ObjectiveTo investigate whether the eight- year program students retain the skills from the endoscopy simulator gastroscopy training.Methods4 trainees accepted virtual reality simulator gastroscopy training and performed a standardized VR gastroscopy scenario at the end of training,and after a median 12 months without practice ( retention ).The intensified training was done by trainees based on the differences between the training end and the retention for a median 12 months and the number of intensified training times was found.ResultsThe significant differences existed in the overinsufflation and opeirational force and time.The score at the training end was better than after retention.Through the average 5.5 times intensified trainings the original levels could be reached.ConclusionThrough Endoscopy Simulator the key skills could be retained well and through a litde training the original levels could also be reached.

OBJECTIVETo investigate the anti-tumor effect of dendritic cells (DC) pulsed with G422 glioblastomas RNA in mice bearing intracranially G422 glioblastomas.

METHODSDCs were pulsed in vitro with glioblastomas G422 cell RNA. The tumor-bearing mice were injected intratumorally or subcutaneously with pulsed DCs, PBS, non-pulsed DCs. The survival duration of mice was recorded. Serum levels of cytokine IFN-gamma, IL-2, IL-10, IL-4 were detected. Pathological examination was performed.

RESULTSThe survival duration of mice with DC-based vaccine increased significantly(P<0.01). The serum IFN-gamma level was increased (P<0.01) and IL-10 level was decreased (P<0.05) after treatment. Pathological examination showed necrotic tumor in the treatment mice.

CONCLUSIONDC vaccination can significantly increase survival duration of mice with intratumoral or subcutaneous administration of vaccines.

Animals ; Brain Neoplasms ; immunology ; therapy ; Cancer Vaccines ; immunology ; Dendritic Cells ; immunology ; transplantation ; Glioblastoma ; immunology ; therapy ; Immunotherapy ; methods ; Mice ; RNA, Neoplasm ; immunology ; Random Allocation ; T-Lymphocytes, Cytotoxic ; immunology

Intraoperative awareness is a very serious complication of general anesthesia.Several studies have evaluated the potential association between bispectral index (BIS) and intraoperative awareness,however,the results obtained were controversial.Therefore,we performed a meta-analysis to further assess the association between the BIS monitoring and the incidence of intraoperative awareness.A comprehensive search was conducted to identify all eligible studies from the online literature databases published prior to Feb.2017.A total of five studies with 17 432 cases and 16 749 controls were included.An odds ratio (OR) and a 95％ confidence interval (CI) were calculated to examine the strength of the association.The results showed that in the overall analysis,the association between the BIS monitoring and the incidence of intraoperative awareness was not significant (OR=0.58,95％ CI=0.22-1.58,P=0.29).A stratified analysis by comparing different anesthesia methods revealed that BIS monitoring group showed a lower incidence of intraoperative awareness in patients with intravenous anesthesia when compared with non-BIS monitoring group (OR=0.20,95％ CI=0.08-0.49,P=0.0004),whereas there was no statistically significant difference in the incidence of intraoperative awareness between BIS and non-BIS monitoring groups in patients with inhalation anesthesia (OR=1.13,95％ CI=0.56-2.26,P=0.73).In conclusion,our meta-analysis showed that BIS monitoring had no appreciable advantage in the reduction of the intraoperative awareness incidence in inhalation anesthesia,while showed a remarkable superiority in intravenous anesthesia.

OBJECTIVE: To investigate the regulation of hepatitis C virus (HCV) core protein on the activity of signal transducers and activators of transcription 3 (stat3). METHODS: A cell line expressing stable HCV core protein-QSG7701-core was constructed by transfecting the pcDNA3. 1-core (expressing HCV core protein) into the human immortalized hepatocyte line QSG7701. The phosphorylation and DNA binding activity of stat3 were detected by immunocytochemistry, Western blot, and electrophoretic mobility shift assay (EMSA). RESULTS: The expression level of phosphorylated stat3 in QSG7701-core cells was significantly lower than that in QSG7701-pcDNA3. 1 cells and untransfected QSG7701 cells, but there were no significant differences in the expression levels of total stat3 among the 3 groups. The positive signal of phosphorylated stat3 in nucleus of QSG7701-core cells was obviously weaker than that in QSG7701-pcDNA3. 1 cells and untransfected QSG7701 cells. EMSA showed that DNA binding activity of stat3 in QSG7701-core cells significantly decreased. CONCLUSION The expressionof HCV core protein in human hepatocyte line may suppress the phosphorylation and DNA binding activity of stat3, which may be one of the causes for resistance against interferon.
Cell Line ; DNA-Binding Proteins ; metabolism ; Hepatocytes ; cytology ; metabolism ; Humans ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Transfection ; Viral Core Proteins ; biosynthesis ; genetics

Most drugs need to interact with cell membrane to reach the biological target, so that membrane affinity assay is an important early screening step in drug discovery. However, at present, the traditional oil-water distribution method is still used, a new, simple and accurate method for membrane affinity assay is urgently needed. In this study, according to the colorimetric principle, a new assay model based on polydiacetylene vesicles was optimized through a series of experiments including different concentrations of vesicle solution, temperature, or pH reaction environment. On this basis, tetracaine hydrochloride, 2-methylimidazole and histamine were used as model drugs to measure the membrane affinity constants and verify the between-batch precision of the optimized assay model (relative standard deviation less than 5%). In addition, polydiacetylene vesicles were stable for up to 180 days, demonstrating the potential application of the assay model. This strategy is simple, stable, reliable, with high reproducibility, low cost and easy to promote, which provided a new tool and a new direction for the high-throughput assay of membrane affinity.

OBJECTIVE: To compare effects of three different methods for mtDNA extraction from common sarcosaphagous insects including cetyl trimethyl ammonium bromide (CTAB) method, sodium dodecyl sulfate-potassium acetate (SDS-KAc) method and sodium dodecyl sulfate-proteinase K (SDS-PK) method. METHODS: Seventy-two insects from four species [Chrysomya megacephala (Fabricius, 1784), Eusilpha bicolor (Fairmaire, 1896), Paraeutrichopus pecoudi (Mateu, 1954), Vespa velutina (Lepeletier, 1836)] were collected from the corpses of the rabbits in Changsha district. The total DNA of above samples was extracted by CTAB, SDS-Kac and SDS-PK methods. The purity and concentration of DNA were examined by protein-nucleic acid spectrophotometry, and mtDNA were amplified by specific primers and PCR products were detected by agarose gel electrophoresis. Then PCR products were sequenced and subsequently up-loaded to GenBank. RESULTS: mtDNA was successfully extracted with three methods from most of the samples. The SDS-PK method was better in DNA purity compared to other methods and the CTAB method was superior in extracting DNA from old samples, while SDS-KAc method showed no significant difference for extraction effects of different samples. CONCLUSION The most appropriate method should be chosen depending on different situations. SDS-PK method is expected to obtain high-quality DNA, while CTAB method is preferred in extracting obsolete samples. SDS-KAc method is low cost and can be used in various kinds of preliminary experiments.
Animals ; Coleoptera/genetics* ; DNA Primers ; DNA, Mitochondrial/isolation & purification* ; Diptera/genetics* ; Electrophoresis, Agar Gel ; Entomology ; Forensic Medicine/methods* ; Gene Amplification ; Insecta/genetics* ; Polymerase Chain Reaction/methods* ; Quaternary Ammonium Compounds/chemistry* ; Rabbits ; Reproducibility of Results ; Sequence Analysis, DNA ; Sodium Dodecyl Sulfate/chemistry*

Objective To investigate the clinical value of detecting the serum level of ryanodine receptor antibodies (RyR-Ab) in the diagnosis and evaluation of myasthenia gravis (MG). Methods Enzyme-linked immunosorbent assay (ELISA) and Western blotting were used to detect the serum level of RyR-Ab in 81 patients with MG, 45 non-MG patients with other neurological disorders and 50 healthy control subjects. Results The results of ELISA showed a serum RyR-Ab positivity rate of 23.4% in the in MG patients, significantly higher than the rates in non-MG patients and the healthy subjects (P<0.05). The serum RyR-Ab positivity rate was 77.2% in MG patients with thymoma (MGT), 14.2% in those with thymie atrophy (MGA), and 6.6% in those with thymie hyperplasia (MGH). A positive correlation was found between the titer of serum RyR-Ab and the severity of muscle weakness (r=0.547, P<0.05). Compared with CT scanning, serum RyR-Ab level had low sensitivity (77.2%) but high specificity (91%, P<0.05) for diagnosis of MGT. According to the results of Western blotting, the serum RyR-Ab positivity rate was 81.8% in MGT patients, significantly higher than that in thymomβ-free MG patients and NMG patients (P<0.05). Conclusion RyR-Ab positivity is mainly found in MGT patients, and detection of serum RyR-Ab may help evaluate the severity of MG and assist in CT-based diagnosis of MGT.

OBJECTIVETo investigate the effects of total saponins of panax ginseng (TSPG) on proliferation and differentiation of human embryonic neural stem cell (NSC) into dopaminergic neuron.

METHODIsolation, cultivation and identification of human embryonic NSC from cerebral cortex of 7-12 week abortus. By using flow cytometry and MTT assay, the effects of various concentration of TSPG and TSPG cooperating with cytokines( EGF, bFGF) in NSC culture media for 3 days on proliferation of human embryonic NSC has studied. By employing immunocytochemistry assay of the expression of tyrosine hydroxylase (TH), the effect of different dilution of TSPG and TSPG cooperating with IL-1 on induced differentiation of human embryonic NSC into dopaminergic neuron has researched.

RESULTTSPG can significantly promote the proliferation of NSC. When TSPG cooperating with EGF and bFGF, the proliferation of NSC is much stronger than that of only using FGF and bFGF. TSPG also induces NSC to differentiate into dopaminergic neuron, especially when TSPG is cooperating with IL-1.

CONCLUSIONTSPG can not only obviously accelerate the proliferation of NSC, but also significantly induce differentiation of NSC into dopaminergic neuron.

Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dopamine ; metabolism ; Drug Synergism ; Embryonic Stem Cells ; cytology ; drug effects ; metabolism ; Epidermal Growth Factor ; pharmacology ; Fibroblast Growth Factor 2 ; pharmacology ; Humans ; Immunohistochemistry ; Interleukin-1 ; pharmacology ; Neurons ; cytology ; drug effects ; metabolism ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Saponins ; isolation & purification ; pharmacology ; Tyrosine 3-Monooxygenase ; metabolism

OBJECTIVETo investigate the expression of occludin, ZO-1, MMP-2, and MMP-9 in cerebral microvasculature following Pulse Electromagnetic Field (PEMF) induced BBB permeability change.

METHODSSprague-Dawley rats were randomized into PEMF and sham exposed groups (n = 8). After exposure to PEMF at 0.5, 1, 3, 6, and 12 h, BBB permeability was measured by Evans-Blue extravasation. The expression of occludin, ZO-1, MMP-2, and MMP-9 were detected by real-time quantitative reverse transcriptase PCR and western blotting. MMP-2 and MMP-9 activity were detected by EnzChek gelatinase assay.

RESULTSCompared with the sham group, PEMF exposure led to increased permeability of the BBB to EB, which was prolonged after exposure. BBB permeability became progressively more severe, and recovered at 6 h. The gene and protein expression of occludin and ZO-1 were significantly decreased, while MMP-2 and MMP-9 expression were significantly increased after exposure to PEMF. All levels of expression recovered 12 h following PEMF.

CONCLUSIONChanges to BBB permeability were related to the alteration expression of tight junction proteins and matrix metalloproteinase after exposure to PEMF.

Animals ; Blood-Brain Barrier ; Electromagnetic Fields ; Male ; Matrix Metalloproteinases ; metabolism ; Proteins ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tight Junctions ; metabolism

Female ; Humans ; Infant ; Propionic Acidemia ; diagnosis