Objective To reconstitute a transactivation system in yeast (yeast model) for screening the pesticides acting on ecdysone metabolism route and eventually influencing the process of ecdysis. Methods The fragment of 5 times repeated EcRE from Drosophila melanogaster was synthesized and the HSP27 promoter from D. melanogaster genome was amplified with PCR. The two sequences were connected and followed by a reporting gene——green fluorescence protein(GFP) gene. The EcRE-HSP27 promoter-GFP fragment was inserted into the expression plasmid pPIC3.5 and integrated into the yeast chromosome to construct yeast A. EcR and USP coding sequences of Aedes albopictus were synthesized, and these two fragments were inserted into Pichia pastoris expression plasmid pGAPZ as two respective reading frames. The two reading frames were integrated into Pichia pastoris chromosome in another recombinant site(pGAPZ and pPIC3.5k share different recombinant sites while being integrated into Pichia pastoris yeast chromosome). EcR and USP were constituted and expressed in the yeast. This recombinant yeast was called yeast B. The model yeast was thus constructed. A known ecdysone agonist-tebufenozide was used to test the yeast model. The effect of tebufenozide on the model yeast was observed under fluorescent microscope. Semi-quantitative RT-PCR was used to test the transcrip-tion level of GFP in the tebufenozide affected yeast and the control. Results In the model yeast, the intracellular expressed EcR and USP constituted EcR/USP heterodimer interacting with EcRE, the expression of GFP was activated, and green fluorescence was observed in model yeast under fluorescent microscope. Tebufenozide affected model yeast showed less fluorescence in comparison to the control model yeast, indicating that the transcription of GFP was suppressed by tubufenozide. Yeast housekeeping gene Actin-1 was used as inner control, semi-quantitative RT-PCR was operated and the result was scanned. The ratio of the brightness of GFP to Actin-1 was calculated automatically, and that of tubufenozide added yeast and the control yeast was 0.614 and 1.134 respectively. This result showed a low transcription level of GFP in tebufenozide affected model yeast, comparing to that of the control. Conclusion The ecdysone-related transacting system in yeast has been constructed, and the model yeast can be used to screen the ecdysone agonists which can act on the ecdysone metabolic route.

Objective To discuss the clinical value of Protein induced by Vitamin K Antagonist-Ⅱ (PIVKA-Ⅱ) and alpha-Fetoproteins (AFP) in diagnosing hepatocellular carcinoma (HCC) and monitoring the treatment effects.Methods Patients were recruited by the Affiliated Hospital of Qingdao University,from August 2013 to March 2014.Serum levels of PIVKA-Ⅱ and AFP were measured by both chemiluminescence assay (CLIA) and electrochemiluminescence assay (ECLA) in patients with HCC (n =148),intrahepatic cholangiocellular carcinoma (n =37),gastric cancer and colorectal cancer (n =44),cirrhosis (n =63),chronic hepatitis B (n =38) and healthy subjects (n =57).To analyze the areas under the receiver operating characteristic curves (ROC-AUC) and to compare the sensitivity and specificity of single PIVKA-Ⅱ or AFP assay,and the combined detection.To analyze the correlation of PIVKA-Ⅱ and both tumor size and TNM staging,so do AFP,respectively.To compare the serum level changes of the two indicators in HCC patients before and after treatment.Results The serum levels of both PIVKA-Ⅱ and AFP in HCC group were higher than that in intrahepatic cholangiocellular carcinoma,gastric cancer and colorectal cancer,cirrhosis,chronic hepatitis B and healthy subjects groups (PIVKA-Ⅱ:U =866.50,424.00,958.00,292.00 and 448.00 ; AFP:U=713.00,440.50,1 182.00,614.00 and 399.00,P ＜0.001).The ROC-AUCs of the single PIVKA-Ⅱ or AFP assay and the combined detection in HCC group were not statistically different (P ＞ 0.05).The sensitivity of PIVKA-Ⅱ (87.16％) was higher than that of AFP (68.92％,x2 =4.73,P ＜ 0.05) in diagnosing HCC ; the sensitivity of the combined detection of PIVKA-Ⅱ and AFP(93.24％) was higher than that of PIVKA-Ⅱ itself (87.16％,adjusted x2 =64.70,P ＜ 0.01) ;while the specificities among them did not show statistical significance (P ＞ 0.05).Tested by Spearman rank correlation,the serum levels of PIVKA-Ⅱ and AFP were both positively related to tumor size (r =0.716,0.475 respectively,P ＜ 0.001).The serum levels of PIVKA-Ⅱ and AFP in HCC patients increased gradually correlated with tumor size (H =72.70,37.02 respectively,P ＜ 0.001) and the positive rates of PIVKA-Ⅱ and AFP were gradually improved (x2 =26.74,21.62 respectively,P ＜ 0.01),too.Based on the International TNM Staging System,the serum levels of PIVKA-Ⅱ and AFP (H =46.63,21.38 respectively,P ＜0.001) and the positive rates of PIVKA-Ⅱ and AFP (PIVKA-Ⅱ:x2 =20.40,P ＜0.01 ;AFP:x2 =8.33,P ＜0.05) in HCC patients from Ⅰ-Ⅳ stages were increased as TNM stages elevated.The serum levels of PIVKA-Ⅱ and AFP in HCC patients were both dropped sharply compared with preoperative levels (Z =-4.59,-4.22 respectively,P ＜ 0.001) and also both dropped in each of the Ⅰ-Ⅳ TNM stages (PIVKA-Ⅱ:Z =-2.85、-2.98、-2.70 respectively,P ＜ 0.05 ; AFP:Z =-2.48、-3.82、-2.50 respectively,P ＜ 0.05) compared with serum levels before treatment.Conclusion PIVKA-Ⅱ and AFP both have high clinical application values in diagnosing HCC and monitoring treatment effects.The sensitivity of PIVKA-Ⅱ in diagnosing HCC is significantly higher than AFP,and the sensitivity can be elevated by the combined detection in diagnosing HCC without reducing the specificity.

In recent years,the research of non-contact biomedical monitoring has continuous development and progress.This review gives an overview of the research status of heart,lung and brain non-contact monitoring methods.The correlation techniques of capacitance electrocardiogram,magnetic induction,radar non-contact monitoring of heart and lung,and non-contact monitoring of brain are analyzed comprehensively.Capacitance electrocardiogram monitors the heart and lung activities useing effect of change in capacitance between the electrodes.Magnetic induction monitors the heart and lung activities useing the Maxwell principle,while radar monitoring the heart and lung activities uses the Doppler effects.Non-contact monitoring of brain adopts the magnetic induction tomography imaging technology.Then elaborate related research at home and abroad,and summarize the advantages and disadvantages of these monitoring methods on the basis of the analysis of monitoring principles.Finally foreground that may dominate this area of new equipment for heart,lung and brain non-contact monitoring in the future is expected.

Objective:To study the characterization of NK cell line co-transfected with hSCF and hIL-15 gene.Methods:pcDNA3-hSCF and pcDNA3-hIL-15 were transfected into NK-92 cell line with LipofectAMINETM,RT-PCR was used to identify NK-92 cell which express hSCF and IL-15,the activity of supernantes was respectively assayed by TF-1 and CTLL-2 cell line. CD3, CD16, CD56, CD25, CD48, CD54, CD69 and CD95 molecules were tested by FACS.Results:We established NK-92S15 cell line which express hSCF and hIL-15 steadily, proliferation ability demonstrated it had more extensive assembling trend,growing with enough cell number,the cytotoxicity of NK-92S15 cell was decreased compared with parental cell line when incubated with rhIL-2.CD56 and CD16 showed on difference while CD25,CD48, CD54 and CD95 decreased significantly.Conclusion:The characterization of NK-92 could be changed by co-transfecting with hSCF and hIL-15 gene, which was benefit to observe the character of NK cell activation and cytotoxicity related molecules. The gene transfection of NK-92 cell made it suitable for further study.

OBJECTIVE: To study the anti-tumor and immunological effect of extracts from Thymus quinquecostatus Celak on mice transplanted S180 tumor cells. METHODS: Different doses of volatile oil and alcohol extracted substances from Thymus quinquecostatus Celak were given to mice bearing S180 tumor for 9 days. Tumor inhibition rates and coefficients of spleen and thymus were determined. RESULTS: Tumor inhibition rates of the groups with alcohol extracts (40 g crude drug.kg(-1).d(-1) and 20 g crude drug.kg(-1).d(-1)) were 51.5% (P<0.01) and 36.4% (P<0.05) respectively, and those of the groups with volatile oil (40 g crude drug.kg(-1).d(-1) and 20 g crude drug.kg(-1).d(-1))were both 39.4% (P<0.05). CONCLUSION: The extracts from Thymus quinquecostatus Celak have anti-tumor activities. The coefficient of spleen in group with alcohol extracts (40 g crude drug.kg(-1).d(-1))was close to normal value, and its coefficient of thymus was between that of the negative control group and the group with cyclophosphamide (0.02 g.kg(-1).d(-1)). The anti-tumor activity of the alcohol extracts was significantly higher than that of the control group and the tumor inhibition rate was depending on drug concentration. Depending on index of immunity,the extracts from Thymus quinquecostatus Celak may have some influences on immunity.

OBJECTIVE: To screen the anti-tumor fraction of ethanol extracts from Thymus quinquecostatus Celak and investigate its anti-tumor effect on human leukemia cell line. METHODS: Ethyl acetate, n-butanol and acetone fractions were separated from the ethanol extracts of wild Thymus quinquecostatus Celak. Growth inhibiting effects of these extracts on human leukemia cell lines K562 and HL-60 were determined by live cell counting and cell growth curve analysis. The possible anti-tumor mechanism was studied by morphological analysis with norcantharidin as a positive control. RESULTS: Ethyl acetate fraction could significantly inhibit the proliferations of K562 and HL-60 cells, and the inhibiting effect depended on the concentration of ethyl acetate fraction. Ethyl acetate fraction could induce apoptosis of K562 and HL-60 cells. The n-butanol and acetone fractions had no significant inhibiting effect on K562 and HL-60 cells. CONCLUSION: Ethyl acetate fraction is the major anti-tumor fraction in ethanol extracts from Thymus quinquecostatus Celak.

Objective To evaluate expression of E-cadherin and COX-2 in placentas of hypertensive disorder complieating pregnancy,and study the relationship between them.Method The E-cadherin and COX-2 of twenty-one cases gestational hypertension,along with 17 cases preeclampsia,and 20 cases normal pregnancy placentas were investigated by immunohistochemistry.Results The expression of E-cadherin was significantly higher and the expression of COX-2 was lower in preeclampsia cases than that in normal pregnancy(P＜0.01).The expression of E-cadherin negatively correlated with the expression of COX-2 (r=-0.371.P＜0.05).Conclusions The alteration of E-cadheria and COX-2 in placentas are correlated with preeelampsia.The up-reguhted expression of E-cadherin and down-regulated expression of COX-2 maybe regulated by the same factor.

The establishment of clinical medicine with professional degree is an important reformation of high-level professionals in medical field. With the expansion of the number of postgraduates of clinical medicine with professional degree, how to establish and perfect the training mode and how to improve the training quality has become an important research for the postgraduate education with professional degree. This paper discusses the establishment and perfection of training mode of full-time postgraduates of clinical medicine with professional degree by combining the documents and development of professional degree in my university.

Objective:To compare the content of total saponins in Paridis Rhizome from Wudang mountain area to explore the cor-relation between the quality of medicinal materials and the production areas and species. Methods: The content of total saponins in Paridis Rhizome was determined by an ultraviolet-visible spectrophotometer at 406nm with perchloric acid as the chromogenic reagent. Results:The saponins content in Paridis Rhizome from Wudang mountain area had obvious differences:the minimum was 1. 29%, and the maximum was up to 10. 22%. The content of total saponins had no obvious correlation with species, production area and altitude. Conclusion:The quality of Paridis Rhizome is unstable in Wudang mountain area, and that will affect the effectiveness and safety of the clinical medication. Only by promoting the standardized planting of Chinese medicine materials, the stable quality of Paridis Rhizo-me can be ensured.

Objective:To investigate the distribution of four polyphyllins in different parts of paris polyphylla by means of compa-ring the contents of relative constituents in paridis rhizome and fibril to provide reference for comprehensive exploitation and utilization of paris polyphylla. Methods:Paris polyphylla samples were collected from Wudang mountain area and Shennongjia forest area. The contents of main secondary metabolites including polyphillinⅠ,Ⅱ,Ⅵ andⅦin paridis rhizome and fibril were evaluated by HPLC. Results:The four polyphyllins contents had obvious differences in different parts of polyphylla. The total content of four polyphillins in fibril was significantly higher than that in rhizome. Diosgenin compositions had no significant difference in the two parts, and the con-tents of pennogenin compositions in fibril showed significantly higher than those in rhizome. Conclusion:As for the main chemical com-positions contained in polyphylla, there is chemistry equality between fibril and rhizome, thus both of them can be used for medicine. However, as for the contents of four steroidal saponins, the distribution of secondary metabolites has obvious difference between fibril and rhizome, and the result may be caused by steroidal saponins transferred to rhizome for storage after the synthesis in fibril.