HER2, a member of epidermal growth factor receptor family proteins, is overexpressed in about 30% of human breast cancer. Increased levels of HER2 are associated with poor patient prognosis and enhanced metastasis. RNA interference (RNAi) is developed recently as a new technique which can inhibit gene expression specifically in mammalian cells. On the basis of previous study,in which two target sequences with favorable RNAi effect on HER2 were identified, a series of dual promoter siRNA-expressing vectors containing two opposing U6 and H1 promoters were constructed. After transfection of HER2-overexpressing SKBR3 breast cancer cells with the siRNA-expressing vectors, downregulation of HER2 was identified quantitatively. Subsequently, the siRNA-expressing cassettes were subcloned into lentiviral vectors by LR recombination reaction and lentivirus was prepared successfully. The results from infection of SKBR3 cells with siRNA-expressing lentivirus demonstrated that lentiviral-mediated RNAi could downregulate HER2 expression efficiently through fluorescent quantitative PCR (FQ-PCR), western blot, and FACS analysis. Furthermore, cell growth was inhibited in cell proliferation assay after treatment with siRNA lentivirus.A new tool for clarifying the function of HER2 in cancer metastasis and developing the gene therapy drug was offered.

Objective To investigate the association between high-frequency oscillatory ventilation (HFOV) and the incidence of brain injury in premature infants(BIPI) with respiratory distress syndrome (RDS).Methods A total of 136 premature infants who were diagnosed as RDS and treated with mechanical ventilation between January 1,2014 and June 30,2016 were enrolled.Atter stratified by gestational age of 36-34 weeks,33-32 weeks,31-28 weeks and ≤ 27 weeks,the neonates were randomly divided into two groups (68 cases each):conventional mechanical ventilation (CMV) and HFOV groups.Duration of ventilation and changes in blood gas parameters following 24,48 and 72 hours of ventilation were monitored and compared between the two groups.Incidences of BIPI and complications in the two groups were calculated and their associations with gestational age and birth weight were analyzed.Moreover,incidences of cure rates in the two groups were comparatively analyzed.Independent samples t-test,two-way analysis of variance,Chi-square test or Fisher's exact test was used for statistical analysis.Results (1)Significant difference was observed neither in the perinatal factors (prenatal glucocorticoid usage,and incidences of premature rupture of membrane and gestational diabetes mellitus),nor in the severity of RDS between the two groups (all P＞0.05).(2) The average duration of ventilation in the CMV group was higher than that of the HFOV group [(68.44±10.3) vs (64.7±8.5) h,t=2.285,P＜0.05].No significant difference in the values of pH,partial pressure of carbon dioxide (PaCO2) or partial pressure of oxygen (PaO2) before and after 24,48 or 72 hours of ventilation treatment was found between the two groups (all P＞0.05).(3) Neither the incidence of hemorrhagic brain injury nor that of non-hemorrhagic brain injury showed any significant difference between the CMV and HFOV groups [36.8％ (25/68) vs 39.7％ (27/68);16.2％ (11/68)vs 14.7％ (10/68),both P＞0.05].The total incidence of BIPI showed no significant difference [44.1％(30/68) vs 45.8％(33/68),22=0.266,P=0.606].The smaller gestational age at birth and the lower birth weight,the higher incidence of BIPI,although no significant difference was shown in the incidence of BIPI when compared among different gestational age groups and different birth weight groups (all P＞0.05).(4) The incidence of complications in the CMV group was higher than that in HFOV group [25.0％(17/68) vs 11.8％(8/68),22=3.970,P=0.044],while the cure rate of RDS was similar [94.1％(64/68) vs 95.6％(65/68),x2=0.151,P=0.703].Conclusions HFOV is a safe and reliable therapy for preterm infants with RDS.Compared with CMV,HFOV can shorten the duration of ventilation and reduce the incidence of complications without increasing the risk of BIPI.However,the cure rate of RDS is not increased by HFOV.

Animal model is an animal material with human mimic performance established in biomedical scientific research. It can be used as experimental basis for studies of experimental hypothesis and clinical hypothesis. It can shorten the research time and observe the whole process of disease occurrence, development or prevention and treatment. Human biomedical research is largely limited by the biological complexity. In order to overcome this limitation, based on the immunosuppressive characteristics of a severely immunodeficient ( SCID) or recombinant activated gene ( Ragnul ) in mice, humanized mouse models of human diseases can be established and have been widely used to study the underlying principles of human immunobiology and complex pathological mechanisms of human diseases. This approach has become one of the important ways to promote the development of medical sciences, with practicality and foresight. In this paper, the application and research progress of humanized mouse models are reviewed.

OBJECTIVETo evaluate the long-term efficacy and safety of endoscopic tissue adhesive(N-octyl-α-cyanoacrylate) injection for the treatment of gastric varices.

METHODSA retrospective study was performed to review the clinical and follow up data of 169 patients with gastric variceal who received tissue adhesive injection at the Fudan University Affiliated Zhongshan Hospital between April 2004 and December 2011.

RESULTSThere were 128 males and 41 females with a mean age of 56.8(37-85) years old. One hundred and thirty-one patients received one injection, 38 received two injections or more with a mean of 1.12 per patient. Volume of injection ranged from 1 to 3 ml(mean, 1.7 ml). Eighty-three patients received adhesive injection alone, 231 received injection combined with ligation, 50 received combined sclerotic agent injection. All the patients had follow up ranging from 1 to 78 months(mean, 3.4 months). The treatment outcome was satisfactory in 130 patients(76.9%), good in 36(21.3%), and ineffective in 3(1.8%). The rate of ectopic embolization was 3.0%, and the rate of early re-bleeding was 1.2%. Postoperatively there were no septic complications or esophageal stricture. There were no deaths within 2 weeks.

CONCLUSIONInjection of tissue adhesive under endoscopic guidance for treatment of gastric varices is convenient, safe and effective.

Adult ; Aged ; Aged, 80 and over ; Esophageal and Gastric Varices ; therapy ; Female ; Follow-Up Studies ; Gastroscopy ; Humans ; Injections, Intralesional ; Male ; Middle Aged ; Retrospective Studies ; Tissue Adhesives ; administration & dosage ; therapeutic use ; Treatment Outcome

·AIM: To observe the therapeutic effect of visual perception training combined with total nutrition meal sequential therapy in the treatment of myopic amblyopia.·METHODS:Totally 73 children ( 135 eyes ) with myopic amblyopia were divided into control group ( 36 cases, 67 eyes) and treatment group (37 cases, 68 eyes) according to random number table. The control group were treated with traditional spectaculars and grating covering combined with fine eyesight training;the treatment group were treated with visual perception training combined with total nutrient meal sequential therapy. The visual acuity, diopter and average diopter of two groups were compared before and after treatment at 3, 6mo and 1a. The curative effect of two groups of children was compared after 1a treatment. And the adverse reactions were recorded in two groups during the treatment period. The recurrence rate of amblyopia in 1a follow-up was compared between two groups.·RESULTS: The difference of visual acuity between two groups was not significant at 3mo (P>0. 05). The visual acuity of the treatment group was significantly higher than that of the control group at 6mo and 1a (P<0. 05). There was no significant difference in diopter between the two groups after 3, 6mo and 1a (P>0. 05), but the average annual refractive changes in the treatment group were significantly lower than that in the control group ( P<0. 05). The basic cure rate and total effective rate of the treatment group were significantly higher than that of the control group (P<0. 05). There were no severe adverse reactions occurred between two groups during the treatment period. The recurrence rate of amblyopia in the treatment group was significantly lower than that in the control group (P<0. 05) after 1a follow-up.· CONCLUSION: Visual perception training combined with total nutrition meal sequential therapy in the treatment of myopic amblyopia in preschool children can significantly improve patients' visual acuity, reduce the average annual diopter changes, improve the therapeutic effect, reduce the recurrence rate of amblyopia.

OBJECTIVETo study the effect of resveratrol bovine serum albumin nanoparticles on SKOV3 cell line and its mechanisms.

METHODSThe morphological changes of the cells exposed to the nanoparticles were observed by apoptotic body/cell nucleus DNA staining under inverted microscope and fluorescence microscope, and the pathway of cell death was determined by phosphatidylserine translocation. Western blotting was performed to detect the activation of cyto.c, caspase-3 and caspase-9.

RESULTSDNA ladder was detected with gel electrophoresis and the cell death was partially inhibited by the pan-caspase inhibitor Z-VAD-FMK. Gel electrophoresis displayed both DNA ladder and smear in RES-BSANP exposed groups, while DNA ladder disappeared in Z-VAD-FMK group and only the smear was left. Cyto.c in the cytoplasm was released at 2 h, while the expression of caspase-9 protein reached the peak level at 4 h and caspase-3 expression was obvious enhanced at 8 h. At 4 h, caspase-9 expression in the cells exposed to 100 µmol/L RES-BSANP was decreased significantly as compared to the cells treated with 50 µmol/L RES-BSANP (P>0.05).

CONCLUSIONRES-BSANP can induce the necrosis and apoptosis of SKOV3 cells via either caspase-dependent or caspase-independent pathways.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Death ; drug effects ; Cell Line, Tumor ; Cytochromes c ; metabolism ; Female ; Humans ; Nanoparticles ; Ovarian Neoplasms ; pathology ; Stilbenes ; administration & dosage ; pharmacology

OBJECTIVETo investigate the role of tissue factor (TF) expression in the invasive and metastatic ability of colorectal carcinoma and explore the influence of TF on the invasive ability of HT-29 cells.

METHODSTF expression of specimens from 85 colorectal carcinomas and 6 colorectal adenomas was observed by immunohistochemistry. The role of TF expression in prognosis and tumor invasion and metastasis was analyzed. The plasmids pcDNA3.1/Zeo bearing either sense or antisense-TFcDNA were transfected into HT-29 cells by the way of Lipofectamine 2000. TF proteins in transfected and untransfected HT-29cells were detected by Western blot. In vitro Matrigel invasion assays were performed to show the invasive ability of those cells.

RESULTSTF expression was positive in 40 (47.1%) of 85 colorectal carcinoma specimens, but negative in normal mucosa and adenoma specimens. TF expression showed significant correlation with tumor invasive depth (r = 0.895, P < 0.01). TF expression showed significant correlation with synchronous and metachronous hepatic metastasis (r = 0.974, P < 0.01 and r = 0.963, P < 0.01 respectively). TF expression was a significant risk factor for hepatic metastasis (P < 0.01) and prognosis (P < 0.01). TF expression in HT-29 cells with sense/antisense-TFcDNA transfection was more/less than that of the cells without transfection. The invasive ability of HT-29 cells with sense-TFcDNA transfection was increased in vitro compared with the untransfected cells, but HT-29 cells with antisense-TFcDNA transfection got the contrary change.

CONCLUSIONSTF may take part in the invasive and metastatic process of primary colorectal carcinoma, and TF expression may be an indicator of hepatic metastasis and prognosis for colorectal carcinoma patients. TF expression may increase the invasive ability of HT-29 cell in vitro.

Blotting, Western ; Cell Movement ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; HT29 Cells ; Humans ; Immunohistochemistry ; Logistic Models ; Multivariate Analysis ; Thromboplastin ; analysis ; genetics

OBJECTIVETo investigate the changes of p38MAPK expression in a rat model of Alzheimer disease (AD).

METHODSSeventy-two adult SD rats were randomized equally into 4 groups, and a single-dose injection of Abeta25-35 (dementia group), normal saline (saline group), SB203580 (inhibitor group), or DMSO (inhibitor control group) was administered into the lateral cerebral ventricle. Y-maze tast was performed to evaluate the behavioral changes of the rats after the injections, and on days 4, 7 and 14 after the injection, p38MAPK expression in the hippocampal CA1 area was measured by means of immunohistochemistry.

RESULTSOn days 7 and 14 following Abeta25-35 injection, the training times, error number and total reaction time were significantly higher in dementia group than in saline group (P<0.05), but all these indices were significantly lowered in the inhibitor group as compared with the dementia group (P<0.05). Immunohistochemistry revealed obvious p38 expression in the dementia group 4 days after Abeta25-35 injection, which increased significantly with the passage of time (P<0.01). The gray scale in the inhibitor group was significantly higher than that in the dementia group (P<0.01).

CONCLUSIONp38MAPK activation in the hippocampal CA1 area is an event that persists during the entire course of Abeta25-35-induced AD in rats, and the inhibitor SB203580 prevents p38MAPK expression and improves the learning and memory abilities of the rats.

Alzheimer Disease ; chemically induced ; enzymology ; metabolism ; Amyloid beta-Peptides ; administration & dosage ; toxicity ; Animals ; Hippocampus ; drug effects ; enzymology ; Immunohistochemistry ; Male ; Maze Learning ; drug effects ; Peptide Fragments ; administration & dosage ; toxicity ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; p38 Mitogen-Activated Protein Kinases ; biosynthesis

A switch thrombin aptamer sensor was constructed based on the host-guest competition mode ofβ-cyclodextrin (β-CD ) . The aptamer that modified with ferrocene ( Fc ) at its terminal was fixed on the surface of the gold electrode via the host-guest recognization with β-CD. When thrombin was present, the configuration of aptamer transformed from vertical linear to "G-quadruplex" and far away from the electrode surface, which resulted in a decrease in the redox current of the aptamer probe and produce "Signal-off"effect. On the basis of this, a high sensitive detection of thrombin was made. The detection result indicated that the thrombin concentration had a consideration linear response to the signal of aptasensor between 5 . 0 × 10-13-5. 0 × 10-9 mol/L, and as low as 2. 0 × 10-13 mol/L thrombin had been detected. This method for thrombin detection showed a higher specificity compared to other protein molecules. Besides, the sensor was constructed easily and possessed excellent regeneration, which provided a significant platform for highly efficient real-time detection of thrombin in biological serum samples.

OBJECTIVETo investigate the role of tissue factor(TF) in hematogenous metastasis of human colorectal carcinoma cells (LOVO) in vivo.

METHODSThe eukaryotic expression vectors pcDNA3.1/Zeo bearing either sense or antisense TFc DNA were transfected into LOVO cells by lipofectamine 2000. TF protein expression in the transfected cells was detected by Western blot. Eighteen nude mice (Babl/c nu/nu) were randomly divided into three groups, and then transfected and untransfected LOVO cells were implanted via tail vein respectively. The nude mice were sacrificed 8 weeks after implantation, and the number of metastatic nodules in the lung was used to assess the metastatic ability of LOVO cells.

RESULTSCompared with the untransfected group, TF expression of LOVO cells and the numbers of metastatic nodules in the lung increased in sense-TF cDNA transfection group (P< 0.05, P< 0.01, respectively), whereas decreased in antisense-TF cDNA transfection group (P< 0.05, P< 0.01, respectively).

CONCLUSIONTF can increase the hematogenous metastatic ability of human colorectal carcinoma cells (LOVO) in vivo.

Animals ; Cell Line, Tumor ; Colorectal Neoplasms ; metabolism ; pathology ; DNA, Complementary ; genetics ; Humans ; Lung Neoplasms ; metabolism ; secondary ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Thromboplastin ; genetics ; metabolism ; Transfection