Background C57BL/6andB10R Ⅲareroutinemurinespeciesusedinexperimental autoimmune uveitis (EAU).The inflammation is light for mouse after immunization whereas it is prominent for B10R Ⅲ.ObjectiveThis study was to observe the killing effect of interphotoreceptor retinoid binding protein (IRBP) 1-20-specific T cells on mouse retinal astrocyte.Th1 and Th17 cells effect in the EAU mechanism was discussed.MethodsB10RllⅢ mice and C57BL/6 mice were immunized with IRBP 161-180 and IRBP 1-20 in complete Freund adjuvant (CFA).The infiltrating cells of diseased B10R Ⅲ eyes were analyzed by flow cytometry.IRBP 1-20-specific T cells were isolated from the drainage lymph node and spleen and cultured in IL-2 or IL-23 for Th1 and Th17 cells polarization,respectively.Th1 and Th17 cells cultured for 5 days were seeded on the mouse retinal astrocyte monolayer pretreated with gamma interferon.Cell interaction was observed and the quantity of TNF-α was tested by ELISA.Every test was repeated 6 times and the mean was calculated.The maintenance of experimental animals complied with the Statement of ARVO.ResultsThere were lots of infiltrating cells in the eyes of B10Rm mice after immunization,including 9.5％ IFNγ+ cells,5.1％ IL-17+cells and 41.4％ CD45+ cells.Six days after IRBP1-20 stimulation and cultured by IL-2 and IL-23,44.0％ and 8.0％ cells were IFNγ+,and 1.0％ and 26.0％ cells were IL17+.Twentyfour hours after the interaction between Th1 or Th17 and retinal astrocyte,retinal astrocyte died and detached.The killing effect of Th17 was stronger than Th1.48 hours after co-culture of Th1 or Th17T cells with astrocytes,the concentrations of TNF-α were ( 500± 10 ) and ( 801 ±24 μg/L) μg/L,respectively,with a significant statistical difference (t =-20.36,P =0.00).ConclusionsBoth Th1 and Th17 can kill retinal astrocyte,but Th17 plays a key role in the EAU pathogenesis process.The killing effect is caused by intercellular contact and interaction under the induction of cytokines.

OBJECTIVE: To prepare inclusion complex of SBE7-β-CD with trimethoprim(TMP) and optimize the preparation process, to evaluate the products by structural characterization and molecular simulation. METHODS: The TMP/SBE7-β-CD inclusion complex was prepared by the ultrasound-freeze-dry method and the preparation process was optimized by Box-Behnken Design-response surface method(BBD-RSM). Inclusion complex was characterized by FT-IR, DSC, XRPD and 1H-NMR. Molecular docking method was used to simulate 3-dimensional conformations of the inclusion complex and the binding energy was calculated. The dissolution and stability were tested. RESULTS: The optimum conditions of TMP/SBE7-β-CD inclusion complex were: temperature(52 ℃), time(45 min), and the ratio of SBE7-β-CD and TMP(mol/mol, 1.7∶1). All characterizations(FT-IR, DSC, XRPD and 1H-NMR) indicated the formation of TMP/SBE7-β-CD inclusion complex. The best 3-dimensional docking conformation was consistent with the characterizations, and the binding energy was -9.015 kcal·mol-1. The TMP dissolution rate of the inclusion complex increased significantly, the hygroscopicity is strong. CONCLUSION: The preparation process of TMP/SBE7-β-CD inclusion complex optimized by BBD-RSM is reasonable and feasible. The characterizations of inclusion complex are reliable. The molecular simulation is corresponded to the characterized results and provided reliable theoretical basis for inclusion experiments.

Abdominal Injuries ; etiology ; prevention & control ; Humans ; Iatrogenic Disease ; Intraoperative Complications ; prevention & control

To develop a LC-MS/MS method for the determination of protocatechuic acid, protocatechuic aldehyde, salvianolic acid A, salvianolic acid B, cryptotanshinone and tanshinone II(A) in rat plasma and brain. The plasma and brain samples were precipitated with ethyl acetate, then were separated on an Agilent eclipse plus-C18 column (2.1 mm x 50 mm, 3.5 microm) using acetonitrile (consisting of 0.1% formic acid) and water (consisting of 0.1% formic acid) as mobile phase in gradient elution mode. The mass spectrometer was operated under both positive and negative ion mode with the ESI source, and the detection was performed by MRM. The transition of 154.3/153.1 m/z for protocatechuic acid, 137.3/108 m/z for protocatechuic aldehyde, 493.0/295.2 m/z for Salvianolic acid A, 718.0/520.0 m/z for salvianolic acid B, 321.4/152.3 m/z for chloramphenicol, 297.4/254.3 m/z for cryptotanshinone, 295.5/249.3 m/z for tanshinone II(A) and 285.2/154.0 m/z for Diazepam. The calibration curves in the range of 0.625-1 000 microg x L(-1) for protocatechuic acid and protocatechuic aldehyde, 1.25-1 000 microg x L(-1) for salvianolic acid A, 2.5-1 000 microg x L(-1) for salvianolic acid B, 0.15-1 000 microg x L(-1) for cryptotanshinone, 0.625-1 000 microg x L(-1) for tanshinone II(A) are with good linearityin rat plasma and brain. The analysis method is sensitive, simple, and suitable enough to be applied in the pharmacokinetic study of the 6 main components. Animal testing gives the lgBB of the drugs and further studies of the 6 components cross the blood-brain barrier can be carried out.
Animals ; Benzaldehydes ; administration & dosage ; blood ; pharmacokinetics ; Benzofurans ; administration & dosage ; blood ; pharmacokinetics ; Blood-Brain Barrier ; metabolism ; Brain ; metabolism ; Caffeic Acids ; administration & dosage ; blood ; pharmacokinetics ; Catechols ; administration & dosage ; blood ; pharmacokinetics ; Chromatography, Liquid ; methods ; Diterpenes, Abietane ; administration & dosage ; blood ; pharmacokinetics ; Hydroxybenzoates ; administration & dosage ; blood ; pharmacokinetics ; Injections, Intravenous ; Lactates ; administration & dosage ; blood ; pharmacokinetics ; Phenanthrenes ; administration & dosage ; blood ; pharmacokinetics ; Plant Preparations ; administration & dosage ; blood ; pharmacokinetics ; Rats ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry ; Tandem Mass Spectrometry ; methods

Air Pollutants, Occupational ; analysis ; Gas Chromatography-Mass Spectrometry ; Halogenated Diphenyl Ethers ; analysis ; Workplace

Carcinoma, Non-Small-Cell Lung ; diagnostic imaging ; Humans ; Lung ; diagnostic imaging ; Lung Neoplasms ; diagnostic imaging ; Perfusion Imaging ; Radiotherapy, Intensity-Modulated ; methods ; Tomography, Emission-Computed, Single-Photon ; methods

Accident Prevention ; Humans ; Malpractice ; Medical Errors ; prevention & control ; Surgical Procedures, Operative

ObjectiveTo explore the influence factors of prognosis of spinal cord injury patients. MethodsFollow-up data of 121 cases with spinal cord injury was analysed with retrospective cohort study.ResultsThe highest recover rate was in the patients who accepted the special rehabilitation therapy,while that of recovering two class was in the patients who accepted the magnetic stimulation.Conclusions Special recovering cure and magnetic stimulation can surely promote recovery of spinal cord injury patients.

Object To study the botanic characteristics of leaves from three plants of Goniothalamus (Bl.) Hook. f. et Thoms. in order to correctly distinguish them from numerous plants of the genus, which are important resource of anticancer medicine.Methods The maceration method and paraffin method were used to study the epidermis and structures of leaves from G. griffithii Hook. f. et Thoms., G. leiocarpus (W. T. Wang) P. T. Li and G. yunnanensis W. T. Wang. Results Three leaves were morphologically similar in the structure, but there were some anatomical differences among them. For example, the absence of druses in the epidermis and the presence of fibrous sclereids in the lamina mesophyll of leaves from G. griffithii, while the presence of druses in epidermis and the absence of fibrous sclereids in lamina mesophyll of the leaves from G. griffithii and G. yunnanensis were observed. In addition, epidermal hairs of G. griffithii were composed of three cells, stomatas were always normal, there were seven oil cells and 25 mucilage cells per mm leaf width in lamina mesophyll and the vascular tissue of the midrib consisting of ten small bundles. However, epidermal hairs of G. yunnanensis were composed of two cells, many abortive stomatas were present at the distal surface, there were only four oil cells and 16 mucilage cells per mm leaf width and the vascular tissue of the midrib consisted of 12 small bundles.Conclusion Three species were easily identified on the basis of epidermal and structural characters of the leaves of them.

OBJECTIVETo investigate extracellular splitting pattern of mitochondria and the depressant effects of CsA on the process and explore the mechanism of post-traumatic SIRS and its therapeutic strategy.

METHODSTen male SD rats with 60 to 70 days age and 240 to 280 g weight were used for mitochondrial isolation. Freshly isolated mitochondria were randomly divided into two groups, which were cultured in blood plasma with or without CsA respectively for 8 h. COX and MDH were assayed by ELISA every 30 min. Meanwhile, Rat macrophage cell line NR8383 were treated as follows, control (group A): cultivation with normal medium; NR8383+CsA co-culture group (group B): culture medium was supplemented with CsA of 10 mmol/L; NR8383+intact mitochondria co-culture group (group C): culture medium was supplemented with intact mitochondria (mtDNA=5 g/ml); NR8383+intact mitochondria+CsA co-culture group (group D): culture medium was supplemented with intact mitochondria (mtDNA=5 μg/ml)and CsA of 10 mmol/L; NR8383+disrupted mitochondria co-culture group (group E): culture medium was supplemented with disrupted mitochondria (mtDNA=5 μg/ml); NR8383+disrupted mitochondria+CsA co-culture group (group F): culture medium was supplemented with disrupted mitochondria (mtDNA=5 μg/ml)and CsA of 10 mmol/L. TNF-α and IL-6 concentrations in supernatant were assessed at 1, 3, 5 h after culture.

RESULTSIn the mitochondria plasma cultures, MDH and COX levels were increased with the time and peaked at about 3 h and 3.5 h; CsA can delay the appearance of peak to 4.5 h. Among different treated groups,there was no significant difference in TNF-α and IL-6 between group A and group B; there was significant difference in TNF-α and IL-6 other groups. After 1 h culture, compared with group C, no significant difference of TNF-α and IL-6 was observed in group D, while TNF-α and IL-6 were significant higher in group E; after 3 h culture, compared with group C, TNF-α and IL-6 were significantly lower in group D, while TNF-α and IL-6 were significantly higher in group E; after 5 h culture, compared with group C, TNF-α and IL-6 were significantly lower in group D, while no significant difference of TNF-α and IL-6 were observed in group E. At each time point, there was no significant difference in TNF-α and IL-6 between group F and group E.

CONCLUSIONMitochondria can split in serum with time, which will further activate macrophages. CsA has depressant effect to mitochondrial splitting on the process and will therefore inhibit the activation of macrophages.

Animals ; Cells, Cultured ; Cyclosporine ; pharmacology ; Interleukin-6 ; secretion ; Male ; Mitochondria ; drug effects ; Prostaglandin-Endoperoxide Synthases ; analysis ; Rats ; Rats, Sprague-Dawley ; Systemic Inflammatory Response Syndrome ; drug therapy ; etiology ; Tumor Necrosis Factor-alpha ; secretion