Objective To explore an ideal approach to establish rabbit model of hepatic fibrosis suitable for radiological and serological research.Methods Totally 40 rabbit models of hepatic fibrosis was established by intraperitoneally injecting 5% carbon tetrachloride in oil solution or 100% carbon tetrachloride(both groups,0.1 ml/kg,once a week),and drinking 5% ethanol water.Another 8 rabbits who drank normal water and received a peritoneal injection at 0.1 ml/kg served as control.The rabbits(5 or 2 per time for model group and control) were killed at 6,8,10 and 12 weeks respectively after first injection.Their livers were resected for gross and morphological observation with HE and Masson staining.Results Death of the models usually happened within 4 weeks after first injection and became stable after 6 weeks.Mortality of 5% carbon tetrachloride group was 60%,but 25% in 100% carbon tetrachloride group.Macroscopy and microscopy indicated that liver fibrosis was observed in rat models.Conclusion Long-term intraperitoneal injection of carbon tetrachloride results in rabbit hepatic fibrosis.Injection of 100% carbon tetrachloride intraperitoneal,at 0.1 ml/kg,once a week,combined with and 5% ethanol as drinking water is a suitable approach to establish rabbit model of hepatic fibrosis with low mortality and high success rate.

Objective To investigate the relationships between the concentration of serum NT-proBNP,New York Heart Association(NYHA) classification,left ventricular ejection fraction((LVEF),)and the value of serum NT-proBNP in the evaluation of cardiac function and the diagnosis of chronic heart failure.Methods One hundred and two subjects were selected,including 30 healthy controls and 72 patients with heart failure.The concentration of serum NT-proBNP was determined by an automated electrochemiluminescence immunoassayan Roche Elecsys 2010.LVEF was measured by the modified Simpson′s equation with echocardiography.The diagnosis of clinical physician was considered to be the golden standard for heart failure.Results The relationships between the concentration of serum NT-proBNP and NYHA,serum NT-proBNP levels(287.7 pg/ml) was significantly higher in heart failure patients(287.7 pg/ml) than that in healthy controls 287.7 pg/ml vs (46.1 pg/ml),P

0.05)],which less than in control group [(4.614?1.683) IU/cm,(0.119?0.068) IU/cm,(564.2?53.8) ?m Pa

ObjectiveTo investigate the concentration of water fluoride and the application of water defluoridation projects in Suizhou,and to provide a basis for water improvement and prevention of the disease.MethodsCross-sectional study was carried out to investigate completely the water defluoridation projects built between 1986 and 2009 in the area under Suizhou's jurisdiction; 3 source water and terminal water samples of the projects in use were collected,respectively; 1 water sample was collected,respectively in the 3 projects with out-ofcommission or discarded water sources that used to be major water supply,for detection of fluoride,chloride,total hardness,heavy metals and other indicators.ResultsA total of 21 projects were investigated,14 projects were in normal use (66.7％),7 projects were out of order and abandoned (33.3％).Of the 14 projects normally used,13projects with water fluoride concentration ≤ 1.0 mg/L(92.9％)and 1 project with water fluoride concentration higher than 1.0 mg/L(7.1％),the concentration of water fluoride in the 7 abandoned projects was higher than 1.0 mg/L(100％).Arsenic and lead levels of all the water samples were normal.There were two water samples with iron content exceeded the standard,and one total hardness of water samples exceeded the standard in the normally used 14 peripheral water sources.One chloride content exceeded the standard,one manganese content exceeded the standard,two iron content exceeded the standard and three total hardness exceeded the standard in the seven abandoned projects.ConclusionsSome defluoridation projects are stopped using and abandoned in Suizhou,and the water fluoride exceeds the standard.

Aim Powdered injections of luotai consist of total saponins of panax notoginseng(PNS) which protect against ischemia injury from myocardial reperfusion injury.Methods ISO-induced acute myocardial ischemia model in rat was made, which decreased S-T sigments in ECG.Luotai after intravenous injection or oral can protect against S-T sigment significant reduction and ∑ST after ISO induced acute myocardial ischemia .Results Luotai 50 mg?kg-1 and 100 mg?kg-1 significantly inhibited S-T sigment decreases. Conclusion Luotai protect ISO induced acute myocardial ischemia and has dose-dependent effect.This ISO-induced acute myocardial ischemia model in rat has some significant advantages,for example,higher stability, good duplication, quickly filtrates, easily masters.

D-lactonohydrolase is useful in the procedure of resolution of racemic pantolactone to produce D-pantolactone, but the activity and stability under low pH of the wild type enzyme is not satisfactory enough to be applied to industrial production. The expected properties of wild type enzyme were enhanced by directed evolution. According to the formation of products and pH indicators, a screening system was designed. After three sequential error prone PCR and one round DNA shuffling followed by screening, Mut E-861, the best mutant with improved activity and stability under low pH situation was obtained. Gene analysis of the Mut E-861 mutant indicated that the mutant enzyme had A352C, G721A mutations and a silent mutation of position 1038. Moreover, the activity and stability of Mut E-861 were determined. The results showed that the activity of this mutant was 5.5-fold higher than that of wild type, and the stability under low pH was improved at no expense of D-lactonohydrolase activity. After incubated at pH 6.0 and pH 5.0 the activity of D-lactonohydrolase could be retained 75% to 50%, however, compared with 40% to 20% for wild type.
Carboxylic Ester Hydrolases ; biosynthesis ; genetics ; DNA Shuffling ; Directed Molecular Evolution ; Enzyme Stability ; Escherichia coli ; enzymology ; genetics ; Mutagenesis, Site-Directed ; Mutant Proteins ; genetics ; metabolism ; Polymerase Chain Reaction ; methods ; Protein Engineering ; Saccharomyces cerevisiae ; enzymology ; genetics

BACKGROUNDOverexpression of G-protein coupled receptor 34 (GPR34) affects the progression and prognosis of human gastric adenocarcinoma, however, the role of GPR34 in gastric cancer development and progression has not been well-determined. The current study aimed to investigate the effect of GPR34 knockdown on the proliferation, migration, and apoptosis of HGC-27 gastric cancer cells and the underlying mechanisms.

METHODSThe expression of GPR34 in gastric cancer cell line HGC-27 was detected by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. HGC-27 cells were employed to construct the stable GPR34 knockdown cell model in this study. Real-time RT-PCR and Western blotting were applied to validate the effect of short hairpin RNA (ShRNA) on the expression of GPR34 in HGC-27 gastric cells. The proliferation, migration of these cells were examined by Cell Counting Kit-8 and transwell. We also measured expression profile of PI3K/PDK1/AKT and ERK using Western blotting.

RESULTSThe ShRNA directed against GPR34 effectively inhibited both endogenous mRNA and protein expression levels of GPR34, and significantly down-regulated the expression of PIK3CB (P < 0.01), PIK3CD (P < 0.01), PDK1 (P < 0.01), phosphorylation of PDK1 (P < 0.01), Akt (P < 0.01), and ERK (P < 0.01). Furthermore, GPR34 knockdown resulted in an obvious reduction in HGC-27 cancer cell proliferation and migration activity (P < 0.01).

CONCLUSIONSGPR34 knockdown impairs the proliferation and migration of HGC-27 gastric cancer cells in vitro and provides a potential implication for therapy of gastric cancer.

Apoptosis ; genetics ; physiology ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; genetics ; physiology ; Humans ; RNA, Small Interfering ; genetics ; Real-Time Polymerase Chain Reaction ; Receptors, Lysophospholipid ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism

OBJECTIVETo recognize the characteristics of desmoplastic small round cell tumor (DSRCT) and improve the standard of diagnosis.

METHODSWe retrospectively reviewed the clinical data on the treatment of 2 patients with DSRCT in terms of their conditions, tissue sources, pathologic characteristics, immunohistochemical methods, clinical manifestation, diagnosis, treatment and prognosis.

RESULTSClinical manifestations were complicated. The 2 patients were mis diagnosed before operation. Their tumors consisted of irregular nests of small and round cells, with nuclear hyperchromatism and scant cytoplasm embedded in a plenty of fibrous connective tissues. The edge of the nest was clear, with different sizes and shapes. Immunohistochemically, the 2 patients were positive for CK or EMA, NSE, des and vim of the epithelium, nerve, muscle and interstitial. They died 9 months after operation.

CONCLUSIONSThe tumor may occur in the abdomen, pelvic cavity and other sites, with different clinical manifestations. Routine examination should be replaced by immunohistochemical test for correct diagnosis of the tumor. Prognosis of most patients is not good.

Adolescent ; Carcinoma, Small Cell ; diagnosis ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasms, Connective Tissue ; diagnosis ; Retrospective Studies

BACKGROUNDCerebral hyperperfusion syndrome is an important complication of carotid endarterectomy (CEA). An >100% increase in middle cerebral artery velocity (MCAV) after CEA is used to predict the cerebral hyperperfusion syndrome (CHS) development, but the accuracy is limited. The increase in blood pressure (BP) after surgery is a risk factor of CHS, but no study uses it to predict CHS. This study was to create a more precise parameter for prediction of CHS by combined the increase of MCAV and BP after CEA.

METHODSSystolic MCAV measured by transcranial Doppler and systematic BP were recorded preoperatively; 30 min postoperatively. The new parameter velocity BP index (VBI) was calculated from the postoperative increase ratios of MCAV and BP. The prediction powers of VBI and the increase ratio of MCAV (velocity ratio [VR]) were compared for predicting CHS occurrence.

RESULTSTotally, 6/185 cases suffered CHS. The best-fit cut-off point of 2.0 for VBI was identified, which had 83.3% sensitivity, 98.3% specificity, 62.5% positive predictive value and 99.4% negative predictive value for CHS development. This result is significantly better than VR (33.3%, 97.2%, 28.6% and 97.8%). The area under the curve (AUC) of receiver operating characteristic: AUC(VBI) = 0.981, 95% confidence interval [CI] 0.949-0.995; AUC(VR) = 0.935, 95% CI 0.890-0.966, P = 0.02.

CONCLUSIONSThe new parameter VBI can more accurately predict patients at risk of CHS after CEA. This observation needs to be validated by larger studies.

Aged ; Blood Pressure ; physiology ; Cerebrovascular Circulation ; physiology ; Cerebrovascular Disorders ; physiopathology ; Endarterectomy, Carotid ; Female ; Hemodynamics ; physiology ; Humans ; Male ; Middle Aged ; Prospective Studies

OBJECTIVETo study the therapeutic effects to block the TGF-beta1 (transforming growth factor beta1) signal transduction by antisense Smad4 gene on experimental fibrotic liver.

METHODSUsing the rat model of liver fibrosis induced by Carbon Tetrachloride (CCl4)/ethanol, we transfected antisense Smad4 gene mediated by adenovirus via portal vein infusion into the liver, and observed the expression of Smad4 by Retro-Polymerase Chain Reaction (RT-PCR) and Western Blot. We also investigated the pathologic features and collagen expression.

RESULTSIn the non-therapeutic cirrhotic liver, the expression of Smad4 mRNA was significantly increased than normal liver, and so was the collagen I. After antisense Smad4 gene being transfected, the expression of Smad4 mRNA and that of collagen I in the therapeutic liver was significantly decreased, compared with the non-therapeutic cirrhotic liver. The fibrous degree of therapeutic liver was also reduced compared with the non-therapeutic fibrous liver.

CONCLUSIONThese results indicate that because antisense Smad4 gene could block TGF-beta1 signal transduction by reducing the expression of Smad4, so it could inhibit the production of extracellular matrix (ECM) and improve hepatic fibrosis.

Adenoviridae ; genetics ; Animals ; Antisense Elements (Genetics) ; therapeutic use ; Collagen Type I ; analysis ; DNA-Binding Proteins ; antagonists & inhibitors ; genetics ; Liver ; pathology ; Liver Cirrhosis, Experimental ; metabolism ; pathology ; therapy ; Male ; Rats ; Rats, Wistar ; Signal Transduction ; Smad4 Protein ; Trans-Activators ; antagonists & inhibitors ; genetics ; Transforming Growth Factor beta ; antagonists & inhibitors ; Transforming Growth Factor beta1