Objective:To explore the effect of infection on autophagy-related proteins,Beclin-1 and LC3,expression in cerebral white matter in newborn rats.Methods: A total of 64 two-day-old Sprague-Dawley(SD) rats were randomly divided into control and experimental groups(n=32 each).At day 2 to 6 after birth,the rats in experimental group were intraperitoneally injected with 0.6 mg/kg of lipopolysaccharide(LPS) once a day to establish a white matter injury induced by infection in neonatal rats while the rats in control group were injected with equal amounts of normal saline.Rats were sacrificed to collect brain tissues at 12 hour,1,3,5 d after model establishment.HE staining was performed to observe the pathological changes.Changes in the expression of Beclin-1 and LC3 in rat white matter were examined by Western blot and RT-PCR.Results: Growth and development of rat in experimental group was slow,cerebral white matter lesions were obvious.Compared with the control group,the experimental group Beclin1 and LC3 protein and mRNA levels in the model after 12 h began to express,1 d reached the peak,and then decreased,each time points were higher than the control group(P<0.05).Conclusion: Early infection in neonatal rats can cause white matter damage;the expression of autophagy-related proteins,Beclin1 and LC3,showed that autophagy may be involved in the pathological process of white matter damage induced by infection.

Antineoplastic Agents/therapeutic use* ; Gastrointestinal Stromal Tumors/drug therapy* ; Humans ; Liver ; Liver Neoplasms/drug therapy* ; Lung ; Naphthyridines/therapeutic use* ; Urea/analogs & derivatives*

Animals ; Ginkgo biloba ; chemistry ; Lung ; blood supply ; Malondialdehyde ; metabolism ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; Superoxide Dismutase ; metabolism

Objective To investigate the efficacy of transplantation of induced bone marrow mesenchymal stem cells(MSCs)via various route for treatment of chronic liver injury.Methods MSCs were isolated and expanded by density gradient centrifugation combined with adherent culture.MSCs were induced to differentiate into hepatocyte-like cells in vitro.The markers of hepatocyte lineage were examined by RT-PCR and immunocytochemistry.The changes of ultramicrostructure of MSCs were observed by transmission electron microscope.One hundred and fourteen Wistar rats were treated with mixture of 40%CCl4 and peanut oil to establish chronic liver injury model and another 6 rats were served as control.Twelve rats in model group were died within 8 weeks,and the rest rats were divided into portal vein,spleen and tail vein transplantation groups.The rats were transplanted with DAPI-labelled MSCs which were induced for 14 days.The distribution of the DAPI-labelled cells were observed by fluorescence microscopy.The liver function and pathology were examined.Results AFP mRNA expression were detected at 7th and 14th day after induction,expressions of ALB mRNA,CK18 and hepatocyte antigen were found at 14th and 2lth day.The ultrastructure examination revealed that MSCs were enlarged with a lot of endoplasmic reticulum,mitochondrion,lysosome and glycogen granule.DAPI labelled cells were found in the spleen and liver transplantion groups.The pathological changes of liver were alleviated in all treated groups,especially in intraspleenic transplantion group(P＜0.05).Conclusions Induced bone marrow MSCs can ameliorate liver function of chronic hepatic injury after transplantation.The intraspleenic transplantation is better than others.

Background Full femtosecond laser refractive surgery has been generally applied,but the improvement of visual quality after surgery is a vital problem.It is well-known that modulation transfer function (MTF) is a major evaluation indicator.Objective To investigate the change of the MTF of operated eyes after the full femtosecond laser refractive surgery.Methods A retrospective series of cases study was adopted.Forty eyes of 23 patients with the spherical equivalent of-2.25 to-5.75 D who received full femtosecond laser refractive surgery were enrolled in Nanjing General Hospital of Nanjing Military Command from July to October,2012.The change of visual acuities was analyzed and compared among before operation and 1 day,1 week and 1 month after operation.MTF value in total aberration,low and high order aberrations under the 3.0 mm and 6.0 mm pupil size conditions were detected with i-Trace Visual Function Analyzer at above time points.The MTF values among various time points,between different pupil diameters and different spatial frequencies (5,10,15,20,25 c/d) were compared respectively.Results Compared with the preoperation,the visual acuity improved in postoperative 1 day,1 week and 1 month,showing a significant improvement among various time points (F =10.341,P =0.000).Corrected diopter was 3.795 D at average.MTF values under the 3 mm pupil diameter gradually elevated with the lapse of time after operation,without significant differences among different time points whatever total aberration,low and high order aberrations(all at P＞0.05).The MTF values under 6 mm pupil diameter of total aberration and low order aberration were significantly higher in postoperation than those in preoperation(all at P＜0.05),but no significant difference was seen in MTF of high order aberration among various time points (F =0.260,P =0.854).Compared with the MTF under the 3 mm pupil diameter,those of low and high order aberrations under the 6 mm pupil diameter considerably declined in preoperation and postoperative 1 month (preoperation:P =0.050,0.001 ; postoperation:P =0.012,0.001).MTF values under the 3 mm pupil diameter were not different at various spatial frequencies (all at P＞ 0.05).However,MTF values under the 6 mm pupil diameter were gradually increased from preoperation through postoperative 1 month at 10,15,20 and 25 c/d (P =0.044,0.043,0.024,0.014).Conclusions Full femtosecond laser refractive surgery can eliminate low aberration,increase the MTF and acuity,and therefore improve the visual quality.MTF is one of the key indicators assessing visual quality after full femtosecond laser refractive surgery.

Objective To investigate the effects of RNA interference(RNAi)targeting human epithelial growth receptor-2(HER-2)gene on the invasive and chemotactic ability of SKOV3 cells. Methods Glyeeraldehyde-3-phosphate dehydrogenase(GAPDH)was used as the positive control. Lipofectamine 2000 mediated transient transfection was conducted to transmit the siRNA into SKOV3 cells. Three pairs of specifically targeted(HER-2 siRNA Ⅰ,HER-2 siRNA Ⅱ,HER-2 siRNA Ⅲ)sequence were selected in the coding region of HER-2 mRNA.Transfection of HER-2 siRNA was conducted with lipofeetamine 2000 in ovarian carcinoma cell line SKOV3.The HER-2 gene expression was assessed by real- time PCR and western blot assays.Changes of invasive and chemotaetie capacity of SKOV3 cells were measured by polycarbonates coated with or without matrigal.Results Western blot results showed that the expression of GAPDH protein was decreased in specifically transfected cells and with the increase of siRNA dose,the expression of GAPDH protein was decreased.GAPDH protein gray value in control group, different doses(0.5,1.0,1.5,2.0 ?g)GAPDH siRNA interference groups were 0.6855?0.0259,0.5698 ?0.0275,0.4542?0.0296,0.3341?0.0178 and 0.1816+0.0180,respectively.There was a significant difference in each group(F=198.126,P

This study was carried out to evaluate the anti-inflammatory and free radical scavenging activities of flavans from flex centrochinensis S. Y. Hu in vitro and their structure-activity relationship. LPS-stimulated RAW 264.7 macrophage was used as inflammatory model. MTT assay for cell availability, Griess reaction for nitric oxide (NO) production, the content of TNF-alpha, IL-1beta, IL-6 and PGE, were detected with ELISA kits; DPPH, superoxide anion and hydroxyl free radicals scavenging activities were also investigated. According to the result, all flavans tested exhibited anti-inflammatory effect in different levels. Among them, compounds 1, 3, 4 and 6 showed potent anti-inflammatory effect through the inhibition of NO, TNF-alpha, IL-lp and IL-6, of which 1 was the most effective inhibitor, however, 2 and 5 were relatively weak or inactive. The order of free radical scavenging activities was similar to that of anti-inflammatory activities. Therefore, these results suggest that 3, 4 and 6, especially of 1, were,in part responsible for the anti-inflammatory and free radical scavenging activity of Ilex centrochinensis. Hydroxyl group at 4'-position of B-ring plays an important role in the anti-inflammatory and free radical scavenging capacities.
Animals ; Anti-Inflammatory Agents, Non-Steroidal ; chemistry ; pharmacology ; Cell Line ; Cyclooxygenase 2 ; immunology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Flavanones ; chemistry ; pharmacology ; Free Radical Scavengers ; chemistry ; pharmacology ; Ilex ; chemistry ; Interleukin-6 ; immunology ; Macrophages ; drug effects ; immunology ; Mice ; Nitric Oxide ; immunology ; Tumor Necrosis Factor-alpha ; immunology

In order to enhance the specificity of TNF-α monoclonal antibody to inflamed site, a bispecific antibody BsDb that targets TNF-α and the extra-domain B (ED-B) of fibronectin (FN) was constructed by covalently linking the anti-TNF-α single chain Fv antibody (TNF-scFv) and the anti-ED-B scFv L19 via a flexible peptide linker deriving from human serum albumin (HSA). ED-B is an antigen specifically expressed at the inflamed site. BsDb is expressed in E. coli, identified by immunoblot, and purified with affinity chromatography. This was followed by further examination of its bioactivities and pharmacokinetics. We demonstrated that BsDb retained the immunoreactivity of its original antibodies as it could simultaneously bind to TNF-α and ED-B and neutralize the biological action of TNF-α. In the collagen-induced arthritis mice model, BsDb selectively accumulate in the inflamed joint with a maximal uptake of (12.2 ± 1.50)% ID/g in a single inflamed paw and retain in the inflamed paw for at least 72 h. In contrast, BsDb showed a short serum half-life of (0.50 ± 0.05) h and a rapid clearance from normal tissues. The findings reported herein indicate that BsDb has good specificity to the inflamed site and low toxicity to normal tissues. BsDb is therefore likely to have greater clinical applications in the treatment of rheumatoid arthritis and other autoimmune diseases. This laid a stable basis for its preclinical study.
Animals ; Antibodies, Bispecific ; chemistry ; Antibodies, Monoclonal ; chemistry ; Arthritis, Experimental ; Escherichia coli ; Fibronectins ; chemistry ; Half-Life ; Humans ; Mice ; Single-Chain Antibodies ; chemistry ; Tumor Necrosis Factor-alpha ; chemistry

OBJECTIVETo explore the underwater decompression schedule for 100 m Trimix conventional diving operations and evaluate its safety through a simulated rabbits Trimix conventional diving.

METHODSAccording to the Haldane theory, the assumed time units, the classification of tissue compartments, the nitrogen super-saturation safety coefficient and the selection of methods used for the calculation of the simulated 100 m Trimix conventional diving schedule were properly selected, and the calculating method for the dive decompression schedule was thus firmly established. In our experiments, five tissue compartments were selected during the calculation of decompression schedule: 5 min, 10 min, 20 min, 40 min and 75 min, and the nitrogen super-saturation safety coefficient was calculated by 1.6. Eight New Zealand rabbits were performed a simulated 100 m Trimix dive program which was established according to the Haldane theory, and eight rabbits for intact group. The tissues wet/dry ratio and ethology were detected and observed before and after the simulated diving to evaluate the safety of decompression schedule.

RESULTSBy using the developed underwater decompression schedule, abnormal ethology changes in rabbits could not be observed after compression and decompression to the surface; and the tissues wet/dry ratio of simulated diving rabbits had no significant changes compared with the intact group (P > 0.05).

CONCLUSIONThe decompression schedule calculated by Haldane theory seemed to be safe and reliable, the diving breathing gas concentration did not cause oxygen toxicity and nitrogen narcosis among the dive rabbits, and dive efficiency was greatly improved by using enriched oxygen gas in UPTD safety range during decompression.

Objective To explore the expression of V-ATPase in colon cancer and its clinical significance. Methods Detecting the expression of V-ATPase mRNA in 20 paired of colon tumor tissues and normal tissues by using reverse transcription-polymerase chain reaction ( RT-PCR) and real-time quantitative polymerase chain reaction( Real-time PCR) , and testing the expression of V-ATPase protein by immu-nohistochemistry of EnVinsion. Results The expression of V-ATPase mRNA in tumor tissues and its paired normal tissues were (5. 37 ± 0. 44) and (2. 03 ± 0. 35)(P<0. 01). The positive immunohistochemistry of V-ATPase in tumor tissues and its paired normal tissues were 69. 1%(47/68) and 5. 8%(4/68) respectively, and the positive expression were primarily in cytoplasm and cytomembrane. Overexpression of V-ATPase was associated with tumor stage (P<0. 05), lymph node metastasis (P=0. 044), distant metastasis (P=0. 049), vessel in-vasion (P=0. 044) and differentiation (P<0. 001). Conclusion Overexpression of V-ATPase plays a significant role in the carcinogene-sis and the progression of colon cancer, which might be an important postoperative therapeutic target.