This paper is to report the development of a rapid and sensitive method for the determination of s-oleylpropanolamide (OPA) in various tissues of rat (brain, heart, lung, liver, spleen, small intestine, kidney, adipose tissue and muscle), and to assess the applicability of the assay to tissue distribution. OPA was extracted by liquid-liquid extraction method with undecylenoylethanolamide as an internal standard. The concentrations of OPA were determined by LC-MS/MS after a single intragastric dose of 50 mg x kg(-1) at 4 time points (5 rats per group). With multiple reactions monitoring mode (MRM) the limit of quantification (LLOQ) was determined at 1 microg x L(-1). The calibration curve was linear from 1 to 2 x 104 microg x L(-1) (r > or = 0.999 0) for tissue homogenates. Validation parameters such as accuracy, precision and recovery were found to be within the acceptance criteria of the assay validation guidelines. The highest concentration was found in small intestine (the highest time point is 15 min) and heart (the highest time point is 90 min). The assay is rapid, sensitive and applicable to studying tissue distribution of OPA in rats.

Objective To establish a learning evaluation indexes systen of flipped teaching method,and provide the basis for evaluating academic performance of each student who majors in nursing.Methods The indexes were determined by Delphi method of two rounds of consultation involving 15 experts,and the weight of every index was set up by analytic hierarchy process.Results The response rates of questionnaire investigation for the two rounds of consultation were 13/15 and 12/13 respectively.The authority value of experts was 0.847.The Kendall concordance coefficients for the indexes of first-level,second-level and third-level were 0.373,0.258 and 0.595 respectively,P＜0.05.Finally,the evaluation indexes system including 3 first-level indexes,5 second-level indexes,and 6 third-level indexes was set up.Conclusions The results of the two rounds of expert consultation are reliable.The consistency of expert opinions is good.The study provides a scientific and systematic evaluation tool for evaluating nursing undergraduates' performance under flipped teaching.

Brucellosis is an infectious-allergic anthropozoonosis caused by invasion of Brucella.It belongs to category B infectious disease in China.The epidemic situation of brucellosis is raging again in China just like the rest of world.Especially after 2000,brucellosis became one of the fastest growing infectious diseases.By 2012,all Chinese administrative units at provincial levels except Taiwan had reported the cases of brucellosis.In this paper,we analyzed the epidemic features,prevention and control of brucellosis.We elucidated deep social reasons and the root problems,which impede the prevention and control of brucellosis.We have also discovered the important problems,which should be focused on and solved in the process of prevention and control of brucellosis.

Objective: To investigate the roles of Bcl-xl and Bcl-xs in the development and progression of endometrial carcinoma, and to explore their correlation.Methods: The expression of Bcl-xl and Bcl-xs in 32 cases of endometrial carci-noma, 12 cases of endometrial atypical hyperplasia, 6 cases of endometrial simple hyperplasia and 10 cases of normal en-dometrial tissues were examined by RT-PCR and Western-blot.Results: The expression of Bcl-xl mRNA and protein was significantly higher in endometrial cancer tissues than in normal endometrial tissues (P<0.05), and was statistically associat-ed with the pathological stage of endometrial carcinoma.(F=5.33, P=0.02).The expression of Bcl-xs mRNA and protein in atypical endometrial hyperplasia and endometrial carcinoma tissues were significantlly lower than that in normal endometri-al tissues (P<0.05), which was also associated with clinical stage and lymph node metastasis of endometrial carcinoma (P<0.05).The expression of Bcl-xl was negatively correlated with the expression of Bcl-xs in different endometrial tissues (r=-0.76).Conclusion: The abnormal expression of Bcl-xs and Bcl-xl was a factor for the pathogenesis and development of endometrial carcinoma.The negative correlation between Bcl-xl and Bcl-xs in different endometrial tissues as well as their relative expression ratio may have certain impact on the genesis of endornetrial cancer.

Objective To describe the feelings,career development needs and the coping styles of stress experience of nurses with master's degree within one year after graduation.Methods The semistructured in-depth interview was conducted in 15 nurses with master's degree individually.The data were analyzed by qualitative content analysis method and the themes were abstractod.Results Three themes were extracted,including feelings,their career development needs and coping styles of stress.The nurses with master's degree reported feelings of heavy work pressure; lack of a sense of accomplishment; strong sense of loss.Their career development needs included improvement of professional knowledge and clinical skills; organizational supports; improvement of professional English and received a doctor' s degree.There were three main coping styles of stress:adjusting attitude and self role orientation; competence improvement; adjusting interpersonal relationship.Conclusions Nursing masters suffer from heavy pressure in clinical practice within one year after graduation.They have some occupation development needs.Nursing managers should explore positive and effective methods to relieve the nurses' stress and ratioually use and actively guide nursing masters,and provide direction and space for achieving their occupation value.

Objective To evaluate the effect of acupuncture point stimulation on the urodynamics of spinal cord injury patients.Methods Sixty spinal cord injury cases were divided into a research group and a comparison group.Both groups were given bladder training, but the research group also received acupuncture point stimulation.The subjects' maximum cystometric capacity, maximum detrusor pressure and residual urine were measured, along with renal function and any urinary tract infections.The scale of activity of their daily lives was compared before and 3 months and 6 months after the treatment.Results There were no statistically significant differences between the groups before the treatment.There were statistically significant differences in maximum cystometric capacity, maximum detrusor pressure, residual urine and Barthel index (BI) after 3 and 6 months of treatment.In the research group, maximum detrusor pressure, residual urine and BI scores had all improved after 6 months compared with the 3 month values.In the comparison group only the BI score had improved significantly between 3 and 6 months, and this was not reflected in a significant change in the scale of activity in daily life.After 3 months, only the average BI differed significantly between the two groups.After 6 months all indexes except the BI and maximum cystometric capacity differed significantly.There were also significant differences between the groups in reflex voiding, percussion voiding, and abdominal pressure voiding.Conclusions Acupuncture point stimulation can improve the bladder function of spinal cord injury patients.It can also decrease the incidence of urinary tract infection and the possibility of renal dysfunction.

Objective To investigate the biological effects of internal radiation and therapeutic effectiveness of 131I-labeled anti-epidermal growth factor receptor (EGFR) in colorectal cancer of model mice.Methods Nano-liposome characterized for EGFR-targeting was constructed.The efficacy of cellular binding and uptake of the liposome was evaluated by the analysis of confocal microscopy observation and the iodide uptake assay.After intra-tumor injections of 74 MBq (740 MBq/ml) 131 I-antiEGFR-BSA-PCL,131 I-BSA-PCL,131 I or an equivalent volume of normal saline.The biological effects of internal irradiation and therapeutic efficacy of the liposomes on colorectal cancer modeled in a male BALB/c mouse were evaluated by means of tumor size,body weight,histopathology,and SPECT imaging.Results The confocal fluorescence images showed that the antiEGFR-BSA-PCL was successfully internalized into LS180 cells.The 131I uptake efficacy of 131I-antiEGFR-BSA-PCL was significantly higher than that of 131I-BSA-PCL in LS180 cells (t =2.77-5.40,P ＜ 0.01).Tumor size measurement showed that tumor growth was inhibited by the treatment with 131 I-EGFR-BSA-PCL and 131I-BSA-PCL,but had no significant differences between these two groups (P ＞0.05).It was found that the 131I-antiEGFR-BSA-PCL was markedly taken up by the tumor and reac hed its uptake value of (21.61 ± 1.0 1) and (20.58 ± 0.65)％ ID/g at 72 h following drug injection,which was higher than the uptake value of 131 I (t =9.36,8.69,P ＜ 0.01).SPECT imaging assay showed that,after being injected into mouse tumor,the 131 I-EGFR-BSA-PCL and 131I-BSA-PCL were uniformly distributed inside the tumor.Conclusions 131 I-antiEGFR-BSA-PCL obviously suppresses the development of colorectal cancer in mice.

Objective To construct 131 I labeled anti?EGFR immunoliposome nanoparticle ( 131 I?Cetuaximab ( C225)?BSA?PCL) , and investigate its inhibitory effect on EGFR?overexpressing cancer cells in vitro. Methods Anti?EGFR liposome nanoparticle C225?BSA?PCL and non?targeted liposomes BSA?PCL were constructed. The products were observed with transmission electron microscopy and dynamic light scat?tering. The EGFR?targeted binding and cellular uptake in EGFR?overexpressing cancer cells were observed with flow cytometry and confocal microscopy. Anti?EGFR and non?targeted liposomes were labeled with 131 I using the chloramine?T method. The targeted cell killing effects of 131 I labeled liposomes were analyzed using MTT assay. The time?dependent cellular uptake analysis was used to evaluate the slow?release effects of the 131 I labeled liposomes. The independent?samples t test was used for data analysis. Results The EG?FR?targeted liposome C225?BSA?PCL and non?targeted liposome BSA?PCL were successfully constructed, and the effective diameters were approximately 130-180 nm. Flow cytometry and confocal microscopy re?vealed significant uptake of C225?BSA?PCL in EGFR?overexpressing tumor cells. BSA?PCL could also bind to cells with minimal and weak tumor retention. The EGFR?targeted radioactive liposome 131I?C225?BSA?PCL showed greater targeted cell killing effect than non?targeted liposome 131I?BSA?PCL,the IC50 values of 131I?C225?BSA?PCL and 131 I?BSA?PCL were 0. 03-1. 32 and 0. 25-12. 19, respectively. The uptakes of 131 I?C225?BSA?PCL was higher than that of 131 I?BSA?PCL ( t=3.03-16.86, all P<0.05) and reached the maxi?mal level at 4 h after incubation. Conclusions The EGFR?targeted liposome C225?BSA?PCL demonstrated superior cellular binding and uptake on EGFR?overexpressing cancer cells compared with BSA?PCL. The EGFR?targeted radioactive liposome 131 I?C225?BSA?PCL had favorable intracellular retention and excellent targeted cell killing effect, and could effectively suppress the growth of EGFR?overexpressing cancer cells.

BACKGROUND:Bone marrow mesenchymal stem cel transplantation has not been thoroughly reported on its effects on apoptosis in hepatoma carcinoma cel s and inflammatory factor level.
OBJECTIVE:To investigate the effect of rat bone marrow mesenchymal stem cel s on dynamic change of inflammatory factors and cel apoptosis during hepatocarcinogenesis.
METHODS:Sixty healthy Sprague-Dawley rats were divided randomly into healthy group (n=30), control group (n=30) and transplantation group (n=30). Healthy group was given ordinary feed and normal water, while other groups were given diethylnitrosamine solution in drinking water to induce liver cancer models. Then, rats in the transplantation group were subjected to bone marrow mesenchymal stem cel transplantation via the tail vein. Two weeks after cel transplantation, CXCL5, interleukin-8 and interleukin-6 levels were tested by ELISA, mRNA level of hepatocyte nuclear factor 1αdetected by RT-PCR, expression of Bcl-2 and Bax in liver tissue measured by immunohistochemical method, and liver cancer cel apoptosis index detected by TUNEL technique.
RESULTS AND CONCLUSION:After modeling, the expressions of CXCL5, interleukin-8 and interleukin-6 in the control group were significantly higher than those in the healthy group (P<0.05), while these indexes were reduced significantly after bone marrow mesenchymal stem cel transplantation (P<0.05) and close to the normal levels (P>0.05). Bone marrow mesenchymal stem cel transplantation significantly up-regulated the mRNA level of hepatocyte nuclear factor 1αin the liver tissue that was decreased obviously after modeling (P<0.05). In addition, the expression of Bcl-2 was reduced, while the expression of Bax and the apoptosis index increased significantly in the transplantation group compared with the control group (P<0.05). These findings indicate that bone marrow mesenchymal stem cel transplantation contributes to hepatocyte differentiation and regeneration in liver cancer rats by reducing serum inflammatory factor levels and promoting apoptosis in hepatoma carcinoma cel s.

Objective To detectthe phenotype and gene mutations underlying aninherited dysplasminogenemia pedigree and search the virulence gene.Methods The peripheral venous blood samples of the proband and his family members (fourteen subjects of three generations in total) were collected,and their prothrombin time(PT),activated partial thromboplastin time(APTF),thrombin time(TT),fibrinogen (FIB),fibrinogen degradation products (FDP),D-dimmer (D-D)weretested on a STAGO analyzer,the plasminogen activity (PLG:A) and plasminogen antigen (PLG:Ag) were analyzedby thechromogenic substrate assay and rocket immunoelectrophoresis,respectively.All 19 exons,5' and 3' untranslated regions of PLGwere amplified with PCR.Direct DNA sequencing was used to analyze the amplified products,which wereconfirmed by backward sequencing.Three bioinformatics online softwares (SIFT,PolyPhen-2 andMutationTaster) were used to forecast the possible impact of the mutations on the protein function.At last,themodel analysis of mutate site was taken on a Swiss-Pdb Viewer software.Results The PLG:Avalue of theproband and other 6 family members were decreased to the half,while the PLG:Ag was normal.The D-Dand FDP value of the proband,his grandma and father were slightly higher.DNA sequencing has revealedthat the proband and the other 6 members of this family had the same mutation of g.38829G ＞ A in exon 15,leading to the missense mutationp.Ala601Thr.The results of bioinformatics softwares showed that themutation could affect the thePLGfunction.Protein model analysis indicated that the hydrophobic interaction force and hydrogen bond between the amino acids were changed,which might affect the stability of the PLG.In addition,all the members of this family take the heterozygous SNP of g.2501C ＞ A in the 5 ＇UTR.Conclusions The p.Ala601Thr found in the inherited dysplasminogenemia pedigree in the exon 15 was responsible for the reduced PLG:A of the family,the dysplasminogenemia and this mutation were both reported for the first time in China.