Objective To investigate the feasibility of Wilms’tumor gene 1 (WT1)-specific CD8+T cells from periph?eral blood for the treatment of breast cancer by detecting the killing activity of WT1 specific CD8+T cells on breast cancer cells. Methods Flow cytometry was used to detect WT1-specific CD8+T cells in the peripheral blood of 20 samples from HLA-A2 seropositive healthy donors, which were isolated by WT1/MHC streptamer magnetic beads and cultured. The func?tion of WT1-specific CD8+ T cells were analysis by cytotoxicity assay. Results Twelve of 20 healthy donors had naive WT1-specific CD8+T-cell frequencies of>0.5%, and 4 of 20 even>1.0%of all CD8+T cells. After positive selection by magnetic cell separation, a purity of up to 80%can be achieved. WT1 specific CD8+T cells can specifically kill breast can?cer cell line with WT1 polypeptide. Conclusion WT1 specific CD8+T cells can be detected in peripheral blood of healthy volunteers. WT1 specific CD8+T cells have killing effect on breast cancer cells, suggesting the feasibility of adoptive immu?notherapy for breast cancer.

Objective To investigate the dynamic changes of dengue viral loads and IgM antibody in patients with dengue fever ( DF) and to analyze their relationships with disease severity. Methods A to-tal of 1 508 serum samples were collected from 1 140 hospitalized patients including 1 050 mild cases and 90 severe cases within 10 days after the onset of DF in Guangzhou in 2014. The viral loads were determined by using fluorescence quantitative RT-PCR. ELISA was performed to measure the dengue virus ( DENV)-spe-cific IgM antibody. Results In general, the DENV viral loads in patients declined gradually from 108 copies/ml on day 1 to 103 copies/ml on day 10 after the onset of DF. The viral loads in severe cases were significantly higher than those in mild cases on days 5 to 7 (P<0. 05). The positive rates of DENV RNA in serum samples also decreased with the disease progression from 100% on day 1 to 40% on day 10. Com-pared with the mild cases, the patients with severe DF showed higher positive rates of DENV RNA on day 6 and day 8 (P<0. 05). The DENV-specific IgM antibody could be detected on day 2 and the secretion of IgM antibody increased gradually with the disease progression. The levels of IgM antibody in mild cases were sig-nificantly higher than those in severe cases (P<0. 001). The positive rates of IgM antibody in patients in-creased form 8% on day 2 to 95% on day 6. Higher positive rates of IgM antibody were detected in mild ca-ses on days 5 and 6 as compared with those in patients with severe DF (P<0. 05). Conclusion High viral load and low level of IgM antibody during the fastigium of DF were closely associated with the disease severity.

OBJECTIVE: To explore the forensic pathological features of death caused by anaphylactic shock. METHODS: One hundred and forty-two death cases of anaphylactic shock were retrospectively analyzed. The IgE level in the serum of anaphylactic shock cases were statistically compared with that of 62 non-anaphylactic shock cases. RESULTS: Most cases (77.46%) of anaphylactic shock death occurred in the medical institutes, with intravenous drug administration accounting for 53.53% of anaphylactic shock death. β-Lactam antibiotics, glucocorticoid and herbal medications were responsible for a significant proportion of such cases. Although characteristic histopathological changes were absent in vast majority of these anaphylactic shock cases, the differences of IgE levels in the serum between anaphylactic shock group and non-anaphylactic shock group were statistically significant (P<0.05). CONCLUSION Combined information including clinical data, autopsy results, IgE level, and other specific test results should be evaluated together in the forensic pathological diagnosis of anaphylactic shock.
Anaphylaxis ; Autopsy ; Cause of Death ; Forensic Pathology ; Humans ; Infusions, Intravenous ; Retrospective Studies ; Serum

Objective To study the effects of the composite of propolis and magnolol on the growth of Streptococcus mutans (Sm).Methods The solution concentration of the propolis (250,125,62.5,31.25,15.625 g/L),magnolol (0.0625,0.0313,0.0156,0.0078,0.0039 g/L) and the composite of those two (propolis,magnolol) A1 (250 g/L,0.0625 g/L),A2(125 g/L,0.0313 g/L),A3 (62.5 g/L,0.0156 g/L),A4 (31.25 g/L,0.0078 g/L),A5 (15.625 g/L,0.0039 g/L) were chosen according to pretested minimum inhibitory concentration (MIC).The solutions were proportionally diluted to the presupposed concentration before operated on the Sm.Meanwhile,the solvent and the bacterial were also operated on the Sm as controls.Slip diffusion method was adopted to measure the inhibitory effect of those solutions on Sm.Results Bacteria inhibition zones were appeared at the concentration of 62.5 g/L prololis and 0.0156 g/L magnolol respectively.A inhibition zone appeared at the lower concentration of A5 (15.625 g/L,0.0039 g/L) of the propolis and magnolol composite.As general,the inhibition effects increased with concentration.The difference were statistically significant,P ＜ 0.05.Conclusions The inhibitory effect of the composite of propolis and magnolol would be much stronger than either of those two separately.

Objective To compare the differences in the evaluation for simulation level of students standardized patients (SSP) between undergraduates and teachers .Methods Eleven teachers and ninety-one undergraduates were selected , and investigated with self -designed questionnaire on the evaluation for simulation level of SSP .Results The aggregate score of the evaluation for simulation level of students standardized patients among undergraduate students was (3.76 ±0.30), which was (3.69 ±0.23) among teachers, and the difference was not statistically significant (t =0.73,P >0.05).But , there were statistically significant differences in the item 11 ( SSP answered the questions pointedly ) , the item 12 ( SSP didn ’ t use inductive language , or medical terms ) and the item 13 ( The scenario and situation simulated by SSP in each experimental unit were consistent ) between undergraduates and teachers [(3.88 ±0.36) vs (3.55 ±0.21), (3.92 ±0.31) vs (3.47 ±0.79), (3.80 ±0.54) vs (3.56 ±0.20); t =2.03,t =5.32,t =2.15, respectively;P<0.05].Conclusions SSP should be assessed from multiple perspectives , so the evaluation subjects should be increased in use of standardized patients assessment , which can reduce the error .

Objective To investigate the pharmacokinetic processes of adriamycin-heparinized mesoporous silica anticancer drug delivery system (AMS-HP) in rats.Methods Six male Sprague-Dawley rats were used.AMS-HP was administered intravenously via a sublingual vein with a dose of 8 mg · kg-1.Blood was sampled from the tail before treatment and 0,0.08,0.17,0.5,1,3,6,12,24,36,48,60,72 h after given drugs.The plasma concentration of adriamycin in AMS-HP was determined by HPLC.The pharmacokinetic parameters were determined by DAS 2.0 software.Results The main pharmacokinetic parameters of adriamycin in AMS-HP were as follows:Cmax,t1/2,tmax,V/F,CL/F,AUC0-t,AUC0-∞ were (5.71 ± 0.46) L · h-1 · kg-1,(15.81±2.01) h,(3.00 ±0.00) h,(140.12 ±17.87) L · kg-1,(118.74±18.04) ng · mL-1,(3599.96 ±881.99) ng · mL-1 · h,(3838.51 ± 817.60) ng · mL-1 · h,respectively.Conclusion The HPLC method of plasma concentration of adriamycin in AMS-HP is specific and has good accuracy,precision and high sensitivity.Adriamycin in AMS-HP has a long half-life and a slow release effect in vivo.

BACKGROUNDHuman group O red blood cells have great benefit in specialized transfusion areas such as armed conflict and natural calamity. The group B antigen differs structurally from group O antigen only by the addition of one terminal alpha-linked galactose residue. In this study we aimed to remove the terminal galactose from group B red blood cell to get group O red blood cell.

METHODSalpha-galactosidase cDNA was cloned by RT-PCR from Catimor coffee beans grown on Hainan Island of China. The vector for alpha-galactosidase cDNA expression was constructed and transferred into Pichia pastoris cells by electroporation. The transgenic cells were cloned by fermentation and the recombinant alpha-galactosidase was purified by ion exchange chromatography. After studying the biochemical characters of alpha-galactosidase, we have used it in converting human erythrocytes from group B to group O.

RESULTSThe purity of recombinant alpha-galactosidase was higher than 96%, which was thought to be suitable for the use of blood conversion. Enzymatically converted human group O red blood cells (ECHORBC) exhibited membrane integrity, metabolic integrity, normal cell deformation and morphology. There were no coagulation between ECHORBC and any group of human blood. The ECHORBC will keep normal structure and function for a period of 21 days at 4 degrees C in monoammoniumphosphate nutrient solution. Experiments with Rhesus monkeys and gibbons showed that transfusion of enzymatically converted erythrocytes was safe.

CONCLUSIONECHORBC can be easily obtained from group B red blood cell by alpha-galactosidase digestion. This study suggests that ECHORBC could be transfused to patients safely and efficiently.

ABO Blood-Group System ; classification ; metabolism ; Animals ; Blood Transfusion ; Cloning, Molecular ; Coffee ; enzymology ; Erythrocytes ; metabolism ; Humans ; Macaca mulatta ; Quality Control ; Recombinant Proteins ; isolation & purification ; pharmacology ; alpha-Galactosidase ; immunology ; isolation & purification ; pharmacology ; toxicity

In order to meet the demand for safe transfusion in special conditions and to utilize the donated blood supply efficiently, technology has been developed to convert erythrocytes from type A, B, or AB to "universal donor" blood. Conversion of blood type B to O was performed by means of recombinant alpha-galactosidase digestion. The results showed that blood type B to O was converted successfully, 1 transfusion unit of red cells of group B (100 ml totally) could converted to universal blood cells in the optimal conditions including pH 5.6, 26 degrees C, 2 hours, obturation and sterilization. It is concluded that the universal red blood cells converted from group B to group O are conformed to demand of identification rules of biological products, no harmful effects of alpha-galactosidase on cell structure and function are observed. The converted red cells can stored in 4 degrees C for 21 days.
ABO Blood-Group System ; classification ; immunology ; Blood Group Incompatibility ; prevention & control ; Blood Transfusion ; methods ; Coffee ; enzymology ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Erythrocytes ; immunology ; metabolism ; Humans ; Isoantigens ; drug effects ; metabolism ; Recombinant Proteins ; metabolism ; pharmacology ; alpha-Galactosidase ; genetics ; metabolism ; pharmacology

Nowadays,with the continuous deepening and development of vocational education teaching reform,medical higher vocational education always takes moral education as the fundamental task.As an independent type of education,vocational education should always deepen the integration of industry and education and the integration of science and education.Through the teaching research of"problem chain+course ideological and political case",this study innovates the coordinated education team of drug nursing curriculum,the collaborative education method and the collaborative education evaluation,and improves the teaching effect.

Aim To investigate the protective effect of 1, 8-Cineole on the injury of human aortic endothelial cells (HAECs) induced by high glucose (HG) via regulating autophagy. Methods Cells were incubated with different doses of 1, 8-Cineole followed by exposing to HG for 60 h, and MTT assay was used to analyse cell viability, lactate dehydrogenase (LDH) was used to detect cytotoxicity, and Western blot was used to detect Beclin1, LC3-II/I, p62, caspase-3 and caspase-9 expressions. Autophagy inhibitor (chloroquine, CQ) was treated on HAECs, and the expressions of Beclinl, LC3-II/I, p62, caspase-3 and caspase-9 were measured by Western blot. Results 1, 8-Cineole increased cell viability, reduced the content of LDH, activated autophagy and inhibited apoptosis. Compared with control group, the expression of Beclinl, LC3-II/I, p62, caspase-3 and caspase-9 in CQ group increased. Simultaneously, the expression of above-mentioned between CQ + HG group and CQ + HG + CH group. Conclusions 1, 8-Cineole has protective effect on the injury of HAECs induced by high glucose, and its effect is related to improving autophagy flux.