Baculovirus has many advantages as vectors for gene transfer. We demonstrated that recombinant baculovirus vectors expressing p35 (Ac-CMV-p35) and eGFP (Ac-CMV-GFP) could be transduced into human kidney 293 cells efficiently. The level of transgene expression was viral dose dependent and high-level expression of the target gene could be achieved under the heterogonous promoter. MTT assay suggested that both Ac-CMV-p35 and Ac-CMV-GFP did not have cytotoxic effect on human embryo kidney 293 cells. Cell growth curve showed the Ac-CMV-p35 and Ac- CMV-GFP transduced and non-transduced cells had similar proliferation rate, so baculovirus-mediated p35expression had no adverse effect on cell proliferation. In addition, baculovirus-mediated p35 gene expression protected human embryo kidney 293 cells against apoptosis induced by various apoptosis inducers such as Actinomycin D, UV or serum-free media. These results suggested that the baculovirus vector mediated p35 gene expression was functional and it could be widely used in molecular research and even gene therapy.

Aged ; Female ; Humans ; Infant ; Lung Diseases ; diagnostic imaging ; Male ; Pulmonary Artery ; diagnostic imaging ; Tomography, X-Ray Computed

Objective To evaluate the protective effect of polysaccharide nucleic acid fraction of bacillus Calmette-Guerin (BCG-PSN) against atopic dermatitis (AD) in Nc/Nga mice,and to explore its possible mechanism.Methods Sixteen Nc/Nga mice were classified into normal control group (n =4),low-concentration BCG-PSN group (n=5) and high-concentration BCG-PSN group (n =7) to be subcutaneously injected with sodium chloride physiological solution,BCG-PSN of 0.1 mg/kg and 0.5 mg/kg respectively,at 1,8,15 and 22 days of age.Dinitrochlorobenzene (DNCB) was repeatedly and topically applied to these Nc/Nga mice to induce AD-like lesions at 49 days of age.The preventive effect of BCG-PSN against AD was evaluated by dermatitis scores,scratching frequency,histopathological manifestations and immunological parameters (including IgE,i nterleukin (IL)-4 and-12,and interferon (IFN)-γ).Results Repeated injection of BCG-PSN within 4 weeks after birth significantly decreased the severity of DNCB-induced AD-like lesions,dermatitis scores and scratching behavior in Nc/Nga mice.There was no statistical difference in scratching frequency between the high-and low-concentration BCG-PSN groups.BCG-PSN treatment reduced the plasma level of IgE in Nc/Nga mice in a dose-dependent manner.BCG-PSN at 0.5 mg/kg increased the number of cells secreting IFN-γ in skin lesions of mice.Both doses of BCG-PSN down-regulated IL-4 level,but up-regulated IL-12 level in the culture supernatant of spleen mononuclear cells from mice.Conclusion Early injection of BCG-PSN could protect Nc/Nga mice against dermatitis by promoting the proliferation of IFN-γ-secreting cells,increasing the synthesis of IL-12,and reducing the levels of IL-4 and IgE.

Objective To observe the perioperative management of cardiac surgery and extracorporeal circulation method in patients with glucose-6-phosphate dehydrogenase deficiency(G6PD).Methods Ten patients with G6PD deficiency underwent uneventful cardiac surgery procedures between January 2005 and December 2010.Twenty patients who had non-G6PD deficiency were as a control group,the selected conditions were the same gender,age,body mass,the risk of heart disease surgery.The preoperative management in patients with G6PD deficiency mainly focused on avoiding the drugs implicated in haemolysis,reducing the surgical stress,using moderate hypothermia extracorporeal circulation and enhancing blood conservation.Observed indicators included the assisted ventilation time,urine volume,the drainage volume of chest tube,the amount transfusion of red blood cells and plasma,the level of hemoglobin and serum total bilirubin in the 2nd day after surgery,ICU stay.Results Compared with the control group,patients with G6PD deficiency had no significant difference in duration of ventilation after the operation,drainage,urine,Hgb,bilirubin levels,and blood transfusion[(9.3 ± 4.5)h vs (8.6 ± 5.7)h,(2100 ±670)ml vs (1950 ± 490) ml,(253 ± 146)ml vs (260 ± 120)ml,(1.3 ± 1.0)U vs (1.8 ± 1.2)U,(96 ± 25)g/L vs (99 ± 12)g/L,and (24 ± 8)μmol/L vs (27 ± 1 l)μmol/L,t =0.978,2.032,1.257,0.891,2.182,2.271,and 1.329,all P ＞ 0.05].The duration of ICU discharge was significantly longer in the glucose-6-phosphate dehydrogenase deficient group[ (2.6 ± 0.6)d vs (1.8 ± 1.5)d,t =2.704,P ＜ 0.05].Conclusions Cardiac surgery can be performed safely in patients with G6PD deficiency with enhanced perioperative management.

Objective To investigate the development of ultrastructure in grafts from a xenograft model of human hemangioma on nude mice.Methods Human infantile hemangioma tissue was obtained in surgery and planted into subcutaneous layer of nude mice in small pieces.The grafts were harvested on the 2~(nd),5~(th),15~(th),30~(th),45~(th) day and then at 2~(nd),3~(rd),4~(th),6~(th) month for transmission electron microscopic view.Results In early stage after transplantation,signs of degeneration and necrosis were more evident.On the 30~(th) day,the number of endothelial cells increased and some of the endothelial cells were larger in size with high nuclear to plasm ratio.At the 2~(nd) month,the number of endothelial cells increased,while some of these cells turned flat.Later,high endothelial cells decreased in number and collapse and pyknosis of plasm,formation of secondary lysosome,margination of chromatin,fragmentation of nucleus were evident with surrounding deposition of adipose and collagen.Conclusions With xenotransplantation of human infantile hemangioma tissue onto nude mice,the grafts may survive ischemia and proliferate,and then gradually involute.The development resembles a natural course of human infantile hemangioma.

Objective To investigate the meaning of expression of apoptosis related molecules NFKBp65 and p53 and GPX1-mRNA in patients with Keshan disease(KSD).Methods Sixteen chronic Keshan Disease patients were enrolled in KSD group according to electrocardiogram,chest X ray film and clinical examinations on 15,September in 2009,and 23 healthy people were included in control group from physical examination taken in The Second Affiliated Hospital of Xi'an Jiaotong University.Fresh blood(5 ml)was collected from antecubital vein of all subjects in the fasting state.Total mRNA and protein of blood sample were isolated using Trizol.GPX Assay Kit was used to detect GPX enzyme activity,and GPX1-mRNA expression was determined by SYBR Real-Time PCR.Meanwhile,expression of apoptosis related molecules NFKBp65 and p53 were determined by Western blot.Results GPX enzyme activity decreased significantly in KSD group[(108.61±14.10)U]compared with control group[(122.78±11.89)U,t=2.874,P<0.05],GPX1-mRNA level of KSD group(0.553±0.299)notably KSD group(0.802±0.057)compared with control group[(1.065±0.355),t=6.829,P<0.01].p53 increased in KSD group(1.604±0.191)compared with control group[(1.137±0.186),t=3.033,P<0.05].Conclusiom Decreased GPX1-mRNA expression may result in lower GPX enzyme activity of patients with KSD.Thus oxidative damage increases and cadioeyte apoptosis is activated by activating apoptosis signal pathway.

Carcinoma ; metabolism ; Cell Differentiation ; genetics ; Gallbladder Neoplasms ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Kangai-1 Protein ; genetics ; metabolism ; NM23 Nucleoside Diphosphate Kinases ; genetics ; metabolism

Humans ; Job Satisfaction ; Occupational Health ; Reward ; Stress, Psychological ; physiopathology ; Surveys and Questionnaires ; Workload

Isolation and purification of chemical constituents from solid culture of endophyte Chaetomium globosum in Imperata cylindrical was performed through silica gel column chromatography, gel filtration over Sephadex LH-20 and preparative HPLC. Nine compounds were obtained and their structures were determined as chaetoglobosin F(1), chaetoglobosin Fex(2), chaetoglobosin E(3) cytoglobosin A(4), penochalasin C(S), isochaetoglobosin D (6), N-benzoylphenylalaninyl-N-benzoyphenylalaninate(7), uracil(8) and 5-methyluracil(9), respectively, based on HR-MS and NMR data and comparison with literatures. Compound 7 was isolated from Chaeeomium sp. for the first time. In vitro cytotoxicity of compounds was evaluated using MTT mothed and 1,3,4 and 5 showed inhibition activity to the human cervical carcinoma cell HeLa with IC50 values of 99.43, 23.77, 97.92, 86.25 micromol x L(-1), while positive cotolocisnin Ad apno1ch alse IC50 24.33 micromol x L(-1).
Biological Factors ; chemistry ; pharmacology ; Cell Line ; Cell Survival ; drug effects ; Chaetomium ; chemistry ; Endophytes ; chemistry ; Humans ; Molecular Structure ; Poaceae ; microbiology ; Spectrometry, Mass, Electrospray Ionization