Objective To establish a method for the determination of 17 kinds of free amino acid in serum by the ultra perform-ance liquid chromatography(UPLC) with AQC pre-column derivatization .Methods After proteins precipitation in serum sample by acetonitrile ,free amino acids with aminoquinoline-N-butyl hydroxy diimide and carbamate(AQC) were reacted to generate the struc-tural stable derivative products with ultraviolet absorption .These products were separated by reversed-phase C18 column ,detected with UV detector and quantitated by the external standard .Results The within-run precision of 17 kinds of free amino acid was 2 . 81% -7 .35% and the between-run precision was 7 .44% -15 .40% .The linear relationship was good in the range of 2 .5-250μmol/L ,the linear correlation coefficient was 0 .9960 -1 .0000 ,RSD<3% .Conclusion Using this method for determining 17 kinds of amino acid in serum ,the complete separation is obtained within 10 min ,the results are accurate ,stable and highly efficient .

OBJECTIVE:To compare the new Prescription Regulatory Regime(try out)with the old“Prescription Reg?ulatory Regime”.METHODS:The2prescription regulatory regimes were analyzed in terms of law,standard operating proce?dure of drug dispensing.RESULTS&CONCLUSION:The new Prescription Regulatory Regime(try out)is operated more within the law than the old one,and in which,pharmacists’positions and responsibilities were clearly stated,the limit of checking and monitoring had been enlarged meanwhile drug dispensing standard operating procedures had been clarified.

Magnetic resonance (MR) images can be used to detect lesions in the brains of patients with multiple sclerosis (MS). An automatic method is presented for segmentation of MS lesions using multispectral MR images in this paper. Firstly, a Pd-w image is subtracted from its corresponding T1-w images to get an image in which the cerebral spinal fluid (CSF) is enhanced. Secondly, based on kernel fuzzy c-means clustering (KFCM) algorithm, the enhanced image and the corresponding T2-w image are segmented respectively to extract the CSF region and the CSF-MS lesions combinatoin region. A raw MS lesions image is obtained by subtracting the CSF region from CSF-MS region. Thirdly, based on applying median filter and thresholding to the raw image, the MS lesions were detected finally. Results were tested on BrainWeb images and evaluated with Dice similarity coefficient (DSC), sensitivity (Sens), specificity (Spec) and accuracy (Acc). The testing results were satisfactory.
Algorithms ; Brain ; pathology ; Humans ; Image Enhancement ; Magnetic Resonance Imaging ; Multiple Sclerosis ; diagnosis ; Sensitivity and Specificity

Objective To systematically evaluate the short-term outcomes of Da Vinci robotic surgical system and laparoscopy in pancreaticoduodenectomy.Methods Literatures were researched using PubMed,Embase,Medline,VIP database,Chinese Journal Fulltext Database from January,2013 to October,2016 with the key words including laparoscopic,robotic,Da Vinci,pancreaticoduodenectomy,腹腔锐,达芬奇机器人,胰十二指肠切除术”.The cohort studies about comparison of short-term outcomes of Da Vinci robotic surgical system and laparoscopy in pancreaticoduodenectomy were received and enrolled.Patients undergoing pancreaticoduodenectomy using Da Vinci robotic surgical system and laparoscopy were respectively allocated into the Da Vinci group and laparoscopy group.Two reviewers independently screened literatures,extracted data and assessed the risk of bias.Count data were described as odd ratio (OR) and 95％ confidence interval (CI).Measurement data were represented as weighted mean difference (WMD) and 95％CI.The heterogeneity of the studies was analyzed using the I2 test.Results Five retrospective cohort studies were enrolled in the Meta analysis,and the total sample size was 364 patients,including 159 in the Da Vinci group and 205 in the laparoscopy group.The results of Meta analysis showed that there were statistically significant differences in the rate,of conversion to open surgery,spleen-preserving rate,operation time and duration of postoperative hospital stay between Da Vinci group and laparoscopy group (OR=0.18,3.80,WMD =-37.54,-4.47,95％CI:0.05-0.60,2.01-7.18,-47.46--27.62,-6.70--2.24,P＜0.05).There were no statistically significant difference in the incidence of pancreatic fistula and overall incidence of complications between Da Vinci group and laparoscopy group (OR=0.95,0.55,95％CI:0.59-1.54,0.29-1.03,P＞0.05).Conclusions Da Vinci robotic surgical system and laparoscopy are safe and feasible in pancreaticoduodenectomy,with the same incidence of postoperative complications.Compared with laparoscopy,Da Vinci robotic surgical system can not only reduce the rate of conversion to open surgery,operation time and duration of postoperative hospital stay,but also increase spleen-preserving rate,meanwhile,it does not increase the incidence of postoperative complications.

Objective According to principle of forming primer dimer,a new cloning gene method was established. Methods Two and more oligonucleotide with complementary partnership in the 3′ end were synthesized by artifical method,oligonucleotide,Taq DNA polymerase and dNTP were mixed in the proportion of reasonable,then expanded and extended by PCR,result was identifed by agarose gel electrophoresis.Results A clear special band was appeared in the agarose gel. Conclusion Short fragment target gene less than 500 bp may be synthesized by this method.

BACKGROUND: Nerve growth factor is secreted and synthetized by a variety of cells, such as inflammatory calls and repairing calls, its biological effects are diverse and closely related to the process of wound repair, but its mechanism is not yet clear.OBJECTIVE: To observe the influence of nerve growth factor on the biological characteristics of scar fibroblasts.METHODS: Eight clinical surgical resection specimens, including 5 face and neck hyperplastic scar or keloid specimens, did not receive any treatment; three were prepuce specimens following circumcision (normal tissue). By use of tissue block method, the scar and normal skin fibroblasts were cultured, followed by digestion passage. The scar tissue and normal tissue flbroblasts at 3-6passages in the logarithmic phase were seeded in 96-well plate and divided into the experimental group (scar flbroblest group) and the control group (normal skin fibroblasts group), with two parallel holes in each group were added with 3,33, 0.33 mg/L nerve growth factor, 50 μL. Inverted microscope was used to observe fibroblast morphology. At 24, 48, 72 hours after culture, the absorbanca value was measured using MTT. Fibroblast DNA content and cell apoptosis were determined by flow cytometry.RESULTS AND CONCLUSION: The fibroblasts were adherent cells, the scar and normal skin tissues were shown to cell free out of tissue block and gradual expansion at 4-6 days after incubation. Compared with normal skin fibroblasts, the pathological scar fibroblasts became larger, irregular shape and arrangement. MTT results showed that nerve growth factor could promote the normal and hypertrophic scar fibroblasts growth, which becomes more apparent. Flow cytometry results showed that by adding nerve growth factor, the percentage of scar fibroblasts at proliferating S-G_2-M phase was higher than that in the control;group; with a Iower level of apoptosis. It is indicated that nerve growth factor plays an obviously promoting role on normal and scar skin fibroblasts growth and proliferation, especially on the scar skin.

s and no restenosis was found in 3 patients according to cerebral angiography. Preliminary results show that PTAS in the management of the vertebrobasilar arterial stenosis is a safe and effective method.

Objective To explore the mechanism of cytokines for the scars,and to study the effect of platelet derived growth factor(PDGF)on the biological behavior of fibroblasts in scars.Methods Fibroblasts of scars and normal skins were cultured in vitro.The results were observed and analyzed by light inverted microscopy(LM),and 3-(4,5-dimethyl-thiazol-2-yl)-2,5 ciphenyl tetrazolium bromide (MTT)assay.The effects of PDGF on the biological behaviors of fibroblasts of scars were also determined. Results In vitro study,using LM,FCM and MTT assay,showed that proliferation of fibroblasts were inereased significantly when PDGF was added to the cultures,as compared to the control groups.Conclusions PDGF can increase fibroblast proliferation.These results demonstrate that PDGF is beneficial for wound healing at early stage.

Objective To separate the CD1c+ MDC and CD304 PDC subsets and analyze their immunobiologic properties.Methods MDC or PDC were separated by immunomagnetic beads and then induced by TNF-α or CpG2006 OND.The phenotype of different DC subsets and allogeneic T-cell-stimulating capacity were detected,respectively.Results The purlty of separated DC was about ninety-five percent.MDC expressed HLA-DR,CD11c,CD4 and CD33,PDC expressed HLA-DR,CD4 and CD45RA,and the percentage of CD40,CD80 were up-regulation after induced to mature.The results of mixed lymphocyte reaction(MLR):MDC had strong potential to stimulate allogeneic T lymphocytes proliferation,while PDC had the weak stimslate potential.The supematant IFN-r in all groups was increased.which the groups of MDC and mMDC were more obviously.The supematant IL-10 was increased obviously in groups of PDC and mPDC.The expression of IFN-r in CD4+ cells were increased obviously after stimulated by MDC or mMDC.PDC and mPDC had the effect of inducing of CIM+ CD25high regulatory T lymphocyte in MLR.The expressing of Foxp3 mRNA had no evident difference in all groups.Conclusion Two different DC subsets in the donors peripheral blood which mobilized with G-CSF were in the state of immature and could be induced to mature in the suitable condition.although expressed high constitutive levels of HLA,DR.No matter how MDC was mature or not,they had strong potential to stimulate allogeneic T lymphocytes proliferation.MDC enhanced the production of IFN-r among T lymphocytes,and the increase of IFN-r was probably the result of TH1 polarization.Unlike MDC,PDC was less efficient present antigens and had poor T-cell-stimulating capacity,probably because of their lesser ability to phagocytose,process and present antigens to the MHC molecules.Although PDC could promote the production of IFN-r,it seemed that did not produced by TH 1.PDC could not promote TH2 cells to product IL-4,and the polarization of TH2 probably reflect the production of IL-10.PDC could induce the generation of CD4+ CD25high regulatory T cells.

ObjectiveToanalyzetheclinicalandimagingcharacteristicsinpatientswithcarotidsteal syndrome ( CSS ) and to investigate its compensatory pathw ays, diagnosis, and treatment. Methods The medical history and imaging data of the patients with CSS were colected. Their vascular lesions, colateral circulation, treatment, and prognosis w ere analyzed. Results A total of 11 patients w ith CSS (8 males and 3 females, mean age 66.7 ±5.1 years) were enroled in the study. Their clinical manifestations were posterior circulation transient ischemic attack (TIA) ( n=9, 81.8%), posterior circulation infarction ( n=1, 9.1%), and anterior circulation TIA ( n=1, 9.1%). A total of 19 pathological arteries w ere found:12 (63.1%) w ith occlusion, 2 (10.5%) w ith subtotal occlusion, 4 (21.0%) w ith severe stenosis and 1 (5.2%) w ith artery dissection. Seven patients (63.6%) w ere bilateral internal carotid artery lesions, 3 (27.2%) w ere unilateral bilateral internal carotid artery lesions, and 1 (9.1%) w as bilateral common carotid artery occlusion. Eleven patients had primary col ateral circulation, including posterior communicating artery patency in 10 patients (90.9%) and anterior communicating artery patency in 1 patient (9.1%). Four patients (36.3%) had secondary col ateral circulation and 1 (9.1%) had tertiary col ateral circulation. Al patients w ere treated w ith medication on the basis of the management of risk factors. Three patients w ere treated w ith stenting and tw o were treated with carotid endarterectomy. No stroke occurred in al patients during folow -up til September 2014. Conclusions The vascular lesions of patients w ith CSS often occur in the extracranial segment of internal carotid artery. Usual y the compensatory blood is through the circle of Wil is. The presentation is ischemia in the stolen arteries. Its diagnosis needs to be examined by digital subtraction angiography. On the basis of medication therapy, some patients may be treated w ith surgery or endovascular intervention.