Based on the necessity of carrying out bioethics education to the probationer nurses, this paper put forward to introduce psychological sitcom to the probationer nurses′ bioethics education and described the practice process in detail. It also discussed the mechanism to improve the effectiveness of bioethics education from the per-spectives of creation, dramatization, and performance of psychological sitcom.

Objective:To explore the effect of group tutoring in the adaptation education for college freshmen.Method:All freshmen that attended Wenzhou medical college in 2008 are enrolled in group tutoring to help them better adapt to the new life in college,and the tutoring effect is appraised by a self-designed questionnaire.Result:The freshmen show a general progress and satisfaction after receiving group tutoring,and regard it quite necessary to conduct activities like group tutoring at the beginning of college life to help make freshmen better adaptive.Conclusion:Group tutoring is very effective in the adaptation education for college freshmen,and has a universal applicability for all college students.

Objective:To explore the effects of the natural plant ingredients lemon essential oil(LEO),limonene(LIM)and tea poly-phenols(TP)on the cell surface hydrophobicity and adherence of Streptococcus mutans(S.mutans).Methods:S.mutans were treated by sub-minimal inhibitory concentration(MIC)levels of LEO,LIMand TP respectively.Adsorption to hexadecane was used to measure the hydrophobic interaction of S.mutans.A classical 96-cell microtitre plate production assay using crystal violet staining was employed to visualize the adherence of S.mutans to hard tissue surface.Results:LEO,LIMand TP at sub-MIC levels could inhibit the cell sur-face hydrophobicity and adherence of S.mutans in a dose-dependent manner(P <0.05).At 1 /2 MIC and 1 /20 MIC,the inhibitary effect of LEO was stronger than that of LIMand TP(P <0.05).Conclusion:LEO may possess anticariogenic potential.

Objective:To explore the antioxidant activity of natural plant ingredients lemon essential oil(LEO),limonene(LIM) and tea polyphenol(TP).Methods:The UV-Vis spectrometry was used to determine the 3 agents on the scavenging activity of 1 ,1 diphenyl-2-trinitrobenzene hydrazine free radicals(DPPH·)and hydroxyl free radical (·OH).Results:LEO,LIM and TP showed scavenging effect on the 2 kinds of free radicals.The (DPPH·)scavenging effect:LEO(IC50 =0.01 4)>TP(IC50 =0.01 4)>LIM (IC50 =0.002);the (·OH)scavenging effect:TP(IC50 =0.079)>LEO(IC50 =0.01 3)>LIM(IC50 =0.004).In a certain con-centration range,scavenging effect was increased with the concentration increase.Conclusion:As a nontoxic natural extract,lemon essential oil has strong antioxidant activity.

Background Retinal pigment epithelial (RPE) cell transplantation is a novel approach to the treatment of hereditary retinal diseases, however, human-derived RPE cell line occurs de-differentiation during in vitro cell culture.Studies showed that early abnormal activation of Akt/mammalian target of rapamycin (mTOR) signal pathway is the primary cause of RPE cell line de-differentiation, therefore, the inhibition of mTOR pathway will be helpful for the retard of de-differentiation of RPE cells.Objective This study aimed to investigate whether rapamycin can suppress the activation of mTOR pathway and promote differentiation of ARPE19 cells.Methods ARPE-19 cells were incubated in 12-well plate and divided into control group and rapamycin-treated group.DMSO or rapamycin with the final concentration of 400 nmol/L was added in the medium of the control group and the rapamycin-treated group, respectively.The cells of each group were collected 24 hours and 48 hours after cultured.The expression of zonula occludens-1 (ZO-1) in the cells was examined by immunofluorescence.The relative expression levels of RPE cell specific genes and proteins were assayed by using real-time quantitative PCR and Western blot, respectively.The detected results were compared between the two groups.Results ZO-1 was expressed in both group,but the fluorescence intensity was evidently enhanced in the rapamycin-treated group.The relative expression levels of RPE65, MERKT and LRAT mRNA in the cells increased by 25.97％ , 29.71％ and 13.00％ in the rapamycin treated group compared with the control group 24 hours after cultured (P=0.04,0.04,0.04) , and the expression levels of RPE65, LRAT, rLBP1, BEST1 , keratin18 and MERKT mRNA elevated by 174.00％ , 88.00％ , 56.18％ ,193.81％ ,10.83％ and 35.02％ in the rapamycin-treated group in comparison with the control group 48 hours after cultured (P =0.00,0.04,0.01,0.04,0.04,0.03).In addition, the expressions of p-mTOR, p-P70S6 and p-S6 protein were weaker in the rapamyein-treated group than those in the control group both 24 hours and 48 hours after cultured.Twenty four hours after cultured,the expression level of ZO-1 protein raised by 40％ in the rapamycin-treated group compared with the control group (P =0.01);while 48 hours after cultured,the expression levels of ZO-1 ,MERKT, catenin and LRAT proteins elevated by 36.00％ ,57.37％, 13.68％ and 41.07％ in the rapamycintreated group in comparison with the control group (P=0.01,0.00,0.04,0.04).Conclusions Rapamycin can suppress the activation of mTOR signaling pathway and up-regulate the expressions of RPE specific genes in ARPE19 cells.Inhibition of mTOR pathway might be an effective way for culturing RPE cells in vitro.

Objective To set up the quality standard of TCM preparation Qingfei pill. Methods TLC methods were used to the qualitative identification of Fructus Termininaliae Immaturus and Radix Sophorae Subprostratae in the preparation. The content of Gardenoside, the effective component of Fructus Gardeniae, was determined by HPLC method, using Agilent Eclipse XDB-C18 (150 mm×4.6 mm, 5 μm) and acetonitrile-water (10∶90) as mobile phase with flow rate at 1.0 mL/min. The column temperature was room temperature, and the detection wavelength was 238 nm. Results The TLC methods for Fructus Termininaliae Immaturus and Radix Sophorae Subprostratae identifications were simple and rapid, and showed good repetitiveness. Gardenoside presented a good linear relationship within the range of 0.238-3.808 μg, and the average recovery rate was 97.24%, RSD was 1.60% (n=6). Conclusion These methods showed strong specificity and good repetitiveness, and can be used to control the quality of the product effectively.

Objective To analyze the effect of GC-rich DNA fragments on the level of transgenic expression in Chinese hamster ovary (CHO) celts and its position effect.Methods The synthetic DNA fragment with GC-rich was cloned into the 5'or 3'or both 5'and 3'ends of expression cassette of expression vector.Three new expression vectors (pIRES-G1,pIRES-G2 and pIRES-G3) which was inserted with the GC-rich DNA fragments in different position were transfected CHO ceils,respectively,and then was observed under fluorescence microscope;the control vector was pIRES-EGFP.Stable transfected cell lines were screened under G418,and enhanced green fluorescent protein(EGFP) expression was analyzed by flow cytometry and the transgenic copy number was detected by quantitative real-time quantitative polymerase chain reaction (qRT-PCR).Results Three expression vectors with a GC-rich DNA fragments in different position were constructed successfully.The insertion of GC-rich DNA fragments at 3'end and both 5',3'ends of the box of expression vector could obviously improve the expression level of vector in CHO cells;and the expression level of the stably transfected CHO cells increased 1.39 fold and 1.32 fold compared to the control vector,respectively;the transgene copy number increased 1.32 fold and 1.24 fold compared with the control vector.While the insertion of GC-rich DNA fragments at 5'end of expression cassette had no obvious effect on the level of gene expression.Conclusion The role of DNA fragment with GC-rich in improving the transgenic expression of CHO cells is related to its position in the vector.The insertion of GC-rich DNA fragments at 3'end and both 5',3'ends of the box of expression vector can improve transgenic expression.

Background Retinal pigment epithelium (RPE) cell transplantation is the primary means of human trial for the treatment of retinal degeneration.Induced pluripotent stem cells (iPSCs) will become an important source for cell transplantation.In addition,iPS-RPE cells may provide a personalized treatment platform for the patient's own cells treatment.Objective This study was to evaluate the feasibility of human fibroblasts differentiate toward iPSCs from retinitis pigmentosa (RP) patients and toward iPSC-RPE cells from non-RP individual by retroviral transfection of Oct4,Sox2,c-Myc and KLF4 genes.Methods Human thigh skin tissues were obtained from a RP patient with hotspot mutation of SNRNP200 p.S1087L and individual without SNRNP200 p.S1087L mutation,respectively,with the size 2 c m×3 cm.Human dermal fibroblasts were isolated and cultured by trypsin digestion and explant method.The fibroblasts were transfected by a series of retrovirus and cultured by human embyonic cellconditioned medium to generate and induce iPSCs,and then the iPSCs were identified by morphology,alkaline phosphatase (AP) staining and immunofluorescence assay of specific markers of pluripotent stem cells.iPSCs suspension were injected into SCID mouse to observe the tumorigenesis.The iPSCs from non-RP subject were induced to differentiate toward iPS-RPE cells by embryonic body (EB) inducing method,and iPS-RPE cells were identified by detecting the expression of RPE65,zonula occludens protein 1 (ZO-1) and lecithin retinol acyltransferase (LRAT).Results Cultured human dermal fibroblasts showed fusiform or polygon shape and intercellular close arrangement,and Vimentin was positively expressed in the cells.Small cell colonies were harvested 5-7 days after infected by retroviruses,and the morphology changed from spindle into round mass.The hESC-like iPSCs clonies appeared 20 days after cultivation,and the positive expressions of hESC-specific surface antigens including SSEA3,SSEA4,TRA-1-60,TRA-1-81 and Nanog were found in the cells 25-30 days after cultivation,and the positive staining of AP was obtained 12 weeks after cultivation.A teratoma was formed at the injection site of iPSCs suspension in SCID mouse.Immunofluorescence technique showed that RPE cell-specific proteins including RPE65,ZO-1 and LRAT proteins were positively expressed in iPS-RPE cells at 30 days after differentiation.Conclusions Mutation SNRNP200 p.S1087L of RP patient-specific iPSCs can be induced from human dermal fibroblast by retrovirus infection method.The function and morphology of the iPSCs were similar to hESCs.Human iPSCs cell line generated from the dermal fibroblasts of non-RP individuals can differentiate into iPS-RPE cells.

Objective To investigate the effect of Shenmai injection on purine content in rat cerebral cortex in order to provide a theoretical basis concerning its brain protective mechanism. Methods Sixteen Sprague-Dawley (SD) rats were randomly divided into two groups:normal saline control group and Shenmai injection group, with 8 rats in each group. Shenmai injection 15 mL/kg was injected intraperitoneally into the rats in Shenmai injection group, while in the normal saline group, an equal volume of normal saline was intraperitoneally injected. After the injection for 24 hours, the rats were sacrificed, and the cerebral cortex was removed on ice, homogenized and its supernatant was extracted;then high performance liquid chromatography (HPLC) was used to detect adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), adenosine and inosine contents in the supernatant of cerebral cortex. Results Compared with normal saline control group, ATP, ADP, AMP, adenosine and creatinine content in the cerebral cortex of Shenmai injection group were significantly higher, the differences being statistically significant [ATP (ng/L): 31.62±5.12 vs. 20.25±4.53, ADP (ng/L): 37.04±6.72 vs. 25.12±7.35, AMP (ng/L): 87.82±20.37 vs. 33.23±10.34, adenosine (ng/L): 2.82±0.15 vs. 1.12±0.61, creatinine (ng/L): 11.72±1.05 vs. 6.05±2.55, P < 0.05 or P<0.01]. Conclusion Shenmai injection can elevate ATP, ADP, AMP, adenosine and creatinine contents in the cerebral cortex of rats, possibly that is the theoretical basis for brain protective mechanism of Shenmai injection.

Objective To explore the clinical characteristics of monochorionic or dichorionic twin pregnancy and the high-risk factors for selective intrauterine growth restriction.Methods 460 women with twin pregnancy were divided into a monochorionic group and a dichorion group.The related clinical features were compared between the two groups.Logistic regression was used to analyze the high-risk factors for selective intrauterine growth restriction.Results The maternal age,conception way,and mode of delivery differed significantly between the two groups (P ＜ 0.05).There were significant differences in the rates of selective intrauterine growth restriction and preterm premature rupture of membranes (P ＜ 0.05).The neonatal weight (large or small) and the rate of neonatal transfer differed significantly (P ＜ 0.05).Logistic regression showed that gestational age and birth weight were the risk factors.Conclusions The chorionic nature plays an important role in the process and outcomes of maternal pregnancy.Monochorionic pregnancy is a high risk factor for selective intrauterine growth restriction,meaning the major cause of selective intrauterine growth restriction may originate from the placenta,with should be a placenta-derived disease.