OBJECTIVE To probe the pathogen′s distribution of ventilator-associated pneumonia(VAP),in order to offer the evidence of clinical therapy,prevent the onset of VAP and apply the antibiotics reasonably.METHODS We applied the methods of etiology,microscopic identification,bacteria culturing etc on 74 mechanical ventilation patients,and analyzed the etiology of early-onset and late-onset VAP in contrast.RESULTS Totally 121 pathogens were cultivated altogether in all 74 VAP patients.In the 36 pathogens which were cultivated from 29 early-onset VAP patients,there were 66.67% of simple culture(24 patients,24 strains) and 33.33% of co-culture(5 patients,12 strains),and in the 85 pathogens which were cultivated from 45 late-onset VAP patients there were 17.64% of simple culture(15 patients,15 strains) and 82.35% of co-culture(30 patients,70 strains).The proportion of co-culture in the late-onset VAP patients was prominantly higher than that in the early-onset ones(?2=27.821,P

Aneurysm, Dissecting ; therapy ; Aortic Aneurysm ; therapy ; Humans

Objective:To identify the healing effect of electrospun silk fibroin-BMP-2 as a biologic pulp capping agent to inflammatory pulp in rat caused by lipopolysaccharide ( LPS) .Methods:A total of 30 healthy adult male Wistar rats were randomly divided into five groups:(1) normal control group without operation;(2) blank control group without capping agents;(3) calcium hydroxide capping group;(4) electrospun silk fibroin capping group;(5) electrospun silk fibroin-BMP-2 capping group .Bilateral up-per first molars of each rat in group 2-5 were drilled to expose the pulp to LPS which was used to estab-lish a model of inflammatory pulp .The exposed pulp was capped with different capping agents or without capping agents.Then the hole was sealed.The animals were sacrificed on days 3, 7, and 14 post-opera-tion and histological analysis was carried out , including HE stain and CD 14 immunohistochemical stain . Results:On day 7 and 14 , the lowest inflammatory reaction score in HE stain among pulp capping groups was that of silk fibroin-BMP-2 group .The next were calcium hydroxide group and silk fibroin group .That of blank control group was the highest .The ranking of reparative dentine scores of those groups was just reversed.The D values of immunohistochemical stain of CD 14 were not significantly different in groups applied pulp capping agents but significantly lower than blank control group on days 3 and 7.However, the D value of silk fibroin-BMP-2 group ( 0 .145 ±0 .011 ) was significantly lower than blank control group (0.287 ±0.019), calcium hydroxide group (0.170 ±0.017) and silk fibroin group (0.175 ± 0 .018 ) on day 14 .Conclusion:Electrospun silk fibroin compounded with BMP-2 promoted wound hea-ling of exposed pulp and had better potential to stimulate formation of reparative dentine to establish a suitable environment for pulp recovery .

Asthma is a common disease with recurrent onset which severely affects patients' quality of life.Acupuncture can improve pulmonary functions in asthma patients and thus treat this disorder.To summarize the status of acupuncture treatment for asthma,we have collected clinical literatures published in the recent 10 years and analyzed the influence of acupuncture on pulmonary functions in asthma patients from the aspects of frequently used points,needling techniques,manipulation and mechanisms to provide references for treating asthma with acupuncture.

ObjectiveTo investigate the feasibility of poly l lysne to preparation microencapsules for cell transplantation therapies.MethodsUsing drop generative technique preparation Alginate poly l lysne Alginate (APA) microencapsules containing nerve cell/tissue. The concentration of nerve growth factor in supernatant was detected by two antibody sandwich method of enzyme linked immunosorbent assay.ResultsThe nerve cell/tissue in microencapsules retain reliable cell viability and function. Conclusions The APA is proved with reliable biocompatibility and strength,would work as an immunoisolation tools to exert important function in nerve renovate.

Objective To study the role of protective antigen(PA)and N-terminal segment of lethal factor (LFn)in the entrance of EGFP(enhanced green fluorescent protein)into HeLa cells. Methods The DNA fragments encoding LFn and EGFP were amplified,respectively,and cloned into the plasmid pET-21 a(+)one after another to construct a recombinant plasmid pET-LFn-EGFP. The plasmid was txansformed into BL21 cells to express LFn-EGFP protein under the induction of IPTG. The protein was purified by Ni chelating chromatography. After incubation with LFn-EGFP in the presence of PA or not, the HeLa cells were analyzed by flow cytometry or laser confocal microscopy. Results The fusion protein LFn-EGFP was purified by over 90% homogeneity and retained the ability of LF to bind with PA when incubated with J774A.1 macrophage cells,and could get into HeLa cells. Conclusion The LFn-EGFP could enter the HeLa cells in a PA independent pathway. But PA could help more LFn-EGFP molecules enter into HeLa cells.

A total of 121 subjects comprising 40 normal subjects,58 patients with overweight or obesity and 23 patients with Cushing's syndrome were recruited in the study.The modified radioimmunoassay (RIA) for salivary cortisol test was established'and its normal range was determined.Then the diagnostic value of the salivary cortisol for the initial diagnosis of Cushing's syndrome was evaluated and single midnight salivary cortisol test demonstrated a sensitivity of 100.0% and specificity of 91.4 %.Salivary cortisol test can be recommended as a first-line diagnostic parameter for Cushing's syndrome.

To establish an HPLC-MS/MS method for the analysis of quercetin, kaempferid and isorhamnetin in rats plasma and study its pharmamacokinetics after an intragastrical administration of Hippophae rhamnoides extracts. Five healthy male Sprague-Dawley (SD) rats were given single doses of H. rhamnoides extracts (quercetin 26.35 mg x kg(-1), kaempferid 4.040 mg x kg(-1), isorhamnetin 31.37 mg x kg(-1)), and then their orbital sinus blood samples were collected at different time points. The drug plasma concentration of the three flavonoids was determined by HPLC-MS/MS method. After that, the main pharmacokinetics parameters were calculated by using Kinetica 5. 0. 11 software. The methodological test showed that the linear concentration ranges of quercetin, kaempferid and isorhamnetin were 7.500-600.0 μg x L(-1) (R2 = 0.998 5), 1.000-80.00 μg x L(-1) (R2 = 0.998 5 ) and 10.00-800.0 μg x L(-1) (R2 = 0.998 0), respectively. The inner and inter-days precisions were both less than 14.0%. The plasma samples showed a good stability and consistency with the requirement of biological sample analysis after the samples were frozen once and placed at - 20 degrees C for 15 d and room temperature for 6 h and the treated analytes were placed at -20 degrees C for 24 h. For quercetin, the pharmacokinetic parameter t(½β), AUC(0-∞), MRT(0.∞), C.(max) and T(max) were (113.3 ± 19.37) min, (12 542.14 ± 3 504.05) μg x h x L(-1), (119.6 ± 13.29) h, (164.6 ± 27.33) μg x L(-1) and (5.199 ± 0.840 3) h, respectively. For kaempferid, the pharmacokinetic parameters t(½β), AUC(0-t), MRT(0-∞), C(max) and T(max) were (79.85 ± 17.15) min, (934.51 ± 94.59) μg x h x L(-1), (81.50 ± 13.75) h, (80.15 ± 14.24) μg x L(-1) and (3.827 ± 0.902 7) h, respectively. For isorhamnetin, the pharmacokinetic parameters t1,2,, AUC(0-t), MRT(0-∞), C(max) and T(max) were (118.3 ± 20.73) min, (26 067.77 ± 4 124.60) μg x h x L(-1), (129.0 ± 16.30) h, (269.6 ± 29.32) μg x L(-1) and (6.513 ± 1.450) h, respectively. The HPLC-MS/MS analysis method established in this study was proved to be sensitive and accurate and could be applied in the pharmacokinetic study of quercetin, kaempferid and isorhamnetin in rat plasma.
Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Hippophae ; chemistry ; Kaempferols ; blood ; pharmacokinetics ; Male ; Quercetin ; analogs & derivatives ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

The present study was aimed at the comparison of the pharmacokinetics of pure chlorogenic acid and extract of Solanum lyratum Thunb. The animals were allocated to two groups, and were administered chlorogenic acid or extract of S. lyratum Thunb. at a dose of 50.0 mg/kg orally. Blood samples were collected up to 8 h post-dosing. Plasma chlorogenic acid analyses were performed using an HPLC method with UV detector. The pharmacokinetic parameters were evaluated using non-compartmental assessment. Significant differences existed in the two groups for AUC0-t, AUC0-∞ and CLz/F. The reliable HPLC method was successfully applied to the determination of chlorogenic acid in rat plasma at dosage of 50.0 mg/kg.

Background The underlying mechanism of choroidal neovascularization(CNV) is multifactorial and complex.Focal adhesion kinase(FAK) plays a crucial role in controlling essential cellular processes and influencing distinct steps of the angiogenic response.But to our knowledge,seldom study on the effect of FAK on CNV formation has been reported previously.Objective In this study,the effect of several CNV risk factors on the expression of FAK in cultured retinal pigment epithelium(RPE) cells was investigated to illuminate effect of FAK on CNV.Methods Human RPE cells were isolated from donor eyes and exposed to H2O2,swallow of outer segment of photoreceptors(POS) and extracellular matrix(ECM) separately with the treating as follows:RPE cells were co-cultured with 10,20,50 and 100μmol/L H2O2 for 20 days;POS(1×106/ml) were co-cultivated with RPE cells for 20 days(setting control group,POS group,hypoxia group with 200μmol/L CoCl2,and POS+hyoxia group);RPE cells were cultured on the plates coated with 100mg/L fibronectin(FN),laminin(LN) or collagen typeⅠfor 30minutes or 1 hour.The expression of FAK and pFAK in RPE cells were examined by Western blot analysis.Results FAK was highly expressed in the 20μmol/L and 50μmol/L H2O2 groups compared with control group(P＜0.01);while he expression level of pFAK was reduced after treated with H2O2 in comparison with the control group(P＜0.01).After cultured with POS for 20 days,the undigested lysosome could be observed in RPE cells.The expressions of FAK and pFAK in RPE cells were not significantly changed between control group and POS groups(P＞0.05),but those in hypoxia group were significantly up-regulated in comparison with control group(P＜0.01).Compared with the hypoxia group,the expression amount of pFAK was elevated in POS+hyoxia group(P＜0.01).In comparison with control group,the increased pFAK expression was seen in FN,LN and collagen typeⅠtreating for 1-hour groups(P＜0.05,P＜0.01).Conclusion FAK pathway participates in several CNV-initiated signaling,such as H2O2,POS and ECM,in cultured RPE cells.It is reasonable to believe that FAK potentially plays an important role in CNV-dependent disorder.