Graphene oxide exacerbates dextran sodium sulfate-induced colitis via ROS/AMPK/p53 signaling to mediate apoptosis.

BACKGROUND: Graphene oxide (GO), a novel carbon-based nanomaterial, has promising applications in biomedicine. However, it induces potential cytotoxic effects on the gastrointestinal (GI) tract cells, and these effects have been largely uncharacterized. The present study aimed to explore the toxic effects of GO on the intestinal tract especially under pre-existing inflammatory conditions, such as inflammatory bowel disease (IBD), and elucidate underlying mechanisms. RESULTS: Our findings indicated that oral gavage of GO worsened acute colitis induced by 2.5% dextran sodium sulfate (DSS) in mice. In vitro, GO exacerbated DSS-induced inflammation and apoptosis in the FHC cell line, an ideal model of intestinal epithelial cells (IECs). Further, the potential mechanism underlying GO aggravated mice colitis and cell inflammation was explored. Our results revealed that GO treatment triggered apoptosis in FHC cells through the activation of reactive oxygen species (ROS)/AMP-activated protein kinase (AMPK)/p53 pathway, as evidenced by the upregulation of cytochrome c (Cytc), Bax, and cleaved caspase-3 (c-cas3) and the downregulation of Bcl-2. Interestingly, pretreatment with an antioxidant, N-acetyl-L-cysteine, and a specific inhibitor of AMPK activation, Compound C (Com.C), effectively inhibited GO-induced apoptosis in FHC cells. CONCLUSIONS: Our data demonstrate that GO-induced IECs apoptosis via ROS/AMPK/p53 pathway activation accounts for the exacerbation of colitis in vivo and aggravation of inflammation in vitro. These findings provide a new insight into the pathogenesis of IBD induced by environmental factors. Furthermore, our findings enhance our understanding of GO as a potential environmental toxin, which helps delineate the risk of exposure to patients with disturbed intestinal epithelial barrier/inflammatory disorders such as IBD.

Viral etiology of influenza-like illnesses in Huizhou, China, from 2011 to 2013.

Little information is available on the etiology and prevalence of viruses other than influenza viruses causing influenza-like illnesses (ILIs) in China. This study was conducted for simultaneous detection and identification of 14 respiratory viruses in Huizhou using real-time PCR. In total, viruses were detected in 48.66 % of ILI patient samples, in which influenza virus (19.98 %) was the most commonly detected, followed by rhinovirus (7.46 %), human coronaviruses (3.63 %), human metapneumovirus (3.06 %), parainfluenza virus (3.06 %), respiratory syncytial virus (2.39 %), adenovirus (2.29 %), and human bocavirus (1.43 %). Co-infections occurred in 5.35 % of all tested specimens and 11.00 % (56/509) of infected patients. Children under 5 years and adults older than 60 years were more likely to have one or more detectable viruses associated with their ILI (OR=1.75, 95 % CI: 1.37; 2.23). Influenza virus was detected during each month of each year, and increased viral activity was observed in 2013. Infections with adenovirus and human metapneumovirus had characteristic seasonal patterns. No significant differences were found in positive the rate between the gender groups, while significantly differences in positive rate were found among the different age groups (P-value<0.001). This study confirmed that multiple respiratory viruses may circulate concurrently in the population and play an important role in the etiology of ILI. The most frequent symptoms associated with respiratory viruses were sore throat, rhinorrhea and headache. This information needs to be considered by clinicians when treating patients presenting with ILI, and it could serve as a reference for government officers when designing and implementing effective intervention plans.

Lyn is involved in CD24-induced ERK1/2 activation in colorectal cancer.

BACKGROUND AND AIM: CD24 expression is associated with human colorectal cancer (CRC). Our previous data indicated that CD24 promoted the proliferation and invasion of colorectal cancer cells through the activation of ERK1/2. Since Src family kinases are frequently deregulated in CRC and closely related to the MAPK signaling pathway, we investigated the impact of Lyn, an important member of SFKs, on CD24-induced ERK1/2 activation in CRC. METHODS AND RESULTS: The interaction of CD24 and Lyn was identified by co-immunoprecipitation (Co-IP) and ectopic expression of CD24-induced Lyn activation. Inhibition of Lyn activation by phosphatase PP2 in SW480CD24cells abrogated CD24-induced invasion. The results of the Co-IP and immunofluorescence assay revealed that overexpression of CD24 enhanced the interaction of Lyn and ERK1/2 and induced the nuclear translocation of Lyn. However, inhibition of Lyn activity attenuated CD24-induced ERK1/2 activation, and depletion of CD24 disrupted Lyn-ERK1/2 interaction. Immunohistochemistry analysis for 202 cases of CRC showed that the expression of both CD24 and Lyn was positively correlated with tumor grade, stage, lymph node and distant metastasis. Patients with lower expression of CD24 or Lyn had a higher survival rate. The Cox multivariate analysis showed that CD24 expression, but not Lyn expression, was an independent prognostic factor of CRC. CONCLUSIONS: Our results suggest that Lyn is involved in CD24-induced ERK1/2 activation in CRC. The expression of CD24 is associated with activation of Lyn and ERK1/2, which might be a novel mechanism related to CD24-mediated regulation of CRC development.

Graphene oxide aggravated dextran sulfate sodium-induced colitis through intestinal epithelial cells autophagy dysfunction.

Graphene oxide (GO) is one of the most promising nanomaterials used in biomedicine. However, studies about its adverse effects on the intestine in state of inflammation remain limited. This study aimed to explore the underlying effects of GO on intestinal epithelial cells (IECs) in vitro and colitis in vivo. We found that GO could exert toxic effects on NCM460 cells in a dose- and time-dependent manner and promote inflammation. Furthermore, GO caused lysosomal dysfunction and then blockaded autophagy flux. Moreover, pharmacological autophagy inhibitor 3-Methyladenine could reverse GO-induced LC3B and p62 expression levels, reduce expression levels of IL-6, IL-8, TLR4, and CXCL2, and increase the level of IL-10. In vivo, C57BL/6 mice were treated with 2.5% dextran sulfate sodium (DSS) in drinking water for five consecutive days to induce colitis. Then, GO at 60 mg/kg dose was administered through the oral route every two days from day 2 to day 8. These results showed that GO aggravated DSS-induced colitis, characterized by shortening of the colon and severe pathological changes, and induced autophagy. In conclusion, GO caused the abnormal autophagy in IECs and exacerbated DSS-induced colitis in mice. Our research indicated that GO may contribute to the development of intestinal inflammation by inducing IECs autophagy dysfunction.

[Screening of high-risk group with colorectal cancer].

OBJECTIVE: To measure the rate of high-risk group and the detection rate of colorectal cancer (CRC) in communities in Guangdong province and to provide scientific rationales for formulating mass screening plans in high-risk group. METHODS: Mass survey was conducted by questionnaire combined fecal occult blood test (FOBT) in Huizhou region, Guangdong Province, to sort out the high-risk population of CRC. Then the high-risk population was screened by colonoscopy and pathology to identify CRC. The differences were compared by direct expenditure which was used to calculate screening cost. RESULTS: A total of 68,953 people were surveyed. There were 940 people in high-risk group (detection rate: 1.36%), 3118 in immunity FOBT positive group (detection rate: 4.52%), Merging aforementioned two groups, there were 3870 in population at risk (detection rate: 5.61%). The CRC detection rate in high-risk group, immunity FOBT positive group, population at risk and average-risk group was 506.3/10(5), 314.3/10(5), 315.9/10(5) and 17.7/10(5) respectively. The positive predictive value of CRC screening scheme by high-risk group questionnaire-colonoscopy was 0.43% while CRC screening scheme by FOBT-colonoscopy 0.22%. In terms of direct expenditure of CRC per case in high-risk group and immunity FOBT positive group was 47,834.5 yuan and 82,303.6 yuan. The latter was 1.7 times than that of the former. CONCLUSIONS: The scheme of questionnaire combined FOBT for CRC is an effective way in mass survey. The scheme by high-risk group questionnaire-colonoscopy has a much better cost-effectiveness than that of the scheme by FOBT-colonoscopy so that it should be one of the preferred methods for individual screening in high-risk group.

[Epidemiology investigation of colorectal cancer on community group in Guangdong province].

OBJECTIVE: To explore the epidemiology characteristics of colorectal cancer on community group in Guangdong province. METHODS: Huidong was took as the research spot to investigate the sickness and incidence of colorectal cancer among a whole community group in Guangdong province, and two projects were used simultaneously: "screening of colorectal cancer in high risk group" and "screening project using fecal occult blood test". The numeration time for population of 10 towns was set at July 1, 2005. Five towns were sampled out from Huidong with 100,859 persons. Meanwhile, the diseases and death cause registering system was established. RESULTS: The incidence of colorectal cancer in Huidong community group was 15.2/100,000 (male 17.2/100,000, female 13.0/100,000), the age-adjusted incidence was 17.6/100,000 (male 19.5/100,000, female 16.7/100,000). The morbidity was 41.9/100,000 (male 46.5/100,000, female 37.2/100,000), the age-adjusted morbidity was 49.0/100,000 (male 54.1/100,000, female 45.6/100,000). The mortality was 5.0/100,000 (male 5.9/100,000, female 4.0/100,000) and the age-adjusted morbidity was 6.4/100,000 (male 7.2/100,000, female 5.7/100,000). CONCLUSION: The incidence of colorectal cancer in Huidong community group is lower than that of high incidence area in China, but gets close to the normal incidence area of China and general level of the world.

[Effect of Tiam1 overexpression on proliferation and metastatic potential of human colorectal cancer].

OBJECTIVE: To confirm the role of Tiam1 (T lymphoma invasion and metastasis 1) gene in the proliferation and metastasis of colorectal cancer. METHODS: Proliferative and metastatic abilities of Tiam1 transfectant were investigated by subcutaneous injection of cells and surgical orthotopic transplantation (SOI) in mice. RESULTS: The expression of Tiam1 led to a pronounced increase in HT29/Tiam1 cell growth starting from day 7, up to 2.5 fold increase of tumor volume at day 20 post injection. Tumors in the HT29/Tiam1 group receiving surgical orthotopic implantation were significantly heavier than those in HT29/mock group (t = -14.916, P < 0.01). In vivo metastasis assay by SOI showed that in HT29/Tiam1 group, 7/7 of mice developed peritoneal metastases and 4/7 had hepatic lesions. In addition, one of the seven HT29/Tiam1 group mice had tumors in lung, spleen and lymph nodes. In the HT29/mock group, only 2/7 of animals had peritoneal metastases and none produced detectable tumor in the liver. CONCLUSIONS: Tiam1 gene plays an important role in the proliferation, invasion and metastasis of colorectal cancer. It may serve as a useful clinical marker for tumor progression and metastasis of colorectal cancer.

[Clinical epidemiological characteristics of 3870 cases of colorectal cancers in Guangdong region].

OBJECTIVE: To explore the clinical characteristics and pattern of incidence of colorectal cancers in Guangdong region. METHODS: Analysis and summary were made for 3870 colorectal cancer patients pathologically confirmed in the Nanfang Hospital and Huizhou Municipal Central Hospital of Guangdong province. RESULTS: Median age of 3870 cases with colorectal cancer was 55.3. The high-risk age ranged from 41 to 70.204 cases among them were young patients (age < or = 30, 5.3%). With increase of age the number of cases with rectal cancers decreased gradually, while cancers occurring in the right hemi colon increased gradually. The ratio between male and female was 1.42:1. There were altogether 3958 colonic cancer lesions found in all the cases. Among them 3783 (97.8%) cases presented with a single lesson, 87 (2.2%) cases presented with multiple lesions. 2243 (56.7%) lesions located in the rectum, 717 (18.1%) in the left hemi colon, 998 (25.2%) in the right hemi colon. Histological types in all the lesions cases were grouped as follows: tubular adenocarcinoma 2943 (76.0%); papillary adenocarcinoma 256 (6.6%); mucinous carcinoma 425 (11.0%); and miscellaneous types 246 (6.4%). Colorectal cancers with poor differentiation occurring in the young were 38.2% while in the middle age and the elderly were 29.9% and 14.6%, respectively. The difference between two groups showed a statistical significance (P < 0.01). The cases with confirmed stage A, B, C and D were 234 (6.0%), 1936 (50.0%), 1310 (33.9%) and 390 (10.1%), respectively, according to Dukes' staging system. The cases with the progressing stages (B, C, D stages) were 3636 (94.0%) among all the cases. CONCLUSIONS: The number of patients with colorectal cancer admitted in hospital increased gradually in the recent 20 years, and showed a trend with the decrease percentage in rectal cancer and the gradual increasing in right hemi colon cancer with increase of patients age. Half of the colorectal cancer occurred in the rectum, the rest occurred in the left and right hemi colon. The three clinical epidemiological characteristics of colorectal cancer, which once existed in Chinese, has disappeared in Guangdong region.

[The risk factors and impact of gastroesophageal reflux disease on quality of life in general population].

OBJECTIVES: To assess the risk factors of gastroesophageal reflux disease (GERD) in the general population and its impact on health-related quality of life (HRQOL). METHODS: A face-to-face interview was carried out in Guangdong Province using a validated Chinese version Reflux Disease Questionnaire (RDQ) to assess the prevalence of GERD. Random clustered sampling of permanent inhabitants aged 18 to 90 years was carried out under stratification of urban and suburban areas. The impact of GERD on HRQOL was evaluated using the Chinese version of SF-36. The statistical analysis was performed with SPSS 10.0 programs. RESULTS: A total of 83 GERD patients were collected and 166 healthy subjects were selected as control. There was no difference in prevalence between male (2.6%) and female (2.4%). There was no significant association between age and prevalence of GERD symptoms. Divorced/widowed/separated subjects (OR 4.61) and subjects with severe working burden (OR 3.43) were significantly more likely to report GERD symptoms. As compared with the general population, subjects with GERD symptoms experienced considerable impairment in quality of life. CONCLUSIONS: Psychosocial factors may play important roles in the production of GERD symptoms. GERD has a negative impact on HRQOL.

[The trend of clinical characteristics of colorectal cancer during the past 20 years in Guangdong province].

OBJECTIVE: To study the trend of clinical characteristics of colorectal cancer during the past 20 years in Gaungdong province. METHODS: The clinical data of 3870 cases of colorectal cancers confirmed pathologically who were hospitalized to Nanfang Hospital and Huizhou Municipal Hospital, Guangdong province, 1985 - 2004 were divided into 5-year groups and analyzed. RESULTS: The hospitalization number of colorectal cancer in the 2000 - 20004 5-year period was 102%, higher than that of the 1985 - 1989 period with an annual increase of 5.1%. The median age of incidence 2000 - 2004 was 58.6 years, 8.4 years higher than that in 1985 - 1989. The male to female ratio of the 3870 patients was 1.42:1. The male to female ratio 2000 - 2004 was 1.35:1, lower than that 1985 - 1989 (1.50:1). The proportion of rectal cancer 2000 - 2004 was 49.7%, significantly lower than that in 1985 - 1989 (64.8%), while the proportion of right hemi-colon cancer 2000 - 2004 was 28.7%, significantly higher than that in 1985 - 1989 (18.0%). The proportion of moderately and well differentiated cancer 2000 - 2004 was 80.6%, higher than that in 1985 - 1989 (70.1%), and the proportion of poorly differentiated cancer 2000 - 2004 was 19.4%, lower than that in 1985 - 1989 (29.9%). The proportion of colorectal cancer at Dukes A stage 2000 - 2004 was 9.8%, higher than that in 1985 - 1989 (3.2%). CONCLUSION: In the past 20 years, the incidence of colorectal cancer has increased in Guangdong province with a n increase of median age of incidence, The male to female ratio has decreased, and the incidence of right hemi-colon cancer, the rates of higher differentiated cancer and Dukes A stage cancer have increased.

Diagnostic Performance of DNA Hypermethylation Markers in Peripheral Blood for the Detection of Colorectal Cancer: A Meta-Analysis and Systematic Review.

DNA hypermethylation in blood is becoming an attractive candidate marker for colorectal cancer (CRC) detection. To assess the diagnostic accuracy of blood hypermethylation markers for CRC in different clinical settings, we conducted a meta-analysis of published reports. Of 485 publications obtained in the initial literature search, 39 studies were included in the meta-analysis. Hypermethylation markers in peripheral blood showed a high degree of accuracy for the detection of CRC. The summary sensitivity was 0.62 [95% confidence interval (CI), 0.56-0.67] and specificity was 0.91 (95% CI, 0.89-0.93). Subgroup analysis showed significantly greater sensitivity for the methylated Septin 9 gene (SEPT9) subgroup (0.75; 95% CI, 0.67-0.81) than for the non-methylated SEPT9 subgroup (0.58; 95% CI, 0.52-0.64). Sensitivity and specificity were not affected significantly by target gene number, CRC staging, study region, or methylation analysis method. These findings show that hypermethylation markers in blood are highly sensitive and specific for CRC detection, with methylated SEPT9 being particularly robust. The diagnostic performance of hypermethylation markers, which have varied across different studies, can be improved by marker optimization. Future research should examine variation in diagnostic accuracy according to non-neoplastic factors.

[Tiam1 expression correlates with the biological behavior of human colorectal carcinoma cells].

OBJECTIVE: To investigate whether the Tiam1 gene expression enhances the invasive and metastatic capabilities of colorectal carcinoma cells. METHODS: Endogenous expression of Tiam1 in five colorectal carcinoma cell lines was investigated by RT-PCR. Tiam1/C1199HA cDNA was transfected into HT29, a colorectal carcinoma cell line without endogenous Tiam1 expression. RNA and protein expression of Tiam1 gene in the transfectants were detected by RT-PCR, immunohistochemistry and Western blot respectively. The biological behaviors of the transfectants were investigated by MTT and in-vitro invasion assays. RESULTS: Tiam1 gene was highly expressed in LoVo and SW620 cells. Low level expression was seen in HCT116 and SW480 and no expression was found in HT29. Transfection of Tiam1 significantly increased the proliferation of HT29 cells along with markedly enhanced in-vitro invasion and metastasis. CONCLUSIONS: Tiam1 gene plays an important role in the invasion and metastasis of colorectal carcinoma. It may be a useful marker for metastasis of colorectal carcinoma.

[Evaluation of the quality of life of patients with irritable bowel syndrome].

OBJECTIVE: To explore and assess the health-related quality of life (HRQOL) of irritable bowel syndrome (IBS) patients in the population. METHODS: Random clustered sampling involving permanent inhabitants aged 18 - 80 yr was carried out under stratification of urban and suburban areas in Guangdong Province. Altogether 231 IBS patients fulfilling the Rome II criteria and 636 Non-IBS as control were collected. The impact of IBS on HRQOL was evaluated using the Chinese version of SF-36. RESULTS: (1) There were no statistically significant differences between IBS and Non-IBS groups in aspects of sex, age educational level, and distribution according to areas (P > 0.05). (2) IBS patients reported significantly poorer HRQOL than controls (Non-IBS) on all SF-36 subscales (P < 0.05). The patients had poorer HRQOL than the Non-patients, but their differences weren't significant (P > 0.05). (3) The scores on all SF-36 subscales were highly associated with the frequency of abdominal pain in IBS patients (P < 0.05); They were also correlated to degree of effects of IBS symptoms on life reported by IBS patients (P < 0.05); The association between the scores and the symptom of fatigue which is the most extra-intestinal symptom in IBS was significant (P < 0.05); (4) Copying style was highly correlated to the eight SF-36 subscales; IBS still had a significant impact on patients after partialing out the effect of copying style. CONCLUSIONS: IBS symptoms had a negative impact on HRQOL and the SF-36 could be adopted to detect the differences between IBS group and Non-IBS group, which may be used as an outcome measure in future treatment studies. However, the development of IBS-specific measures of quality of life is necessary.

[A population-based epidemiologic study of irritable bowel syndrome in Guangdong province].

OBJECTIVE: To explore the prevalence of irritable bowel syndrome (IBS) and its risk factors in Guangdong province. METHODS: A questionnaire was used to screen IBS by face-to-face interviews according to Manning (modified including symptoms of constipation) and Rome II criteria. Random clustered sampling involving permanent inhabitants aged 18 - 80 years was carried out under stratification of urban and suburban areas. Potential risk factors were explored by comparing the frequencies among IBS group and non-IBS group using chi(2) and logistic analysis of multivariate adjusted for age and gender. RESULTS: A total of 4178 residents (male 1907, female 2271) were investigated. Mean age among the responders was (43 +/- 14) years. The response rate was 98%. The adjusted prevalence of IBS in Guangdong province is 5.67% according to the modified Rome II criteria, and is 11.50% according to Manning criteria. There is no difference between urban and suburban areas. The female was predominant in IBS, and the ratio of male to female was 1:1.25 (Rome II) and 1:1.34 (Manning). The age was poorly correlated with the prevalence. Events including history of analgesic use such as NSAID (OR = 3.83), history of food allergies (OR = 2.68), psychological distress (OR = 2.18), life events (OR = 1.89), history of dysentery (OR = 1.63) and negative coping style (OR = 1.28) are highly associated with IBS (P < 0.05). CONCLUSION: IBS is a common disorder in Guangdong Province which deserves greater care and further investigation.

Circulating microRNAs as a novel class of diagnostic biomarkers in gastrointestinal tumors detection: a meta-analysis based on 42 articles.

OBJECTIVE: MicroRNAs (miRNAs) have become the focus of most recent efforts in cancer research. However, there have been inconsistencies in the literature regarding the suitability of circulating miRNAs for early detection of gastrointestinal cancers. This study aims to assess the diagnostic performance of circulating miRNAs in detection of gastrointestinal cancer through a meta-analysis. METHODS: Eligible studies were selected by conducting a systematic literature search of public databases. The sensitivity and specificity were used to plot the summary receiver operator characteristic (SROC) curve and calculate the area under the SROC curve (AUC). The between-study heterogeneity was evaluated by Q test and I2 statistics. Subgroup analyses and meta-regression were further performed to explore the potential sources of heterogeneity. All analyses were performed using the STATA 12.0 software. RESULTS: A total of 107 studies from 42 articles were included for the meta-analysis according to the inclusion criteria. The overall analysis of all gastrointestinal cancers showed that circulating miRNAs have a relatively good diagnostic performance in gastrointestinal cancers, with a sensitivity of 0.75, a specificity of 0.81 and an AUC of 0.85. In addition, subgroup analyses based on different type of miRNA assay suggested that single-miRNA assay displayed a relatively low diagnostic performance with the AUC values of 0.84 for gastric cancer (GC) and 0.79 for colorectal cancer (CRC), while multiple-miRNAs assay significantly improved the diagnosing accuracy with AUC rising to 0.92 for GC and 0.89 for CRC. Another interesting finding was that plasma-based miRNA assay reach a higher accuracy compared with serum-based one for GC, while opposite conclusion was drawn for CRC. CONCLUSIONS: In conclusion, circulating miRNAs, particularly the combination of multiple miRNAs, may present as promising biomarkers for the diagnosis of gastrointestinal cancers. Further large-scale prospective studies are necessary to validate their potential applicability in human cancer diagnosis.

Validation of methylation-sensitive high-resolution melting (MS-HRM) for the detection of stool DNA methylation in colorectal neoplasms.

BACKGROUND: Methylation-sensitive high-resolution melting (MS-HRM) is a new technique for assaying DNA methylation, but its feasibility for assaying stool in patients with colorectal cancer (CRC) is unknown. METHODS: First, the MS-HRM and methylation-specific PCR (MSP) detection limits were tested. Second, the methylation statuses of SFRP2 and VIM were analyzed in stool samples by MS-HRM, and in matching tumor and normal colon tissues via bisulfite sequencing PCR (BSP). Third, a case-control study evaluated the diagnostic sensitivity and specificity of MS-HRM relative to results obtained with MSP and the fecal immunochemical test (FIT). Finally, the linearity and reproducibility of MS-HRM were assessed. RESULTS: The detection limits of MS-HRM and MSP were 1% and 5%, respectively. The diagnostic sensitivities of MS-HRM (87.3%, 55/63) in stool and BSP in matching tumor tissue (92.1%, 58/63) were highly consistent (κ=0.744). The MS-HRM assay detected 92.5% (37/40) methylation in CRCs, 94.4% (34/36) in advanced adenomas, and 8.8% (5/57) in normal controls. The results of MS-HRM analysis were stable and reliable and showed fairly good linearity for both SFRP2 (P<0.001, R(2)=0.957) and VIM (P<0.001, R(2)=0.954). CONCLUSIONS: MS-HRM shows potential for CRC screening.

Aberrant activation of Sonic hedgehog signaling in chronic cholecystitis and gallbladder carcinoma.

Sonic hedgehog (Shh) signaling has been extensively studied and is implicated in various inflammatory diseases and malignant tumors. We summarized the clinicopathological features and performed immunohistochemistry assays to examine expression of Shh signaling proteins in 10 normal mucosa, 32 gallbladder carcinoma (GBC), and 95 chronic cholecystitis (CC) specimens. The CC specimens were classified into three groups according to degree of inflammation. Compared with normal mucosa, CC, and GBC specimens exhibited increased expression of Shh. The immunoreactive score of Shh in the GBC group was higher than that in the mild to moderate CC groups but lower than that in the severe CC group (P < .05). Expression of Patched (Ptch) and Gli1 gradually increased from non-malignant cholecystitis to malignant tumors. Compared with CC specimens, GBC specimens showed higher cytoplasmic and membranous expression for Ptch (P < .05). Gli1 staining showed cytoplasmic expression of Gli1 in both CC (60% for mild, 77% for moderate, and 84% for severe) and GBC specimens (97%). Nuclear expression of Gli1 was detected in 16% of severe CC specimens with moderate to poor atypical hyperplasia, and in 62.5% of GBC specimens. Shh expression strongly correlated with expression of Ptch and Gli1. Furthermore, patients with strongly positive Gli1 staining had significantly lower survival rates than those with weakly positive staining. Our data indicate that the Shh signaling pathway is aberrantly activated in CC and GBC, and altered Shh signaling may be involved in the course of development from CC to gallbladder carcinogenesis.

microRNA-16 represses colorectal cancer cell growth in vitro by regulating the p53/survivin signaling pathway.

Dysregulated expression of microRNAs (miRNA) is a hallmark of cancer. miR-16 has been reported to be downregulated and to act as a tumor suppressor in different cancer types. In the present study, we sought to investigate the possible roles and mechanisms of miR-16 and its relationship with p53 and survivin in CRC cells. We showed that miR-16 was downregulated in 67% of CRC tissues and was correlated with the degree of histological differentiation. Experiments in vitro showed that overexpression of miR-16 inhibited the proliferation and induced apoptosis of CRC cells through the intrinsic apoptosis pathway. We further showed that miR-16 repressed survivin expression at both the mRNA and protein levels and the survivin gene was a direct target of miR-16. In addition, miR-16 reduced p53 expression and p53 increased miR-16 levels, with downregulation of miR-16 targets survivin, cyclin D1 and CDK6. Our findings suggest that miR-16 represses colorectal cancer cell growth in vitro by regulating the p53/survivin signaling pathway. Our findings provide further evidence for the involvement of dysregulated miRNAs in CRC, and miR-16 could serve as a molecular target for CRC therapy.

Diagnostic value of stool DNA testing for multiple markers of colorectal cancer and advanced adenoma: a meta-analysis.

BACKGROUND AND OBJECTIVES: The diagnostic value of stool DNA (sDNA) testing for colorectal neoplasms remains controversial. To compensate for the lack of large-scale unbiased population studies, a meta-analysis was performed to evaluate the diagnostic value of sDNA testing for multiple markers of colorectal cancer (CRC) and advanced adenoma. METHODS: The PubMed, Science Direct, Biosis Review, Cochrane Library and Embase databases were systematically searched in January 2012 without time restriction. Meta-analysis was performed using a random-effects model using sensitivity, specificity, diagnostic OR (DOR), summary ROC curves, area under the curve (AUC), and 95% CIs as effect measures. Heterogeneity was measured using the χ(2) test and Q statistic; subgroup analysis was also conducted. RESULTS: A total of 20 studies comprising 5876 individuals were eligible. There was no heterogeneity for CRC, but adenoma and advanced adenoma harboured considerable heterogeneity influenced by risk classification and various detection markers. Stratification analysis according to risk classification showed that multiple markers had a high DOR for the high-risk subgroups of both CRC (sensitivity 0.759 [95% CI 0.711 to 0.804]; specificity 0.883 [95% CI 0.846 to 0.913]; AUC 0.906) and advanced adenoma (sensitivity 0.683 [95% CI 0.584 to 0.771]; specificity 0.918 [95% CI 0.866 to 0.954]; AUC 0.946) but not for the average-risk subgroups of either. In the methylation subgroup, sDNA testing had significantly higher DOR for CRC (sensitivity 0.753 [95% CI 0.685 to 0.812]; specificity 0.913 [95% CI 0.860 to 0.950]; AUC 0.918) and advanced adenoma (sensitivity 0.623 [95% CI 0.527 to 0.712]; specificity 0.926 [95% CI 0.882 to 0.958]; AUC 0.910) compared with the mutation subgroup. There was no significant heterogeneity among studies for subgroup analysis. CONCLUSION: sDNA testing for multiple markers had strong diagnostic significance for CRC and advanced adenoma in high-risk subjects. Methylation makers had more diagnostic value than mutation markers.