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 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.

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 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.

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 vitamin K3 (VK3) effect on apoptosis of human liver cancer cells and its mechanism. Methods The SMMC-7721 cells were cultured in the experiment. The inhibitory effects of VK3 on SMMC-7721 cells were tested by CCK-8. Morphological evaluation of apoptosis was performed by Hcechst33342 staining. The distribution of cell cycle and apoptosis, and the expression of Fas were assayed by flow cytometry. Fas mRNA expression were detected by RT-PCR. And the concentration of soluble Fas(sFasL) in the culture supernatant were measured by EIJSA. Results The inhibitory rates ofVK3 (at concentrations of 2,5,10,20,25 umol/L for 48 h) on SMMC-7721 growth were 33.8% ,50.1%,63.9% ,78.5% and 84.7%, respectively. Compared with the control group, the number of cells in the G0/G1 phase increased, while that of S phase decreased. The apoptotic cell rates were 18.75%, 25.80%,38.80% ,29.92% and 26.18% ,respectively. The apoptosis cells were strongly stained by Hcechst33342.On exposure to VK3 at the concentration of (2,5,10 umol/L) after 48 h, the mean fluorescence intensity ofFas on cell surface and the expression of fas mRNA and the concentration of FasL in the culture supematantin SMMC-7721 were increased, but they all decreased at the high concentration of VK3. Conclusion VK3can inhibit the proliferation of SMMC-7721 cells and induce apoptosis via up-regulating expression of Fas and sFasL.

Objective To evaluate the international blood smear review criteria for improvement using Beckman-Coulter hematology analyzers and find out proper slide review criteria suitable for Chinese population. Methods 3 600 random-selected blood samples were tested in three hospitals using MAXM, GENS and LH750 5-diff automated analyzers and a manual differential with a smear review was performed on all samples in the study. True positive rate, true negative rate, false positive rate and false negative rate were calculated according to the international blood smear review criteria. We have set up smear review criteria for Chinese population by analyzing false positive and false negative cases according to Chinese clinical conditions. Another 240 blood samples were tested in three hospitals using the same analyzers to verify the new slide review rules. Results According to international blood smear criteria, the true positive rate was 4.9%, false positive rate was 24.2%, true negative rate was 70.4% and false negative rate was 0.5%. The international smear review criteria were modified into 23 criteria by analysis of above statistical data. In addition, we added four WBC differential ratio rules in positive smear criteria. After modification, the true positive rate was 9.9% (355/3 600) ,false positive rate was 17.1% (617/3 600), true negative rate was 71.2 % (2 563/3 600) and false negative rate was 1.8% (65/3 600). No blast cell was missed using both smear review criteria. A little higher false negative rate after modification was caused by supplemented differential ratio roles in positive smear criteria. Verification results were satisfactory;The tree positive rate was 13.7% (33/240) ;false positive rate was 15.8% (38/240) ;false negative rate was 2.5% (6/240) and true negative rate was 68.0% (163/240). Conclusions Although the smear review criteria suggested by the International Consensus Group is clinically important, the false positive rate increases when they are used in Chinese population. The modified slide review criteria used on Beckman-Coulter hematology analyzers in this study are more suitable for Chinese laboratories.

Objective To identify gene mutations and explore the molecular mechanism of a pedigree with inherited coagulation F Ⅶ deficiency. Methods The levels of F Ⅶ: Ag in the proband and other family members were measured by ELISA assay. The values of PT, F Ⅶ: C and other coagulant parameters were determined by one-stage clotting for laboratory phenotype diagnosis. All the exons,exon-intron boundaries and 5',3' untranslated sequences of F7 gene were amplified by direct sequencing. The detected mutations were further confirmed by sequencing the other stand. The CLC Protein Workbench software was used to analyze the species conservation of the mutated site and the protein secondary structure. 100 healthy individuals were selected to exclude gene polymorphism. Results PT, FⅦ∶C and FⅦ: Ag in the proband and his sister were abnormal, which were 36. 3 s, 5.0%, 40. 7% and 33.4 s,5. 0%, 37.4%, respectively. Both PT and FⅦ∶C in the proband's father, mother, daughter and niece were slightly abnormal, which were 14.9 s, 14. 6 s, 15.5 s, 14. 6 s and 70%, 85%, 59%, 79%, respectively.The heterozygous mutations c. 784T ＞ C and c. 964T ＞ G in exon 8 of F7 gene were found in the proband,resulting in the substitutions of Ser269Pro and Cys329Gly respectively. Compound heterozygous mutations c. 784T ＞ C and c. 964T ＞ G were found in the proband's sister. The proband's mother was heterozygous for c. 784T ＞ C. His father, daughter and niece were heterozygous for c. 964T ＞ G. The protein biological characteristics analysis revealed that the Cys329Gly caused the change of spatial configuration, and Ser269Pro led to the change of amino acid polarity and hydrophobicity. Conclusion Compound heterozygous mutations of Cys329Gly and Ser269 Pro in F7 gene may be the underlying molecule mechanism of FⅦ deficiency in this pedigree.

Objective To explore the association between the C46T polymorphism of coagulation factor Ⅻ (FⅫ) gene and the involvement of FⅫ activity (FⅫ:C) in patients with unexplained recurrent spontaneous abortion (URSA), and to elucidate its role in the pathogenesis of URSA. Methods This study included 203 patients with URSA (URSA group) and 171 healthy women with at least one child and no history of infertility or miscarriage (control group) in the southern area of Zhejiang Province. The C 46T polymorphism of the FⅫ gene was analyzed with matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) in all subjects. The values of prothrombin time, activated partial thromboplastin time (APTT), fibrinogen, FⅫ:C and other coagulant parameters were determined. The frequency distribution of the wild-type (CC), heterozygote (CT), homozygote (TT) genotypes and C and T alleles were compared between the patients and controls. A comprehensive analysis of association was conducted between C46T genotypes and the FⅫ:C levels in URSA patients. Results The CC, CT, TT genotypes of the FⅫgene were observed in 7 (3.4%, 7/203), 83 (40.9%, 83/203) and 113 (55.7%, 113/203) patients with URSA versus 7 (4.1%, 7/171), 46 (26.9%, 46/171) and 118 (69.0%, 118/171) controls. The frequency of CT in the patients with URSA was significantly higher than that in controls, but the frequency of TT in the patients was lower than that in controls (χ2=7.939, OR=1.884, 95%CI:1.210-2.935, P<0.05). The frequencies of allele C and allele T were observed in 97 (23.9%, 97/406) and 309 (76.1%, 309/406) patients with URSA versus 60 (17.5%, 60/342) and 282 (82.5%, 282/342) controls. The distribution frequency of allele T in URSA group was lower than that in control group (χ2=4.510, OR=1.475, 95%CI:1.029-2.115, P<0.05). The FⅫ:C levels in the patients were (102±13)%in CC genotype, (78±11)%in CT genotype and (59± 9)%in TT genotype, respectively. The differences of the FⅫ:C levels between the CC and CT, CT and TT, CC and TT genotypes in the patients were significant (all P<0.05). Conclusions The low level of FⅫ:C maybe result from the T allele of the FⅫgene in URSA patients. The CT genotype might be relative to the pathogenesis of URSA in a Chinese Han female population from the southern area of Zhejiang province.