Adult ; Case-Control Studies ; Glutathione Peroxidase ; blood ; Humans ; Middle Aged ; Occupational Exposure ; Oxidation-Reduction ; Silicosis ; blood ; Superoxide Dismutase ; blood

Objective To investigate the effect of fusidic acid cream on inflammatory reaction caused by skin barrier damage.Methods Eight male SKH-1 hairless mice were included in this study.The back of each of these mice were equally divided into six regions measuring 1 cm × 2 cm in size,which were then assigned into six groups:blank control group remaining untreated,barrier-impaired group,barrier-impaired and fusidic acid-treated group,barrier-impaired and vehicle-treated group,barrier-unimpaired and fusidic acid-treated group,barrierunimpaired and vehicle-treated group.Stratum corneum was removed by adhesive tape stripping to establish an animal model of acute skin barrier damage in the corresponding skin regions of these mice,and fusidic acid cream or vehicle was topically applied to the corresponding regions once.Twelve hours later,skin surface swab samples were collected from the back of these mice followed by bacterial culture and colony counting.Mice were then sacrificed,and skin tissue specimens were resected from these mice,and subjected to real-time fluorescence-based quantitative PCR for the measurement of the mRNA expressions of myeloid differentiation factor 88 (MyD88),interleukin-1α (IL-1α),IL-6,epidermal antibacterial peptides S100a8 and S100a9.Statistical analysis was carried out by repeated-measures analysis of variance (ANOVA) and least significant difference (LSD) test.Results The mRNA expressions of MyD88,IL-1α,IL-6,S100a8 and S100a9 were all significantly higher in the barrier-impaired group than in the blank control group (all P ＜ 0.05).Specifically,the mRNA expression level of MyD88 in the barrier-impaired group was 8 times that in the blank control group (8.3 ± 3.0 vs.0.8 ± 0.4).Compared with the barrier-impaired group,the barrier-impaired and fusidic acid-treated group showed a significant decrease in the mRNA expressions of IL-1α (2.8 ± 0.3 vs.20.1 ± 10.0,F =47.11,P ＜ 0.01),IL-6 (1.6 ± 2.3 vs.9.4 ± 4.0,F =16.18,P＜ 0.01),S100a8 (1.5 ± 1.4 vs.5.0 ± 1.6,F=59.71,P＜ 0.05) and S100a9 (1.2 ± 0.7 vs.3.4 ± 1.6,F=21.94,P ＜ 0.05).Conlusions Fusidic acid cream could attenuate the inflammatory reaction caused by acute skin barrier damage,which might partly explain its action mechanism in the treatment of inflammatory skin diseases.

Objective To evaluate the inhibitory effect of butyl flufenamate (BT) on ultraviolet (UV)-induced acute skin phototoxic reaction,and to investigate its possible mechanisms.Methods Eight SKH-1 hairless mice were included in this study.The back of each SKH-1 hairless mouse was divided into six regions,which were then randomly classified into six groups:blank group receiving no treatment,UV group receiving UV radiation only,BT + UV group and vehicle + UV group topically treated with BT ointment and vehicle respectively followed by UV radiation,UV + BT group and UV + vehicle group topically treated with BT ointment and vehicle respectively after UV radiation.Skin samples were obtained from these mice at 24 hours after treatment.Subsequently,hematoxylin-eosin (HE) staining was performed,real-time PCR was carried out to detect mRNA expressions of caspase-3,p53,COX-2,PGER1,interleukin (IL)-1β,IL-6,and an immunofluorescence assay was conducted to observe the expression of caspase-3.Statistical analysis was carried out by repeated-measures analysis of variance (ANOVA).Results Compared with the UV group,both BT + UV group and UV + BT group showed a decrease in the degree of skin edema and number of apoptotic cells at 24 hours after UV radiation.Real-time PCR showed that the mRNA expressions of caspase-3,p53,COX-2,PGER1,IL-l β and IL-6 were significantly higher in the UV group than in the blank group (all P ＜ 0.05),but significantly lower in the BT + UV group than in the UV group (all P ＜ 0.05),and only the expressions of caspase-3 and p53 mRNAs were significantly decreased in the UV + BT group compared with the UV group (both P ＜ 0.05).The immunofluorescence assay revealed that the expression of caspase-3 increased in the UV group compared with the blank group,but decreased in both BT + UV group and UV + BT group compared with the UV group.Conclusion BT could partially inhibit UV-induced acute skin phototoxicity in SKH-1 hairless mice.

We wished to study the intracellular transport of adenoviruses. We constructed a novel recombinant adenovirus in which the structural protein IX was labeled with a mini-singlet oxygen generator (miniSOG). The miniSOG gene was synthesized by overlapping extension polymerase chain reaction (PCR), cloned to the pcDNA3 vector, and expressed in 293 cells. Activation of miniSOG generated sufficient numbers of singlet oxygen molecules to catalyze polymerization of diaminobenzidine into an osmiophilic reaction product resolvable by transmission electron microscopy (TEM). To construct miniSOG-labelled recombinant adenoviruses, the miniSOG gene was subcloned downstream of the IX gene in a pShuttle plasmid. Adenoviral plasmid pAd5-IXSOG was generated by homologous recombination of the modified shuttle plasmid (pShuttle-IXSOG) with the backbone plasmid (pAdeasy-1) in the BJ5183 strain of Eschericia coli. Adenovirus HAdV-5-IXSOG was rescued by transfection of 293 cells with the linearized pAd5-IXSOG. After propagation, virions were purified using the CsC1 ultracentrifugation method. Finally, HAdV-5-IXSOG in 2.0 mL with a particle titer of 6 x 1011 vp/mL was obtained. Morphology of HAdV-5-IXSOG was verified by TEM. Fusion of IX with the miniSOG gene was confirmed by PCR. In conclusion, miniSOG-labeled recombinant adenoviruses were constructed, which could be valuable tools for virus tracking by TEM.
Adenoviruses, Human ; chemistry ; genetics ; metabolism ; Arabidopsis Proteins ; chemistry ; genetics ; metabolism ; Flavoproteins ; chemistry ; genetics ; metabolism ; Humans ; Phototropins ; chemistry ; genetics ; metabolism ; Singlet Oxygen ; chemistry ; Staining and Labeling ; Transfection

Human adenovirus type 41 (HAdV-41) is considered to be a "fastidious adenovirus". E1-deleted HAdV-41 cannot be rescued or amplified in 293 cells. To propagate recombinant HAdV-41 in 293 cells, the backbone plasmid pAdbone41 was reconstructed. That is, the E3 coding sequence of HAdV-41 was deleted and replaced with the HAdV-5 E4orf6 gene; and the E1A enhancer of HAdV-5 was inserted upstream of the E4 promoter of HAdV-41. Novel adenoviral plasmid pAd41E4EE-GFP was generated by homologous recombination of the shuttle plasmid pSh41-GFP with the modified backbone plasmid in the Escherichia coli BJ5183 strain. Adenovirus HAdV-41-E4EE-GFP was rescued by transfecting 293 cells with linearized pAd41E4EE-GFP. After seven rounds of propagation, viruses were purified by the CsCl ultracentrifugation method. HAdV-41-E4EE-GFP in 1.0 ml with a particle titer of 8 x 10(10) vp/mL was obtained which had a particle-to-infectious ratio of 50 : 1. The genome of HAdV-41-E4EE-GFP was confirmed by restriction analyses and polymerase chain reaction. These results showed that a novel HAdV-41 vector system was established in which recombinant HAdV-41 could be constructed and packaged in 293 cells.
Adenoviruses, Human ; genetics ; growth & development ; physiology ; Cell Line ; Genetic Vectors ; genetics ; physiology ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Plasmids ; genetics ; metabolism ; Recombination, Genetic ; Transfection ; Virus Assembly

As a novel targeting drug delivery system, acid-sensitive micelles have many advantages, such as increasing solubility of lipophilic drugs, acid-sensitive release, high drug loading, etc. They can load drugs though non-covalent encapsulation and covalent conjugation methods. In tumor tissues, drugs are released quickly from the depolymerized micelles with lipophilic copolymer protonation or lypohydrophilic copolymer hydrolysis and covalent conjugated drugs are released when the acid-sensitive covalent linkage breaks. This review mainly advances acid-sensitive micelles for the tumor targeting drug delivery systems from drug-loaded methods and release mechanisms.
Animals ; Antibiotics, Antineoplastic ; administration & dosage ; pharmacology ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Cell Line, Tumor ; Doxorubicin ; administration & dosage ; pharmacology ; Drug Carriers ; chemistry ; Drug Delivery Systems ; Humans ; Hydrogen-Ion Concentration ; Lactates ; chemistry ; Micelles ; Paclitaxel ; administration & dosage ; pharmacology ; Polyethylene Glycols ; chemistry ; Polymers

Adult ; Aorta, Thoracic ; abnormalities ; surgery ; Heart Arrest, Induced ; Humans ; Male

Objective:To investigate the role of transcription factor SP1 decoy oligodeoxynucleotides(ODN) on expression of ? Gal in SV-40-PED cells.Methods:Immortalized porcine aortic endothelial cells of the PED line were cultured and transfected with ?1,3galactosyltransferase(?1,3GT) specific decoy ODN.Cells transfected with mismatch ODN was used as negative controls.Twenty-six hours later the cells were collected.The expression of ? Gal was determined with fluorescence microscope and Western blot.The expression of ?1,3GT mRNA was examined by RT-PCR.Results:Fluorescence microscopy observed the decreased fluorescence of ? Gal after decoy ODN transfection.Western blot showed that the average absorbance of the PED cells transfected with decoy ODNs was(48.2?0.9).It is 52.6% of the mock group(P0.05).Conclusion:?1,3GT gene reduce actually occurs following transfection of decoy ODN.Porcine endothelial cells can be the targets of decoy ODN.

Objective To evaluate the efficiency of transcatheter closure of secundum artrial septal defect (ASD) using Amplatzer occluder device in 5 years.Methods There were 18 children with ASD.Each ASD was occluded using Amplatzer occluder device through the percutaneous procedure.The closure procedure was guided by fluoroscopy and transthoracic echocardiography.All patients were followed up by physical examination,color flow echocardiography,chest radition,electrocardiography at 48 h,1month,3 month,6 month,one year,two years,three years,four years and five years.Results Cardiac murmur disappeared or lightened.Arrhythmia and right heart volume in all patients were improved after the procedure.A case revealed a trival shunt of short duration and vanished after one month color flow echo.A case has “spring′s beat” in heart when she goes to bed in three years after the procedure.Another patient had serious headach after closure and recoved after heparin therapy.Conclusions Transcatheter closure of secundum ASD using amplatzer occluder device is safe and efficient by follow-up.In order to prevent cerebral embolism,heparin should be injected in vein in two days.

Objective To investigate the efficacy of spectral entropy measurement in reflection of depth of anesthesia and noxious stimulation. Methods Forty-five patients of ASAⅠorⅡ were randomly divided into three groups(n=15).Group A,B and C received fentanyl 1,3 and 5 ?g/kg,respectively,3 min before target controlled infusion(TCI) of propofol.Intubation was performed when the effect-site concentration(CE)reached 3.5 ?g/mL,which was maintained until 5 min after incision.Response entropy(RE),State entropy(SE) as well as heart rate(HR),mean arterial pressure(MAP) were measured at the time points of before fentanyl and 2,3 min after fentanyl,every CE of propofol steps,before intubation,immediately and 1,3,5 min after intubation,before skin incision,and 0.5,1,3,5 min after skin incision,respectively. Results Three minutes after receiving fentanyl,the values of RE and SE in the three groups decreased significantly in a dose-dependent manner,and increased obviously at the same degree during intubation and after skin incision.The values recovered to the level before stress stimulation 1 min after intubation and 5 min after skin incision.There were no differences in the fluctuation of RE and SE among the three groups when the CE of propofol reached 1.0 ?g/mL.Conclusion Spectral entropy may effectively reflect the depth of anesthesia,but not analgesia during anesthesia.