in 115 (75.7%) patients (102 overt, 13 occult). The more common abnormalities detected were small bowel tumors (39.1%) and angioectasia (30.4%). DBE altered management in 83.5% of patients with positive findings. Follow-up was obtained on 119 patients (mean 16 months, range 8-23 months). Of the 95 patients with follow-up and a positive DBE finding, 85 (89%) had no further rebleeding. The procedure was well tolerated with 23 patients (15.1%) experiencing mild self-limited bleeding during the procedure.DBE appears to have a high diagnostic yield and therapeutic impact in patients with OGIB with previously negative upper endoscopy and colonoscopy.

OBJECTIVE: To study: (i) the induction of apoptosis in gastric cancer cells by trichosanthin; and (ii) the relationship between apoptosis and the expression of bcl-2. METHODS: During in vitro experiments, morphological studies and the terminal deoxynucleotidyl transferase-mediated dUTP–digoxigenin nick end-labeling (TUNEL) method were used to detect apoptosis in gastric adenocarcinoma cell line SGC-7901 before and after trichosanthin treatment. An immunohistochemical staining method and northern blot hybridization were used to detect the expression of the apoptosis-related gene bcl-2 before and after trichosanthin treatment. RESULTS: When SGC-7901 cells were treated with trichosanthin (0.1 μg/mL, 36 h), they presented some typical apoptotic morphological changes that were observed by fluorescent staining. These morphological changes included nuclear condensation and nucleosomal fragments forming a lunate body under the nuclear membrane. When SGC-7901 cells were treated with trichosanthin (0.1 μg/mL) for 36, 42 or 48 h, TUNEL staining revealed a significant increase in the apoptotic index (AI), from 3.78 ± 1.11%, 3.98 ± 1.12% and 3.85 ± 1.08%, to 11.30 ± 2.33%, 10.22 ± 2.00% and 11.18 ± 1.85% (P < 0.01), respectively. When SGC-7901 cells were treated with trichosanthin (0.1 μg/mL, 32 h), immunohistochemical staining revealed a decreased expression of the bcl-2 protein product: the staining density decreased from ++/+++ to –/+ (P < 0.01). When SGC-7901 cells were treated with trichosanthin (0.1 μg/mL, 24 h), northern blot hybridization showed a decreased expression of bcl-2 RNA: hybridization decreased from 35.19 ± 2.34 to 22.27 ± 3.90 (P < 0.01). CONCLUSIONS: Trichosanthin is able to induce apoptosis in gastric cancer. The apoptosis may be mediated by the downexpression of the apoptosis-related gene bcl-2.

Bone marrow-derived myofibroblasts (BMFs) have been shown to promote tumor growth. Here, we found that BMFs or BMF conditioned medium (BMF-CM) induced cancer stem cell-like sphere formation of colon cancer cells. The co-cultured BMFs, but not co-cultured cancer cells, expressed higher levels of IL-6 than BMFs or cancer cells cultured alone. Anti-mouse IL-6 neutralizing antibody, JAK2 inhibitors and STAT3 knockdown in mouse cancer cells reduced BMF- and BMF-CM-induced sphere formation of colon cancer cells. When co-injected, BMFs significantly enhanced tumorigenesis of colon cancer cells in mice. Our results demonstrate that BMFs promote tumorigenesis via the activation of the IL-6/JAK2/STAT3 pathway.

Background The rising incidence and mortality of colorectal cancer (CRC) in China highlights the pressing need for population-based screening. Flexible sigmoidoscopy (FS) is becoming increasingly popular as a primary screening tool; and in many regions of China, colonoscopy capacity is limited. However, FS is preferred for patients with a low risk of advanced proximal neoplasia (APN; ie, higher numbers needed to screen [NNS], since the test predominantly examines the distal colon. Tailoring endoscopic screening based on the risk of APN is necessary, especially in resource-deprived regions. Since old age and female sex are two important CRC risk factors, we hypothesised that younger Chinese women aged 50–59 years had a significantly lower APN rate than did older Chinese men. Methods In 2013–15, we recruited 5833 asymptomatic participants aged 50–75 years who were registered for CRC screening in a large hospital-based endoscopy unit in Shanghai, China. Those with poor bowel preparation and whose colonoscopy failed caecal intubation were excluded. All polyps were removed as deemed appropriate and examined by histopathologists who were masked to the patients' data. We used χ 2 test to compare APN among different age and gender groups. The respective NNS were evaluated. Institutional ethics board approval was obtained by the hospital while informed consent was sought before subject recruitment. Findings For men, the prevalence of APN was 2·5% (31/1264, 95% CI 1·7–3·5) in those aged 50–59 years, 4·1% (30/735, 2·9–5·8) in those aged 60–64 years, and 5·2% (38/727, 3·8–7·1) in those aged 65–75 years. In women, the APN prevalence was 0·7% (11/1607, 0·4–1·2) in those aged 50–59 years, 0·8% (6/766,0·3–1·8) in those aged 60–64 years, 4·8% (35/734,3·4–6·6) in those aged 65–75 years in those aged 66 years or older (p Interpretation These findings supported the application of sigmoidoscopy for CRC screening among female individuals younger than 65 years, given their low risk of APN and high NNS. Colonoscopy could be favoured for Chinese individuals aged 65 years or older. The feasibility and cost-effectiveness of this simple, tailored screening strategy in different Chinese populations should be further examined. Funding None.

In this paper, developed is an effective solution to selective etching of ultra-thick copper sacrificial layer for the realization of 3-D suspended metal microstructures, in which the thickness of sacrificial Cu laminations can reach more than 100 microns. The proposed effective etching solution is based on an additive complex in the ammonia liquor with the assistance of oxidant. The primary purposes of the additive complex catalyst account for strengthening the complexing capabilities of the etching liquor as well as increasing the selective etching ratio of Cu. The effects of process conditions on etching rate and surface morphology were investigated. Therefore, the complex-based etching solution to ultra-thick Cu SL possesses unique features in high selective etching ratio, low-cost, high efficiency, time-saving, and compatible with MEMS process. The novel etching solution has a potential application in fabricating the complicated 3-D MEMS devices with movable suspensions.

LMX1A is epigenetically inactivated in cervical cancer. However, the expression and methylation status of LMX1A in gastric cancer tissues remains unknown. In the present study, we found that the expression of LMX1A was significantly decreased in gastric cancer tissues compared with normal tissues. A statistically significant inverse association was found between the LMX1A methylation status and expression of LMX1A in tumor tissues (P = 0.008). Restoration of LMX1A induced cell apoptosis and suppressed anchorage-independent growth, suggesting LMX1A may be a potential biomarker for gastric cancer.

Interleukin-33 (IL-33) is a novel member of the IL-1 family of cytokines, and it is closely related to IL-18, one of the best characterized members of the IL-1 family. It’s been demonstrated that elevated levels of IL-18 are involved in a wide variety of tumors, especially in gastric cancer. The purpose of this study was to determine the correlations between serum IL-33 levels and the clinicopathologic features in gastric cancer patients. Serum samples were collected from 68 patients with gastric cancer and 57 controls. Serum IL-33 levels were measured by ELISA. Classical tumor markers of CEA and CA19-9 levels were routinely detected by chemiluminescence immunoassay. Western blot analysis was used to detect IL-33 expression in gastric cancer tissue samples and cell lines. The relationship between serum levels of IL-33 and clinical characteristics in patients was analyzed. IL-33 levels in the serum of gastric cancer patients were significantly elevated in comparison with that of healthy volunteers. Furthermore, higher serum levels of IL-33 in gastric cancer patients were found to correlate with several poor prognostic factors like depth of invasion, distant metastasis and advanced stage (stage III/IV). On the other hand, serum IL-33 levels did not correlate with CEA and CA19-9. The expression of IL-33 protein was upregulated in carcinoma tissues in comparison with matched normal tissues, and no statistically significant difference was found between the four gastric cancer cell lines and human gastric epithelial cell line GES-1. Serum IL-33 may be a useful biomarker for predicting the prognosis of gastric cancer.

Hepatocyte growth factor (HGF) activator inhibitor type 2 (HAI-2/SPINT2) encodes Kunitz-type protease inhibitor that regulates HGF activity. Inspection of the human HAI-2/SPINT2 locus uncovered a large and dense CpG island within the 5' region of this gene. Analysis of cultured human gastric tumor lines indicated that HAI-2/SPINT2 expression is either undetectable or in low abundance in several lines; however, enhanced gene expression was measured in cells cultured on the DNA demethylating agent 5-aza-2'-deoxycytidine. Bisulfite DNA sequencing confirmed the densely methylated HAI-2/SPINT2 promoter region. Forced expression of HAI-2/SPINT2 induced cell apoptosis, suppressed anchorage independent growth in vitro and tumor growth in vivo. We investigated HAI-2/SPINT2 aberrant methylation in patients with gastric cancer. The HAI-2/SPINT2 methylation was found preferentially in cancerous tissues (30 of 40, 75%) compared with nontumor tissues (no methylation was detected), indicating that this aberrant characteristic is common in gastric malignancies. In conclusion, epigenetic inactivation of HAI-2/SPINT2 is a common event contributing to gastric carcinogenesis and may be a potential biomarker for gastric cancer.

Telomere repeat binding factor 2 (TRF2) plays a key role in the protective activity of telomere and is overexpression in several kinds of solid cancer cells. However, the role of overexpressed TRF2 in colorectal carcinoma remains unclear. The aim of this study was to determine the expression of TRF2, address the mechanism of TRF2 overexpression in human colorectal carcinoma. In present study, we examined the expression of TRF2 in colorectal cancer tissues from 39 patients, peritumoral normal tissues from 21 patients, and colon carcinoma SW480 cell line by quantitative PCR, immunohistochemistry and western blot. After siRNA silencing TRF2 expression in SW480, tumorigenesis of TRF2 was tested by cell proliferation, soft agar assay, cytofluorimetric analysis and cytogenetic analysis. To discover transcription factor that mediated TRF2 expression, Chromatin Immunoprecipitation (Chip) Assay and Electrophoretic mobility shift assays (EMSA) were employed.Overexpression of TRF2 protein was detected in SW480 cells and 19 of 39 colorectal carcinoma tissues (49%), no overexpression was observed in 21 of 21 adjacent peritumoral normal colorectal tissues. After siRNA silencing TRF2 expression, the proliferation and colony formation of SW480 cells were significantly inhibited. Defective TRF2 induced apoptosis and increased chromosomal instability in SW480 cells, in which there were more end-to-end fusions and ring chromosomes. Chip assay and EMSA showed that transcription factor Sp1 is involved in upregulation of TRF2. These results indicate that TRF2 is overexpressed in colorectal carcinoma, Sp1 upregulates TRF2 expression, TRF2 inhibition reduces tumorigenesis of colorectal cancer, which suggests that TRF2 and SP1 may become new targets for the development of anti-cancer therapy in colorectal carcinoma.

The BTG4 gene belongs to the BTG family of genes endowed with antiproliferative properties. In this study, we have found that BTG4 undergoes promoter CpG island hypermethylation-associated inactivation in gastric cancer and 5'-aza-2'-deoxycytidine (DAC) treatment restores BTG4 expression. We also found BTG4 levels were significantly reduced in primary gastric cancer but not in normal gastric tissues. BTG4 reexpression in gastric cancer causes growth inhibition of colony assays and nude mice. Taken together, our data support BTG4 as a candidate tumor suppressor gene that is epigenetically silenced in the majority of gastric cancers.

To evaluate clinical feasibility and efficacy of endoscopic metal stenting combined with radiotherapy for treatment of medium and late stages of esophageal carcinoma.Thirty-four patients of late stage esophageal carcinoma were treated with endoscopic stent implantation in combination with radiotherapy. Evaluations of CES stainless steel metal stent on the effect of radiation, clinical symptom alleviation and complication and survival rates in both groups were made.(1) CES stainless steel metal stent had no effect on radiation dosage distribution. (2) Dysphagia was markedly alleviated in both groups and different complication rates were observed between the two groups. (3) Six, nine and twelve month survival rates of inpatients treated with combined therapies were significantly higher than in simple stent implantation patients.Endoscopic metal stenting in combination with radiotherapy was a feasible and practical management in treating medium and late stages esophageal carcinoma and was superior to simple metal stent implantation.

Pancreatic intraepithelial neoplasia (PanIN) is the most common premalignant lesion of the pancreas. Further understanding of the biological behavior and molecular genetic alterations in the stepwise progression of PanINs is necessary toward the development of pancreatic ductal adenocarcinoma (PDAC) interventions. In this study, we analyzed the morphological characteristics, molecular alterations, and biological behavior of pancreatic wild-type and neoplasia tissues, including analysis of PanIN cell line SH-PAN (isolated from Pdx-1-Cre; LSL-KrasG12D/+ mouse) and PDAC cell line DT-PCa (isolated from Pdx1-Cre; LSL-KrasG12D/+; LSL-Tp53R172H/+ mouse). Results show that KrasG12D induces ductal lesion PanINs. Increased expression of EGFR, Her-2/Neu, p-MAPK and β-Catenin was observed in low-grade PanINs. Tp53 was not expressed in wild-type and low-grade PanINs, however, increased expression was observed in high-grade PanINs. Furthermore, SH-PAN cells did not exhibit any colony formation and showed significantly lower migration and invasion ability compared with DT-PCa cells. Notably, we first found PPP2R2A (protein phosphatase 2, regulatory subunit B, alpha) expression was significantly higher in SH-PAN cells than DT-PCa cells, and was high in 96 of 172 peritumoral normal human pancreatic tissues and 20 of 36 human low- or middle-grade PanIN tissues, whereas, was weak or negligible in 12 of 20 human high-grade PanIN tissues and 124 of 172 human PDAC tissues post-operation. The expression of PPP2R2A appears to be correlated with clinical survival. Taken together, KrasG12D – driven PanIN showed the tumorigenic ability, however, did not undergo a malignant transformation, and decreased expression of PPP2R2A in PDACs may provided a new target for pancreatic carcinoma intervention.