Objective:To establish a method for the determination of oleanolic acid in She medicine Clematis florida var. plena. Methods:The HPLC method was carried on a Lanbo-Kromasil C18 chromatography column using acetonitrile-water ( 86∶ 14 ) as the mobile phase. The detection wavelength was 205 nm, the flow rate was 1. 0 ml·min-1 , and the column temperature was 25 ℃. Re-sults:The calibration curve of oleanolic acid was linear within the range of 0. 52-15. 60μg (r=0. 999 9). The average recovery of oleanolic acid was 92. 13%(RSD=3. 0%, n=9). Conclusion:The method is simple, rapid, accurate and reproducible, which can be used as an ideal method for the quality control of She medicine Clematis florida var. plena. .

Objective To establish a gas chromatographic method for the determination of 41 organophosphorus pesticide retention in ginseng. Methods Samples were extracted with acetonitril, purified by PSA solid phase extraction column, eluted by acetone-normal hexane, detected by FPD, and separated by ZB-1701 capillary column. Temperature for the samples was increased through program and the samples were determined by gas chromatography. Results 41 organophosphorus pesticides showed good linear relation in the range of 5-1000 μg/kg, and correlation coefficient was 0.991-0.998. The average recoveries at the three fortification levels (50, 100, 500μg/kg) were between 69.8% and 99.7% with the RSD between 1.9% and 15.3%. The limits of quantification (LOQ) of the 41 pesticides were between 9μg/kg and 45μg/kg. Conclusion The method developed in this study is with high degree of accuracy, good sensitivity, easy and simple to handle.

ObjectiveTo investigate the expression characteristics of Caspase-8 in gastric carcinoma tissues and its correlation with clinical pathological features.Methods The mRNA and protein of Caspase-8 in gastric carcinoma and its surrounding tissues were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry,respectively.ResultsThe Caspase-8 mRNA in gastric carcinoma tissues was obviously lower than that in its surrounding ones (0.154±0.065 vs 0.394±0.107,t =14.04,P ＜0.01),which was correlated with lymph node metastasis,tumor stage,differentiation grade (P＜0.01).The Caspase-8 protein in gastric carcinoma tissues was also significantly lower than that in its surrounding ones (31.48％ vs 85.19％,x2 =32.04,P ＜ 0.01),which was correlated with tumor stage,differentiation grade (P＜0.05).ConclusionsCaspase-8 is low-expressed in gastric carcinoma,which may play an important role in the occurrence and progression of gastric carcinoma.

Objective To observe the clinical effect and adverse reaction of oxycodone sustained-release tablet and morphine tablet in dose titration therapy on moderate and severe chronic cancer pain. Methods Sixty patients suffering from moderate and severe cancer pain, without using opioid drugs, were divided into oxycodone sustained-release tablet group and morphine tablet group by random digits table method with 30 cases each. The patients in oxycodone sustained-release tablet group were administered 10 mg oxycodone sustained-release tablet every 12 h, and the patients in morphine tablet group were administered 5 or 10 mg morphine tablet whenever needed. The total dose of opioid drugs was acquired after 24 h, and was converted into equal dose of oxycodone sustained-release tablet. The condition of pain control and adverse reaction were observed and recorded in a week. Results During the titration, the number of daily outbreak pain and daily medication in oxycodone sustained-release tablet were significantly lower than those in morphine tablet:(1.27 ± 1.53) times vs. (4.87 ± 1.98) times and (3.37 ± 1.78) times vs. (5.10 ± 2.20) times, and there were statistical differences (P<0.05). At the first day after titration, the pain relief rate of oxycodone sustained-release tablet was significantly higher than that of morphine tablet:83.33%(25/30) vs. 60.00%(18/30), and there was statistical difference (P<0.05). While at the third day after titration, there was no statistical difference in the pain relief rate between 2 groups (P>0.05). At the first day after titration, the incidence of daily outbreak pain in oxycodone sustained-release tablet was significantly lower than that in morphine tablet:23.33%(7/30) vs. 53.33% (16/30), the rate of reaching steady pain control state was significantly higher than that in morphine tablet: 86.67% (26/30) vs. 63.33% (19/30), and there were statistical differences (P<0.05). There were no statistical difference in the incidence of daily outbreak pain, rate of reaching steady pain control state at the third day after titration, time of reaching steady pain control state, and incidence of adverse reaction (P>0.05). Conclusions The pain relief rate and side effect of oxycodone sustained-release tablet is similar to that of morphine tablet in dose titration therapy on moderate and severe chronic cancer pain, but analgesic effect is faster than morphine tablet. Oxycodone sustained-release tablet decreases the number of outbreak pain and relieves patients′ pain in the titration process. Oxycodone sustained-release tablet may have advantage of time and effect, which is worth to be widely used in clinical therapy.

Objective To examine the complement component 1 Q subcomponent-binding protein (C1QBP) gene expression in human resistance choriocarcinoma cell lines and its parental cell line JeG-3,and to investigate whether silence C 1QBP by small interference RNA could reverse the resistance of human resistance choriocarcinoma cell lines to its relevant chemotherapy drugs.Methods Expression of C1QBP mRNA and protein in cells were detected by real-time fluorogenic quantitative PCR and western blot,respectively.The difference of C 1QBP expression was compared between human resistance choriocarcinoma cell lines and its parental cell line JeG-3.Sub-cellular location was proved by confocal immunofluorescence microscopy.A lentiviral vector containing short hairpin RNA (shRNA) targeting C 1QBP was constructed and cotransfected with the packaging plasmid mixture into 293T cells by lipofectamine 2000.The human resistance choriocarcinoma cell lines were infected with the packaged lentivirus.Real-time fluorogenic quantitative PCR and western blot were used to validate whether the C 1QBP gene expression was silenced.The cell counting kit 8(CCK8)was used to determine the drug sensitivity.Results (1)The C1QBP mRNA expression levels among four human resistance choriocarcinoma cell lines[JeG-3/floxuridiuum (FUDR),JeG-3/methotrexate (MTX),JeG-3/etoposide (VP),JeG-3/dactinomycin (KSM)] were 2.520±0.680,1.770±0.230,1.940±0.090 and 1.740±0.350 folds compared to that in JeG-3 cells.The C1QBP protein was higher expression level in human resistance choriocarcinoma cell lines than that in JeG-3.The immunofluorescence methods and confocal analysis showed that C1QBP localized predominantly in the mitochondrial matrix.(2)The C1QBP mRNA expression in JeG-3/FUDR cells after infected with lentiviral vector were decreased by 93.1％ (P＜0.01).The protein expression of C 1QBP in JeG-3/FUDR cells after infected with lentiviral vector were almost completely suppressed.The resistance indexes of four human resistance choriocarcinoma cell lines(JeG-3/FUDR,JeG-3/MTX,JeG-3/VP,JeG-3/KSM) were respectively 86.3％,93.9％,92.8％ and 89.9％,which were decreased remarkably by knockdown the C 1QBP expression (P＜0.05).Conclusions C1QBP is overexpressed in human resistance choriocarcinoma cell lines compared with parental cell line JeG-3.Inhibition of C 1QBP by lentivirus-mediated small interference RNA could effectively reverses the resistance of human resistance choriocarcinoma cell lines to its relevant chemotherapy drugs.

Objective To study the correlation between 8-Iosmerie Porastglnadin-2a(8-iso-PGF2α) 、hypersensitive C-reactive protein(hs-CRP) and coronary heart disease(CHD). Methods 153 CHD patients were divided into 3 groups,including 52 cases of acute myocardial infarction(AMI) ,50 cases of unstable angina(UAP) ,51 cases of stable angina(SAP) and control group consisted of 50 healthy people. The levels of hs-CRP and 8-iso-PGF2α were measured. Person correlation analysis was used to analyze the relationship between the level of hs-CRP and 8-isoPGF2α. Results The levels of hs-CRP and 8-iso-PGF2α were significantly higher in AMI, UAP and SAP group than those in control group(all P ＜0.05). Compared with SAP group,the levels of hs-CRP and 8-iso-PGF2α were increased in AMI and UAP groups (all P ＜ 0. 05) . The level of hs-CRP was positively associated with the level of 8-iso-PGF2α. Conclusion hs-CRP and 8-iso-PGF2α should be the markers of coronary atherosclerosis and involved in the process of CHD. The levels of serum hs-CRP and 8-iso-PGF2α were correlated with the severity of CHD.

Objective To determine the pathway of macrophage metalloelastase (MME)generate active angiostatin by decomposing plasminogen and its effect on inhibiting growth of tumor and microvessel density (MVD) in vivo in mouse models. Methods The recombined plasmid pEGFPC1-MME was constructed. Thirty mice were subcutaneously inoculated with CT-26 cells that were stably transfected with pEGFP-C1-MME (MME-transfected group), 30 with CT-26 cells transfected with empty vector pEGFP-C1 (vector-transfected group) and 30 with CT-26 cells (non-transfected group). Radioiodination and radioisotope tracer were used to explore the pathway of angiostatin generation in vivo. Results SDS-PAGE electrophoresis analysis revealed that, in the PAGE gel contained the protein with molecular weights of 35 000 and 38 000, radioactivity in MME-transfected group was significantly higher than vector-transfected and non-transfected groups (P = 0. 00).Western blotting analysis demonstrated two bands containing 35 000 and 38 000 fragments in three groups. Quantification of the protein signals by image analysis revealed that the levels of 35 000 and 38 000 fragments were obviously increased in MME-transfected group (9.32±1.52 and 5.61±2.24,respectively) than those in vector-transfected (2.47 ± 0.23 and 0. 67 ± 0. 12, respectively) and nontransfected (1.21±0. 69 and 0. 86 ± 0.44, respectively) groups (P= 0.00). The average value of MVD and fluorescent express of vascular endothelial growth factor (VEGF) were lower in MMEtransfected group when compared with those in vector-transfected and non-transfected groups (P =0.00). The average tumor size in MME-transfected group was small in comparison with vectortransfected and non-transfected groups (P= 0.00). Conclusions MME is demonstrated to be one of matrix metalloproteinase that closely related with angiostatin production and has inhibitory effect on tumor growth in tumor-bearing mice.

Objective To investigate clinical presentations,laboratory examinations,magnetic resonance imaging (MRI) appearances and treatment of hypertrophic cranial pachymeningitis (HCP).Methods The clinical data of 13 patients with HCP receiving comprehensive therapy in Huashan Hospital from January 2007 to January 2013 were analyzed retrospectively.Results The onset of HCP was mostly chronic with an average duration of 26.7 months.The main clinical manifestations of the 13 patients were chronic headaches (12/13) and cranial nerve paralysis (12/13).Inflammation markers and cerebro-spinal fluid (CSF) protein levels increased in patients with HCP and gradually became normal after the treatment.The MRI demonstrated local or diffused thickened dura located in tentorium (10/13),falx cerebrum (5/13),frontal lobe (4/13),temporal lobe (7/13) and parietal lobe (4/13).The signal intensity was isointense on T1-weighted MR images and hypointense on T2-weighted MR images.Enhanced MR images showed conspicuous enhancement of the dural edges.Corticosteroid therapy improved the clinical symptoms in 12 of 13 patients.Conclusions HCP typically causes headache and paralysis of multiple cranial nerves.Enhanced MRI shows characteristic manifestations.At present corticosteroid therapy is the treatment of choice followed by immunosuppressive agent and radiotherapy.

Objective To investigate the most sensitive methods for diagnosing spontaneous internal carotid artery dissection (sICAD) and spontaneous vertebral artery dissection (sVAD) respectively,for the sake of earlier and more accurate diagnosis.Methods Consecutive patients with sICAD and sVAD who visited the Department of Neurology and Radiology,Huashan Hospital Affiliated to Fudan University during 2008-2013 were retrospectively reviewed and the sensitivity of CT angiography (CTA),magnetic resonance T1-weighted fat-suppressed images (MR T1-FS) and digital subtraction angiography (DSA) for the diagnosis of sICAD and sVAD was compared.Results Eighty patients (62 male,18 female; mean age (45.7 ± 11.9) years) were included in the study.There were 99 arterial dissections in total,45 cases of sICAD,52 cases of sVAD and 2 cases of spontaneous middle cerebral artery dissections.The sensitivity of CTA,DSA and MR T1-FS for diagnosing sICAD was 97.5％ (39/40),90.0％ (36/40) and 69.6％ (16/23) respectively,while for sVAD was 89.8％ (44/49),84.6％ (44/52) and 100.0％ (27/27) respectively.Conclusions sICAD and sVAD have significant differences in many aspects including diagnostic strategies.CTA and MR T1-FS seem to be the most sensitive methods for the diagnosis of sICAD and sVAD respectively.Although DSA has been considered as the gold standard for the diagnosis of artery dissection,this imaging technique does not allow analysis of artery wall thickness,thus also has limitations.It is likely that the diagnostic sensitivity will be improved by combining CTA and MR T1-FS.

Objective To study the effect of ING5 gene on growth inhibition of gastric cancer. Methods ING5 expressing plasmid was transfect?ed into SGC?7901 cells. The cell viability was assessed by CCK?8,cell apoptosis and cycle was detected by flow cytometry analysis,the migration and invasion was evaluated by scratch and transwell,the expressions of mRNA and protein were determined by real?time quantitative PCR and West?ern blot respectively. Nude mice were implanted subcutaneously with SGC?7901 cells and tumor size and related protein were analyzed. Results After transfection of pEGFP?N1?ING5,the proliferation of gastric cancer SGC?7901 cells was significantly inhibited(P<0.05),the apoptosis was decreased(P<0.05),the percentage of G1 phase was increased and G2 phase was decreased(P<0.05),the ability of migration and invasion was reduced(P<0.05),the expression of NF?κB,PI3K,p?Akt1/2/3,Bcl?2,XIAP,β?catenin,c?myc mRNA,and protein levels were significantly in?creased(P<0.05),and the expression of Akt1/2/3、MMP9 mRNA and protein levels were significantly decreased(P<0.05). Conclusion The ING5gene inhibits the SGC?7901 cell proliferation,induces G1 arrest,inhibits the migration and invasion,and effectively regulates the related genes and proteins about cell proliferation,cell cycle and adhesion,but reduces apoptosis. ING5 can inhibit the evolution and development of gastric can?cer.