Utilizing standardized patient(SP) for classroom simulation is common in current medical education. In this paper, incentive measures and combination of SP with theoretical examination, simulated people and clinical practice were proposed after in-depth analysis of advantages and disadvan-tages of using medical students as SP in terms of recruitment, training, and application. All these mea-sures were intended to promote the development of simulative medical education that in turn to cultivate students to be competent in practice.

Objective To evaluate the design formulas of the optimal location for splitting pectoralis major in terms of making type Ⅰ or Ⅱ dual-plane implant pocket.Methods Sixty-five patients with micromastia were collected.Breasts were divided into two types according to the soft-tissue pinch thickness of the lower pole:type Ⅰ (thickness＞2 cm;34 cases) and type Ⅱ (thickness ≤2 cm;31 cases).The optimal levels at which the pectoralis major (PM) was severed were 3/4 or 2/4 of the distance of new inframammary fold for type Ⅰ or Ⅱ dual-plane pockets,respectively.All patients completed the pre-and post-operative BREAST-Q augmentation modules before and 6 months after surgery.The scores were changed into hundred-mark system by QScore software.Results The recovery processes were well-off.The breasts contour was good.Patients reported higher scores of satisfaction with breasts,psychosocial well-being and sexual well-being after surgery than before surgery (62.0±8.9 vs 27.9± 4.3,65.0± 17.2 vs 17.4±8.3,60.5± 14.2 vs 30.3±5.5,P＜0.05).The mean satisfaction score for the overall outcome was 90.6±5.4.However,there was no significant difference in physical well-being between before operation and aftre operation (85.3±9.5 vs 84.7± 10.6).No complications such as severe capsular contracture,or displacement occurred.Conclusions The design formulas make the determining procedure of the optimal location for pectoralis major splitting for two types dual-plane implant pockets easier and more exactly.Our modified design method can provide the implant with the optimal soft tissue coverage,and bring desired and stable breast aesthetic outcomes.The higher satisfaction and quality of life reported by patients indicate that the formulas are feasible and worth to recommend.

Objective To design a test phantom for clinical routine quality control of magnetic resonance imaging (MRI).Methods The commonly used phantom such as Magphan SMR170 and Victoreen 76-903 were used for experiments with the same parameters,and the test data of all the modules were compared.The advantages and disadvantages of all the modules were analyzed,and an optimization scheme was proposed accordingly.Results The phantom could be applied to the detection of high-image-quality MRI machine,and solved the problems in positioning,size matching and etc.Conclusion The new phantom meets the demand of magnetic resonance imaging clinical routine testing,and provides a certain reference for phantom optimization design.

Cardiac output (CO) monitoring is a crucial part of the hemodynamic status monitoring. So far, thermodilution method, which is clinically recognized as the gold standard method to monitor cardiac output, still has irreplaceable advantages. This paper mainly introduces the use of platform for cardiac output measurement based on thermodilution method, mainly including three parts: the hardware platform, software design and algorithm process. A large amount of test data of this system has been got by CO simulator testing in the laboratory and preliminary clinical tests in the hospital. The testing result showed that using the proposed system can achieve good accuracy and repeatability.
Algorithms ; Cardiac Output ; Hemodynamics ; Humans ; Monitoring, Physiologic ; instrumentation ; methods ; Thermodilution

A new method of using photoactivable ester with azido group was described to immobilize urease on polyether sulfone(PES) film surface. The effects of photoactive enzyme concentration, temperature, pH, irradiation time on the activity of immobilized urease were investigated. Reused times and storage stability were also studied. The results showed that the surface concentration of urease immobilized on PES surface was about 0.33 mg/cm2. When the irradiation time was 5 minutes, the relative activity of immobilized urease was the highest and the activity increased with the increase of the concentration of photoactive urease solution. The optimum pH and temperature of immobilized urease were 7 and 50 degrees C respectively. The relative activity of immobilized urease was stable (50%) after 12 times reused at 50 degrees C.
Enzyme Stability ; Enzymes, Immobilized ; metabolism ; radiation effects ; Membranes, Artificial ; Photochemistry ; Polymers ; Sulfones ; Urease ; metabolism ; radiation effects

OBJECTIVETo explore the HSVtk gene expression mediated by the retroviral vector and to obtain high titer recombinant retroviral virus.

METHODSThe recombinant vector pRevTRE/HSVtk was constructed by inserting HSVtk gene into pRevTRE. The recombinant retrovirus, which was produced from cloned PA317 cells screened by hygromycin B after "micro-pingpong" technique transferring with pRevTRE/HSVtk plasmids DNA by using modified calcium phosphate precipitation method. HSVtk gene expression was performed on target cells and virus titers were detected in different cultured temper, time and sodium butyrate concentration.

RESULTSThe recombinant retroviral vector pRevTRE/HSVtk was constructed and HSVtk gene expression was detected on target cells after they were infected with the recombinant retrovirus.

CONCLUSIONHigh titer of retroviruses could be obtained in the culture medium of PA317 cell line through "micro-pingpong" technique at 30 hours and 10 mmol/L sodium butyrate concentration followed by frozen ultrafiltration.

Animals ; Breast Neoplasms ; enzymology ; pathology ; virology ; Cell Line ; Cell Line, Tumor ; Female ; Gene Expression Regulation, Enzymologic ; Genetic Vectors ; Humans ; Mice ; NIH 3T3 Cells ; Recombination, Genetic ; Retroviridae ; genetics ; Simplexvirus ; enzymology ; genetics ; Thymidine Kinase ; biosynthesis ; genetics ; Titrimetry ; Transfection

In order to investigate the application of recombinant adeno-associated virus (rAAV) vector containing Tet regulation system and HSVtk gene in cancer gene therapy, pAAV/TRE/HSVtk/Tet-On was constructed and identified with PCR and restriction enzyme digestion. Packaging cells HEK293 were cotransfected with plasmids pAAV/TRE/HSVtk/Tet-On, pAAV-RC and pAAV-helper to produce infectious rAAV, and CsCl2 densitygradient centrifugation method was performed for purification and concentration of rAAV. The viruses were then transduced into MCF-7 cells. The results of dot blot hybridization indicate that the rAAV can transfer the target gene into MCF-7 cells. MTT assay showed that GCV could kill AAV-infected MCF-7 cells under the induction of Dox. The data demonstrated that rAAV containing Tet regulation system and HSVtk gene was successfully obtained, and could be used for further investigation of in vivo and in vitro experiments.
Cell Line, Tumor ; Dependovirus ; genetics ; metabolism ; Doxycycline ; pharmacology ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Simplexvirus ; enzymology ; genetics ; Thymidine Kinase ; genetics ; Transfection

BACKGROUNDRevTet-On gene expression system was used to deliver the suicide gene tk to human breast cancer cell line MCF-7 and control the tk gene expression level. The animal model of human breast cancer on severe combined immune deficiency (SCID) mice was set up to explore the suicide gene therapy by the regulation of Tet-On.

METHODSHerpes simplex virus-thymidine kinase (HSVtk) gene was inserted into the plasmid pRevTRE and the recombinant retroviral vector pRevTRE/HSVtk was constructed. Using modified calcium phosphate co-precipitation method, two transfections, pRevTRE/HSVtk and pRevTet-On were performed for MCF-7 cell line and selected by hygromycin B and G418. MCF-7 cell line that stably expressed Tet-regulated tk gene was established. HSVtk gene expression in the MCF/TRE/tk/Tet-On cell line was under the control of Doxycycline (Dox). Cell viability was also determined by MTT assay, whereas HSVtk gene expression was analyzed by reverse transcription-PCR (RT-PCR).

RESULTSMCF/TRE/tk/Tet-On cell survival rate was decreased from 100% to less than 20% when ganciclovir (GCV) concentration was increased from 0 to 1000 microg/ml at 1 microg/ml of Dox after 72 hours of GCV administration. At 1 microg/ml of GCV concentration, the cell numbers decreased from 7 x 10(4) cells/ml to 2 x 10(4) cells/ml when Dox concentration was increased from 0 to 1500 ng/ml after 72 hours culture. In addition, bystander effects were generated in vitro when 10% - 25% of transduced MCF-7 cells were mixed in untransduced MCF-7 cells. On the other hand, the human breast cancer models in SCID mice were set up. The tk gene was expressed with the regulated character after MCF/TRE/tk/Tet-On cells were implanted into the female SCID mice 7 days after Dox induction followed by intraperitoneally administration of GCV for 23 days. Subcutaneous tumors in SCID mice that were implanted with MCF/TRE/tk/Tet-On cells shrank remarkably after Dox and GCV administration as compared with the control.

CONCLUSIONThe human breast tumor cells (MCF-7) expressing HSVtk gene can be eradicated by administration of GCV and induced with tetracycline or its derivative Dox in vitro and in vivo.

Animals ; Breast Neoplasms ; therapy ; Bystander Effect ; Cell Line, Tumor ; Cell Survival ; Doxycycline ; pharmacology ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; Genetic Therapy ; methods ; Genetic Vectors ; Herpesviridae ; genetics ; Humans ; Mice ; Mice, SCID ; Retroviridae ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Thymidine Kinase ; genetics ; Transfection

OBJECTIVE: To investigate the expression pattern of adapter protein with a Src-homology 2 domain (SH2B1), the suppressor of cytokine signaling-3 (SOCS3), protein-tyrosine phosphatase 1B (PTP1B) and neturopetide Y (NPY) in obese and normal mice hypothalamus and its relation with serum leptin and insulin levels. METHODS: The obesity animal model was prepared with healthy C57/bl6 mice. Lee's index and Homeostasis model assessment-insulin resistance (HOMA-IR) were calculated. The mRNA levels of SH2B1, SOCS3, PTP1B and NPY were measured by fluorescent quantitation RT-PCR. The SH2B1 and NPY protein expressions were detected by Western blot. RESULTS: Compared with the normal mice of the same age, SH2B1 mRNA expression in the obese mice hypothalamus decreased. SOCS3 and PTP1B mRNA expression increased. Western blot showed that SH2B1 protein expression decreased, while NPY protein expression increased in the obese mice. Linear correlation analysis showed that the serum leptin and fasting insulin levels were negatively correlated with SH2B1mRNA expression and positively correlated with SOCS3 and PTP1B mRNA expression. CONCLUSION SH2B1, SOCS3, PTP1B and NPY are key factors for obesity development.
Adaptor Proteins, Signal Transducing ; metabolism ; Animals ; Hypothalamus ; metabolism ; Insulin ; blood ; Insulin Resistance ; Leptin ; blood ; Mice ; Mice, Inbred C57BL ; Neuropeptide Y ; metabolism ; Obesity ; metabolism ; Protein Tyrosine Phosphatase, Non-Receptor Type 1 ; metabolism ; RNA, Messenger ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins ; metabolism

italic>Artemisia argyi (A. argyi) is a Chinese herbal medicine in China. The main active components are volatile oils, flavonoids, and other compounds, which have various pharmacological activities. Methoxylated flavonoids are the main active ingredients in A. argyi. Flavonoid O-methyltransferase (FOMT) is a key enzyme in the O-methylation of flavonoids. In order to further understand the function and characteristics of FOMT proteins, this paper carried out the whole genome mining and identification of FOMT genes in A. argyi and performed phylogenetic, chromosomal localization, gene sequence characterization, subcellular localization prediction, protein structure, gene structure analysis, and expression pattern analysis. The results showed that a total of 83 FOMT genes were identified in the genome of A. argyi. The phylogenetic tree shows that FOMT genes are divided into two subgroups, CCoAOMT (caffeoyl CoA O-methyltransferase) subfamily (32 genes) and COMT (caffeic acid O-methyltransferase) subfamily (51 genes). Gene sequence analysis showed that the number of amino acids encoded by FOMT was 70-734 aa, the molecular weight was 25 296.55-34 241.3 Da, and the isoelectric point was 4.51-9.99. Compared with 32 members of the CCoAOMT subfamily, nearly 1/3 of the 51 members of the COMT subfamily were hydrophobic proteins and 2/3 were hydrophilic proteins. Subcellular localization prediction showed that more than 80% of CCoAOMT subfamily members were located in the cytoplasm, and 96% of COMT subfamily members were located in the chloroplast. COMT subfamily members have more motifs than CCoAOMT subfamily members. The N-terminal motifs of COMT subfamily proteins are relatively variable, while the C-terminal motifs are relatively conserved. Expression pattern analysis showed that CCoAOMT subfamily members were mainly expressed in roots, while COMT members were mainly expressed in leaves. Some FOMTs showed the tissue expression specificity by real-time quantitative PCR analysis, especially in leaves. In this study, we identified and analyzed the FOMT gene family in A. argyi, and provided a theoretical basis for further research on the function of FOMTs and the biosynthesis of methylated flavonoids in A. argyi.