A rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometric (RRLC-Q-TOF-MS) method was established and optimized for the analysis of pharmacokinetic behavior of ginsenoside Rb2 in rats by intravenous injection administration.The metabolism of ginsenosides Rb2 in vivo rat was also explored.In the experiment,Agilent SB C18 column was selected for the sample separation with 0.1％ aqueous formic acid solution as mobile phase (A) and acetonitrile as mobile phase (B) at a flow rate of 0.2 mL/min,and the injection volume was set to 5 μL.Q-TOF-MS was carried out in electron pray ionization (ESI) negative ion mode.The limit of quantification (LOQ,S/N =10) and limit of detection (LOD,S/N=3) were 0.10 and 0.08 μg/mL,respectively,and the linear range was 0.1-1.26 μg/mL.The experiment results showed that the concentration-time profile of ginsenoside Rb2 conformed to a two-compartment pharmacokinetic model after intravenous administration for rats.The mean plasma elimination half-lives were (23.58±1.10) min (t1/2α),(1306.55±147.23) min (t1/2β) for Rb2.By analyzing the urine of rats after intravenous administration and the fecal samples after oral administration of ginsenoside Rb2,it was found that the metabolites were M6,M2 (CY),F2,and C-K.

Cell Adhesion ; Cell Proliferation ; Cells, Cultured ; Chitosan ; chemistry ; Coated Materials, Biocompatible ; Collagen ; chemistry ; Humans ; Materials Testing ; Polyethyleneimine ; chemistry ; Static Electricity ; Surface Properties ; Titanium ; chemistry

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Objective To examine the relationship between CCT 5 ,γδ T cell and autoimmune diseases .Methods Recombinant CCT5 protein was cloned , expressed and purified in E.coli.Three peptides of CCT5 protein were used to prepare for anti-CCT5 monoclonal antibodies .Purified CCT5 protein was used to expand γδT cells from pe-ripheral blood mononuclear cells (PBMC).Plasma level of CCT5 in healthy donors, patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) were detected by ELISA assays .The correlation analysis between plasma CCT5 concentration and the percentage of different subtypes of γδT cells measured by flow cytometry was made . Results The CCT5 gene was amplified by PCR and the length of the target fragment was 1 750 bp.The expressed 65 ku CCT5 protein was purified and validated by SDS-PAGE.Two paired monoclonal anti-CCT5 antibodies were screened to detect CCT5 protein in plasma.Immobilized recombinant CCT5 protein was able to induce specific sig-nificant amplification of peripheral γδT cells.Correlation analysis of 10 healthy donors indicated significant corre-lation between the plasma CCT 5 concentration and the proportion of Vγ9 and Vδ2 γδ T cells.The plasma CCT5 concentration significantly decreased in autoimmune diseases patients , including RA and SLE .Conclusions These data suggest that CCT 5 could be a novel Vγ9δ2 γδT cell-related factor in autoimmune diseases , which deepen the understanding of Vγ9δ2 γδT cell function in autoimmune diseases .

Objective To explore the value of 1H-MRS on the evaluation of Alzheimer's disease (AD) with neural stem cells (NSCs) transplantation in an APP-PS1 double transgenic (tg) AD mouse model.Methods NSCs from C57BL/6 mice were cultured and amplified.APP-PS1 tg mice (n =30) aged 12 months were used as the study group,and mild-type mice (n =15) were used as the control group.Animals in the study group were randomized into two subgroups,the AD mice in one subgroup received NSCs transplantation (NSCs group) and in another subgroup received phosphate buffer saline (PBS,PBS group)in bilateral hippocampal CA1.Animals in the control group were not treated.Using a 7.0 T high-fieldstrength MR imager,1H-MRS was performed before and 6 weeks after transplantation to measure the area under the peak of n-acetyl aspartate (NAA),glutamate (Glu),myo-inositol ( mI),choline (Cho) and creatine (Cr) in the hippocampal area,NAA/Cr,Glu/Cr,mI/Cr and Cho/Cr ratio were calculated and compared with histopathological results (including Nissl's staining and electron microscope examination).Comparisons among NSCs,PBS and control groups were conducted by one-way ANOVA.Results NSCs from C57BL/6 mice were cultured successfully. Before transplantation,the mean NAA/Cr,Glu/Cr and mI/Cr in NSCs,PBS and control groups were 0.89 ± 0.05,0.88 ± 0.04 and 1.15 ± 0.05,0.40 ± 0.03,0.39 ± 0.03 and 0.45 ± 0.05,0.67 ± 0.05,0.67 ± 0.05 and 0.52 ± 0.04,respectively,and differences were statistically significant (F =148.918,7.529,59.468,P ＜ 0.01 ). There were no significant differences in NAA/Cr,mI/Cr and Glu/Cr ratios between NSCs and PBS groups before transplantation (t =0.147,0.096,0.207,P ＞ 0.05 ),but the differences were significant compared with the control group (t =0.255,0.467,0.171 and t =0.269,0.527,0.151,P ＜0.05).Six weeks after transplantation,the mean NAA/Cr,Glu/Cr and mI/Cr in three groups were 1.13 ±0.07,0.86 ±0.05 and 1.14 ±0.05,0.45 ± 0.04,0.38 ± 0.02 and 0.44 ± 0.03,0.58 ± 0.04,0.67 ± 0.04 and 0.53 ± 0.04,respectively,and differences were statistically significant ( F =112.092,23.076,44.367,P ＜ 0.01 ).NAA/Cr and Glu/Cr ratios were increased and mI/Cr was decreased in NSCs group,and the difference was significant compared with PBS group at the same time point ( t =0.271,0.071,0.089,P ＜ 0.05 ).There were no significant differences in NAA/Cr and Glu/Cr ( t =0.013,0.012,P ＞ 0.05 ),but there was a significant difference in mI/Cr between NSCs and control groups ( t =0.046,P ＜ 0.05).There were no significant differences in Cho before and after transplantation among the three groups (P ＞ 0.05 ). Nissl's staining showed that the number of neurons in the hippocampal area increased more significantly in tg mice receiving NSCs than that without receiving NSCs.Electron microscopy showed that most hippocampal NSCs in NSCs group were morphologically normal with abundant organelles,while hippocampal NSCs in PBS group were swollen with sparse synapses.Conclusion 1H-MRS is able to display intracranial metabolite changes before and after NSCs in APP-PS1 double transgenic AD mice and has an applicable value in evaluating the therapeutic effect of NSCs on AD.

Objective To explore the prevalence of polycystic ovary syndrome (PCOS) in nurse of reproductive age and compare the characteristics of four phenotypic subgroups.Methods A cross-sectional survey was carried out in nurses aged 18-45 years in Renji Hospital in 2011.Questionnaire and anthropometric and biochemical assessments were made.Pelvic ultrasound evaluations were made and blood androgen levels were determined.Diagnosis of PCOS was based on Rotterdam 2003 criteria,consisting of anovulation/oligo-ovulation (ANOVU),clinical and/or biochemical hyperandrogenism (HA) and polycystic ovaries (PCO).Results There were 520 participants and finally 486 individuals finished questionnaire and androgen level determination.283 subjects were totally normal,48 suffered from PCO,129 HA,and 89 ANOVU.54 out of 486 women were diagnosed as PCOS,a prevalence of 11.1％.A significant difference exited only in age among four phenotypic subgroups (P＜0.05).There was no statistic difference in other parameters.Conclusion Establishing an explicit definition of each condition in PCOS criteria has important investigational implications and increase the comparability of published researches.Application of Rotterdam criteria is feasible for earlier diagnosis and timely intervention in order to prevent serious complications.

BACKGROUND:Micro-arc oxidation technique is used to modify the surface properties of titanium and titanium al oy.
OBJECTIVE:To explore the surface properties of micro-arc oxidation film and its effect on the attachment, proliferation and alkaline phosphatase activity of MC3T3-E1.
METHODS:Forty-six pure titanium discs, 10 mm in diameter and 2 mm in thickness, were randomly divided into two groups. The discs of the experimental groups were treated by micro-arc oxidation technique in an electrolytic solution containing 0.02 mol/L sodiumβ-glycerophosphate and 0.2 mol/L calcium acetate;while the discs of the control group was machine-polished. The surface appearance of the discs was observed by a scanning electron microscopy, the ratio of calcium to phosphorus on the coating surface was detected by X-ray spectroscopy, and the crystal ine phase composition of the coating was detected by X-ray diffraction analysis. cellular morphology in the process of attachment was observed under the scanning electron microscope. cellproliferation was determined by cellcounting kit-8 at 2, 4, 74 days, while alkaline phosphatase activity were determined at 7 and 14 days.
RESULTS AND CONCLUSION:After micro-arc oxidation treatment, a rough and porous calcium-phosphate film was formed on the surface of titanium. The elements of micro-arc oxidation coating main mainly included Ca, P, O and Ti, and the micro-arc oxidation film was mainly composed of titanium oxide, calcium titanate, calcium phosphate and calcium metaphosphate. Under the scanning electron microscope, pseudopods appeared to grow out of the cells on the surface of micro-arc oxidation coating after 1 hour culture, and the typical morphology of the MC3T3-E1 cells could be observed at 4 hours. MC3T3-E1 proliferation (4 and 7 days of culture) and alkaline phosphatase activity (7 and 14 days of culture) were enhanced significantly in the micro-arc oxidation group compared with the control group. These findings indicate that the rough and porous calcium-phosphate coating produced by micro-arc oxidation technique on pure titanium surface could promote the early attachment, proliferation and osteogenic differentiation of MC3T3-E1.

Objective To explore changes of metabolites in APP/ PS1 double transgenie mice of Alzbeimer disease (AD) by 1H-MR spectroscopy (1H-MRS) and the application value of in early diagnosis of AD.Methods 1H-MRS was performed in 35 APP/PS1 transgenie mice of AD ( study group) and 20 wild type mice ( control group) at age of 3, 6 and 9 months using a 7.0 T MR system.Sub-peak areas of N-acetyl aspartate (NAA), myo-inositol (mI) and creatine (Cr) in the cerebral cortex and hippocampus were measured, and the NAA/Cr and mI/Cr ratios were calculated.The changes in pathology between the two groups were compared.Using the lower limit of 95% confidence interval (CI) of the ml/Cr ratio and the upper limit of 95% CI of the NAA/Cr ratio of AD mice as the threshold, their influences on sensitivity,specificity and accuracy of various age groups of AD animals were compared.Comparison of the 1H-MRS indexes between study mice and wild type mice at each time point were conducted by a two-sample t test.Results The mean mI/Cr ratios of AD mice were 0.68± 0.03, 0.72± 0.04, and 0.77 ± 0.04 respectively at 3, 6 and 9 months of age; while they were 0.63 ± 0.04, 0.64 ± 0.03, and 0.64 ± 0.04 respoetively in control group, the difference was significant ( t = 2.814, 5.146, 14.437, P < 0.01 ).Compared with the control group, the mI/Cr ratio of the 3-month-old AD mice of the study group was significantly increased,and histological examination showed proliferation and activation of neuroglial cells in the cerebral cortex and hippoeampus.The mean NAA/Cr ratio were 1.17 ±0.08, 1.04 ±0.05, and 0.90 ±0.05 respectively at 3,6 and 9 months of age in study group, while they were 1.18 ±0.07, 1.16 ±0.07, and 1.18 ±0.08respectively in control group.There were no significant difference ( t = 0.752, P > 0.05 ) between the study group and control group at 3 months of age, and the NAA/Cr ratio decreased significantly only at 6 and 9 months of age ( t = - 8.514, - 5.646, P < 0.01 ).The immunohistochemical exam demonstrated the appearance of Aβ plaque.According to threshold of mI/Cr, the sensitivity of AD mice of 3, 6 and 9 months of age was 80% (28/35), 84% ( 26/31 ) and 85% ( 23/27 ), and the specificity was 85% ( 17/20 ),94% (17/18) and 100% ( 16/16), and the accuracy was 82% (45/55), 88% (43/49) and 91% (39/43),respectively.For NAA/Cr, the sensitivity of AD mice of 6 and 9 months of age was 84% (26/31) and 89% (24/27), and the specificity was 89% (16/18) and 100% (16/16), and the accuracy was 86% (42/49) and 93% (40/43), respectively.Conclusions NAA and mI are the most sensitive and specific markers for early assessment of AD, and change of mI is earlier than that of NAA.Quantitative analysis of mI may provide important clues for early diagnosis of AD.

Objective To explorer the application in detection the position accuracy of the multileaf collimator of Varian accelerator with dynamic therapy log files. Methods A pre-designed MLC format files named PMLC for two Varian accelerators, the dynamic treatment log files were recorded 10 times on a different date, and be converted into the MLC format files named DMLC, compared with the original plan PMLC, so we can analysis two files for each leaf position deviation. In addition, we analysis the repeatability of MLC leaves position accuracy between 10 dynalog files of two accelerators. Results No statistically significant difference between the average position of the 10 times leaf position of the two accelerators,their were 0. 29 -0. 29 and 0. 29 -0. 30(z = -0. 77, P=0. 442). About 40% ,30% ,20% and 10% of the leaf position deviation was at ≤0. 2 mm, 0. 3 mm,0. 5 mm and 0. 4 mm,respectively. the maximum value was 0. 5 mm. More than 86% of the leaf position are completely coincident between 10 dynamic treatment files of two accelerators,The rate of position deviation no more 0. 05 mm was 96. 6% and 97.3%, respectively.And the maximum value was 0. 09 mm. Conclusions Dynamic treatment log file is a splendid tool in testing the actual position of multi-leaf collimator. The multi-leaf collimator of two accelerators be detected are precise and stabilized.