The medical law science interdisciplinary talents play an crucial role in solving medical service dispute and alleviating contradictory. Based on the questionnaire survey and the interview to the court and law offices as well as the analysis of the society's demand for the knowledge structure and ability of medicine law professionals, we proposed the construction of the training mode for medicine law professionals with the ability to adapt to the social demand.

The primary structure of TNK-tissue plasminogen activator (TNK-tPA) was characterized using liquid chromatography-mass spectrometry (LC-MS). Firstly, the molecular mass of deglycosylated protein was measured. Then peptide mass mapping and MS/MS of the reduced, alkylated and trypsin-digested sample were tested and analyzed so as to verify its amino acid sequence and identify post-translational modifications. Results show that the amino acid sequence was consistent with designed structure; about 5% of M207 was oxidized; T61 was fucosylated with -80% occupancy; N103, N448 and N184 (-15% occupancy) were glycosylated with complex-type oligosaccharides. LC-MS coupled with proper sample pretreatment is approved to be a rapid and powerful approach to characterize the primary structure of TNK-tPA.

Objective This study is to investigate the inhibitory effect of Arsenic Trioxide ( As2O3 ) on salivary adenoid cystic carcinoma -2(ACC-2)cell xenografts and angiogenesis in nude mice .Methods The nude mice with ACC-2 cell xenografts were randomly distributed into four groups ,control normal saline ( NS) ,5-Fu group[5-Fu10 mg/(kg· d)],low-dose group[As2O32.5 mg/(kg· d)],and high-dose group[As2O3 5.0 mg/(kg· d)].0.2 mL NS or drugs were given by intraperitoneal injection for 10 days.Tumor volume and weight were measured ,the microstructure was observed by HE staining ,the expression of VEGF and MVD were detected by immunohistochemistry .Results The average tumor volume was significantly smaller and the average tumor weight was significantly lighter in As 2 O3 groups than in control groups ,especially in the high-dose group, the inhibitory rate were 52.17%and 56.04%(P<0.01).Compared with the control group,the MVD of treat-ment group was markedly decreased ,the pipe diameters were smaller in treatment group .The expression levels of VEGF and MVD were significantly lower in As 2 O3 groups than in control group , especially in the high -dose group.Conclusion As2O3 has obviously inhibitory effects on growth of ACC -2 cell xenografts and angiogenesis in nude mice by suppressing the expression of VEGF and MVD .The anticancer effect of As 2 O3 is related to the As2O3 concentration .

Objective To investigate the effects of fasudil on expressions of Bcl-2 and Bax in cerebral cortex of model rats with subarachnoid hemorrhage (SAH). Methods Thirty rats were divided into sham operation group, SAH group and SAH+fasudil group, 10 rats in each group. Double injection of blood into occipital cistern method was used for SAH model in SAH group and SAH+fasudil group. In the sham operation group, the blood injection was instead by normal saline. In the SAH+fasudil group, at 30 min after the second injection of blood, rats were administrated with intraperitoneal injection of fasudil at a dose of 3 mg/kg. The general situation, neurological score, TUNEL staining for cortex cell apoptosis, immune histochemical staining and Western blotting assay for Bcl-2 and Bax protein expression were compared 24 h after the operation between the three groups. Results Compared with the sham operation group, rats in SAH group and SAH +fasudil group appeared obvious neurological deficits. The neurological score was significantly lower in SAH group ( 2.68 ± 0.31) than that of sham operation group (5.00±0.00). The neurological score was significantly higher in SAH+fasudil group (3.27 ± 0.35) compared with that of SAH group (2.68 ± 0.31, P<0.05). There was obvious cell apoptosis in SAH group and SAH+fasudil group, and the apoptosis was less in SAH+fasudil group than that of SAH group (P<0.05). The level of Bcl-2 expression was significantly lower in SAH group than that of sham operation group, and Bax expression was significantly higher in SAH group than that of sham operation group (P<0.05). The level of Bcl-2 expression was significantly higher in SAH+fasudil group than that of SAH group, but Bax expression was significantly lower in SAH+fasudil group than that of SAH group (P<0.05). Conclusion Fasudil can improve the neurological damage in rats with SAH, which may be related with the regulation of apoptosis related proteins Bcl-2 and Bax.

The main purpose of ideology and morality course is not only to pass on knowledge and theory to students but also to teach them ideology and conduct norms on the basis of knowledge and theory. The reform and exploration of teaching patterns of ideology and morality course should concentrate upon the three ways of "theory teaching, case analysis and classroom discussion", so that teachers and students can interact and cooperate with each other, serve the same teaching theme and bring into fullplay teachers' leading function and students' initiative in classroom teaching.

Objective To investigate the expression and function of thymus and activation regulated chemokine (TARC) and its special receptor CCR4 at placenta villous in the first trimester placenta villous.Methods Placenta villous was collected from healthy women undergoing artificial abortion at 6 to 8 weeks of gestation,mRNA levels of TARC,CCR4 were analyzed using semi-quantitative reverse transcription (RT)-PCR methods.Immunohistochemistry assay was used to assess the protein localization and expression of TARC,CCR4.Additionally,extravillous cytotrophoblasts were isolated and cultured.Expression of TARC and CCR4 was measured by immunofluorescence assay.Invasion of cell line HTR8/SVneo was analyzed by transwell assay at concentration of 10,25,50 and 100 ng/ml of TARC matched with RPMI 1640 fetal bovine serum free eulture medium as control group.In the mean time,blocking experiment was also added to detect TARC regulating cell invasion,which were classified into four groups:control,100 ng/ml rhTARC,20 μg/ml anti-TARC + 100 ng/ml rhTARC,100 ng/ml rhTARC + 20 μg/ml IgG.The influence of 100 ng/ml TARC on expression level of integrin-α5 and integrin-β1 were measured by using western-blot assay.Results (1)In vivo assay:expression of TARC and CCR4 mRNA were detectable in first trimester placenta villous,TARC protein was localized in cytotrophoblasts,syncytiotrophoblasts and cell column especially on the distal portion,while CCR4 protein was localized on invading interstitial cytotrophobalsts.(2)In vitro assay:①TARC,CCR4 was also expressed in primary isolated extravillous cytotrophoblasts by immunofluorescence assay; ②Matrigel invasion assay demonstrated that TARC had specific dose dependent stimulatory effects on the cells invading through the matrigel precoated filter,the number of cells migration into the lower chamber were:142 ± 31 at 10 ng/ml group,161 ±46 at 25 ng/ml group,201 ±30 at 50 ng/ml group,312 ±48 at 100 ng/ml group,117 ± 33 at control group,the significant response observed from 25 ng/ml (P ＜ 0.05)and reached a peak effect at 100 ng/ml (P ＜ 0.01); ③Blocking experiment demonstrated that when trophoblast invasion was monitored in response to TARC neutralizing antibody (15 μg/ml) together with rhTARC 100 ng/ml.The stimulatory activity of rhTARC was completely overcome,with the cells invasion into the lower chambers were 100 ng/ml rhTARC,20 μg/ml anti-TARC + 100 ng/ml rhTARC,100 ng/ml rhTARC +20 μg/ml IgG,control:313 ±47,113 ±41,287 ±75 and 128 ±23,respectively;④Western-blot assay demonstrated that if cells were treated with 100 ng/ml rhTARC,the expression of integrin-α5 were significantly increased(P ＜0.01),integrin-β1 level also increased when compared with control(P ＜0.05).Conclusion TARC was expressed specifically at human fetal-maternal interface.Trophoblast invasion and migration mainly was regulated by up-regulation integrin-α5 and integrin-β1,which plays an role in trophoblasts differentiation and placentation.

Aim To study the inhibitory effect of chitosan on peroxidation and monocytes adhesion to vascular endothelial cells induced by high concentration of glucose. Methods Human umbilical vascular endothelial cells (HUVEC) were treated with high glucose, and high glucose with different concentrations of chitosan for 24 h. Hydroxyl radicals (OH?) and malon-dialdehyde (MDA) were measured. Monocytes Raw 264.7 were pre-incubated with Rhodamin123, and then co-cultured with HUVEC for 30 min, followed bymicroscope observation and determination of the monocytes adhesion. Finally, the mRNA expression of vascular cell adhesion molecular-1 (VCAM-1) was evaluated by reverse transcription polymerase chain reaction (RT-PCR). Results Concentrations of OH? and MDA in HUVEC increased after incubation with high glucose. Both of the amount of adhesive monocytes and mRNA expression level of VCAM-1 in HUVEC were induced by high glucose. Inversely, chitosan inhibited these changes in a dose-dependent manner without any cytotoxicity to cells. Conclusion Chitosan can scavenge free radicals and prevent peroxidative injury on vascular endothelial cells, which further down-regulates the expression of VCAM-1 and consequently inhibits the adhesion of monocytes to endothlial cells.

The amino acid sequence of the fusion protein FP3 was measured by two types of LC-MS/MS and its primary structure was confirmed. After reduction and alkylation, the protein was digested with trypsin and glycosyl groups in glycopeptide were removed by PNGase F. The mixed peptides were separated by LC, then Q-TOF and Ion trap tandem mass spectrometry were used to measure b, y fragment ions of each peptide to analyze the amino acid sequence of fusion protein FP3. Seventy-six percent of full amino acid sequence of the fusion protein FP3 was measured by LC-ESI-Q-TOF with the remaining 24% completed by LC-ESI-Trap. As LC-MS and tandem mass spectrometry are rapid, sensitive, accurate to measure the protein amino acid sequence, they are important approach to structure analysis and identification of recombinant protein.

Structure of a recombinant chimeric anti-CD20 IgG1 monoclonal antibody was verified by the application of high-performance liquid chromatography-mass spectrometry (HPLC-MS)and N-terminal sequencer. Molecular masses, N-terminal sequences and peptide maps of the antibody treated with different reagents and enzymes were measured. Results indicate that the amino acid sequences of light and heavy chains and 10 disulfide bonds were consistent with theoretical structure. By comparison of molecular masses and peptide maps for the fully glycosylated and deglycosylated samples, the N-linked glycosylation site was identified. The method is simple, rapid, precise, and could be referred to the quality control and structure determination of other IgG1 products.

Accurate determination of biological activity is essential in quality control of recombinant human brain natriuretic peptide (rhBNP). In previous study, we successfully developed a genetically modified cell line 293GCAC3-based ELISA assay for rhBNP. But ELISA procedure is still tedious, so this study was aimed to develop a rapid and simple bioassay for rhBNP using GloSensor technology, which provides a platform of flexible luciferase-based biosensors for real-time detection of signaling events in live cells, including cGMP production. A reporter cell line 293GCAGlo-G1 was constructed by transfecting pGloSensor?40 F plasmid into 293GCAC3. The reporter assay based on 293GCAGlo-G1 showed high precision with intra-assay CV being 8.3% and inter-assay CV being 14.1%; high accuracy with 80%, 100% and 120% recovery rate being 99.2%, 102.4% and 99.0% respectively; and great linearity with R2of linear fitting equation being 0.99. Besides, no significant difference was found in test results of reporter assay and 293GCAC3-based ELISA assay (paired t test, p＝0.630). All these results suggested that the reporter assay was a viable assay for biological determination of rhBNP.