4-Hydroxycoumarins ; blood ; Animals ; Chromatography, High Pressure Liquid ; methods

AIM:To construct eukaryotic expression vector expressing double shRNA sections targeting Survivin gene.METHODS: Eukaryotic expression vector expressing double shRNA sections targeting Survivin gene were designed and chemically synthesized.They were directionally inserted into plasmid pGenesil-1 with respectively U6 promoter and termination code,the common green fluorescence protein(EGFP) gene and Neo gene. In this way,the vector of pGenesil-1 shRNA containing 2 sections of Survivin shRNA were constructed and they were transfected into the pancreatic cancer cell Bx-PC3.Transfection was detected by fluorescence microscope.The inhibition expression of Survivin mRNA was measured by RT-PCR.RESULTS: HE1 and HE2 plasmids were identified by the biocatalyst cut which confirmed the exactitude and were analyzed by the sequence analysis which verified the perfect clone plasmid inserted by them.CONCLUSION: A eukaryotic expression vector of double short hairpin RNA for Survivin gene is successfully constructed.The pancreatic cancer cells Bx-PC3 succeed to be transfected and expression of Survivin mRNA is inhibited obviously.

Objective:To explore the effect of P-glycoprotein ( P-gp) activity on its mediated imatinib mesylate accumulation and intracellular drug membrane permeability. Methods: 1199G/wt ABCB1 and 1199A/mut recombinant plasmids were transferred into HEK293 cells, respectively, and the expression levels of mRNA in different cell models were investigated by RT-PCR. Cell Counting Kit-8(CCK-8) assay was used to detect the drug toxicity in different cell models, HPLC was applied to determine the drug concentra-tion in different cell models and evaluate the intracellular accumulation, and transmembrane resistance experiment was employed to de-tect the transmembrane permeability and evaluate the effect of P-gp activity on drug transportation. Results: Cytotoxicity test showed that the drug concentration in the transferred cells was lower than that in the control group, which proved that P-gp had the function of mediating drug out of cells. HPLC and transepithelial electrical resistance experiment showed that compared with the wild type of AB-CB1 (1199G) cells, mutation of ABCB1 1199A cells had stronger effect on P-gp mediated mesylate imatinib accumulation and drug membrane permeability. Conclusion:The experiment manifested that ABCB1 (1199G/A) site mutation can change the coding protein P-gp activity and the polymorphisms will lead to the increase of mesylate imatinib clearance rate and the decrease of effective drug con-centration in target cells. Meanwhile, the clarification of ABCB1 genetic types in clinics can guide the individualized medication of imatinib mesylate.

Objective Little is known about the role of low oxygen concentration in the formation of the capsule of Cryptococ -cus neoformans .This study was to investigate the effect of hypoxic condition on the capsule size of Cryptococcus neoformans . Methods We cultured 39 Cryptococcus neoformans strains in liquid Sabouraud medium and then incubated them at the normoxic (21%) and hypoxic concentration (1%), respectively.After 72 hours of incubation, we observed the capsules formed at different ox-ygen conditions under the Olympus microscope and the measured their sizes using the Axiovision software . Results Of the total number of Cryptococcus neoformans strains , 30 showed larger capsules in the normoxic than in the hypoxic condition , 17 with statisti-cally significant differences in their diameters (P<0.05), while the other 9 strains exhibited just the opposite results , 4 with statisti-cally significant differences in their diameters ( P<0.05). Conclusion Hypoxia might inhibit the growth of the Cryptococcus neoformans capsule.However, further studies are needed on its correlation with the serum or gene types of the Cryptococcus neofor-mans strain.

The study of protein-surface interactions represents one of the most important topics in the field of biomaterials.It is believed that blood cells do not actually contact the biomaterial surface directly,but rather interact with the molecular structure of the adsorbed protein layer.Therefore,protein plays a key role in regulation and induction of cells.It provides ideas for biomaterials designing that exploring the molecular mechanism of controlling protein adsorption and regulating cell response.This paper reviews the research progress of the interactions of protein and surfaces,and prospects the future research direction.

Objective To develop an accurate,reliable method without the need of initialization in animal PET modeling for estimation of the tracer kinetic parameters based on the artificial immune network.Methods The hepatic and left ventricular time activity curves (TACs) were obtained by drawing ROIs of liver tissue and left ventricle on dynamic 18F-FDG PET imaging of small mice.Meanwhile,the blood TAC was analyzed by sampling the tail vein blood at different time points after injection.The artificial immune network for parametric optimization of pharmacokinetics (PKAIN) was adapted to estimate the model parameters and the metabolic rate of glucose (Ki) was calculated.Results TACs of liver,left ventricle and tail vein blood were obtained.Based on the artificial immune network,Ki in 3 mice was estimated as 0.0024,0.0417 and 0.0047,respectively.The average weighted residual sum of squares of the output model generated by PKAIN was less than 0.0745 with a maximum standard deviation of 0.0084,which indicated that the proposed PKAIN method can provide accurate and reliable parametric estimation.Conclusion The PKAIN method could provide accurate and reliable tracer kinetic modeling in animal PET imaging without the need of initialization of model parameters.

Objective: To observe the effects of nerve block on MA and EA analgesia and mast cells degranulation in adjuvant arthritis rats in order to analyze the difference between the underlying peripheral mechanism of MA and EA analgesia. Methods: Zusanli (ST 36) was chosen as the acupuncturing point on adjuvant arthritis rats. Eighty SD rats were randomized into 10 groups, including control, model, normal+Lido, EA, Lido+EA, DLido+ZEA, Xlido+ZEA, MA, Lido+MA, Dlido+ZMA groups, with 8 rats in each group. Paw Withdraw Latencies and mast cells degranulation ratios were recorded and computed. Results: Compared with model group, the pain threshold (PT) and mast cells degranulation ratios of EA and MA groups were obviously increased (P<0.05 or P<0.01). The analgesia effects of both MA and EA were obviously attenuated after nerve block by pretreated with Lidocaine hydrochloride at the acupuncturing point or at the acupiont of proximal end in the same neural stem while they were not by the same manipulations at the acupoint of distal end. Degranulation ratios of mast cells caused by MA or EA were not obviously affected by nerve block. Conclusion: The effective signal of nerve conduction of MA analgesia is generated after the degranulation of mast cells in the process of acupoints activation or needle sense and it is the direct cause of generation of nerve signal. In the condition of EA, the acupoints activation is caused by the stimulation of electric signal on nerve receptor while the degranulation of mast cells is concomitant or feedback effect.

Objective To assess the diagnostic value of various preoperative imaging examinations for the point of the lateral femoral circumflex artery perforators through the deep fascia,in order to provide reasonable evidence for the design of location and protocol of anterolateral thigh perforator flap.Mehtods From September,2013 to December,2013,23 patients (including 1 patient with bilateral anterolateral thigh perforator flap) preparing anterolateral flap surgery were randomly divided into 4 groups(every group was 6 side).The patients in different groups were using different imaging techniques respectively,such as handheld doppler (HHD),Color doppler sonography (CDS),MDCT angiography (MDCTA),CDS and MDCTA united image location technology.The flap sizes varied from 8 cm × 5 cm to 28 cm × 12 cm.Moreover the coincidence rate between preoperative and intraoperative location of the perforator vessel was evaluated.Results Ali flaps were survived after the operation.The donor site were primarily closed while partial skin grafting was performed in 8 cases.The appearance and functional resuhs were satisfactory with following up for 3 to 9 months.The rates of coincidence between the preoperative and intraoperative location of perforating branches were HHD 61.53％,CDS 87.50％,MDCTA 85.71％,CDS and MDCTA united image location technology 93.33％ respectively.The HHD group showed significant decrease compared with the other groups (x2 =7.92,P ＜ 0.01; x2 =6.15,P ＜ 0.05; x2 =12.54,P ＜ 0.01,respectively).The CDS group had no statistically difference with the MDCTA group (P ＞ 0.05).The united CDS and MDCTA group showed significant increase compared with the other groups (x2 =4.22,P ＜ 0.05; x2 =3.94,P ＜ 0.05).Conclusion CDS and MDCTA united image location technology are more accurate for perforator preoperative location in auterolateral thigh flap,and should be widely used.

Objective Trichophyton rubrum strains can cause superficial infection and also deep tissue infection, but relevant animal model has not been reported yet.The aim of this study was to construct an animal model of granuloma infected by T.rubrum. Methods Three T.rubrum strains isolated from clinical granuloma tissues, 2 T.rubrum strains isolated from tinea infection and a standard strain of ATCCMYA4438 were selected.Corticosteroids were given to the Balb/C mice before and after the injection of the T. rubrum and mucin suspension and the mice model of granuloma was established.Direct microcopy, culture and histopathologic method were adopt to verify the infection effects. Results The mice inoculated with the T.rubrum granuloma strains with mucin suspension were examined after 21 days in the condition of applying appropriate dose of glucocorticoids.Direct microscopic examination showed the slender mycelium, fungal culture showed the growth of colony and histopathology showed excessive keratinization of foot tissue, formation of granuloma in the dermis with inflammatory cell infiltration of neutro-philic granulocyte and lymphocytes.However, the mice inoculated with the T.Rubrum tinea strains with mucin suspension showed the negative result. Conclusion The rubrum granuloma mice model can be es-tablished using the clinical isolates of T.rubrum granuloma strains with the mucin and glucocorticoids interventions.

Social governance is a novel form of public administration made based on the analysis and judgment for the power pattern in the course of social administration, a model advocated in the premise of a clearly positioned relationship among the government, marketplace, society and citizens.Social evaluation is an effective carrier and means leveraging the social governance theory, which has found extensive and outstanding use in such issues as expression of public interests and responsibilities,and in tackling sharp social problems. The authors called into play the scenario analysis based on public hospital management reforms with the social governance theory. In addition, they explored social governance at home and abroad as well as the theories, meaning and progress of social evaluation, and analyzed the pathways and probes in social evaluation and governance in the fields in question. On such basis, the paper proposed to explore and develop the social evaluation strategies and study framework for public hospitals in terms of healthcare management, aiming at building the social governance system and policies for public hospitals.