Objective To study the process of foam fractionation of polyphyllin in semi-batch mode. Methods Taking enrichment ratio, recovery rate of polyphyllin, and purity ratio as the performance criteria and using single examining method to examine the operational parameters, i.e. operation mode, air flow rate, initial feed concentration, solution pH value, initial feed height and temperature on separation performance. The optimal conditions of the process were obtained finally. Results The separation performance is good when gas flow rate is 400 mL/min, initial feed concentration (polyphyllins content) is 0.3 mg/mL, pH value is 7, feed height is 30 cm, and feed temperature is 40 ℃. The enrichment ratio is 25.7, recovery ratio is 42.1%, and the foam liquids purity of total polyphyllin is 41.4%, which is 4.5 times higher than that in feed purity. Conclusion Foam fractionation technique could be applied to separate polyphyllin.

Objective To explore a safe and high efficiency way of gene transfection of autocrine motility factor(AMF) in order to provide experimental basis for transplantation of myoblasts carrying AMF gone. Methods Sprague Dawley rat myoblasts were cultured, purified, proliferated and immunohisto-chemically verified. Then, the myoblasts were transfected with AMF and eGFP (enhanced green fluores-cent protein) gene by FIV (feline immunodeficiency virus). Fluorescence microscope and laser scanning confocal microscope were employed to detect eGFP so as to verify positive transfection rate. Expression of AMF was detected by immunohistochemical method. Results Myoblasts with 98% purity could he ob-tained after two weeks of primary culture and purification. Positive transfection rate reached 90.4% when MOI (multiplicity of infection) was 100 (P <0.01). The transfected AMF gene could express normally. Conclusions Explant culture is a proper way in rat myoblast culture. Meanwhile, AMF gene can he effectively transfected into rat myoblast and well expressed via FIV.

Objective To compare the clinical efficiency of two methods in the open surgery treatment of seconda-ry common bile duet stones: transeytie common bile duet exploration with eholedoehoscope (TCBDE),or traditional eholedoehotomy with T-tube drainage.Methods Retrospectively analysis the treatment of 113 eases with secondary bile duet stones (TCBDE group 58 cases ,T-tube group 45 cases) ; Compare the achievement ratio,operating time; complications,average hospital stay and cost between two groups.Results All procedure is successful,without any severe complications.There are no significant differences of operating time.The TCBDE group has sig-nificantly shorter hospital stay and lower cost.Conclusion TCBDE has significant advantages than the traditional choledochotomy,and should be the predominant method in the open surgery of secondary common bile duet stones.

Objective　To explore the effect of hypoxia on the apoptosis of cultured human umbilical vein endothelial cells（HUVECs） and the role of vascular endothelial growth factor（VEGF） in inhibition of apoptosis． Methods　①Culture and identification of HUVECs．②Establishment of hypoxic model（0，12，24，48 h）in HUVECs．③Incubation of HUVECs with VEGF(0 ng, 100 ng) under hypoxic condition for 24 h. ④Detection of apoptosis of HUVECs with TUNEL method． Results　The percentages of apoptosis were different under different hypoxic conditions． The longer hypoxic time was，the higher apoptosis percentage was．VEGF reduced the apoptosis of HUVECs induced by hepoxia． Conclusion　Over-apoptosis EVCs in one of the important factors for the impairment of endothelial function. HEGF inhibits the apoptosis of HVCs and having a pretive function on them.

In recent years, the biopharmaceutical industry has grown rapidly, and the market size of monoclonal antibody drugs has increased significantly. Accurate structural characterization and quality control are the supporting technologies for the development of monoclonal antibody drugs. As a significant post-translational modification of antibody drugs, glycosylation has an important influence on its efficacy, stability, and immunogenicity. The existing literature usually uses liquid chromatography-mass spectrometry to perform major glycosylation modifications of monoclonal antibody drugs. Characterization, there are few studies on low-abundance glycosylation, but the characterization and control of low-abundance glycosylation cannot be ignored. In this study, we have established a qualitative and quantitative analysis technology for N-glycans based on RapiFluor-MS reagent-labeled monoclonal antibody drugs. This method has a short sample processing time and high sensitivity. It can not only characterize the main glycoforms of three monoclonal antibody drugs (adalimumab, bevacizumab, and trastuzumab) but also can quantify low-abundance N-glycans. The results of the study showed that the main glycoforms specified in the Pharmacopoeia could be detected in different batches of monoclonal antibody drugs, but the content of N-glycans in different batches of samples is not identical. After that, we analyzed the N-glycans connection sites and glycoforms at the intact glycopeptide level, further enriching the N-glycans structure information of the monoclonal antibody. The qualitative and quantitative analysis technology of N-glycans based on RapiFluor-MS reagent-labeled monoclonal antibody drugs can realize the in-depth characterization and control of glycosylation modification of monoclonal antibody drugs.

Objective:To quantify the tryptase and T cell immunoglobulin mucin domain-3(TIM-3)double positive mast cells in hu-man chronic periodontitis tissue using double immunofluorescence staining.Methods:25 healthy controls,28 chronic mild periodontitis and 30 chronic advanced periodontitis patients were included.The gingival specimens were stained with HE for histology,and with double immunofluorescence staining for the identification of tryptase and TIM-3 double positive mast cells in gingival tissue.Results:In chronic periodontitis tissue the degree of gingival inflammation was significantly increased,the densities(cells/mm2 )of tryptase and TIM-3 double positive mast cells were significantly increased(P<0.05),in addition,that in chronic advanced periodontitis group was significantly higher than in the chronic mild periodontitis group(P<0.05).Conclusion:Tryptase and TIM-3 double positive mast cells has the similar tendency as the severity of periodontitis inflammation in human periodontitis tissue.Tryptase and TIM-3 double positive mast cells may play an important role in human chronic periodontitis.

Objective To investigate the protective effects of ethanol-extract of Halenia elliptica D. Don on chemical hepatic injury in mice. Methods Mouse models of chemical hepatic injury were induced by intraperitoneal injection of 0.12 %CCL4. Extract of Halenia elliptica D. Don by alcohol was administered,and serum ALT,AST activities and liver glycogen level were measured in mice. Results Compared with the models,the enzyme activities of ALT and AST were significantly reduced and the content of liver glycogen was significantly increased in the ethanol-ex tract of Halenia elliptica groups. Conclusion It is indicated that ethanol-extract of Halenia elliptica D. Don has an effect in protecting the liver.

AIM:To determine the relationship between ischemia,hypoxia and the production of vascular endothelial growth factor in rat myocardium and its basic mechanism. METHODS:(1) 28 Wistar rats were randomly divided into 4 groups: group A,normal control;group B,1 day's acute myocardial infarction;group C,3 day's acute myocardial infarction;group D, 7 day's acute myocardial infarction. (2) Rat cardiac myocytes cultured were primarily divided into some groups, hypoxia incubated 24 hours; PMA groups, hypoxia incubated 24 hours with PKC activator (PMA), A 0 ng/mL; B 10 ng/mL; C 100 ng/mL; D 1 000 ng/mL; Chelerythrine groups, hypoxia incubated 24 hours with PKC inhibitor (chelerythrine), A 0 nmol/L; B 10 nmol/L. (3) By computer scanned and quantitated, vascular endothelial growth factor (VEGF) protein was detected with immunohistochemical technique. RESULTS: The longer time of ischemia and hypoxia was, the higher the VEGF production. The relationship was found between the time of ischemia or hypoxia and the production of VEGF. The production of VEGF protein was further promoted by PMA with different concentration, decreased by chelerythrine.CONCLUSION:Ischemia or hypoxia strongly stimulated the production of VEGF in myocardium, which played an important role in autoprotecting of ischemic or hypoxic myocardium. Hypoxia-induced PKC activation is one kind of basic mechanisms in this course.

?AIM: To explore the effects of “one week with one case” teaching method with college - enterprise cooperation in the theoretical classes of contact lens courses, which provide the basis for teaching reform.?METHODS: Fifty-six students in optometry major of Grade 2012 from Xi’an Medical College were divided into 2 groups randomly. The experimental group of 28 students used “one week with one case” teaching method with college-enterprise cooperation; the control group of 28 people used traditional “one week with one case”teaching method. The examination scores and questionnaire were used to evaluate the teaching effects.?RESULTS:The students of experimental group acquired higher test scores in short-answer questions and the case analysis questions compared with students of control group (P<0. 05). The questionnaire survey showed the“one week with one case” teaching method with college-enterprise cooperation acquired higher scores in 6 items which include the intensity of learning interest, information acquisition ability, team cooperation ability, communication skills, oral communication ability and the satisfaction of teaching method (P<0. 05). Four items included participation, preparation, communication ability and team spirit of experimental group students were significantly superior to those of the control group students (P<0. 05).?CONCLUSION:“One week with one case” teaching method with college-enterprise cooperation can improve comprehensive ability of students. It is an effective teaching method with the characteristics of the contact lens courses.