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

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.

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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.

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.

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.

BACKGROUND: Under indirect co-culture conditions, chondrocytes can induce bone marrow mesenchymal stem cells to differentiate into chondrocytes. Osteocytes are the seed cells of chondrocytes, but co-culture of osteocytes with chondrocytes is rarely reported. OBJECTIVE: To induce the chondrogenic differentiation of periosteum-derived cells (PDCs) by indirect co-culture with chondrocytes. METHODS: Chondrocytes were isolated from the rabbits by trypsin and collagenase Ⅱ digestion. Rabbit PDCs were obtained by the explants culture method. Passage 2 chondrocytes and PDCs underwent indirect co-culture at 1:1 in Transwell system as experimental group. PDCs cultured alone were as control group. After 2 weeks of culture, the cellular morphological changes were observed by inverted contrast phase microscope. The expression levels of collagen type Ⅱ and proteoglycan were detected by collagen type Ⅱ immunohistochemistry and toluidine blue staining. The mRNA expression levels of proteoglycan, collagen type Ⅱ and SRY-related protein-9 were detected by RT-PCR. RESULTS AND CONCLUSION: After 2 weeks of culture, PDCs in the experimental group were gradually transformed to chondrocyte-like cells, while PDCs in the control group still remained long spindle-shaped. Collagen type Ⅱ immunohistochemistry staining and toluidine blue staining were positive in the experimental group, while the results were negative in the control group. Additionally, RT-PCR results indicated that the relative mRNA expression levels of proteoglycan, collagen type Ⅱ and SRY-related protein-9 in the experimental group were significantly higher than those in the control group. Therefore, chondrocytes can induce PDCs to differentiate into chondrocytes under indirect co-culture conditions.

Objective To investigate the value of MRI and~1H-MRS in diagnosis of early stage of diabetic encephalopathy by detecting regional metabolite in cynomolgus diabetes models. Methods Five pathogen-free male adolescent cynomolgus were made type 1 diabetes mellitus models (T1DM) by intravenous injection of streptozotocin (STZ) (100 mg/kg), and the reliability and stability of the modes were assessed with long term follow-up of blood glucose and intravenous glucose tolerance tests. MRI and ~1H-MRS were performed to evaluate the volume, signal intensity and metabolic ratios of NAA/Cr, mI/Cr and Cho/Cr at hippocampus, lateral temporal lobe and occipital lobe 3 years after model establishment. Cortisol in serum was detected with immunoradiometric assay. In addition, 5 normal adult cynomolgus monkeys were selected in the control group and accepted the same examination above. Results ①Intravenous administration of STZ could made stable T1DM monkey model. ②Only mI/Cr ratio increased at hippocampus of diabetic monkeys compared to the control group (P<0.05). ③There was no statistical difference of cortisol in serum between the diabetic group and the control group (P＞0.05). Conclusion ~1H-MRS may detect the metabolic changes of the hippocampus in STZ-induced diabetic adolescent cynomolgus monkeys and may contributes to the early diagnosis of diabetic encephalopathy.

Objective To study the effects of abiotic elicitors methyl jasmonate (MeJA) and salicylic acid (SA) on the alkaloids accumulation and related enzymes metabolism inPinellia ternata suspension cell cultures. Methods Using the leaf petioles-derived suspension cell cultures as the study object, the culture duration, concentrations of MeJA and SA were determined to get the optimal alkaloids accumulation, and the activities of metabolic enzymes IMP dehydrogenase and sAMP synthase were also measured.Results A 9-fold of dried biomass and a 3-fold of alkaloids accumulation were observed inP. ternata suspension cell cultures after culture for 21 d. Both MeJA and SA could significantly promote the accumulation of alkaloids inP. ternata suspension cells. 150 μmol/L MeJA enhanced alkaloids content (4.7 mg/gDW) by 3.6 folds in comparison with control group, whereas 50 μmol/L SA showed a 2.5-fold increase. Meanwhile, 100 μmol/L MeJA and 50 μmol/L SA promoted the increase in IMP dehydrogenase activity by 3.0 and 3.7 fold respectively, and 150 μmol/L MeJA and 100 μmol/L SA showed the increase by 2.6 and 4.4 fold respectively.Conclusion Proper adding exogenous MeJA and SA can promote the accumulation of alkaloids inPinellia ternata suspension cell cultures.