Objective To observe the effects of dexmedetomidine on inflammatory responses and insulin resistance during cardiac operations with cardiopulmonary bypass(CPB).Methods Fifty patients scheduled for elective cardiac valve replacement with CPB were equally and randomly divided into two groups using a random number table:observation group and control group(n =25).A loading dose of dexmedetomidine 1.0 μg/kg was injected intravenously over 1 5 min after induction,followed by continuous infusion at 0.4 μg·kg-1 ·h-1 until the end of CPB in doservation group.While the e-qual volume of normal saline was given in control group.After induction(T1 ),at 30 min after the be-ginning of CPB(T2 ),at the end of CPB(T3 ),and at 2 h after the end of CPB(T4 ),the jugular venous blood samples were taken for determination of serum concentrations of TNF-a,IL-6,insulin and blood glucose.The insulin sensitivity index (ISI ) = 1/(glucose × insulin )was obtained. Results Compared with control group,the serum TNF-a and IL-6 concentrations at T2-T4 were sig-nificantly decreased in observation group(P <0.05),the serum insulin and blood glucose concentra-tions at T2 ,T3 were significantly decreased in observation group(P <0.05 ),the ISI at T2 ,T3 were significantly increased in observation group(P <0.05).Conclusion Dexmedetomidine can reduce in-flammatory responses,thus reducing insulin resistance and blood glucose during cardiac operations with CPB.

Objective To explore the in-patients' evaluation and the correlative requirement for the bachelor degree nursing students. Methods Investigated 10 patients by using the self-developed interview outline, and then summarized the key points of these talking. Results The evaluation from patients to bachelor degree nursing students had included 6 aspects: attitude, clinical nursing skills, knowing about the concrete condition of patients, the wide of nursing students' knowledge, the health education for patients and the ability of communication and cooperation. Conclusion Different have their own different emphasis when evaluate the nursing students, but the evaluation from patients were praiseful.

Objective To observe the effect of silencing Snail gene on the invasion and proliferation ability of human pancreatic cancer cell line PANC1.Methods Lentiviral vectors that can express small hairpin RNA(shRNA) targeting human Snail gene(shRNA-Snail) or shRNA sequence that did not match any known mRNA(shRNA-NC) were constructed,and transfected into PANC1 cells.Untransfected cells served as control.mRNA and protein expression of Snail,α-smooth muscle actin (α-SMA) and E-cadherin was determined by real time quantitative PCR and Western blotting,respectively.In vitro invasion ability was tested by Transwell model.Proliferation ability was measured by CCK-8 assay.Results Compared with those in shRNA-NC group,Snail mRNA (0.27 ± 0.02 vs 0.92 ± 0.03) and protein level (0.26 ± 0.02 vs 0.80 ± 0.02),and α-SMA mRNA (0.33 ±0.04 vs 0.97 ±0.07) and protein level (0.31 ±0.04 vs 0.74 ±0.06) in shRNA-Snail group were obviously decreased,but E-cadherin mRNA (1.57 ± 0.45 vs 0.95 ± 0.08) and protein level (0.86 ± 0.03 vs 0.20 ± 0.03) were greatly increased.The number of cells permeating the septum of transwell [(6.80 ± 0.73)/400 magnification vs (26.80 ± 2.52)/400 magnification,P ＜0.01] was significantly decreased,and cell proliferation was inhibited (0.74 ± 0.05 vs 1.47 ± 0.04,P ＜ 0.01).All the differences above were statistically significant (all P ＜ 0.01).No significant differences were observed between shRNA-NC and normal control group.Conclusions Silencing Snail gene may restrain the invasion and proliferation ability of PANC1 cells to a certain degree.

OBJECTIVE:To provide reference for developing the implementation path of patent standardization for pharmaceuti-cal industry. METHODS:For the situation that pharmaceutical industry is still in the exploratory stage and lacks of viable experi-ence,the mature experiences of electronic communication industry in patent leading standardization implementation,authoritative organizations helping implementing patent standardization and industry alliance to promote the development of patent standardiza-tion were used as reference,combining with standard development cycle,trying to develop the implementation path of patent stan-dardization that met characteristics of pharmaceutical industry. RESULTS & CONCLUSIONS:The implementation path of patent standardization for pharmaceutical industry was divided into standard formation stage,promotion stage and upgrade stage. The main path in each stage was from patent to registration standards officially formed,guiding by policies and regulations of Food and Drug Administration to conduct standard promote by drug generation,and upgrading the drug standard by patent replacement.

Objective To construct a recombinant adenovirus vector containing the gene of major histocompatibility complex(MHC)class Ⅱ transactivator(C Ⅱ TA).Methods The restriction fragment of CIITA was inserted into pUC57 vector with EeoR Ⅰ and Xho Ⅰ.Then,recombinant plasmid pShutde-GFP-CMV CⅡTA was constructed with EcoR Ⅰ and Sal Ⅰ,and was confirmed by restriction enzyme digestion and sequeneing.After the treatment with Ⅰ-Ceu Ⅰ and Ⅰ-See Ⅰ,the fragment C Ⅱ TA from recombinant plasmid DShuttle-GFP-CMV.CⅡTA Was inserted into vector pAdxsi.And the pAdxsi-GFP-C Ⅱ TA wag packed into liposome,and was transfected to 293 cens.Results Recombinant plasmid pShuttle-GFP-CMV-C Ⅱ TA Was constructed successfully. After packed into vector pAdxsi, and transfected to 293 cells, significant virus Dlaques were observed,which showed the successful homologous recombination.The titer of the purified AdC Ⅱ TA was 2.0×10~(11) PFU/ml.Conclusions Recombinant adenovirus AdC Ⅱ TA containing gene of MHC class Ⅱ transactivator was established successfully.

Objective To observe the effect of triptolide combined with gemcitabine on proliferation and apoptosis of pancreatic cancer cells ,and to analyze the relevant mechanisms .Methods After treated with TPL ,GEM or TPL combined with GEM in vitro , PANC-1 cells proliferation was accessed by MTT assay and the interaction between the two drugs was calculated .Apoptotic mor-phological changes and apoptosis rate of the cells were investigated by Hoechst 33258 staining and flow cytometry ,respectively .The expression of signal transduction and transcription factor 3(STAT3) ,cysteine aspartate specific proteases-3(caspase-3) protein were detected by Western blot analysis .Results TPL ,GEM or TPL combined with GEM could significantly inhibit the prolifera-tion of PANC-1 cells ,and the combination of the two drugs had a synergistic effect .The cells of the TPL group ,GEM group ,as well as the combined group showed typical apoptotic morphological changes .Compared with the TPL group and GEM group ,the number of apoptotic cells of the combined group increased significantly .Compared with the control group ,the cells apoptosis rate of the TPL group ,GEM group and combined group was significantly increased (P< 0 .05) ,and the apoptosis rate of the the combined group was significantly higher than that of the monotherapy group(P<0 .05) .TPL combined with GEM synergistically inhibited p-STAT3 protein expression and activated caspase-3 protein expression .Conclusion TPL combined with GEM can synergistically in-hibit proliferation and induce apoptosis of pancreatic cancer PANC-1 cells ,its mechanism is related to the inhibition of STAT 3 sig-naling pathway ,promotion the expression of caspase-3 protein .

Objective:To establish an HPLC method for the determination of genistein in genistein-loaded Eudragit nanoparticles. Methods:The HPLC analysis was carried out on a DiamonsilTM C18(250 mm ×4.6 mm,5μm)column with the mobile phase of meth-anol-water (85∶15) at the flow rate of 1. 0 ml·min-1 . The detection wavelength was set at 261nm and the column temperature was 25℃. Results: A good separation was displayed between the drug peak and the excipient peaks. An excellent linearity range for genistein was obtained from 1.0 μg·ml-1 to 100.0 μg·ml-1(r =0.999 8). The recovery of genistein was 99.85%(RSD =0. 65%,n=9). Conclusion:The method is simple, rapid, accurate and specific, which is suitable for the determination of content and encapsulation efficiency of genistein-loaded nanoparticles.

AIM:To investigate the effect of ?-cyclodextrin inclusion on transdermal absorption of volatile oil in lilac-cassiabark Gel. METHODS: The release test of two kinds of lilac-cassiabark Gel(gel A without ?-cyclodextrin inclusion and gel B with ?-cyclodextrin inclusion) was assayed with Franz diffusion cell with the help of the cumulative amount of eugenol in unit area in vitro. RESULTS: The percutaneous absorption process of eugenol in two kinds of gels were conformed to zero-order releasing model,and the transdermal rates were 16.212?4.128 and 11.344?1.929 ?g/(cm~2?h) respectively.Compared with gel A,the transdermal rate of eugenol was slower in gel B(P

Objective: To construct HBV adr gene vaccine pCMVS 2-S and to observe the specific humoral immune responses in C57BL/6 mice after gene immunization. Methods: The HBV gene vaccine was injected into tibialis anterior muscles of C57BL/6 mice. ELISA was used to detected the anti-HBs antibody in mice sera at various time points after gene transfer. Results: Anti-HBs in sera of mice was tested positive 1 week after plasmid injected, the level of antibody peaked 4 weeks later, and kept high titer for at least 2 months. Conclusion: The good response of humoral immunity can be induced by injection of pCMVS 2-S in C57BL/6 mice.

Objective To determine the lower limit of detection (LLOD) and cut off values of K-ras mutation detection by peptide nucleic acid (PNA) clamping-PCR.Methods The genomic DNA of pancreatic cancer cell lines (PANC1 and SW1990) with codonl2,13 mutation and the genomic DNA of placenta with K-ras wild type were mixed and diluted serially into samples with different mutation rate (0,0.1％,0.2％,0.4％,0.8％,1.6％,3.1％,6.25％,12.5％,25％,50％),PANC1 cells with 1％ mutation rate and SW1990 cells with 30％ mutation rate and 4 samples with the quantity of DNA was 50,20,5,1 ng and 50,10,5,1 ng was prepared.Codon 12,13 mutation of K-ras was determined by PNA-PCR,and the mutation Ct values,overall Ct values were collected,and the △Ct values (mutation Ct values-overall Ct values) were calculated,and the tests were repeated for 10 times.ROC curve was used to analyze the △Ct values and determine the best cut off values for K-ras mutation,and the positive diagnostic rate,LLOD was evaluated.Results The mutation Ct,△Ct values of codon 12 mutation of PANC1 and codon 13 mutation of SW1990 of all the different mutation rates were statistically significantly different (P ＜ 0.05) when compared with negative control group,but the overall Ct values were not statistically significantly different from that of negative control group.For detection of K-ras codon 12 mutation by ROC curve,the relevant area of ROC curve (AUC) was 0.926,the optimum cut off value of △CT was 11,the sensitivity and specificity were 84％ and 100％,respectively,and the LLOD was 0.4 ng.For detection of K-ras codon 13 mutation by ROC curve,the relevant AUC was 0.906,the optimum cut off value of △CT was 9.5,the sensitivity and specificity were 71％ and 100％,respectively,and the LLOD was 1.5 ng.The mutation detection results of fixed rate further confirmed the LLOD.Conclusions This study successfully defines LLOD and cut off value of PNA clamping-PCR/K-ras method in detection of K-ras 12 and 13 codon mutations.This method meets the requirement of clinical application.