ObjectiveTo evaluate the neuroprotective effect and possible mechanisms of edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one) in neonatal Harlequin (Hq) mutant mice brain after hypoxia-ischemia brain injury(HIBD) insult.MethodsThe nine-day-old male Hq mutant mouse pups were assigned randomly either edaravone (n=16) and vehicle (n=17) treatment group. The Hq mice were subjected to left common carotid artery occlusion combined with inhalation 100 mL/L oxygen for 45 minutes. The mice were injected intraperitoneally either with edaravone (10 mg/kg) or equivalent volume of saline immediately after artery occlusion and after hypoxia. Nitrotyrosine and lipid peroxidation formation were evaluated at 3 h and 24 h after hypoxia-ischemia(HI) by using immunohistochemistry staining. Nitrotyrosine formation and caspases activation were evaluated either by immunoblotting or fluorogenic activity measurement at 24 h after HI. Brain injury was evaluated at 72 h by neuropathological score and calculating the infarct volume.ResultsBrain injury encompassed cortex, hippocampus, striatum and thalamus. Edaravone treatment reduced brain injury significantly in all the brain regions. The total infarct volume was reduced 52.8% in edaravone treatment group compared with vehicle group (P<0.001). The edaravone treatment reduced nitrotyrosine formation as well as lipid peroxidation formation significantly, but without obviously effect on caspases activation.ConclusionEdaravone affords neuroprotection after neonatal HI insult, which correlated with the reduction of free radical formation.

Objective To evaluate the neuroprotective effect and possible mechanisms of edaravone(3-methyl-1-phenyl-2-pyrazolin-5-one) in neonatal Harlequin(Hq) mutant mice brain after hypoxia-ischemia brain injury(HIBD) insult.Methods The nine-day-old male Hq mutant mouse pups were assigned randomly either edaravone(n=16) and vehicle(n=17) treatment group.The Hq mice were subjected to left common carotid artery occlusion combined with inhalation 100 mL/L oxygen for 45 minutes.The mice were injected intraperitoneally either with edaravone(10 mg/kg) or equivalent volume of saline immediately after artery occlusion and after hypoxia.Nitrotyrosine and lipid peroxidation formation were evaluated at 3 h and 24 h after hypoxia-ischemia(HI) by using immunohistochemistry staining.Nitrotyrosine formation and caspases activation were evaluated either by immunoblotting or fluorogenic activity measurement at 24 h after HI.Brain injury was evaluated at 72 h by neuropathological score and calculating the infarct volume.Results Brain injury encompassed cortex,hippocampus,striatum and thalamus.Edaravone treatment reduced brain injury significantly in all the brain regions.The total infarct volume was reduced 52.8% in edaravone treatment group compared with vehicle group(P

Objective To establish brain hypoxic-ischemic (HI) model using postnatal day 3(P3) SD rats and evaluate the apoptotic neuronal cells in cerebral cortex and neurofunctional development.Methods The P3 rats were randomly divided into HI group (n=35) and control group(n=18).HI was induced in P3 rats with right carotid artery ligation followed by 2.5 h hypoxia in 60 mL?L-1 oxygen at 37 ℃.The injury of neural cells in the cerebral cortex was evaluated by tumor necrosis factor receptor-1(TNF-R1),Caspase-3 immunostaining and Hematoxylin-Eosin (HE) staining in 24 h and 7 d after HI,respectively.Furthermore,the neurofunctional development was evaluated by negative geotaxis reflex and eye opening time.The data were analyzed by SPSS 12.0 software.Results Caspase-3 and TNF-R1 positive cells were abundant in the ipsilateral cortex at 24 h after HI,compared with contralateral part and control group(P

The preterm birth has been increasing for the last decade. With the development of neonatal intensive care techniques, the survival rate of preterm infants is increased markedly. However, the brain of preterm infants is so vulnerable to injury that preterm brain injury has become an enormous public health problem. Hypoxia-ischemia and infection/inflammation are two main perinatal risk factors causing premyelinating oligodendrocyte and cortical neuron injury. Encephalopathy of prematurity is characterized by diffuse white matter injury and neuronal/axonal disruption, leading to neurological disabilities such as cognitive impairment and cerebral palsy. The advancement in imaging techniques, especially magnetic resonance imaging, provides more information for preterm brain injury and brain development, which contributes to the diagnosis and follow-up of the preterm infants. This article reviews the progress in encephalopathy of prematurity in order to open a new window to prophylaxis and management of this disease.
Brain Diseases ; diagnosis ; pathology ; therapy ; Humans ; Infant, Newborn ; Infant, Premature ; Infant, Premature, Diseases ; diagnosis ; pathology ; therapy ; Leukomalacia, Periventricular ; diagnosis ; Magnetic Resonance Imaging ; Neurons ; pathology ; Tomography, X-Ray Computed

Acupuncture Therapy ; California ; International Cooperation ; Medicine, Chinese Traditional ; methods ; trends ; United States

Objective: To explore the high-efficiency green environmental extraction method of total flavones from Microcos paniculata (MPTF), and investigate its lipid-lowering activity. Methods: The ionic liquid was used to assist the ultrasonic extraction of MPTF, and the extraction process was investigated by single factor experiment and orthogonal test. Wistar rats were randomly divided into normal control group and positive control group (Resuvastatin Calcium Tablets 5.2 mg/kg) by high fat diet, high-fat model group and MPTF low, medium and high dose groups (ig dose MPTF 300, 600, 900 mg/kg), with 10 rats in each group. The total cholesterol (TC), glycerol lipid (TG), high density lipoprotein (HDL-C), low density lipoprotein (LDL-C) levels were determinated. Results: The optimum process conditions for ultrasonic-assisted extraction of MPTF by ionic liquid were as follow: ionic liquid was [BMIM]Cl at concentration of 0.30 mol/L; ratio of material to liquid was 1:40, extraction solvent was 60% ethanol aqueous solution with extraction time of 30 min at 50 ℃. The verification test results showed that the extraction rate of total flavonoids obtained from the extraction process was high and the process was stable. The results of lipid-lowering test showed that the levels of TC, TG and LDL-C were decreased in all doses of MPTF groups, and the level of HDL-C was increased (P < 0.05). With the increase of MPTF dose, the indicators showed obvious trends in a dese-dependent manner. Conclusion: The ionic liquid combined with ultrasonic- assisted extraction of MPTF is stable and feasible, which provides reference for ionic liquid synergistic ultrasound-assisted extraction of poorly soluble active ingredients in Chinese materia medica. MPTF extract has better lipid-lowering effect.

Technological improvement for microorgnism isolation is important since isolation provides substantial materials for the exploitation of new microbial resources. In this study, a new approach, dispersion and differential cetrifugation (DDC), was applied in the isolation of acidophilic and acidoduric streptomycetes from 12 acid soil samples. Contrast with traditional method, the new approach yielded satisfying results with 2 - 20 times isolation efficiency and good selectivity. 45 representatives out of 249 streptomycetes isolates, which belonged to 12 color groups, showed morphology and cell wall type consistent with streptomycetes. The optimum pH range for their growth were between pH 4.5 - 5.5. It is proved that we succeeded in the rare-streptomycetes isolation using DDC approach.

Objective - - To analyze the prevalence and influencing factors of multi site work related musculoskeletal disorders （ ） Methods WMSDs in surgeons. A total of 102 surgeons from four hospitals were selected as study subjects by convenient sampling method. The Chinese version of Musculoskeletal Disorders Questionnaire was used to investigate the prevalence of ， Results WMSDs in the past one year the related individuals and occupational factors. The total prevalence of WMSDs among （ ）， （ ） （ ） surgeons was 54.9%. The top three sites were neck 48.0% lower back 35.3% and shoulder 32.4% . The prevalence of （ vs ，P ） WMSDs in multiple sites was higher than that in a single site 43.1% 11.8% <0.01 . Multivariate logistic regression ， ， analysis showed that surgeons who smoked were tired at work and had a bent back had a higher risk of developing WMSDs ［ （ - ）， （ - ）， （ - ）， P ］ odds ratios and 95% confidence intervals were 3.66 1.41 9.46 8.33 2.15 32.20 and 18.74 2.14 166.77 all <0.01 Conclusion - after excluding the influence of confounding factors. The prevalence rate of multi site WMSDs among surgeons is ， high and the influencing factors include bad living habits and occupational factors such as working load and working posture.

Interleukin-1 receptor antagonist (IL-1ra), a member of IL-1 family, is a naturally occurring IL-1 inhibitor as "receptor antagonist", which blocks biological responses mediated by IL-1. Recombinant human IL-1ra (rhIL-1ra, Kineret) was introduced in clinical trials involving patients with RA. Between 2001 to approximately 2002, rhIL-1 ra was approved by the US Food and Drug Administration and the European Agency for the Evaluation of Medicine Procedure. Unfortunately, 10,000 to 100,000-fold excess amounts of IL-1ra are needed to relieve disease because minimal IL-1 can induce complete biological responses, and the dosage of 100 to approximately 150mg/day in a RA patient is so big that it greatly influence patients' physical, psychological and economical situation. In this study, IL-1ra mutants were established by site-specific mutagenesis to improve its stability. The sites of mutagenesis included R6 K7-AA,R93 K94-AA and K97 R98-AA. IL-1ra and its mutants were expressed in E. coli BL21 (DE3) using pTIG-Trx expressing system with the induction of IPTG. The recombinant proteins were purified by Ni2+ chelate chromatography and Sephadex G75 gel filtration chromatography. The activity of mutants is as high as IL-1ra. We characterized the pharmacokinetic profile of IL-1ra and its mutants. The third mutant's half life is 2.26 times than wt IL-1ra. The study has provided some approaches and experience for further research to improve the metabolism stability of IL-1ra.
Animals ; Escherichia coli ; genetics ; metabolism ; Female ; Humans ; Interleukin 1 Receptor Antagonist Protein ; biosynthesis ; genetics ; pharmacokinetics ; Mutagenesis, Site-Directed ; methods ; Mutant Proteins ; biosynthesis ; pharmacokinetics ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacokinetics

OBJECTIVEAntigen-presenting cells such as monocytes and dendritic cells (DCs) stimulate T-cell proliferation and activation during adaptive immunity. This cellular interaction plays a role in the growth of atherosclerotic plaques. Tanshinone II A (TSN) had been shown to decrease the growth of atherosclerotic lesions. We therefore investigated the ability of TSN to inhibit human monocyte-derived DCs and their T-cellstimulatory capacity.

METHODSDCs derived from human monocytes cultured with recombinant human interleukin (IL)-4 and recombinant human granulocyte-macrophage colony-stimulating factor were co-cultured with TSN and lipopolysaccharide for 48 h. Phosphate-buffered saline was used as a negative control. Activation markers and the capacity of DCs for endocytosis were measured by flow cytometry, and proinflammatory cytokines were measured by enzyme-linked immunosorbent assays. DCs were co-cultured with lymphocytes to measure T-cell proliferation and IL-2 secretion by mixed lymphocyte reactions.

RESULTSTSN dose-dependently attenuated DC expression of costimulatory molecules (CD86), and decreased expression of major histocompatibility complex class II (human loukocyte antigen-DR) and adhesion molecules (CD54). Moreover, TSN reduced secretion of the proinflammatory cytokines IL-12 and IL-1 by human DCs, and restored the capacity for endocytosis. Finally, TSN-preincubated DCs showed a reduced capacity to stimulate T-cell proliferation and cytokine secretion.

CONCLUSIONSTSN inhibits DC maturation and decreases the expression of proinflammatory cytokines, while impairing their capacity to stimulate T-cell proliferation and cytokine secretion. These effects may contribute to the influence of TSN on the progression of atherosclerotic lesions.

Antigen-Presenting Cells ; drug effects ; Atherosclerosis ; immunology ; pathology ; B7-2 Antigen ; metabolism ; Cell Membrane ; drug effects ; metabolism ; Cytokines ; secretion ; Dendritic Cells ; drug effects ; immunology ; secretion ; Diterpenes, Abietane ; pharmacology ; Endocytosis ; drug effects ; Flow Cytometry ; Humans ; Immunity, Cellular ; drug effects ; Inflammation Mediators ; metabolism ; Lymphocyte Activation ; drug effects