Child, Preschool ; Heart Septal Defects, Ventricular ; complications ; Humans ; Leriche Syndrome ; complications ; Male

Objective To compare the esophageal acid and alkaline exposure characteristics of patients with cardia carcinoma after proximal gastrectomy or total gastrectomy.Method A total of 77 patients of cardia carcinoma who underwent radical resection from Sep 2007 to Sep 2011 in our hospital were retrospectively reviewed.24 hour esophageal pH monitoring were performed in all patients.Result Patients were divided into three groups:group TG had total gastrectomy (n =25),group PP had proximal gastrectomy with pyloroplasty (n =33),group NP had proximal gastrectomy (n =19).It revealed that indicators of acid reflux including the overall time length of acid episodes,＞ 5 min times of acid episodes,duration of longest acid episodes,time length of pH ＜ 4.00 and the DeMeester Scores in group NP were significantly higher than in group PP(U =32,P ＜ 0.01 ; U =35,P ＜ 0.01 ; U =23,P ＜ 0.01 ; U =39,P ＜0.01 ;U =49,P ＜0.01 respectively).Only alkline reflux was observed in group TG.The total times of alkaline episodes in PP group was significantly lower than in group TG(U =52,P ＜0.01) and group NP (U =182,P ＜0.01).＞5 min times of alkaline episodes in group TG was larger than in group PP,and that in group PP was larger than in group NP(P ＜0.01).Duration of longest alkaline episodes and total period of pH ＞7.00 in group PP was significantly higher than in group TG(U =125,P ＜ 0.01 ; U =143.5,P ＜ 0.01),and that in group TG was higher than in group NP(U =23.5,P ＜ 0.01 ; U =14,P ＜ 0.01).Conclusions Alkaline reflux deserves more attention in evaluating esophageal reflux in patients with cardia carcinoma after resection.Pyloroplasty is not helpful to relieving esophageal acid episodes while causing severe alkaline reflux.

Objective To screen proteins binding with interferon?(IFN?)from human hepatic cDNA libraty by yeast-two hybrid technique.Methods The IFN?gene was amplified by polymerase chain reaction(PCR)and constructed into pGBKT7 vector as the bait plasmid in yeast-two hybrid system3,pGBKT7-IFN?was then transfected into yeast AH109.The transfected yeast were mated with yeast Y187 containing liver cDNA library plasmid in 2?YPDA medium.Diploid yeast was plated on synthetic dropout nutrient medium(SD/-Trp Leu-His-Ade)and synthetic dropout nutrient medium(SD/-Trp-Leu-His-Ade)containing X-?-gal for selecting.After plasmid extracting and en- zyme cutting analysis,the blue colonies were performed sequence analysis,the results were analyzed by bioinformatics.Results IFN?gene was successfully cloned and expressed in yeast cells.Thirty- four positive colonies were obtained using yeast-two hybrid technique.After sequence analysis,eight clones were found may have a binding effect with IFN protein.Conclusions IFN?genes was success- ful cloned and eight proteins that could bind with IFN?protein were also screened.

OBJECTIVETo investigate the methylation status of zonula occludens-1 (ZO-1) gene promoter and its clinical significance in children with stage IV non-Hodgkin lymphoma (NHL) and to provide a basis for further etiological study and early diagnosis of this disease.

METHODSFifty-five children with a confirmed diagnosis of stage IV NHL (40 cases of T-NHL and 15 cases of B-NHL) were selected as the case group, and 20 children with diseases other than hematologic malignancies were selected as the control group. Bone marrow samples were collected from these subjects. Methylation-specific PCR (MS-PCR) was applied to evaluate the methylation status of ZO-1 gene promoter, and the integrated optical density (IOD) was determined. RT-PCR was used to measure the mRNA expression of ZO-1.

RESULTSMS-PCR showed that the methylated bands of ZO-1 gene promoter were found in 39 (70.9%) of 55 patients in the case group before treatment, while no ZO-1 gene promoter methylation was detected in the control group. With close tracking of 47 cases in the study group, consisting of 32 cases of T-NHL and 15 cases of B-NHL, the rates of ZO-1 gene promoter methylation prior to treatment were 72% and 67%, respectively, (P>0.572). The cases of T-NHL and B-NHL showed no significant changes in methylation rate in the early and middle phases of chemotherapy (P>0.05), but they showed significant changes in methylation rate in the late phase of chemotherapy (P<0.05). RT-PCR showed that the NHL cases carrying methylated ZO-1 gene had no mRNA expression of ZO-1, while all children in the control group had mRNA expression of ZO-1. There was no linear relationship between the total number of peripheral blood leukocytes and ZO-1 gene IOD (r=0.093, P=0.575); a positive correlation was found between the number of malignant cells in bone marrow and ZO-1 gene IOD (r=0.669, P<0.001).

CONCLUSIONSZO-1 gene shows a hypermethylation status in children with NHL, and the methylation level is positively correlated with the number of malignant cells in bone marrow. ZO-1 may be used as a novel molecular marker in early diagnosis, outcome assessment, prognostic evaluation, and detection of minimal residual disease.

Adolescent ; Child ; Child, Preschool ; DNA Methylation ; Female ; Humans ; Infant ; Lymphoma, Non-Hodgkin ; genetics ; Male ; Promoter Regions, Genetic ; Zonula Occludens-1 Protein ; genetics

To investigate the morphogenetic process of human adenovirus type 41 (HAdV-41), 293TE cells were infected with purified wild-type HAdV-41, and ultrathin sections of infected cells were prepared and observed under a transmission electron microscope. Results showed that HAdV-41 entered host cells mainly through three ways: non-clathrin-coated pit, clathrin-coated pit, and direct penetration of plasma membrane. In addition, cell microvilli might help HAdV-41 enter cells. After entering into cells, HAdV-41 virus particles could be found in vacuoles or lysosomes or be in a free state in cytoplasm. Only free virus particles could be found near nuclear pores (NP), suggesting that the virus needed to escape from lysosomes for effective infection and viral nucleoprotein entered the nucleus through NP. Progeny viruses were as-sembled in the nucleus. Three types of inclusion bodies, which were termed as fibrillous inclusion body, condense inclusion body, and stripped condense inclusion body, were involved in HAdV-41 morphogenesis. In the late phase of viral replication, the membrane integrity of the infected cells was lost and viral particles were released extracellularly. This study reveals the partial process of HAdV-41 morphogenesis and provides more biological information on HAdV-41.
Adenovirus Infections, Human ; virology ; Adenoviruses, Human ; genetics ; growth & development ; physiology ; ultrastructure ; Cell Membrane ; virology ; Cell Nucleus ; virology ; Humans ; Virus Release ; Virus Replication

Multimedia computing technology teaching is a new kind teaching method. It conquers many shortcomings such as poor video, poor expression in traditional teaching of microbiology. But it can also strangle the improvising creation of different teachers in teaching and then lead it to be typical " computer teaching" . So we should obey the teaching discipline in making multimedia computing technology courseware. It can lead many sections part from the class teaching if just emphasize the full use of video and audio. We should make the purpose obviously, and make it as easy as possible in teaching, and we should also explore the fixed regularity to make the teaching and studying integrated perfectly.

OBJECTIVETo compare the efficacy and safety between cryoablation (Cryo) and radiofrequency (RF) ablation for treating patients with atrioventricular nodal reentrant tachycardia (AVNRT).

METHODSPatients with AVNRT (n = 304) were divided into Cryo group (n = 67) and RF group (n = 237). The procedure success rate, complete slow pathway block rate, atrioventricular block rate and relapse rate were compared between two groups.

RESULTSThere was no statistically difference between 2 groups in the success rate (Cryo group 98.5% vs RF group 97.0%, P = 0.820), complete slow pathway block rate (Cryo group 98.5% vs RF group 91.6%, P = 0.088), atrioventricular block rate (Cryo group 0 vs RF group 2.5%, P = 0.413), relapse rate (Cryo group 0 vs RF group 1.7%, P = 0.643). But Cryo group had more advantage than RF group.

CONCLUSIONEfficacy and safety were comparable between cryoablation and radiofrequency ablation for treating patients with AVNRT.

Adolescent ; Adult ; Aged ; Catheter Ablation ; methods ; Child ; Cryosurgery ; methods ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tachycardia, Atrioventricular Nodal Reentry ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the effect of Hedgehog (HH) pathway on proliferation and in vitro tumorigenicity of gastric cancer cell lines.

METHODSThe expression of SHH, PTCH, SMO, SUFU and GLI1 in seven cell lines were tested by RT-PCR. siRNA targeting GLI1 mRNA was transfected into MKN28 cells. Cell proliferation and in vitro tumorigenicity were examined by CCK8 and soft agar colony formation test.

RESULTSSHH in six gastric cancer cell lines was up-regulated. Expression of PTCH in KATOIII cell lines and expression of SUFU in MKN28 and KATOIII were reduced. GLI1 siRNA significantly inhibited the expression of GLI1 in MKN28 cell line. Growth rate and colony formation rate of MKN28 cells treated with GLI1 siRNA were significantly lower than those of control cells (all P <0.001).

CONCLUSIONSHH signaling pathway is widely activated in gastric cancer cell lines. The activation of HH signaling pathway promotes the growth of MKN28 cells.

Cell Line, Tumor ; Cell Proliferation ; Gastric Mucosa ; cytology ; Hedgehog Proteins ; metabolism ; Humans ; Oncogene Proteins ; metabolism ; RNA, Small Interfering ; Signal Transduction ; Stomach Neoplasms ; metabolism ; pathology ; Trans-Activators ; metabolism ; Zinc Finger Protein GLI1

Three-dimensional (3D) culture of cells could closely mimic the in vivo situation with regard to cell function and microenvironment compared with plane monolayer cultured cells. In this paper, we established 3D culture of rat WB-F344 cells with rotary cell culture system (RCCS) to simulate microgravity environment, and examined cells proliferation, morphology, microstructure, E-cadherin protein quantity and mRNA expression of adhesion molecules by count the number of cells, optical microscope, transmission electron microscope and reverse transcriptase-polymerase chain reaction (RT-PCR). The results demonstrated that cells were polyhedron with lots of micovilli and mitochondria, which grow well and packed together densely to form irregular aggregates. Adjacent cells were connected with desmosome and tight junction. With the regard, the aggregates behaved 3D growth characteristics. Moreover, compared with control, mRNA level of Fibronectin and E-cadherin protein were increased, the changes maybe is the part mechanism in this microgravity simulated cells culture models which strengthened cells junction. This rotating 3D model might facilitate the study of interactions of cell-cell, cell-matrix and the mechanisms.
Animals ; Cadherins ; genetics ; Cell Adhesion ; Cell Culture Techniques ; methods ; Cell Proliferation ; Fibronectins ; genetics ; Rats ; Spheroids, Cellular ; ultrastructure ; Weightlessness Simulation

OBJECTIVETo explore the effects of hepatitis B virus X gene and p53 on hepatocellular growth.

METHODSTwo kinds of plasmids containing sense and antisense human wild p53 gene respectively were constructed. SMMU-7721 cells were transfected with HBx, sense-wtp53 antisense-wtp53 separately or cotransfected with either HBx and sense-wtp53 or HBx and antisense-wtp53. Flow cytometry was adopted to measure the apoptosis rates and the effects of HBx on cell cycle progression. The activity of p21(Waf1) promoter-luciferase construct was detected. Growth curves for SMMU-7721 stably transfected with pcDNA3 and pcDNA3HBx were analyzed.

RESULTSAfter doxorubicin administration, HBx was noticed able to initiate apoptosis of the liver cells. The apoptosis rate was 5.32% in the pcDNA3 transfected and 12.66% in the pcDNA3HBx transfected groups respectively. HBx could also abrogate p53-mediated apoptosis. The apoptosis rate in groups transfected with pcDNA3, pcDNA3wtp53 and pcDNA3HBx + pcDNA3wtp53 was 5.32%, 11.72% and 4.67% respectively. In compared with the normal group, the number of cells in transiently HBx-expressed group and HBx-transfected group decreased 4.79% and 10.25% respectively. HBx inhibited the activity of p21(Waf1) promoter-luciferase constructed (P < 0.05) and promoted cell growth. The growth rate of HBx expression cells was faster.

CONCLUSIONUnder DNA damage, HBx reduced expression of p21(Waf1) by repressing the activity of p53 protein, followed by disturbing the regulation of G(0)-G(1) cell cycle checkpoint, and promoted the growth rate of hepatoma cells.

Apoptosis ; Carcinoma, Hepatocellular ; pathology ; virology ; Cell Division ; Cell Line, Tumor ; Genes, p53 ; Hepatitis B Antigens ; biosynthesis ; genetics ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; pathology ; virology ; Trans-Activators ; biosynthesis ; genetics ; Transfection ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics