Animals ; Genes, BRCA2 ; Genes, erbB-1 ; Genes, erbB-2 ; Genes, p16 ; Genes, p53 ; Genes, ras ; Genetic Predisposition to Disease ; genetics ; Humans ; Pancreatic Neoplasms ; classification ; genetics ; pathology

Adolescent ; Blood Vessel Prosthesis Implantation ; Child ; Child, Preschool ; Ductus Arteriosus, Patent ; therapy ; Equipment and Supplies ; Female ; Humans ; Infant ; Male ; Video Recording

OBJECTIVETo establish a stable high red fluorescent protein (RFP)-expressing orthotopic transplantation nude mice spontaneous metastasis model of pancreatic cancer.

METHODSStable high RFP-expressing cells SW1990-RFP were injected subcutaneously into mice to establish subcutaneous implantation model. Fluorescent tumor piece from subcutaneous was transplanted into the body of the pancreas to establish surgical orthotopic implantation model. The growth of primary tumor, metastasis and micrometastasis were assessed by whole-body fluorescence imaging system.

RESULTSTwelve RFP orthotopic transplantation nude mice metastasis models of pancreatic cancer were established successfully, the percentage of success rate was 100%. RFP-labeled pancreatic cancer growth could be monitored in real time way. The micrometastasis of primary lesions were detected in early stage with whole-body fluorescence imaging system.

CONCLUSIONSThe RFP orthotopic transplantation nude mice metastasis model of pancreatic cancer is stable and reliable, and can be observed dynamically in vitro in a noninvasive way, with much higher sensitivity and specificity.

Animals ; Disease Models, Animal ; Female ; Luminescent Proteins ; Male ; Mice ; Mice, Nude ; Neoplasm Metastasis ; Pancreas ; pathology ; Pancreatic Neoplasms ; pathology ; Xenograft Model Antitumor Assays

Objective:To improve the genetic stability of HBV gene in transgenic mice.Methods:HBV transgenic mice were bred by backcross and double cross.The HBV gene expression and replication were studied with real-time PCR,ELISA and chemiluminescence.Results:The HBV transgenic mice have stably bred to 23rd generation.The serum HBsAg level is 4122.31?2044.74IU/ml;The rate of HBV transgenic mice whose serum HBV DNA reach 104~106copies/ml was 93.93%.The HBV replication and expression were improved markedly.There is no difference between male and female mice about serum HBsAg level.Conclusion:After breeding the HBV gene was expressed stably with high-level in transgenic mice.

Objective To investigate the influence of iodine excess on expression of TRAIl/TRAIL-sR1 in NOD and Balb/c mice and to study the effect of TRAIl/TRAIL-sR1 on the pathogenesis of experimental autoimmune thyroiditis(EAT). Methods Both Balb/c and NOD mice were divided randomly into control and iodine excess group by feeding with water containing no NaI or 0.05% Nal. The mice were sacrificed after 8 weeks. TRAIL and TRAIL-sR1 mRNA levels were detected by RT-PCR. The function, morphology and apoptosis of thyroids were also observed by ELISA and Tunnel stain. Results Treated by HI, enlarged follicles and flattened epithelium by accumulation of colloid were found in thyroids of both NOD and Balb/c mice. But significant lymphoid cell infiltration and local fibrosis were only found in thyroids of NOD HI group. The relative weight of thyroids of NOD mice in HI group[(104.8±14.5)mg/kg]was heavier than that of control group [(71.8±20.4)mg/kg]. The level of TT4 declined in HI group[(30.77±3.59)mmol/L]compared with control group[(36.43±2.66)mmol/L], meanwhile, the level of TSH was higher in HI group[(6.98±0.66)μg/L]than that in control group [(5.55±0.56)μg/L]. The difference being statistically significant(t=7.773,-9.526,-4.458, all P < 0.05). The relative weight of thyroids of Balb/c mice of HI group[(155.8±20.8)mg/kg]also heavier than that of control group [(105.1±22.0) mg/kg]. The level of TT4 droped in HI group [(19.75±3.32) mmoL/L]was higher than that in control group[(23.46±6.21)mmoL/L], the level of TSH in HI group[(4.14±1.71)μg/L]was higher than that in control group[(3.55±1.41)μg/L], the difference being statistically significant(t=7.554,-7.239,3.140, all P< 0.05). A great deal of apoptotie ceils observed in NOD (3.97±0.91) and Balb/c mice (1.05±0.45) by Tunnel stain were greater than control groups (0.21±0.15, 0.10±0.03), the difference being statistically significant in beth of the two species(t=-7.167,-17.772, both P < 0.05). The apoptosis index of thyroid follicular epithelium in NOD was obviously higher than Balb/c(t=-7.625, P<0.05). The level of TRAIL mRNA did not remarkably change in Balb/c between control group(0.000 59±0.000 39) and HI group(0.001 24±0.000 46, t=-1.940, P>0.05), but it increased apparently in NOD mice HI group(0.018 88±0.005 77) than that of control group(0.009 61± 0.00591, t=-2.71, P<0.05). The level of the expression of TRAIL-sR1 mRNA increased in HI groups of NOD (0.000 53±0.000 15) and Balb/c mice(0.000 42±0.000 09) than that in control groups of NOD(0.000 28± 0.000 05) and Balb/c mice (0.000 17±0.000 06) and the differences were statistically significant between the two species(t=3.050,3.990, all P<0.05). The differences of the expression of TRAIL and TRAIL-sR1 mRNA between the two species were significant(t=-3.37,-4.76, all P<0.05). Conclusions Iodine excess induces colloid goiter in beth species of mice and thyroiditis in NOD mice. The increase of TRAIL and TRAIL-sR1 influenced by iodine excess is one of the molecular bases of follicular epithelium apoptosis and inflammation in thyroids. Genetic factor is a key factor in the pathogenesis of thyroiditis.

Objective To probe into the application effect of quality control circle (QCC) activity in reducing the incidence rate of fistula puncture bleeding in hemodialysis patients.Methods Thirty-six hemodialysis patients in our hospital from January to June 2012 were chosen as the control group (before the QCC activity) ; 36 hemodialysis patients with the same rule in 1 group from July to December 2012 were chosen as the experiment group (after the QCC activity).Six nurses from nephrology volunteer to group a "circle" in order to build-up the QCC activity with the subject of reducing the blood exudation rate during hemodialysis fistula piercing.Before the QCC activity,the data of blood exudation rate was calculated,and the root cause of blood exudation was analyzed during hemodialysis fistula piercing; then the improvement plan was made-up,and the counter measure was implemented.The blood exudation rate during hemodialysis fistula piercing was compared before and after the QCC activity.Results The incidence rate of fistula puncture bleeding after QCC activity was 3.76％,was lower than 7.66％ before the QCC activity,and was better than the origin target,and the differences were statistically significant (x2 =20.634,P ＜ 0.05).Conclusions The implementation of QCC activity is not only benefit to reducing the incidence rate of fistula puncture bleeding in hemodialysis patients,but also is good at circle members' professional technical level and quality management capability,promote the quality improvement of hemodialysis.

OBJECTIVETo investigate the pathogenesis of gastrointestinal motility dysfunction as a result of scald and lipopolysaccharide (LPS) challenge in guinea pigs.

METHODSThirty guinea pigs were enrolled in the study and were randomly divided into 3 groups:i. e. control (n = 10, with intraperitoneal injection of isotonic saline), scald (n = 10, with 30% TBSA deep partial thickness burn) and LPS (n = 10, with intraperitoneal injection of LPS) groups. Thirty minutes after treatment, all animals were gavaged with carbolic ink. The propelled distance of the ink within the gastrointestinal tract was measured. The intestinal tissue was harvested and homogenized, and the contents of CGRP, Na+-K+-ATP enzyme, Mg2+-ATP enzyme, Ca2+-ATP enzyme, Ca2+-Mg2+-ATP enzyme were determined, and the delta phim of haustra coli smooth muscular cell mitochondria was assessed.

RESULTSThe propelled distance of the ink in the gastrointestinal tract in scald (53 +/- 9 cm) and LPS (91 +/- 10 cm) groups was obviously shorter than that in control group (142 +/- 11 cm, P < 0.01). Furthermore, the distance in scald group was shorter than that in LPS group (P < 0.01). The CGRP content in scald and LPS groups [52.0 +/- 39.0 microg/L and 20.0 +/- 23.0 microg/L] was obviously higher than that in control group (0.8 +/-2.0 microg/L, P <0.05 or 0.01), especially in scald group ( P < 0.05). The Na+-K+-ATP enzyme, Mg2+-ATP enzyme, Ca2+-ATP enzyme, Ca2+-Mg2+-ATP enzyme and the delta phim in scald and LPS groups were remarkably lower than those in control group (P <0.005), but there was no difference between scald and LPS groups (P > 0.05).

CONCLUSIONThe gastrointestinal motility of guinea pigs could obviously be inhibited by scald and LPS, especially by scald. LPS might be the key factor to produce change in the membrane potential of mitochondria of intestinal smooth muscle after severe scald.

Animals ; Burns ; pathology ; physiopathology ; Calcitonin Gene-Related Peptide ; metabolism ; Disease Models, Animal ; Gastrointestinal Motility ; drug effects ; Guinea Pigs ; Lipopolysaccharides ; adverse effects ; Myoelectric Complex, Migrating ; drug effects

OBJECTIVETo investigate 18q21 LOH in human pancreatic ductal adenocarcinomas and chronic pancreatitis by fluorescence in-situ hybrydization (FISH) technique, and to analyze the relationship between 18q21 LOH and clinicopathologic characteristics.

METHODSRP11-729G3 and RP11-850A17, the regions on 18q21, were selected as the target fragments, the region RP11-621L6, close to the centromere of chromosome 18, was selected as the reference fragment. The specific BAC clones were used to isolate and purify the corresponding genomic DNA, which were labeled with biotin or DIG by nick translation into dual color probes. 18q21 LOH was assessed by dual-color FISH in 30 cases of pancreatic ductal adenocarcinoma and 10 cases of chronic pancreatitis. All samples were 10% formalin fixed and paraffin embedded. The relationship between 18q21 LOH and clinicopathologic characteristics was analyzed.

RESULTSAmong 30 cases of pancreatic ductal adenocarcinoma, 25 cases showed LOH at the region RP11-729G3 (83.3%), and 26 cases showed LOH at the region RP11-850A17 (86.6%). Among these, 25 cases with LOH at both regions, 1 case showed LOH only at the region of RP11-850A17. No LOH was found in 10 cases of chronic pancreatitis.

CONCLUSIONS18q21 LOH is a high-frequency event in human pancreatic ductal adenocarcinomas. LOH at the regions RP11-729G3 and RP11-850A17 demonstrates a high concordance. 18q21 may play an important role during pancreatic carcinogenesis and tumor progression. 18q21 LOH may be used as a diagnostic marker for pancreatic ductal adenocarcinoma.

Adenocarcinoma ; classification ; genetics ; Adult ; Aged ; Carcinoma, Pancreatic Ductal ; classification ; genetics ; Chromosome Mapping ; Chromosomes, Human, Pair 18 ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Loss of Heterozygosity ; genetics ; Male ; Middle Aged ; Pancreatic Neoplasms ; classification ; genetics ; Pancreatitis, Chronic ; classification ; genetics

OBJECTIVETo investigate the changes of topoisomerase IIalpha (TOP2A) and HER2/neu genes in pancreatic ductal adenocarcinomas of Chinese patients, and to determine their roles during carcinogenesis and tumor progression.

METHODSExpressions of TOP2A and HER2/neu proteins were detected by using immunohistochemistry, while gene amplifications of TOP2A and HER2/neu were assessed by using multi-color fluorescence in situ hybridization (FISH). All the samples were of paraffin embedded and 10% formalin fixed tissue, including 26 cases of pancreatic ductal adenocarcinomas with adjacent non-neoplastic pancreatic tissues, 10 cases of chronic panreatitis, and 10 cases of normal pancreas. The correlation between TOP2A and HER2/neu gene status was analyzed.

RESULTSBy immunohistochemistry, the nuclear positive index of TOP2A in pancreatic ductal adenocarcinomas varied from 0.5% to 70%, and the positive rate of HER2/neu in pancreatic ductal adenocarcinomas was 46.2% (12/26). By FISH, 9/10 TOP2A amplified adenocarcinomas showed TOP2A and HER2/neu gene coamplification, while one case with HER2/neu gene amplification adenocarcinoma showed no TOP2A amplification. No expression of TOP2A, HER2/neu proteins and no amplification of TOP2A and HER2/neu gene were detected in adjacent non-neoplastic pancreatic tissues, chronic pancreatitis tissues and normal pancreas. No relationship was found between protein expression and gene amplification of TOP2A and HER2/neu (P > 0.05). TOP2A gene amplification was significantly correlated with HER2/neu gene amplification (P < 0.01).

CONCLUSIONSProtein expression of TOP2A and HER2/neu are not associated with the gene amplification. There is a significant correlation between TOP2A amplification and HER2/neu gene amplification. Co-amplification of TOP2A and HER2/neu may play an important role in the carcinogenesis and progression of pancreatic carcinoma. Evaluation of the status of TOP2A and HER2/neu may be helpful to achieve target therapy of pancreatic carcinoma.

Adenocarcinoma ; genetics ; metabolism ; pathology ; secondary ; Adult ; Aged ; Antigens, Neoplasm ; genetics ; metabolism ; Carcinoma, Pancreatic Ductal ; genetics ; metabolism ; pathology ; secondary ; DNA Topoisomerases, Type II ; genetics ; metabolism ; DNA-Binding Proteins ; genetics ; metabolism ; Female ; Gene Amplification ; Gene Expression Regulation, Neoplastic ; Genes, erbB-2 ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Liver Neoplasms ; metabolism ; secondary ; Lymphatic Metastasis ; Male ; Middle Aged ; Pancreatic Neoplasms ; genetics ; metabolism ; pathology ; Poly-ADP-Ribose Binding Proteins ; Receptor, ErbB-2 ; genetics ; metabolism

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