The Expression of MCM7 is a Useful Biomarker in the Early Diagnostic of Gastric Cancer.

The aim of this study was to investigate the expression of minichromosome maintenance complex component 7 (MCM7) in gastric mucosal lesions, further to find its potential effect as a biomarker to distinguish intraepithelial neoplasia from gastric mucosal lesions. MCM7 and Ki67 were detected in 93 cases of gastric mucosal lesions by immunohistochemistry. MCM7 and Ki67 expression in GT were lowest compared with other groups (P<0.001), meanwhile there were significant differences compared with Group IM and other groups in MCM7 and Ki67 expression (P<0.001). MCM7 and Ki67 expression in GSC were highest (P<0.05). Groups of LGN, HGN and GIC had no significant differences in MCM7 expression (P>0.05), but there was significant difference compared with Group LGN and Group GIC in Ki67 expression (P<0.05). MCM7 expression elevated with tumor grade increasing and had positive correlation with Ki67 significantly (r=0.940, P<0.001). Furthermore, in some cases, some tumor cells were immunoreactive to MCM7 but negative to Ki67. So we concluded that MCM7 is helpful for us to make differential diagnosis in pathological grade, MCM7 combination of Ki67 may serve as more sensitive proliferation markers for evaluation of gastric carcinoma and precancerous lesions.

Correlation of endoscopic images and histological findings of a high grade dysplasia developed in a gastric xanthoma.

Gastric neoplasia developed in a xanthoma is very rare. We herein report a high grade dysplasia (HGD) arising in a gastric xanthoma removed by endoscopic submucosal dissection (ESD). A 57-year-old man was referred to our hospital for removal of rectal polyps. During surveillance esophago-gastro-duodenoendoscopy before polypectomy, an irregularly shaped gastric xanthoma with unusual color was found in the stomach. Although, magnifying narrow band imaging showed no typical neoplastic vessel or surface pattern on the surface and endoscopic biopsies revealed no tumor, diagnostic ESD was performed because of its irregular shape and unusual color for a commonly seen xanthoma. Histologically, a high grade dysplasia, 6 mm×6 mm in size, was detected within a gastric xanthoma. This is the first report of correlation of endoscopic images and histological findings of a HGD in a gastric xanthoma.

CEACAM1 is overexpressed in oral tumors and related to tumorigenesis.

Carcinoembryonic antigen-related adhesion molecule 1 (CEACAM1) is a type 1 transmembrane glycoprotein belonging to the CEA family, which has been known to exist as either soluble forms in body fluids or membrane-bound forms on the cell surface. Aberrant CEACAM1 expression is associated with tumorigenesis and has been reported in a variety of human tumors, especially malignancies. The aim of this study is to determine the expression of CEACAM1 in oral tumors, trying to study CEACAM1 different expressions as a function of histotype. CEACAM1 expression was observed by immunohistochemistry (IHC) with mouse anti-human antibody for CEACAM1. IHC was performed using avidin-biotin-diaminobenzidine staining. The results were expressed as average score ± SD (0 = negative/8 = highest) for each histotype. Oral tumors expressed more CEACAM1 than normal tissues including squamous and salivary epithelia (P < 0.05). In malignancies, the squamous cell carcinoma overexpressed CEACAM1, compared to well-differentiated squamous cell with more membranous expression; the intermediately and poorly differentiated squamous cell carcinoma showed more cytoplasmic expression (P < 0.05). In addition, the salivary tumors significantly expressed more CEACAM1 than squamous cell carcinoma (P < 0.05). So, we thought oral tumors overexpressed CEACAM1 and the cytoplasmic CEACAM1 might be involved in tumorigenesis, and also CEACAM1 might be regarded as a marker of salivary glandular tumors.

Carcinoembryonic antigen-related cell adhesion molecule 1 is expressed and as a function histotype in ovarian tumors.

Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is a cell-cell adhesion receptor and is implicated in several cellular functions. It is rarely reported in ovarian tumors. The aim of this study is to determine the expression of CEACAM1 in ovarian tumors, trying to see whether CEACAM1 has different expression patterns as a function of histotype. Antigen expression was examined by immunohistochemistry with mouse anti-human antibody for CEACAM1. Immunohistochemistry was performed using avidin-biotin-diaminobenzide staining. The results were expressed as average score ± SD (0, negative; 8, highest) for each histotype. In ovarian tumors, the benign serous and mucinous cystadenoma negatively or weakly expressed CEACAM1, the malignant epithelial tumors strongly expressed CEACAM1, and there was significant difference between benign epithelial tumor and adenocarcinoma (P < .05). The well-differentiated serous adenocarcinoma expressed CEACAM1 mainly with membrane pattern, and the intermediately and poorly differentiated serous adenocarcinomas expressed CEACAM1 mainly with cytoplasmic pattern (P < .05). In addition, CEACAM1 expression is elevated in solid tumors of ovary but variable as a function of histotype. Compared with membranous expression, the cytoplasmic expression was observed almost in metastatic carcinoma that might decrease the adhesive interactions of the carcinoma cells with the surrounding cells, especially with tumor cells, and this could facilitate the tumor cells to metastasize to distant regions. So, we thought that cytoplasmic CEACAM1 might play an important role in tumor progression, especially in tumor metastasis.

DACH1 Expresison in Osteosarcoma and Its Relationship with Proliferation and Angiogenesis.

The aim of this study was to investigate the expression of DACH1 in osteosarcoma as well as its relationship with cell proliferation and angiogenesis in the tumor. DACH1 expression was detected by immunohistochemical staining in the serial sections of the osteosarcoma. The microvessel density (MVD) was counted by CD34 immunohistochemical staining, and immunohistochemical staining of PCNA staining showed the cell proliferation. The impacts of DACH1 expression on tumor proliferation and angiogenesis were evaluated by statistics. The DACH1 had different expression patterns in different osteosarcoma. Conventional osteosarcoma showed stronger DACH1 staining (conventional vs. parosteal: P = 0.037; conventional vs. periosteal: P = 0.028) and more PCNA-positive tumor cells than parosteal and periosteal osteosarcoma (conventional vs. parosteal: P = 0.041; conventional vs. periosteal: P = 0.045), the difference was significant. In addition, conventional osteosarcoma showed more cytoplasmic staining of DACH1 than parosteal and periosteal (conventional vs. parosteal: P = 0.023; conventional vs. periosteal: P = 0.030). Parosteal and periosteal osteosarcoma showed no significant difference in DACH1 expression and cell proliferation index. On the other hand, DACH1 different expression patterns showed significantly different impacts on angiogenesis. In spite of the different subtypes of osteosarcoma, the MVD showed a significant difference in cytoplasmic and nuclear expression patterns of DACH1 (nuclear expression vs. cytoplasmic expression: 5.72 ± 1.19 vs. 9.65 ± 1.24, P = 0.042). Moreover, in the conventional osteosarcoma, the MVD also showed a significant difference in DACH1 cytoplasmic and nuclear staining (nuclear expression vs. cytoplasmic expression: 5.58 ± 0.71 vs. 13.65 ± 1.30, P = 0.019). However, the DACH1 expression intensity showed no significant different impacts on MVD of all kinds of osteosarcoma. DACH1 had different expression patterns and intensity. Cytoplasmic and nuclear expression of DACH1 might play different roles in cell proliferation and angiogenesis of osteosarcoma. Cytoplasmic DACH1 might promote cell proliferation and be associated with angiogenesis.

Regulation Roles of MICA and NKG2D in Human Renal Cancer Cells.

OBJECTIVE: Our aim was to investigation the roles of MHC class I chain-related gene A(MICA) and natural killer cell group 2D(NKG2D) in human renal cancer cells. MATERIALS AND METHODS: The expression of membrane MICA (mMICA) on renal cells and NKG2D on NK cells were detected by flow cytometry (FCM); the content of sMICA were detected by enzyme linked immunosorbent assay (ELISA) and the distribution of mMICA on renal tumor tissues by immunohistochemistry; the interaction between MICA and NKG2D was observed by antibody closed method. RESULTS: Our results showed that the expression of mMICA in renal cancer tissues was significantly higher than in controls, where the soluble MICA was not expressed. Cytotoxic activity of NK cells was significantly reduced after exposure to NKG2D and MICA antibodies (P<0.05), and serum containing sMICA can obviously lower the function of NKG2D (P<0.05). CONCLUSIONS: The interaction of mMICA and NKG2D play important roles in mediation of cytotoxicity of NK cells in RCC. On the other hand, sMICA may mediate tumor immune escape through down- regulated NKG2D expression.

Expression of growth arrest and DNA damage inducible 45a in human oral squamous cell carcinoma is associated with tumor progression and clinical outcome.

OBJECTIVES: The aim of this study was to examine the growth arrest and DNA damage-inducible (Gadd45a) expression and its role in tumor progression, invasion and metastasis in oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: Growth arrest and DNA damage-inducible 45a distribution was detected by immunohistochemistry in tumor sections of 106 patients with primary OSCC and sections of adjacent pericancerous tissues from 60 patients among the 106. The association between the Gadd45a expression and clinical prognosis of OSCC were performed by statistical analysis. Gadd45a gene knockdown was performed in Tca8113 cells by small interfering ribonucleic acid treatment and its effects on cell cycle, and migration were detected by Flow Cytometric (Becton Dickinson, USA) and transwell chamber assay respectively. RESULTS AND CONCLUSION: The results showed that Gadd45a was redistributed to cytoplasm in poorly differentiated carcinoma from its nucleus location in normal tissue (P < 0.05). The expression of Gadd45a was significantly associated with lymph node metastasis, tumor stage and tumor histological grade (P < 0.05). Knockdown of Gadd45a gene abolished the G2/M arrest and increased migrating ability of Tca8113 cell (P < 0.05). The results indicate that Gadd45a play an important role in OSCC metastasis by affecting the bioactivity of the tumor cells, and its distribution may serve for the prediction of clinical outcome of OSCC.

Absence of gamma-interferon-inducible lysosomal thiol reductase (GILT) is associated with poor disease-free survival in breast cancer patients.

Tumor immunosurveillance is known to be of critical importance in controlling tumorigenesis and progression in various cancers. The role of gamma-interferon-inducible lysosomal thiol reductase (GILT) in tumor immunosurveillance has recently been studied in several malignant diseases, but its role in breast cancer remains to be elucidated. In the present study, we found GILT as a significant different expressed gene by cDNA microarray analysis. To further determine the role of GILT in breast cancer, we examined GILT expression in breast cancers as well as noncancerous breast tissues by immunohistochemistry and real-time PCR, and assessed its association with clinicopathologic characteristics and patient outcome. The absence of GILT expression increased significantly from 2.02% (2/99) in noncancerous breast tissues to 15.6% (34/218) in breast cancer tissues (P<0.001). In accordance with its proliferation inhibiting function, GILT expression was inversely correlated with Ki67 index (P<0.05). In addition, absence of GILT was positively correlated with adverse characteristics of breast cancers, such as histological type, tumor size, lymph nodes status, and pTNM stage (P<0.05). Consistently, breast cancers with reduced GILT expression had poorer disease-free survival (P<0.005). Moreover, significantly decreased expression of GILT was found in both primary and metastatic breast cancer cells, in contrast to normal epithelial cells. These findings indicate that GILT may act as a tumor suppressor in breast cancer, in line with its previously suggested role in anti-tumor immunity. Thus, GILT has the potential to be a novel independent prognostic factor in breast cancer and further studies are needed to illustrate the underlying mechanism of this relationship.

Learning curve and interobserver agreement of confocal laser endomicroscopy for detecting precancerous or early-stage esophageal squamous cancer.

BACKGROUND: Confocal laser endomicroscopy (CLE) can provide in vivo subcellular resolution images of esophageal lesions. However, the learning curve in interpreting CLE images of precancerous or early-stage esophageal squamous cancer is unknown. The goal of this study is to evaluate the diagnostic accuracy and inter-observer agreement for differentiating esophageal lesions in CLE images among experienced and inexperienced observers and to assess the learning curve. METHOD: After a short training, 8 experienced and 14 inexperienced endoscopists evaluated in sequence 4 sets of high-quality CLE images. Their diagnoses were corrected and discussed after each set. For each image, the diagnostic results, confidence in diagnosis, quality and time to evaluate were recorded. RESULTS: Overall, diagnostic accuracy was greater for the second, third, fourth set of images as compared with the initial set (odds ratio [OR] 2.01, 95% CI 1.22-3.31; 7.95, 3.74-16.87; and 6.45, 3.14-13.27), respectively, with no difference between the third and fourth sets in accuracy (p = 0.67). Previous experience affected the diagnostic accuracy only in the first set of images (OR 3.70, 1.87-7.29, p<0.001). Inter-observer agreement was higher for experienced than inexperienced endoscopists (0.732 vs. 0.666, p<0.01). CONCLUSION: CLE is a promising technology that can be quickly learned after a short training period; previous experience is associated with diagnostic accuracy only at the initial stage of learning.

High expression of MICA in human kidney cancer tissue and renal cell carcinoma lines.

The overall incidence and mortality of renal cell carcinoma (RCC), the most common kidney cancer, are steadily increasing for reasons that are not fully explained. Our aim was to explore the expression of membrane MHC class I chain-related gene A (mMICA) in human RCC cell lines and tissue specimens, and to determine expression of soluble MICA (sMICA) in serum of patients with renal cell carcinoma, we used flow cytometry (FCM) and immunohistochemistry as well as an enzyme linked immunosorbent assay (ELISA) . The results showed that percentage of mMICA expression was significantly increased in human kidney cancer tissues and RCC cell lines (786-O and Ketr-3) than that in healthy adults and human embryonic kidney 293 (HEK293) cell line individuality (P<0.05). sMICA content in healthy adults was negative, but in renal cancer patients was significantly elevated (P<0.05). Our research showed that high expression of MICA in human kidney cancer, this results show that MICA might serve as potential tumor-associated antigen (TAA) in RCC.

Confocal laser endomicroscopy for in vivo detection of gastric intestinal metaplasia: a randomized controlled trial.

BACKGROUND AND STUDY AIMS: Gastric intestinal metaplasia (GIM) is associated with a risk for development of intestinal-type gastric cancer. This study aimed to compare the diagnostic yield of GIM from confocal laser endomicroscopy (CLE) and white light endoscopy (WLE). PATIENTS AND METHODS: In a prospective, double-blind, randomized study, patients were randomly assigned to receive either CLE with targeted biopsies (group A) or WLE with a standard biopsy protocol (group B). RESULTS: A total of 168 patients were finally analyzed (group A 85, group B 83). On a per-patient analysis, the diagnostic yields of GIM (including GIM with gastric intraepithelial neoplasia [GIN]) for groups A and B were 44.71 % and 31.33 %, respectively (P = 0.074). On a per-biopsy analysis, CLE-targeted biopsy gave a significantly higher diagnostic yield of GIM compared with WLE and standard biopsy, at 65.70 % (113 /172 biopsies) versus 15.73 % (81 /515 biopsies) (P < 0.001). Moreover, the diagnostic yield of the operative link on gastric intestinal metaplasia (OLGIM) assessment stages III and IV was higher at 20.93 % (36 /172 biopsies) in group A versus 4.08 % (21 /515 biopsies) in group B (P < 0.001). In addition, use of CLE-guided biopsy significantly decreased by 68 % (P < 0.001) the mean number of biopsies required per patient. CONCLUSIONS: CLE with targeted biopsies is superior to WLE with standard biopsies for the detection and surveillance of GIM. The number of biopsies needed to confirm GIM is about one third of that needed with WLE with standard biopsies.

Expression and significance of Musashi-1 in gastric cancer and precancerous lesions.

AIM: To investigate expression of stem cell marker Musashi-1 (Msi-1) in relationship to tumorigenesis and progression of intestinal-type gastric cancer (GC). METHODS: Endoscopic biopsy specimens and surgical specimens were obtained, including 54 cases of intestinal-type GC, 41 high-grade intraepithelial neoplasia, 57 low-grade intraepithelial neoplasia, 31 intestinal metaplasia, and 36 normal gastric mucosa. Specimens were fixed in 10% paraformaldehyde, conventionally dehydrated, embedded in paraffin, and sliced in 4-µm-thick serial sections. Two-step immunohistochemical staining was used to detect Msi-1 and proliferating cell nuclear antigen (PCNA) expression. Correlation analysis was conducted between Msi-1 and PCNA expression. The relationship between Msi-1 expression and clinicopathological parameters of GC was analyzed statistically. RESULTS: There were significant differences in Msi-1 and PCNA expression in different pathological tissues (χ² = 15.37, P < 0.01; χ² = 115.36, P < 0.01). Msi-1 and PCNA-positive cells were restricted to the isthmus of normal gastric glands. Expression levels of Msi-1 and PCNA in intestinal metaplasia were significantly higher than in normal mucosa (U = 392.0, P < 0.05; U = 40.50, P < 0.01), whereas there was no significant difference compared to low or high-grade intraepithelial neoplasia. Msi-1 and PCNA expression in intestinal-type GC was higher than in high-grade intraepithelial neoplasia (U = 798.0, P < 0.05; U = 688.0, P < 0.01). There was a significantly positive correlation between Msi-1 and PCNA expression (r(s) = 0.20, P < 0.01). Msi-1 expression in GC tissues was correlated with their lymph node metastasis and tumor node metastasis stage (χ² = 12.62, P < 0.01; χ² = 11.24, P < 0.05), but not with depth of invasion and the presence of distant metastasis. CONCLUSION: Msi-1-positive cells may play a key role in the early events of gastric carcinogenesis and may be involved in invasion and metastasis of GC.

Expression and significance of IGF-2, PCNA, MMP-7, and α-actin in gastric carcinoma with Lauren classification.

BACKGROUND/AIMS: By detecting the expression and distribution of insulin-like growth factor-2, proliferating cell nuclear antigen, and matrix metalloproteinase-7 in cancer cells and the expression of α-actin in interstitial myofibroblasts, we studied their differences and their relationship in intestinal type and diffuse type gastric cancer with Lauren classification. MATERIALS AND METHODS: Clinical and pathological data of 50 patients with gastric adenocarcinoma who underwent primary surgical resection between 2003 and 2008 in Qianfoshan Hospital were collected. The cancer was classified as intestinal or diffuse type with Lauren classification. Immunohistochemical technique was used to detect the protein expression of insulin-like growth factor-2, proliferating cell nuclear antigen, matrix metalloproteinase-7, and α-actin in both gastric cancer tissue and normal gastric mucosa. RESULTS: The expression of insulin-like growth factor-2, proliferating cell nuclear antigen, matrix metalloproteinase-7, and α-actin in the gastric tissue was significantly higher than in the normal gastric mucosa. Insulin-like growth factor-2 was mainly expressed in the nucleus in diffuse gastric cancer and in the cytoplasm in intestinal type. The expression of insulin-like growth factor-2 in the nucleus was correlated with depth of tumor invasion, lymph node metastasis and staging. Proliferating cell nuclear antigen and matrix metalloproteinase-7 showed higher expression in diffuse than in intestinal type gastric cancer. The overexpression of proliferating cell nuclear antigen was relevant to lymph node metastasis in gastric cancer. The overexpression of matrix metalloproteinase-7 was relevant to invasion, lymph node metastasis, distant metastasis, and staging in gastric cancer. There was no significant difference in α-actin expression between the intestinal type and diffuse type gastric cancer. The overexpression of α-actin was relevant to cancer invasion and lymph node metastasis. CONCLUSIONS: The expression patterns of insulin-like growth factor-2 and the expression intensity of proliferating cell nuclear antigen and matrix metalloproteinase-7 were significantly different between diffuse type and intestinal type gastric cancer cells, but the expression pattern and intensity of the interstitial myofibroblast marker showed no significant difference. The clinical pathology distinction between intestinal type and diffuse type gastric cancer may be due mainly to the change in the genetic structure and the phenotype of epithelial cells, and interstitial myofibroblasts and cancer cells jointly promote the invasion and metastasis of gastric cancer.

Magnified and enhanced computed virtual chromoendoscopy in gastric neoplasia: a feasibility study.

AIM: To evaluate the feasibility of a new computed virtual chromoendoscopy (CVC) device (M i-scan) in the diagnosis of gastric neoplasia. METHODS: Patients with superficial lesions no larger than 1.0 cm found during high definition endoscopy were included. Those with advanced or obviously protruded or depressed lesions, lesions larger than 1.0 cm and/or lesions which were not amenable to observation by zoom function were excluded. The endoscopist was required to give the real-time descriptions of surface pit patterns of the lesions, based on surface pattern classification of enhanced magnification endoscopy. According to previous reports, types I-III represent non-neoplastic lesions, and types IV-V represent neoplastic lesions. Diagnosis with M i-scan and biopsy was performed before histopathological diagnosis. Magnified images of gastric lesions with and without enhancement were collected for further analysis. The diagnostic yield of real-time M i-scan and effects on magnification image quality by tone enhancement (TE), surface enhancement (SE) and color enhancement (CE) were calculated. The selected images were sent to another endoscopist. The endoscopist rated the image quality of each lesion at 3 levels. Ratings of image quality were based on visualization of pit pattern, vessel and demarcation line. RESULTS: One hundred and eighty-three patients were recruited. Five patients were excluded for advanced gastric lesions, 1 patient was excluded for poor preparation and 2 patients were excluded for superficial lesions larger than 1.0 cm; 132 patients were excluded for no lesions found by high definition endoscopy. In the end, 43 patients with 43 lesions were included. Histopathology revealed 10 inflammation, 14 atrophy, 10 metaplasia, 1 low grade dysplasia (LGD), 5 high grade dysplasia (HGD) and 3 cancers. For 7 lesions classified into type I, histopathology revealed 6 atrophy and 1 metaplasia; for 10 lesions classified into type II, histopathology revealed 2 inflammation, 7 atrophy and 1 metaplasia; for 10 lesions classified into type III, histopathology revealed 1 inflammation, 8 metaplasia and 1 LGD; for 9 lesions classified into type IV, histopathology revealed 4 inflammation, 1 atrophy and 4 HGD; for 7 lesions classified into type V, histopathology revealed 3 inflammation, 1 HGD and 3 cancers. A total of 172 still images, including 43 images by white light (MWL) and 129 images by M i-scan (43 with TE, 43 with SE and 43 with CE), were selected and sent to the endoscopist who did the analysis. General image quality of M i-scan with TE and SE was significantly better than that of MWL (TE, 4.55 ± 1.07; SE, 4.30 ± 1.02; MWL, 3.25 ± 0.99; P < 0.001). Visualization of pit pattern was significantly improved by M i-scan with SE (1.93 ± 0.25 vs 1.50 ± 0.50, P < 0.001). Microvessel visualization was significantly improved by M i-scan with TE (1.23 ± 0.78 vs 0.76 ± 0.73, P < 0.001). Demarcation line visualization was improved by M i-scan with both TE and SE (TE, 1.75 ± 0.52; SE, 1.56 ± 0.59; MWL, 0.98 ± 0.44; P < 0.001). M i-scan with CE did not show any significant improvements of image quality in general or in the 3 key parameters. Although M i-scan with TE and SE slightly increased the diagnostic yield of MWL, there was no significant difference (P > 0.1). CONCLUSION: Although digital enhancement improves the image quality of magnification endoscopy, its value in improving the diagnostic yield seems to be limited.

High-definition magnifying endoscopy with i-scan in the diagnosis of Helicobacter pylori infection: a pilot study.

OBJECTIVE: This study aimed to compare the diagnostic value and image quality of high-definition magnifying white light endoscopy (M-WLE) and high-definition magnifying endoscopy with i-scan (M-i-scan) for Helicobacter pylori (H.pylori) infection. METHODS: Patients complaining of dyspepsia who underwent esophagogastroduodenoscopy at the Endoscopy Unit of Qilu Hospital (Jinan, Shandong Province, China) from January to March, 2011 were prospectively recruited. The greater curvature of the stomach was carefully observed using both M-WLE and M-i-scan. The gastric mucosal classification and image quality scores were recorded. The relationship between gastric mucosal classification and final diagnosis of H.pylori infection was determined. RESULTS: A total of 84 patients were recruited in this study. The accuracy and specificity of the combination of types 2-3 patterns predicting H.pylori infection were significantly higher for M-i-scan than those for M-WLE (accuracy: 94.0% vs 84.5%, P=0.046; and specificity: 93.5% vs 80.6%, P=0.032), while the sensitivity of the two modes was the same. The M-i-scan provided a better image quality for H.pylori infection than M-WLE (P<0.001). CONCLUSION: The M-i-scan may be superior to M-WLE for the prediction of H.pylori infection, but its diagnostic sensitivity needs to be improved.

Surface maturation scoring for oesophageal squamous intraepithelial neoplasia: a novel diagnostic approach inspired by first endomicroscopic 3-dimensional reconstruction.

OBJECTIVE: Loss of surface maturation and cytonuclear atypia have been regarded as the pathological 'gold standard' for the diagnosis of oesophageal squamous cell intraepithelial neoplasia. However, there has been no satisfactory endomicroscopic method similar to this pathological approach to detect surface maturation and screen for oesophageal squamous cell intraepithelial neoplasia. The aim of this study was to apply a 3-dimensional (3D) confocal endomicroscopic imaging technique to investigate the surface maturation of the oesophageal epithelium and develop new 2-dimensional confocal endomicroscopic criteria based on surface maturation. DESIGN: In the 3D reconstruction phase, intrapapillary capillary loops were reconstructed to demonstrate the stereo configuration of the oesophageal epithelium, and a novel surface maturation scoring (SMS) method for plane confocal images was developed based on the interpretation of the 3D microstructure. In the SMS diagnostic phase, 1214 patients were screened and confocal images from 64 non-invasive oesophageal lesions were independently evaluated using SMS and previous methods. RESULTS: We successfully obtained and interpreted 3D confocal images of the human oesophageal epithelium for the first time. The sensitivity (81.0%, 95% CI 58.1% to 94.6%) and specificity (90.7%, 95% CI 77.9% to 97.4%) of the newly established SMS were superior to previous confocal approaches in distinguishing squamous intraepithelial neoplasia from other non-invasive lesions. CONCLUSIONS: 3D confocal endomicroscopic imaging provides valuable insight into the stereo configuration of the human oesophageal epithelium. SMS is a novel and promising diagnostic method to distinguish neoplasia during ongoing endoscopy.

[Analysis of the status of DACH1 gene promoter methylation in endometrial carcinoma and its clinical significance].

OBJECTIVE: To analyze the status of DACH1 gene promoter methylation and explore its association with the expression of DACH1 gene promoter methylation and clinical significance of endometrium carcinoma (EC). METHODS: From February 2004 to August 2008, a total of 80 EC tissue samples with comprehensive surgical pathology staging were collected and used for this study. Twenty normal endometrium tissues in 2008 were abstained from the fractional curettage because of dysfunctional uterine bleeding as control. All samples were confirmed pathologically. Methylation specific PCR (MSP) was performed to detect the promoter methylation of DACH1 gene, and analyze its influence on the expression of DACH1 and the relationship between DACH1 promoter methylation and clinicopathological factors in EC. DACH1 protein expression was detected by western blot. Chi-square test and Pearson test were used for statistical analysis. RESULTS: The rate of promoter methylation of DACH1 gene in the EC tissues was significantly higher than that in the normal endometrium issues (30% vs. 5%, P < 0.05). There was an association between the expression of DACH1 and DACH1 gene promoter methylation (r = -0.30, P < 0.01). There was statistical difference between the methylation of DACH1 and the pathological grade (P < 0.05) or histological type (P < 0.05). But DACH1 gene methylation was not related with the age, stage, myometrial invasion depth and lymphnode metastasis (P > 0.05). CONCLUSIONS: DACH1 gene promoter methylaion could lead to a decrease or absence in the DACH1 expression in EC. The promoter methylation of DACH1 gene may induce the inhibition of DACH1 expression, which might be one of the mechanisms of DACH1 gene inactivation in human EC.

Propofol vs midazolam plus fentanyl for upper gastrointestinal endomicroscopy: a randomized trial.

AIM: To compare the endomicroscopic image quality of integrated confocal laser endomicroscopy (iCLE) and sedation efficacy of propofol vs midazolam plus fentanyl (M/F). METHODS: Consecutive outpatients undergoing iCLE were prospectively recruited and randomized to the propofol group (P group) or M/F group. The patient, performing endoscopist and endoscopic assistant were blinded to the randomization. The quality of endomicroscopic images and anesthetic efficacy outcomes were blindly evaluated after iCLE examination. RESULTS: There were significantly more good quality endomicroscopic images in the propofol group than in the M/F group (72.75% vs 52.89%, P < 0.001). The diagnostic accuracy for upper gastrointestinal mucosal lesions using confocal laser endomicroscopy favors the P group, although this did not reach statistical significance. Adverse events and patient assessment were not significantly different for M/F vs propofol except for more frequent intraprocedural recall with M/F. Procedure duration and sedation times were significantly longer in the M/F group, while the scores of endoscopist, anesthetist and assistant assessment were all significantly better in the P group. CONCLUSION: Sedation with propofol might increase the proportion of good quality endomicroscopic images, and may result in improved procedural efficacy and diagnostic accuracy during iCLE examination.

Microalterations of esophagus in patients with non-erosive reflux disease: in-vivo diagnosis by confocal laser endomicroscopy and its relationship with gastroesophageal reflux.

OBJECTIVES: Objectively diagnosing non-erosive reflux disease (NERD) is still a challenge. We aimed to evaluate the use of in-vivo confocal laser endomicroscopy (CLE) to examine the microalterations of the esophagus in patients with NERD and its relationship with reflux episodes monitored by multiple intraluminal impedance-pH (MII-pH). METHODS: Patients with gastroesophageal reflux symptoms completed reflux disease questionnaires. NERD was determined by negative gastroscopy. Patients without reflux symptoms were recruited as controls. Pilot clinical study was followed by prospective controlled blinded study. All subjects were examined by white-light mode of the endoscopy followed by the standard CLE mode and then MII-pH monitoring. The microalterations seen on CLE images and the correlation between CLE features and reflux episodes were evaluated, the correlation between CLE and transmission electron microscope (TEM) data was also analyzed. RESULTS: On CLE images, NERD patients had more intrapapillary capillary loops (IPCLs) per image than did controls (8.29 ± 3.52 vs. 5.69 ± 2.31, P=0.010), as well as the diameter of IPCLs (19.48 ± 3.13 vs. 15.87 ± 2.21 µm, P=0.041) and intercellular spaces of squamous cells (3.40 ± 0.82 vs. 1.90 ± 0.53 µm, P=0.042). The receiver operating characteristic analysis indicated that IPCLs number (optimal cutoff >6 per image, area under the curve (AUC) 0.722, 95% confidence interval (CI) 0.592-0.853, sensitivity 67.7%, specificity 71.6%), IPCLs diameter (optimal cutoff >17.2 µm, AUC 0.847, 95% CI 0.747-0.947, sensitivity 81%, specificity 76%), and the intercellular spaces of squamous cells (optimal cutoff >2.40 µm, AUC 0.935, 95% CI 0.875-0.995, sensitivity 85.7%, specificity 90.5%) diagnosed NERD with reasonable accuracy. Combined features of dilatation of intercellular space plus increased IPCLs provided 100% specificity in the diagnosis of NERD patients. The intercellular spaces of squamous cells observed on CLE were highly related to that on TEM findings (r=0.75, P<0.001). Multivariate progressive regression analysis showed that acidic reflux, especially in the supine position, was related to the increased number and dilation of IPCLs in the squamous epithelium (ß=0.063, t=2.895, P=0.038 and ß=0.156, t=1.023, P=0.04). CONCLUSIONS: CLE represents a useful and potentially significant improvement over standard endoscopy to examine the microalterations of the esophagus in vivo. Acidic reflux is responsible for the microalterations in the esophagus of patients with NERD.

Mucosal barrier defects in gastric intestinal metaplasia: in vivo evaluation by confocal endomicroscopy.

BACKGROUND: Helicobacter pylori infection and intestinal metaplasia (IM) are associated with gastric cancer. An impaired gastric mucosal barrier could be involved in this carcinogenesis. OBJECTIVE: To evaluate laser confocal laser endomicroscopy (CLE) for in vivo functional imaging of mucosal barrier defects in patients with IM. DESIGN: Prospective, controlled study. SETTING: A tertiary-care academic center. PATIENTS: This study involved patients with IM of the gastric mucosa who underwent CLE for surveillance. INTERVENTIONS: Specific IM mucosa and non-IM mucosa in patients were identified by CLE, and targeted biopsy samples were taken for histopathology and electron microscopy. MAIN OUTCOME MEASUREMENTS: Post-CLE assessment of paracellular fluorescein leakage was devised and validated by electron microscopy. We also evaluated the effect of H pylori eradication on the mucosal barrier. RESULTS: Forty-two patients were included. Of non-IM samples, the paracellular permeability was significantly increased in H pylori-positive samples compared with H pylori-negative controls (54 ± 31% vs 3 ± 6%, P < .05). Of IM samples, the permeability was significantly increased in both H pylori-negative and H pylori-positive samples (67 ± 34% and 72 ± 28% vs 3 ± 6%, both P < .05). The results of post-CLE assessment correlated well with the electron microscopy findings (R(2) 0.834, P < .0001). After the eradication of H pylori, the paracellular barrier dysfunction of non-IM mucosa was significantly improved as shown by electron microscopy and CLE (both P < .001). However, there was no significant change in IM mucosa. LIMITATIONS: Single-center study. CONCLUSIONS: CLE allows functional imaging of mucosal barrier defects. Gastric IM is associated with an impaired paracellular barrier irrespective of H pylori eradication.