Objective To screen proteins binding with interferon?(IFN?)from human hepatic cDNA libraty by yeast-two hybrid technique.Methods The IFN?gene was amplified by polymerase chain reaction(PCR)and constructed into pGBKT7 vector as the bait plasmid in yeast-two hybrid system3,pGBKT7-IFN?was then transfected into yeast AH109.The transfected yeast were mated with yeast Y187 containing liver cDNA library plasmid in 2?YPDA medium.Diploid yeast was plated on synthetic dropout nutrient medium(SD/-Trp Leu-His-Ade)and synthetic dropout nutrient medium(SD/-Trp-Leu-His-Ade)containing X-?-gal for selecting.After plasmid extracting and en- zyme cutting analysis,the blue colonies were performed sequence analysis,the results were analyzed by bioinformatics.Results IFN?gene was successfully cloned and expressed in yeast cells.Thirty- four positive colonies were obtained using yeast-two hybrid technique.After sequence analysis,eight clones were found may have a binding effect with IFN protein.Conclusions IFN?genes was success- ful cloned and eight proteins that could bind with IFN?protein were also screened.

Objective To evaluate the diagnostic value of velocity parameters of Tardus-Parvus for the detection of renal artery stenosis ( RAS) ( diameter reduction ≥50%) and to determine the useful cutoffs for these parameters .Methods A study group was composed of 221 renal arteries that were detected Tardus-Parvus by color Doppler flow imaging and were referred to abdomino-aorto-re-nal arteriography afterwards .Five Doppler parameters including the peak systolic velocity ( PSV ) , end-diastolic minimum velocity (EDV), resistance index (RI), acceleration time (AT), and accelerated velocity (AC) from each location including renal aorta ( MRA) , segmental artery ( SRA) , interlobar artery ( IRA) , and arcuate artery ( ARA) were archived and compared among the differ-ent groups .Renal artery angiography showed arterial canon reduced 50% or higher RAS .Arteries were considered stenosed on renal arteriography if there was a diameter reduction of greater than 50%.Statistical analysis to determine the best parameter for predicting a RAS was performed with the receiver operating characteristic ( ROC) curves.The sensitivity, specificity, and negative and positive predicting values at various cutoffs were calculated .Results Renal artery stenosis degree was less than 50% in 16 cases, 50%~99%in 197 cases, completely occluded in 2 cases, and no stenosis in 6 case by renal arteriography .For RAS with inner diameter re-duction of 50%or more, the ROC curve analysis showed renal artery flow velocity had a good sensitivity and specificity , 50%~99%of the RAS had optimal threshold value in PSV 20 cm/s, EDV 10 cm/s, RI 0.50, AT 0.09 s, and AC 1.5 m/s2.Conclusions Tardus-Parvus had high specificity and sensitivity for diagnosis of stenosis rate in more than 50% of the RAS, low sensitivity for the narrow degree in 0~49%, and no clinical value for the renal artery occlusion .

Background Intracameral or intracorneal administration of amphotericin B (AMB) can achieve significant therapeutic efficacy in the treatment of recalcitrant fungal keratitis in cases that do not respond to conventional antifungal therapy. However, the ocular pharmacokinetics of the two routes of administration is unclear.Objective The goal of this study was to investigate the level of amphotericin B in cornea and aqueous humor of rabbits after administration of AMB via three different routes. Methods Forty-five healthy domestic rabbits were randomly divided into three groups. 1% amphotericin B of 10 μg was intrastromally or intracamerally injected into 15 rabbits, respectively,in group A and group B. Topical 0. 25% amphotericin B was topically administered to the eyes with corneal epithelial debridement (group C). Experimental animals were sacrificed and the corneas and aqueous humor samples were obtained for the detection of levels of amphotericin B at 30 minutes,6 hours, 1 day,3 and 7 days by high-performance liquid chromatography (HPLC). Results Calibration curves were linear over the range of 0. 10-100. 00 mg/L. The concentration of 0. 10 mg/L was the lowest quantifiable limit. The recovery of amphotericin B ranged from 89. 1% -95.7% from aqueous humor samples and 81.4% -83.6% from the cornea samples. After a single injection,effective drug levels were achieved and maintained for 7 days in cornea in group A, exceeding the minimum inhibitory concentration at which 90% of isolates are inhibited (MIC90) for a wide spectrum of fungi and molds with significant differences in comparison with group B and group C ( P＜0. 05 ). Effective drug levels were achieved in the aqueous humor in group B at 30 minutes after a single injection, but drug levels decreased dramatically within 6 hours. The evident differences were found between group B and group A or group C (P＜ 0.05). A considerable amount of amphotericin B was detected in the cornea and aqueous humor in group C within 1 day.Conclusion Effective high drug levels can be reached in rabbit cornea and aqueous humor after intrastromal and intracameral injection, respectively. Penetration of topical amphotericin B was greatly elevated after epithelial debridement.

Animals ; Aorta ; pathology ; Atherosclerosis ; blood ; etiology ; metabolism ; Chemokine CCL2 ; metabolism ; Cholesterol ; blood ; Cholesterol, Dietary ; administration & dosage ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Dehydroepiandrosterone ; pharmacology ; Diet, Atherogenic ; Immunohistochemistry ; Rabbits ; Random Allocation ; Triglycerides ; blood ; Vascular Cell Adhesion Molecule-1 ; metabolism

The establishment of high specificity and sensitivity method of small molecule monoclonal antibody-based immunoassay has a great importance in the study of small molecule compounds in Chinese medicine, wherein synthesis of small molecule artificial antigen is a critical step in the preparation of small molecule antibodies. Oxidation method using sodium iodide was used to synthesize immunogenic antigen (FRn-BSA) and coating antigen (FRn-OVA) of forsythin. UV spectroscopy and thin layer chromatography showed that forsythin was successfully conjugated with BSA and OVA. After immuned FRn-BSA, the mice could specifically produce anti-forsythin antibodies with titer up to 1:8 000, and the linear range was from 1 mg x L(-1) to 100 mg x L(-1). In this paper, the artificial antigen of forsythin was successfully synthesized, which can be applied for preparation of monoclonal antibodies and establishment of appropriate immune method.
Animals ; Antibodies ; immunology ; Antigens ; chemistry ; immunology ; Bridged Bicyclo Compounds, Heterocyclic ; chemistry ; immunology ; Drugs, Chinese Herbal ; chemistry ; Furans ; chemistry ; immunology ; Male ; Mice ; Mice, Inbred BALB C

Objective To analyze the difference between nurses' emotional labor and job burnout in demography variables.Methods 310 nurses were evaluated with the Style Scale For Emotional Labor and Job Burnout Inventory (MBI-GS Scale).Differences of emotional labor using three different emotional labor strategies of surface process,deep process and automotive regulation and job burnout of nurses with different marital status,education,age,working experience and job title were analyzed.Results Unmarried nurses use surface process more frequently (t=4.475,P =0.000) than married nurses.Deep process was used less frequent by unmarried nurses than married nurse (t =-3.221,P =0.001).There was no significant difference in the usage of automotive regulation and job burnout situation among nurses with different marital status(P ＞ 0.05).Nurses with junior college degree or lower used surface process more frequent (t =2.215,P =O.028) and automotive regulation less frequently (t =-0.022,P =0.044) than nurses with bachelor degree or above.There is no significant difference in the usage of deep process strategy and job burnout situation among nurses with different education(P ＞0.05).Significant difference was found in the usage of deep process strategy and job burnout situation among nurses with different age (F =6.84,5.69,respectively; P ＜ 0.01).Elder nurses (36 years or older) used deep process more frequently.The job burnout feeling decreases with the increasing of the age.There is no significant difference in the usage of surface process and automotive regulation strategies among nurses with different ages (P ＞ 0.05).There was significant difference in the usage of deep process emotional labor strategy and job burnout situation among nurses with different working experiences (F =7.816,3.198,respectively; P ＜ 0.01 or 0.05).Nurses with 1-2 year working experience used surface process more frequently.As the working year grows,nurses tended to use deep process strategy more frequently and the job burnout feeling reduced.There is no significant difference in the usage of surface process and automotive regulation strategies among nurses with different working years (P ＞ 0.05).Nurses with title as head nurse or above used surface process strategy less frequently (t =1.407,P =0.046) and deep process more frequently (t =-2.229,P =0.014) than other nurses.Conclusions There is significant difference in the job burnout feeling and using of three different emotional labor strategies of surface process,deep process and automotive regulation among nurses with different marital status,education,age,working experience and job title.

Laser-induced breakdown spectroscopy (LIBS) was used for analysis of the distribution of S,Mn,Fe,Cr,Mo,Si,Al in a 34CrNiMo6 steel sample cut from a main shaft of wind driven generator.The MnS inclusion area in each ablation craters cover zone was extracted in the way of comparing the metallograph captured by optical microscopy before and after LIBS scanning ablation.The statistic relation between MnS inclusion area and signal intensity of S and Mn was analyzed.The result showed that the abnormal signal of S and Mn occurred at the same position with the existence of MnS inclusion,and their signal intensity showed linear relationship.The abnormal signal of S and Mn were triggered mainly by MnS inclusion.The statistic result also showed linear relationship between signal intensity and MnS inclusion area both for S and Mn.It was possible to determine the inclusion type,size and distribution by analyzing abnormal signal.A simplified ablation model was established to calculate the relation of S and Mn content to MnS inclusion area.The arithmetic result showed a linear relation between the content and MnS inclusion area both for S and Mn.The calculation confirmed the linear relationship between signal intensity and inclusion area observed in experiment.The linear relationship could be interfered by macro-segregation,micro-segregation,deviation in measuring inclusion area,and inclusion spatter in pre-ablation.

Objective To discuss the application of Six Sigma management in quality reinforcement of coronary artery CTA,in order to improve patients satisfaction regarding medical nursing.Methods 1 388 cases who had received coronary artery CTA from July to December 2011 in Peking University Shenzhen Hospital were randomly divided into the observation group of 456 cases and the control group of 476 cases.The patients in control group received conventional CT examination while the observation group received Six Sigma management CT examination.Two groups' score of image quality,defect rate and satisfaction were compared.Results The score of image quality was (4.17 ±0.72) in the observation group and (3.04 ± 1.07) in the control group.The difference was statistically significant (t =8.674,P ＜ 0.01).The defect rate was 0.74％ in observation group,lower than 4.86％ in the control group,and the difference was statistically significant (x2 =373.14,P ＜0.01).Patients' satisfaction was 95.60％ and doctors' satisfaction was 98.00％ in the observation group,both were higher than those in the control group (88.60％ and 92.00％,respectively),and the differences were statistically significant (x2 =11.91,7.58,respectively; P ＜ 0.01).Conclusions Application of Six Sigma management can effectively enhance the quality of coronary artery CTA and increase the quality and efficiency of work.

Sustained activation of β adrenergic receptor (βAR) leads to pathologic cardiac hypertrophy. However, the related mechanisms still remain unclear. In this study, we observe how N-acetylcysteine (NAC) affects the oxidative stress and calcium/calmodulin-dependent protein kinase II (CaMKII) expression in heart of isoproterenol (ISO)-stimulated rats, and investigate whether oxidative stress and CaMKII contribute to the development of sustained βAR-stimulated cardiac hypertrophy. Healthy male Wistar rats were randomly separated into 4 groups: control (CTRL), ISO-treated (ISO), control with NAC supplement (CTRL+NAC) and ISO-treated with NAC supplement (ISO+NAC) groups (6 rats in each group). Systolic blood pressure (SBP) was measured in awake rats with the tail-cuff method every week for two weeks. Heart weight/body weight ratio (HW/BW) and HE staining were used for the detection of myocardial hypertrophy. Myocardial mitochondrial reactive oxygen species (ROS) levels were measured by DCF fluorometry. The expressions of activated-CaMKII (p-CaMKII/CaMKII) and NADPH oxidase 4 (NOX(4)) were determined by Western blot analysis. The results showed that ISO-treated (i.p., daily 3 mg/kg, 2 weeks) rats developed an obvious cardiac hypertrophy as expressed by increases of HW/BW and myocyte cross-section area. Cardiac mitochondrial ROS level was significantly enhanced in ISO group as compared to CTRL group (P < 0.05). The expressions of NOX(4) and p-CaMKII in ISO group were also up-regulated as compared to CTRL group (1.4 and 1.6 times of CTRL, respectively, P < 0.05). NAC supplement significantly suppressed the hypertrophic development of heart in ISO-stimulated rats. The cardiac mitochondrial ROS level showed a significant decrease in rats of ISO+NAC group (P < 0.05 vs ISO). In accordance with this, ISO+NAC group rats also showed marked reductions in the expressions of NOX(4) and p-CaMKII/CaMKII compared to ISO group rats (P < 0.05). There were no significant differences of the detected indices between the rats from CTRL+NAC and CTRL groups. SBP showed no differences among four groups. These results suggest that both oxidative stress and CaMKII play important roles in sustained βAR-stimulated cardiac hypertrophy. NAC may suppress ISO-induced cardiac hypertrophy by down-regulating the expression of activated-CaMKII, and by reducing the level of oxidative stress originated from mitochondria and NADPH oxidase pathways.
Acetylcysteine ; pharmacology ; Animals ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; metabolism ; Cardiomegaly ; physiopathology ; Isoproterenol ; pharmacology ; Male ; Mitochondria, Heart ; metabolism ; Myocardium ; pathology ; NADPH Oxidase 4 ; NADPH Oxidases ; metabolism ; Oxidative Stress ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Receptors, Adrenergic, beta ; metabolism

Arsenic compounds are widely used for the therapeutic intervention of multiple diseases.Ancient pharmacologists discovered the medicinal utility of these highly toxic substances,and modern phar-macologists have further recognized the specific active ingredients in human diseases.In particular,Arsenic trioxide(ATO),as a main component,has therapeutic effects on various tumors(including leukemia,hepatocellular carcinoma,lung cancer,etc.).However,its toxicity limits its efficacy,and con-trolling the toxicity has been an important issue.Interestingly,recent evidence has pointed out the pivotal roles of arsenic compounds in phase separation and membraneless organelles formation,which may determine their toxicity and therapeutic efficacy.Here,we summarize the arsenic compounds-regulating phase separation and membraneless organelles formation.We further hypothesize their potential involvement in the therapy and toxicity of arsenic compounds,highlighting potential mecha-nisms underlying the clinical application of arsenic compounds.