Objective:To prepare superparamagnetic iron oxide(SPIO) nanoparticles and to observe its acute toxicity on mice,so as to pave a way for further study on its long-term toxicity and on its role as a carrier in magnetic resonance gene imaging.Methods: The SPIO nanoparticle was obtained by means of co-precipitation,and its physical and chemical parameters were determined by transmission electron microscope,atomic force microscope,and 1.5 T super conduct MR,etc.According to the administration pathway and doses of SPIO,90 mice were divided into oral administration(with a total dose of 2 104.8 mg/kg and a volume of 40 ml/kg,n=30),intravenous injection(a total dose of 438.5 mg/kg and a volume of 25 ml/kg,n=30) and intraperitoneal injection(with a total dose 1 578.6 mg/kg and a volume of 30 ml/kg,n=30) groups.Another 10 mice in each group receiving the same dose of normal saline via the same pathway served as the controls(n=10).The general condition,the major serologic parameters,and the pathological changes of major organs were observed 14 d after administration in each group.Results: We have successfully prepared SPIO,and its core component was Fe3O4 crystal,with a size of 20-35 nm,a T2 relaxivity of 0.155?106 mol-1?sec-1,a specific saturated magnetization of 68.395 68 emu/g,and a retentivity of 21.463 74 Gs.There was no death of mice during the observation.There was no significant difference in serological parameters between mice of different groups and between each experiment group and their corresponding control group.No edema,degeneration and necrosis were seen in the liver,spleen,kidney,heart,and lungs by H-E staining and marrow by Wright staining;only a few blue particles were observed in the liver and spleen in the administration groups by Prussian blue staining,none observed in the control groups.Conclusion: SPIO prepared in the present study meets the requirement of MR imaging,with no acute toxicity to mice,and warrants further study for future MR gene imaging.

BACKGROUND: Changes of elastic fibers in middle cerebral artery(MCA)is close related withthe aged cerebrovascular disease.OBJECTIVE: To observe the changes of elastic fibers of MCA in different aging rats.DESIGN: A descriptive and controlled study based on experimental animals.SETTING: Department of anatomy and central laboratory in a university.MATERIALS: Totally 36 healthy Wistar rats with either gender, weighing 200 - 280 g, were selected from the Animal Institute of the third medical military university of Chongqing[certification SCXX (army) 2002-007].INTERVENTIONS: Changes of elastic fibers of MCA of different aging rats were observed with light microscope, transmission electron microscope and image analysis system. MAIN OUTCOME MEASURES: ①) Major outcome: changes of elastic lamella in MCA of different aging rats; ②) Secondary outcome: ultramicrostructural changes of internal lamella under the transmission electron microscope. RESULTS: With the increase of age, the folded extent and quantity of internal elastic lamella were decreased, and the content of elastic fibers were also decreased significantly( P ＜ 0.01 ). However, the ratio of collagen fibers to elastic fibers was increased significantly( P ＜ 0.01 ) . In the aging group above 24 months, the internal elastic lamina thinned, delaminated and disrupted, and the lipid deposited in it. Endothelial cells and smooth muscle cells passed through the internal elastic lamina. CONCLUSION: Changes of elastic fibers may be related with the increased susceptibility to the cerebrovascular disease in aged people.

A selective precolumn derivatization liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of glucosamine in human plasma and urine has been developed and validated.Glucosamine was derivatized by o-phthalaldehyde/3-mercaptopropionic acid.Chromatographic separation was performed on a Phenomenex ODS column (150 mm × 4.6mm,5μm) using linear gradient elution by a mobile phase consisting of methanol (A),and an aqueous solution containing 0.2％ ammonium acetate and 0.1％ formic acid (B) at a flow rate of 1 mL/min.Tolterodine tartrate was used as the internal standard (IS).With protein precipitation by acetonitrile and then the simple one-step derivatization,a sensitive bio-assay was achieved with the lower limit of quantitation (LLOQ) as low as 12 ng/mL for plasma.The standard addition calibration curves suitable for clinical sample analysis showed good linearity over the range of 0.012-8.27 μg/mL in plasma and 1.80-84.1 μg/mL in urine.The fully validated method has been successfully applied to a pharmacokinetic study of compound glucosamine sulfate dispersible tablets in health Chinese volunteers receiving single oral doses at 500,1000 and 1500 mg of glucosamine sulfate,as well as multiple oral doses of 500 mg t.i.d.for 7 consecutive days.

Objective:To explore the correlation between the characteristics of contrast-enhanced sonography of intraoperative glioblastoma multiform (GBM) and molecular markers of isocitrate dehydrogenase-1(IDH1).Methods:A retrospective analysis were performed in 30 patients who underwent neurosurgery and pathologically confirmed to be GBM at Beijing Tiantan Hospital from May 2018 to April 2019. All neurosurgical glioblastoma patients after craniotomy underwent conventional ultrasound and contrast-enhanced ultrasound(CEUS) guided navigation. The characteristics of the ultrasound imaging (whether the tumor involves the structure of the corpus callosum, the clarity of the tumor boundary after enhanced ultrasound and whether the tumor has necrotic areas with enhanced ultrasound images) were analyzed. The ratio between tumor necrosis area and whole tumor area (N/W) was measured, and the correlation with IDH1 gene expression was analyzed.Results:There were statistical differences in clarity of tumor boundary after CEUS and tumor necrosis after CEUS between positive IDH1 and negative IDH1 groups(all P<0.05). The positive expression of IDH1 was negatively correlated with the N/W area of the contrast-enhanced ultrasound mode( r=-0.756, P<0.05), suggesting that the expression level of IDH1 gene was negatively correlated with the area of tumor necrosis. Conclusions:Ultrasound contrast agent examination can more accurately distinguish the active proliferation area, hemorrhagic necrosis area and peripheral edema area of glioblastoma. Accurately identifying the extent of tumor necrosis area through ultrasound contrast agent examination can predict expression of IDH1.

OBJECTIVETo observe the regulatory effects of psoralen, oleanolic acid, and stilbene glucoside, three active components of psoralea fruit, glossy privet fruit and tuber fleeceflower root respectively, on Aβ25-35induced self-renewal and neuron-like differentiation of neural stem cells (NSCs).

METHODSEmbryonic NSCs werein vitro isolated and cultured from Kunming mice of 14-day pregnancy, and randomly divided into the control group, the Aβ25-35 group, the Aβ25-35 +psoralen group, the Aβ25-35 +oleanolic acid group, and the Aβ25-35 + stilbene glucoside group. The intervention concentration of Aβ25-35 was 25 µmol/L, and the intervention concentration of three active components of Chinese medicine was 10(-7)mol/L. The effect of three active components of Chinese medicine on the proliferation of NSCs was observed by counting method. The protein expression of Tubulin was observed by Western blot and immunofluorescence. The ratio of Tubulin+/DAPI was caculated. Results Compared with the control group, the sperical morphology of NSCs was destroyed in the Aβ25-35 group, the counting of NSCs, the expression of Tubulin protein, and the ratio of Tubulin /DAPI all decreased (P <0.01, P <0.05). Compared with the Aβ25-35 group, the counting of NSCs, the expression of Tubulin protein, and the ratio of Tubulin + /DAPI all increased in the three Chinese medicine treated groups (P <0. 01, P <0. 05).

CONCLUSIONS25 µmol/L Aβ25-35 could inhibit self-renewal and neuron-like differentiating of NSCs. But psoralen, oleanolic acid, and stilbene glucoside could promote self-renewal of NSCs and neuron-like differentiation.

Amyloid beta-Peptides ; physiology ; Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Embryo, Mammalian ; Female ; Mice ; Neural Stem Cells ; Neurogenesis ; drug effects ; Neurons ; cytology ; Peptide Fragments ; physiology ; Pregnancy

· ATM:To analyze bacterial spectrum and drug sensitivity in conjunctival sac of cataract patients of Kazak.
· METHODS:A total of 538 cases of conjunctival sac secretion in cataract patients of Kazak were collected.The samples were cultured and their sensibilities to antibiotics were tested.
· RESULTS: The bacterial culture was positive in 214 cases.The positive rate was 39.8%. The variety of pathogenic bacteria were mainly made up of gram positive cocci ( 88.3%), and most of them were Staphylococcus epidermidis ( 66.4%), followed by Micrococcus(9.8%).Sex had no effect on conjunctival bacteria rate in the cataract patients of Kazak, while age, place of residence had an effect on camier rate. The camier rate of conjunctival bacteria was significantly higher in people over 60 years old than that in people with age between 40 to 59 years old.And the people from city had a significant lower bacteria positive rate than those from countryside and pastoral. Most of grams were sensitive to Vancomycin, Teicoplanin, Rifampicin, Duly cloth mildew mutual and Amikacin, the tolerance was less than 20%, and they usually had higher tolerance to Penicillin, Erythromycin, Tetracycline and Chloramphenicol (>70%) .
·CONCLUSlON:Gram positivecocci is the most common bacteria in conjunctival sac in cataract patients of Kazak. Staphylococcus epidermidis was most common, followed by Micrococcus.The germ-carrying rate of conjunctival SAC in Kazakh population is associated with the patient’s age and area of residence.The clinical use of antibacterial drugs should be strictly grasp the indications, to reduce the incidence of bacterial resistance.

Objective To explore the feasibility of establishment of NOD/SCID-mouse model with multiple myeloma by using plasma cells from myeloma patients.Methods The femurs and tibias were removed from the New Zealand white rabbits;the muscles,periosteum and cartilage tissues were cleared.Then each bone was cut into two pieces gently along its middle.The NOD/SCID mice weighing 25 - 30 g (4 - 6 weeks)were anesthetized by intraperitoneal injection;rabbit bone was inserted into the right side of the mouse back and engraftment of the bones was allowed to take place after 4 weeks.The 5000 000 purified plasma cells which expressed CD38 +/CD45 - were immunofluorescence labeled and then injected slowly into the implanted rabbit bone through the distal end.The mice were observed weekly;the plasma cells growth in mice was screened by the living-imaging system and the tumor from the mice was determined by biopsy.Results The implanted rabbit bone survived after 4 weeks.The tumor in mice was observed 2 weeks after the purified myeloma cells were injected into the rabbit bone,and it reached 100 mm3 after 8 weeks.Results of the living-imaging system showed that the myeloma cells had uptake in the rabbit bone after 2 weeks of injection and this phenomenon was more pronounced after 8 weeks of injection (2.4×10 4 vs .1.5× 10 5 ,P < 0.05 ).The tumor infiltrated with numerous plasma cells and osteoclasts increased upon the biopsy. Conclusion Rabbit bone marrow implanted into NOD/SCID mice can effectively support local injection of plasma cells of multiple myeloma patients,and the NOD/SCID-mouse model of myeloma has been established.This model can be used to study in vivo experiments related to myeloma and clinical therapeutic approaches for this disease.

Objective To determine the function of conserved amino acids in the fusion-promoting domain of Newcastle disease virus (NDV) hemagglutinin-neuraminidase (HN) protein,clearly understanding mechanism of cell fusion.MethodsUsing a PCR-based site-directed mutagenesis method and the method of homology recombination occurred in vivo to change six conservative amino acids into alanine respectively.Wild type (WT) and all mutant HN proteins were exepressed in BHK-21 cells by the vacciniaT7 RNA polymerase expression system.The amount of each HN protein at the cell surface was determined by fluorescence-activated cell sorter (FACS).Cell fusion efficiency,hemadsorption activity (or receptor binding activity) and neuraminidase activity were determined.Results There was no statistic difference of cell surface expression among WT and each mutant HN protein ( P＜0.05 ).Cell fusion efficiency of each mutant protein decreased to some extent,especially 1103A decreased to 14.2％ in head.Hemadsorption activity of mutant proteins were reduced in different extent,the maximum reduction of which was also 1103A,28.2％ of wt NDV HN.There was different neuraminidase activity among each mutant HN protein.L74A increased slightly to 118.6％.L110A decreased most to 5.2％.I103A decreased second most to 5.7％.Conclusion Conserved amino acids in fusion-promoting domain of NDV HN played an important role in cell fusion.I103 was identified as a key amino acid in this domain.

Objective To synthesize N-(4-fluorophenyl)-N-(4-phenoxyphenyl) cyclopropane-1,1-dicarboxamide (NFNPDB) as small molecular c-Met kinase inhibitor analogue.Methods N-( 4-fluorophenyl )-N-( 4-phenoxyphenyl ) cyclopropane-1, 1-dicarboxamide was synthesized by nucleophilicsubstitution, amidation, etherification, reduction and condensation from diethyl malonate.Results The total yield of target compound was 3.79%, its structure was confirmed by 1 H-NMR.Conclusion The synthesis method of NFNPDB in our research can be easily operated with lost cost and short direction, which lays the foundation for designing the synthetic process of newly small molecular c-Met kinase inhibitor.

Objective To investigate the role of lysyl oxidase (LOX) in synovitis and cartilage destruction by comparing the expression of LOX in synovial fluid and synovium of active rheumatoid arthritis (RA) and active osteoarthritis (OA).Methods LOX in the synovium was detected by immunohistochemistry from 14 patients with active RA,24 patients with active OA and 20 patients with knee injury (the control group).LOX in the synovial fluid was measured by enzyme linked immunosorbent assay (ELISA) from 14 patients with active RA and 24 patients with active OA.T-test was used for statistical analysis.Results The level of LOX expression in active RA synovium (0.012±0.007) was similar to that in active OA synovium (0.013±0.011,P＞0.05).But the expression of LOX in synovium of active RA and active OA was significantly higher than that in synovium of the control group (0.003±0.004,P＜0.01).The amount of LOX in the synovial fluid of active RA [(1.9±1.4) μg/ml] was significantly higher than that of active OA [(1.0±0.4) μg/ml,P＜0.05].Conclusion High expression of LOX in the synovial fluid and synovium of active RA and active OA suggest that LOX may be involved in chronic synovitis and cartilage destruction,and may be related with the extent of synovitis and cartilage destruction.