IL-22 is related to development of human colon cancer by activation of STAT3.

BACKGROUND: It has been previously reported that IL-22, one of the cytokines secreted by Th17 cells, demonstrates both a protective and inflammatory promotion effect in inflammatory bowel disease (IBD) through STAT3 signaling activation. We sought to investigate the role of IL-22 expression in colon cancer (CC). METHODS: The expression of IL-22 and related molecules were detected in human CC, the detail function and mechanism of IL-22 were investigated by in vivo and in vitro model. RESULTS: Our results demonstrated significant upregulation of IL-22 in human CC tumor infiltrated leukocytes (TILs) compared to peripheral lymphocytes. Moreover, our findings demonstrated that IL-22 expression was significantly higher in ulcerative colitis (UC) tissues versus normal colon tissues. Both IL-22 receptor α1 (IL-22RA1) and IL-23 were highly expressed in CC and UC tissues compared to normal controls. TILs exhibiting various IL-22 expression levels isolated from CC patients were demonstrated to enhance tumor growth and metastasis co-transplanted with Hct-116 cells underwent subcutaneous transplantation in mice model. Tumor growth and metastasis was promoted by STAT3 phosphorylation and upregulation of its downstream genes such as Bcl-xl, CyclinD1, and VEGF. In vitro studies confirmed the anti-apoptotic and pro-proliferation effect of IL-22 according to the BrdU cooperation assay and peroxide induced apoptosis analysis with or without the presence of IL-22. CONCLUSION: In this study we demonstrated that excessive IL-22 in the CC and UC microenvironment leads to tumor growth, inhibition of apoptosis, and promotion of metastasis depend on STAT3 activation.

Long-term efficacy of pneumatic dilation and esophageal stenting for the treatment of achalasia.

AIMS: To compare the efficacy and safety of pneumatic dilation with stenting for the treatment of achalasia. METHODS: Achalasia patients treated with pneumatic dilation or stenting were included in this analysis. Clinical symptoms were assessed by the Eckardt score. An esophagram and esophageal manometry were performed at the pretreatment and posttreatment follow-up visits. Data such as patient demographics and complications were collected. A drop in the Eckardt score to ≤3 was defined as treatment success. RESULTS: There were 151 patients treated with pneumatic dilation (n = 76) or stenting (n = 75). The 1-, 12- and 24-month therapeutic success rates were 100, 92 and 79%, respectively, in the dilation group and 100, 93 and 87%, respectively, in the stenting group. The decrease of Eckardt score in the stenting group was significantly notable (p < 0.05) compared to that of the dilation group at the long-term follow-up visits. The maximum esophageal diameter was comparable at baseline and became statistically significant [25 mm (22-30) vs. 22 mm (19-27), p = 0.004] at posttreatment month 24. The recurrence rate was 21% in the dilation group and 13% in the stenting group. The complications of either treatment were usually mild, transient and statistically insignificant. CONCLUSION: Esophageal stenting had a comparable short-term but better long-term efficacy in comparison with pneumatic dilation.

Endoscopic therapy in the treatment of caustic esophageal stricture: a retrospective case series study.

BACKGROUND: Endoscopic procedures, such as balloon/bougie dilation and stent implantation, have gained increasing potential as the treatment of corrosive esophageal stricture. The purpose of the present retrospective case series study was to assess clinical outcomes of endoscopic therapy of esophageal strictures after caustic injury. PATIENTS AND METHODS: Between January 2003 and December 2009, 13 admitted patients that developed esophageal stricture after caustic agent ingestion underwent endoscopic therapy at the First Affiliated Hospital of Nanjing Medical University. Data such as age, gender, caustic agents, site of burn, type of treatment, effectiveness and outcome of endoscopic therapy were recorded. RESULTS: The average follow up was 39 months with a range of 29-70 months. Successful endoscopic therapy was achieved in 12 of 13 patients (92%). Duration of stricture resolution was between 4 and 48 months (mean, 15 months). Among them, seven patients required dilations only, whereas the other five patients received both dilations and stent implantation. There were no severe complications in these patients. CONCLUSIONS: These data suggest that endoscopic therapy is feasible, less invasive and effective for the management of caustic esophageal stricture. After repeat dilation and stenting, patients can achieve stricture resolution in approximately 15 months and avoid surgery.

[Prevention of restenosis with a new modified fully-covered retrievable esophageal stent].

OBJECTIVE: To observe and evaluate the effectiveness of a new modified fully-covered retrievable esophageal stent in preventing restenosis at the proximal end of the stent. METHODS: From January 2008 to October 2011, 380 consecutive patients who underwent placement of a conventional stent or a new modified stent for benign or malignant dysphagia were divided into two groups: conventional stent group 193 patients (male 137, female 56) and modified stent group 187 patients (male 125, female 62). The granulation formation and restenosis rate one month after stenting were evaluated. Data such as patient demographics, outcomes and complications were collected. The results were statistically analyzed by Student t test, chi-squared test, Fisher's exact probability or rank sum test. A P-value less than 0.05 was considered statistically significant. RESULTS: All stents were successfully implanted. They were highly effective in palliating dysphagia. The dysphagia score decreased from 3 (1) to 0 (1) in conventional stent group (P < 0.01), and that from 4 (1) to 0 (1) in modified stent group (P < 0.01). The modified stent group were superior to the conventional stent group in severe granulation formation rate (0 vs 4.7% (9/193), P = 0.004) and restenosis rate (2.7% (5/187) vs 7.3% (14/193), P = 0.041) within one month after stenting, and the modified stent was easier to retrieve. Postoperative remission rate of dysphagia, and complications such as chest pain, bleeding, perforation, stent migration had no statistical differences between the two groups (all P > 0.05). CONCLUSIONS: The new modified fully-covered retrievable esophageal stent can significantly reduce granulation formation at the proximal end of the stent. Using of this stent seems to be a better choice in treating patient of dysphagia, with lower restenosis rate and easier to retrieve.

Metabolomic phenotype of gastric cancer and precancerous stages based on gas chromatography time-of-flight mass spectrometry.

BACKGROUND AND AIM: To study the low-molecular-weight metabolites in blood plasma of patients with the progressive disease, gastric cancer, and to characterize different stages from chronic superficial gastritis (CSG) to chronic atrophic gastritis (CAG), intestinal metaplasia (IM), gastric dysplasia (DYS) and finally gastric cancer (GC). METHODS: We applied gas chromatography time-of-flight mass spectrometry (GC/TOF-MS) to determine metabolites levels in plasma obtained from 80 patients including 19 with CSG, 13 with CAG, 10 with IM, 15 with DYS and 22 with GC (nine preoperation and 13 postoperation). Principal component analysis (PCA) and statistics were used to differentiate the stages and to identify the markers of gastric cancer. RESULTS: Totally, 223 peaks were detected in GC/TOF-MS and 72 compounds were authentically identified. CSG showed distinct difference from the other groups of CAG, IM, DYS and GC, whose plots clustered closely. IM clustered closely to GC, suggesting similar metabolic patterns of them. Fifteen identified metabolites contributed most to the differentiating between CSG and GC, and characterized different stages of GC. Statistics revealed elevated levels of 2-Hydroxybutyrate, pyroglutamate, glutamate, asparagine, azelaic acid, ornithine, urate, 11-eicosenoic acid, 1-monohexadecanoylglycerol and γ-tocopherol, while downregulation of creatinine, threonate in GC group, indicating that GC patients were obviously involved in oxidative stress, and perturbed metabolism of amino acids and fatty acids. CONCLUSION: The metabolic phenotype of CSG is significantly different from GC, while that of IM is similar to it. The discriminatory metabolites characterizing progressive stages from CSG to GC might be the potential markers to indicate a risk of GC.

[Effects of tumor necrosis factor-alpha inducing protein-alpha secreted by Helicobacter pylori on human gastric epithelial cells].

OBJECTIVE: To investigate the effects of tumor necrosis factor-alpha-inducing protein-alpha (Tipalpha) released from Helicobacter pylori (Hp) on human gastric epithelial cells. METHODS: Human gastric epithelial cells of the line GES-1 were cultured. Tipalpha gene located in Hp 0596 was extracted from the genome sequence of the Hp strain 26695 and its open reading frame were cloned into the eukaryotic expressing vector pcDNA3.1. The recombinant plasmid pcDNA3.1-Tipalpha thus obtained and the blank plasmid pcDNA3. 1 were transfected into the GES-1 cells, and the G418-resistent clones were screened. The GES-1 cells transfected with blank vector pcDNA3.1 and normal GES-1 cells were used as controls. RT-PCR and Western blotting were used to detect the expression of Tipalpha. The influences of Tipalpha protein on the cells proliferation, apoptosis and cell cycles, concentration of related cytokines such as TNF-alpha, IL-1beta, and IL-8, and the expression of Bcl-2 and P53 genes were respectively observed by MTT assay, flow cytometry, ELISA, and Western blotting. RESULTS: 1. A eukaryotic expression vector of Hp Tipalpha was successfully constructed. Steadily transfected strains were screened by G418. 2. MTT method showed that the growth curve of the GES-1 cells transfected with GpcDNA3.1-Tipalpha was higher, showing a faster growth. 3. Flow cytometry showed an increase in the proportion of the S-phase and a decrease in the G1-phase in the GpcDNA3.1-Tipalpha-transfected cells [(45.33 +/- 1.03)% vs (38.24 +/- 1.5)%, (33.94 +/- 1.67)%, (41.39 +/- 0.08)% vs (49.74 +/- 0.12)%, (49.27 +/- 0.15)%], and the apoptotic rate of the GpcDNA3.1-Tipalpha-transfected cells was 0.76 +/- 0.04, significantly lower than those of the GpcDNA3.1-transfected cells and non-transfected cells (16.84 +/- 2.16 and 8.36 +/- 1.07 respectively, both P < 0.05). 4. ELISA showed that there was no significant difference in intracellular TNF-alpha concentration among the 3 GES-1 cell groups (all P > 0.05), however, the extra-cellular TNF-alpha level of the GpcDNA3.1-Tipalpha-transfected cells was significantly higher than those of the 2 control groups (both P < 0.05), and that the intra- and extra-cellular IL-1beta and IL-8 concentrations of the GpcDNA3.1-Tipalpha-transfected cells were both significantly higher than those of the 2 control groups (all P < 0.05). 5. The expression level of Bcl-2 gene of the GpcDNA3.1-Tipalphac-transfected cells was obviously higher than those of the 2 control groups, and the expression of P53 gene of the GpcDNA3.1-Tipalphac-transfected cells was lower than those of the 2 control cells. CONCLUSION: The GpcDNA3.1-Tipalpha-transfected cells steadily and highly express Tipalpha protein, that induces the high expression of TNF-alpha, IL-1beta, and IL-8, proinflammatory cytokines, enhances cell proliferation, and upregulates Bcl-2 gene and down-regulates P53 gene. Tipalpha may play an important carcinogenic role in gastric cancer progression.

Metabolic Perturbation and Potential Markers in Patients with Esophageal Cancer.

Clinical diagnosis of esophageal cancer (EC) at early stage is rather difficult. This study aimed to profile the molecules in serum and tissue and identify potential biomarkers in patients with EC. A total of 64 volunteers were recruited, and 83 samples (24 EC serum samples, 21 serum controls, 19 paired EC tissues, and corresponding tumor-adjacent tissues) were analyzed. The gas chromatography time-of-flight mass spectrometry (GC/TOF-MS) was employed, and principal component analysis was used to reveal the discriminatory metabolites and identify the candidate markers of EC. A total of 41 in serum and 36 identified compounds in tissues were relevant to the malignant prognosis. A marked metabolic reprogramming of EC was observed, including enhanced anaerobic glycolysis and glutaminolysis, inhibited tricarboxylic acid (TCA) cycle, and altered lipid metabolism and amino acid turnover. Based on the potential markers of glucose, glutamic acid, lactic acid, and cholesterol, the receiver operating characteristic (ROC) curves indicated good diagnosis and prognosis of EC. EC patients showed distinct reprogrammed metabolism involved in glycolysis, TCA cycle, glutaminolysis, and fatty acid metabolism. The pivotal molecules in the metabolic pathways were suggested as the potential markers to facilitate the early diagnosis of human EC.

Expression of COX-2 proteins in gastric mucosal lesions.

AIM: To investigate the expression of COX-2 proteins in gastric mucosal lesions and to assess the relationship between COX-2 expression and type, pathologic stage, differentiation, or lymph node metastasis in gastric cancer and the relationship between COX-2 expression and H pylori infection in gastric mucosal lesions. METHODS: Thirty patients with gastric carcinoma underwent surgical resection. Samples were taken from tumor site and paracancerous tissues, and ABC immunohistochemical staining was used to detect the expression of COX-2 proteins. H pylori was determined by rapid urea test combined with pathological stating/14C urea breath test. RESULTS: The positive rate and staining intensity of mutant COX-2 gene expression in gastric cancer were significantly higher than those in paracancerous tissues (66.7% vs 26.7%) (P<0.01, P<0.001). There was a significant correlation between COX-2 and pathologic stage or lymph node metastasis type of gastric carcinoma (76.0% vs 20.0%, 79.2% vs 16.7%) (P<0.05). No correlation was found between COX-2 expression and type or grade of differentiation (P>0.05). COX-2 expression of intestinal metaplasia (IM) or dysplasia (DYS) with positive H pylori was significantly higher than that with negative H pylori (50.6% vs 18.1%, 60.0% vs 33.3%) (P<0.05). CONCLUSION: COX-2 overexpression was found in a large proportion of gastric cancer tissues compared with matched non-cancerous tissues and was significantly associated with advanced tumor stage and lymph node metastasis. Overexpression of COX-2 plays an important role in tumor progression of gastric cancer. COX-2 may also play a role in the early development/promotion of gastric carcinoma and is associated with H pylori infection.

Endolumenal endoscopic full-thickness resection of muscularis propria-originating gastric submucosal tumors.

BACKGROUND: This study retrospectively reviewed 48 cases of gastric submucosal tumors (SMTs) treated by endolumenal endoscopic full-thickness resection (EFR) microsurgery in our gastrointestinal endoscopy center. PATIENTS AND METHODS: From November 2009 to October 2012, 48 cases underwent endolumenal EFR for resection of muscularis propria-originating gastric SMTs. Characteristics of the 48 patients, clinical efficacy, safety of EFR, and post-EFR pathological diagnoses were evaluated retrospectively. RESULTS: EFR was successfully performed in 48 cases with 52 lesions. The median operation time was 59.72 minutes (range, 30-270 minutes; standard deviation, 39.72 minutes). The mean tumor size was 1.59 cm (range, 0.50-4.80 cm; standard deviation, 1.01 cm). During the EFR process, dual-channel gastroscopy was applied in 20 cases of SMTs, and paracentesis during the EFR process was applied in 9 cases. EFR for larger SMTs and gastric corpus-originating SMTs had longer operative times. Pathological diagnosis included 43 gastrointestinal stromal tumors, 4 leiomyomas, and 1 schwannoma. A larger tumor size was associated with higher risk of malignancy. No severe postoperative complications were observed. No tumor recurrences were confirmed in follow-up gastroscopy. CONCLUSIONS: The endolumenal EFR technique proved to be feasible and minimally invasive, even for the resection of large gastric tumors originating from the muscularis propria. However, more data on EFR must be obtained and analyzed.

Expression of interleukin-22/STAT3 signaling pathway in ulcerative colitis and related carcinogenesis.

AIM: To investigate the expression of interleukin (IL)-22 and its related proteins in biopsy specimens from patients with ulcerative colitis (UC) and UC-related carcinogenesis. METHODS: Biopsy specimens were obtained from patients with inactive (n = 10), mild-to-moderately active (n = 30), severely active (n = 34), initial (n = 30), and chronic UC (n = 44), as well as UC patients with dysplasia (n = 10). Specimens from patients without colonic abnormalities (n = 20) served as controls. Chronic colitis in experimental mice was induced by 2.5% dextran sodium sulfate. The expression levels of IL-22, IL-23, IL-22R1 and phosphorylated STAT3 (p-STAT3) were determined by immunohistochemistry. Bcl-2, cyclin D1 and survivin expression was detected by Western blotting. RESULTS: Patients with active UC had significantly more IL-22, IL-23, IL-22R1 and p-STAT3-positive cells than the patients with inactive UC and normal controls. Furthermore, IL-22 and related proteins were closely related to the severity of the colitis. The expression of IL-22 and IL-22R1 in the tissue of initial UC was stronger than in that of chronic UC, whereas the expression of p-STAT3 was significantly increased in chronic UC tissues. In dysplasia tissues, the expression level of IL-22 and related proteins was higher compared with controls. Mouse colitis model showed that expression of IL-22, IL-22R1 and IL-23 was increased with time, p-STAT3 and the downstream gene were also remarkably upregulated. CONCLUSION: IL-22/STAT3 signaling pathway may be related to UC and UC-induced carcinogenesis and IL-22 can be used as a biomarker in judging the severity of UC.

Multiple genetic alterations and behavior of cellular biology in gastric cancer and other gastric mucosal lesions:H.pylori infection, histological types and staging.

AIM:To investigate the expression of multiple genes and the behavior of cellular biology in gastric cancer (GC) and other gastric mucosal lesions and their relations to Helicobacter pylori (H. pylori) infection, tumor staging and histological subtypes.METHODS:Three hundred and twenty seven specimens of gastric mucosa obtained via endoscopy or surgical resection, and ABC immunohistochemical staining were used to detect the expression of p53, p16, Bcl-2 and COX-2 proteins.H. pylori was determined by rapid urea test combined with patholo-gical staining or 14 Curea breath test. Cellular image analysis was performed in 66 patients with intestinal metaplasia (IM) and/or dysplasia (Dys). In 30 of them, both cancer and the paracancerous tissues were obtained at the time of surgery. Histolo-gical pattern, tumor staging, lymph node metastasis, grading of differentiation and other clinical data were studied in the medical records.RESULTS:p16 expression of IM or Dys was significantly lower in positive H. pylori chronic atrophic gastritis (CAG) than those with negative H. pylori (CAG: 54.8% vs 88.0%, IM:34.4% vs 69.6%, Dys: 23.8% vs 53.6%, all P < 0.05), Bcl-2 or COX-2 expression of IM or Dys in positive H. pylori cases was signi-ficantly higher than that without H. pylori (Bcl-2: 68.8% vs 23.9%, 90.5% vs 60.7%; COX-2: 50.0% vs 10.8%, 61.8% vs 17.8%; all P <0.05). The mean number of most parame-ters of cellular image analysis in positive H. pylori group was significantly higher than that in negative H. pylori group (Ellipser: 53 plus minus 14, 40 plus minus 12&mgr;m, Area(1): 748 plus minus 572, 302 plus minus 202&mgr;m(2), Area(2): 3050 plus minus 1661, 1681 plus minus 1990&mgr;m(2), all P< 0.05; Ellipseb: 79 plus minus 23, 58 plus minus 15&mgr;m, Ratio-1: 22% plus minus5%,13% plus minus4%,Ratio-2:79% plus minus17%,53% plus minus20%,all P<0.01). There was significant correl-ation between Bcl-2 and histologic pattern of gastric carcinoma, and between COX-2 and tumor staging or lymph node metasta sis (Bcl-2: 75.0% vs16.7%; COX-2: 76.0% vs 20.0%, 79.2% vs 16.7%; all P< 0.05).CONCLUSION:p16, Bcl-2, and COX-2 but not p53 gene may play a role in the early genesis/progression of gastric carcinoma and are associated with H. pylori infection. p53 gene is relatively late event in gastric tumorigenesis and mainly relates to its progression. There is more cellular-biological behavior of malignant tumor in gastric mucosal lesions with H. pylori infec-tion. Aberrant Bcl-2 protein expression appears to be preferentially associated with the intestinal type cancer. COX-2 seems to be related to tumor staging and lymph node metastasis.