AIM:To evaluate the influence of scutellarin on the expression of vascular endothelial growth factor ( VEGF) in high glucose-treated human retinal pigment epithelial cell line ARPE-19 and to observe the effects of scutellarin on the protein expression of VEGF, p-ERK and VEGFR2 in the retinas of type II diabetic rats.METHODS: Cultured ARPE-19 cells were divided into normal control group, scutellarin group, high glucose group and high glucose+scutellarin group.The protein levels of VEGF, p-ERK and VEGFR2 were measured by Western blot.The VEGF release in ARPE-19 cells was detected by ELISA.Normal rats were randomly divided into normal control group and scutellarin group.Diabetic rat model was established by feeding with high-fat diet and injecting with streptozocin, and randomly divided into diabetes group and diabetes treated with scutellarin group.After 16 weeks, the eyes were removed.The morphological changes of the retinas were observed under light microscope with HE staining, and histopathological score was recorded.The expres-sion of VEGF in the retinas was observed by the method of immunohistochemistry.RESULTS:Compared with normal con-trol group, the protein levels of VEGF, p-ERK and VEGFR2 in the ARPE-19 cells decreased in scutellarin group, but in-creased in high glucose group.The histopathological score of the retinas showed significant difference among diabetes group, diabetes treated with scutellarin group and normal control group, and no significant difference between normal con-trol group and scutellarin group was observed.The expression of VEGF increased in diabetic group and was significantly higher than that in scutellarin treatment group (P<0.05).CONCLUSION:Scutellarin inhibits the increased protein le-vels of VEGF, p-ERK and VEGFR2 in ARPE-19 cells, and decreases the expression of VEGF in the retinas of diabetic rats.The suppression of the diabetic retinopathy development by scutellarin may be partly involved in the ERK/MAPK pathway.

Detection of the possible role of ERp46,new endoplasmic reticulum protein,on palmitic acid-inducedendoplasmic reticulum stress-apoptosis pathway in βTC6 cells for the new treatment of type 2 diabetes.Results showed that ERp46 played a protective role in palnutic acid-induced cell apoptosis by decreasing the endoplasmic reticulum stress response through three pathway.

Objective The aim of this study was to investigate the association between methylation of ppENK and S100A4 and pancreatic carcinoma. Methods 31 samples of human tissues of pancreatic cancer and adjacent tissues, five pancreatic carcinoma cell lines and one sample of normal pancreas were collected. ppENK methylation status was detected by MSP and S100A4 methylation status was detected by COBRA. The expression of S100A4 and ppENK protein was investigated by immunohistochemistry and RT-PCR. The association of methylation status of ppENK gene or S100A4 with clinical parameters of pancreatic carcinoma was analyzed. Results No methlation of ppENK was detected in normal pancreatic tissue. SI00A4 gene was highly methylated. Methylation of ppENK was detected in 90.3% of the pancreatic carcinoma tissue in 31 patients; there were no correlation between the status of ppENK methylation with clinical parameters of pancreatic carcinoma. The hypomethylation rate of S100A4 gene was 71%, and it was only related to the serum level of CA 19-9 (P=0.011, OR=0.05) ; the expression rate of S100A4 protein in pancreatic cancer tissue was 87.1%, and the expression rates were correlated with the differentiation of the tumor, namely, poorly differentiated tumor highly expressed S100A4 protein. S100A4 hyomethylation was highly correlated with ppENK methylation (P=0.019). Methylated ppENK was detected in five pancreatic cancer cell lines, and there was no ppENK mRNA expression; S100A4 gene was hypomethylated, while S100A4 mRNA was highly expressed in five pancreatic cancer cell lines. Conclusions ppENK was hypermethylated, while S100A4 was hypomethylated in pancreatic cancer tissue.

Objective To detect the methylation status of ppENK and its role in the pathogenesis of pancreatic carcinoma.Methods The ppENK methylation status in human tissues of pancreatic cancer,pancreatic carcinoma cell lines and normal pancreas was detected by methylation-specific RT-PCR(MSP).The association of methylation status of ppENK gene with clinicopathological parameters was analyzed. The expression of ppENK mRNA was detected by RT-PCR.Two pancreatic carcinoma cell lines (PANC1,AsPC1 ) were treated with demethylating agent (5-aza-dC).The cell growth was measured by MTT.Apoptosis and cell cycle was analyzed by flow cytometry.The expression of DNMT3a was measured by Western blot.Results ppENK mRNA was expressed in normal pancreas.And methylation of ppENK was not detected in normal pancreas.Methylation of ppENK was detected in 90.3％ (28/31) of pancreatic carcinoma tissue,and there were no correlation between methylated ppENK with clinicopathological features of pancreatic carcinoma.There was no ppENK mRNA expression in SW1990,PANC1,PC3,AsPC1,PuPan-1,and ppENK was methylated.Methylated ppENK was associated with no ppENK mRNA expression.After 5-Aza-dC treatment,PANC1,AsPC1 was demethylated and ppENK mRNA expression was reversed.The proliferation of PANC1 and AsPC1 was inhibited in a dose dependent manner.The apoptotic rates of PANC1 and AsPC1 were increased [ (31.57 ± 6.76)％ts (3.21 ±1.43)％,P =0.002,(16.6 ±8.22)％ vs (3.82 ±1.71)％,P=0.058];the expression of DNMT3a protein was decreased; the PANC1 cells of G1 phase significantly increased [ (67.87 ± 2.72 ) ％ vs (54.57 ± 7.18) ％,P =0.040 ],but PANC1 cells of S phase significantly decreased [ ( 22.37 ± 4.31 )％ vs (33.73 ± 4.63)％,P =0.036 ].But the percentage of G1,S phase in AsPC1 cell line was not significantly changed ( P =0.236,0.075 ).ConclusionsppENK demethylation is an important molecular event in inducing ppENK expression inhibition,which can inhibit pancreatic cancer proliferation,promote apoptosis,arrest cell cycle at G1 and decrease the expression of DNMT3a protein.

Objective To evaluate the clinical value of the three-category classification of severe acute pancreatitis (SAP).Methods Clinical data of 337 traditional SAP patients,who were admitted to Peking Union Medical College Hospital (PUMCH)from January 2001 to December 2012,were retrospectively studied.These patients were classified into moderately severe acute pancreatitis (MSAP) and severe acute pancreatitis (SAP) according to the latest 2013 Atlanta Classification.SAP patients were further categorized as critical acute pancreatitis (CAP) and non-CAP.Disease severity,therapy and prognosis among three groups were compared.Results Among the total 337 traditional SAP patients,253 were classified as MSAP and 84 as SAP.In the group of SAP,40 patients were categorized as CAP and 44 as non-CAP.Compared with non-CAP patients,CAP patients had significantly higher mortality rate which was 70％ (28/40).Other results were all significantly higher in CAP group rather than non-CAP group,including ICU admission rate 77.5％(31/40),length of ICU stay (15.5 ± 20.6) days,Ranson,APACHE Ⅱ,BISAP,MCTSI,modified Marshall scores 4.6 ± 1.4,16.8 ± 5.8,3.0 ± 1.0,8.6 ± 1.7,and 7.4 ± 2.9,respectively (P ＜ 0.01 in each endpoint).These parameters of SAP group were also significantly higher than those of MSAP group (P ＜0.01).Conclusions Using the new three-category classification to distinguish traditional severe acute pancreatitis,namely MSAP,SAP,and CAP,can better reflect the severity of disease,predict outcome and guide clinical management.

Objective To explore the clinical features and possible carcinogenesis mechanism of the ulcerative colitis (UC)-associated colorectal cancer. Methods From 1984 to 2008, 6 clinical cases of UC-associated colorectal cancer were collected at Peking Union Medical College Hospital. The characters of morbidity, clinical features, pathology type, treatment and prognosis were analyzed.Immunohistochemistry(IHC) was performed to study the protein expression of adenomatous polyposis coli protein(APC), β-catenin, P53 and Wnt-1 in the specimens. Results The canceration of UC was 1.1 %, higher in female cases (5/6), the average duration was 14.3 years. All cases presented typical UC manifestation, most involved total colon (5/6) and none of them complicated with primary sclerosing cholangitis (PSC). There were 4 rectal cancers and 2 descending colon caners the UC cases collected. The major pathology type was adenocarcinoma with poor prognosis. The positive protein expression ratio of APC, β-catenin, P53 and Wnt-1 were 6/6, 6/6, 5/6 and 6/6 respectively.Conclusion The rectal cancer should be monitored and prevented in UC patients with total colon involved and long disease duration. Multi-pathway may possibly be involved in the carcinogenesis.

Objective To evaluate the significance of detecting plasma methylated Septin9 (SEPT9) alone and combined with fecal immunochemical test (FIT) in screening colorectal cancer (CRC) and adenoma.Methods From October 2014 to April 2015,outpatients received CRC and adenoma screening were enrolled.Colonscopy examination and pathological diagnosis were taken as gold standard.The sensitivity and specificity of plasma methylated SEPT9 and FIT in CRC and adenoma were calculated.The diagnostic value of combined examination was evaluated.Receiver operating characteristic (ROC) curve of SEPT9 in CRC diagnosis was drawn.Results A total of 300 outpatients were enrolled.Among them,CRC was detected in 45 patients (15.0％) and adenoma was detected in 68(22.7％) patients,including 37(12.3％) cases of advanced adenomas.The sensitivity of SEPT9 and FIT for CRC diagnosis was 80.0％ and 88.9％,and the specificity was 95.3 ％ and 54.1％,respectively.The area under concentration curve (AUC) of methylated SEPT9 in CRC diagnosis was 0.923.The sensitivity of SEPT9 and FIT in advanced adenoma diagnosis were 10.8％ and 62.2％,and the specificity were 83.3％ and 49.0％,respectively.The sensitivity of combined examination in CRC diagnosis was 97.8％ and the specificity was 52.9％;the sensitivity in advanced adenoma diagnosis was 67.6％ and the specificity was 47.4％.Conclusions Plasma methylated SEPT9 is helpful in CRC screening,moreover,when combined with FIT,it can increase detection rate of colorectal adenoma.

ObjectiveTo evaluate the application of immunochemical fecal occult blood test for colorectal cancer screening in a gastroenterology clinic. MethodsTotal 512 outpatients received immunochemical fecal occult blood tests and colonoscopy for screening of colorectal cancer from January 2009 to October 2010 in Gastroenterology Clinic of Peking Union Hospital.The application of occult blood test was retrospectively evaluated using colonoscopy and pathological examination as the gold standard. Results Among 512 patients,203 were found positive for immunochemical fecal occult blood. According to the colonoscopy and histological study,115 patients had polyps,9 with high-grade dysplasia and 21 with colorectal cancer.The sensitivity,specificity,positive likelihood ratio and negative likelihood ratio of fecal occult blood to detect colorectal adenomatous polyps was 42.8％,60.9％,1.09 and 0.93,respectively.And those for detection of colorectal cancer and high-grade dysplasia were 76.6％,62.5％,2.05 and 0.37,respectively.ConclusionsImmunochemical fecal occult blood test was useful in the screening for colorectal tumors among gastroenterology clinic patients.More large-scale prospective studies are needed to establish a screening model for colorectal tumors.

ObjectiveTo summarize the clinical features of ulcerative eolitis (UC) complicated by toxic megacolon for early diagnosis and proper treatment. MethodsSix cases of toxic megacolon in the patients suffered from UC in Peking Union Medical College Hospital from 1983 to 2010 were analyzed,and related literature was searched and reviewed.ResultsThe incidence of the toxic megacolon in the patients with UC in our center was 0.7％ (6/824),which was lower than those reported in the literature.There were always risk factors triggering the disease.The prognosis of the patients was poor,even after medical care and surgery intervention.Evaluation of the patients and making right timing to perform the surgery would improve the prognosis of the patients in foreign literature. ConclusionIt's crucial to make early diagnosis of the toxic megacolon in the patients suffered from UC. The right choice and timing to perform urgent surgery or selective surgery may improve their prognosis.

BACKGROUND:Hyperbranched cationic amylopectin is a kind of nonviral gene vectors with low toxicity and good transfection efficiency. However, searching for more efficient methods to delivery it into the body and making the genes expressed are being explored.
OBJECTIVE:To study the expression of DMAPA-Amp wrapped green fluorescent protein (GFP) transferred by modified carotid injection into cerebral ischemic area.
METHODS:Male Sprague-Dawley rats subjected to middle cerebral artery infarction were randomly divided into two groups after 24 hours:experimental group (injected with GFP entrapped DMAPA-Amp via the internal carotid artery) and control group (injected with GFP entrapped DMAPA-Amp via the tail vein). These rats were put to death and their brain tissue was removed after 7 days. The expression of GFP was detected by quantitative PCR and western blot assay, and immunofluorescence staining was performed to detect the expression of GFP located near cerebrovascular endothelial cel s by frozen section.
RESULTS AND CONCLUSION:Compared with the control group, the expression of GFP was much higher in the experimental group detected by quantitative PCR and western blot (P< 0.05). Additional y, the expression of GFP located near cerebrovascular endothelial cel s by frozen section was also higher than that in the control group. Modified carotid injection could significantly promote the expression of hyperbranched cationic amylopectin derivates and GFP in the brain tissue of Sprague-Dawley rats undergoing middle cerebral artery infarction compared with tail vein injection, which indicates DMAPA-Amp and modified carotid injection may cast new lights on the therapy for angiogenesis of ischemic stroke.