Adult ; Aged ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; diagnosis ; epidemiology

Animals ; Hydroxy Acids ; toxicity ; Lacquer ; toxicity ; Mice ; Mice, Inbred ICR ; Superoxide Dismutase ; metabolism

Objective: To establish the plant tissue culture system of Cistanche tubulosa, and determine the effect of drought stress on accumulation of two respective phenylethanoid glycosides in it. Methods The major chemical constituents of C. tubulosa by tissue culture were analyzed by HPLC-UV and HR-MS. The cell growth curves were also determined. In addition, the effects of drought stress on the phenylethanoid glycosides (echinacoside and acteoside) content in the tissue culture system of C. tubulosa were also studied by using NaCl, mannitol and PEG6000 as osmotic regulators, respectively. Results:Chemical constituents analyses revealed that callus and suspension cultures of C. tubulosa could produce the respective phenylethanoid glycosides of echinacoside and acteoside as in wild plant; Cell growth curves indicated that 30 d were the optimum culture period of callus culture; The cell growth rate and the accumulation of echinacoside and acteoside were mostly inhibited when the callus cells were under drought stress induced by NaCl or mannitol. Meanwhile, the accumulation of echinacoside and acteoside in cell suspension culture of C. tubulosa could be effectively enhanced by treatment with PEG6000. The maximum biomass of echinacoside and acteoside could reach to (1.07 ± 0.10) g/L and (0.12 ± 0.01) g/L 15 d after induction, respectively. And their contents were 20.94% and 2.27% separately based on the cell dry weight (DW) after 15 d of treatment with 6% PEG6000, which were 1.29 and 1.19 fold higher than the control group. Conclusion:Drought stress induced by PEG6000 could effectively enhance the accumulation of echinacoside and acteoside in cell suspension culture of C. tubulosa.

In this study the effects of bone marrow stromal cells conditioned medium on the expansion of mature megakaryocytes and colony forming unit-megakaryocyte (CFU-Meg) in vitro were investigated. The serum-free bone marrow fibroblast conditioned medium (F-CM), bone marrow endothelial cell conditioned medium (E-CM) and bone marrow macrophage conditioned medium (M-CM) were collected and ultrafiltrated by using Centriprep-10. F-CM, E-CM, M-CM, the retentate (>10 kDa F-CM, >10 kDa E-CM and >10 kDa M-CM) contained substances whose molecular weight was more than 10 kDa and the filtrate (<10 kDa F-CM, <10 kDa E-CM and <10 kDa M-CM) contained substances whose molecular weight was less than 10 kDa were added in liquid culture system respectively. The results showed that F-CM, >10 kDa F-CM and E-CM, >10 kDa E-CM significantly promoted the expansion of mature megakaryocytes in liquid culture system, the percentage of mature megakaryocytes compared with 0 h control were (287.33-/+16.77)%, (236.67-/+39.72)%, (141.21-/+17.47)% and (179.03-/+30.98)%. But <10 kDa F-CM, <10 kDa E-CM, M-CM, >10 kDa M-CM and <10 kDa M-CM had no positive effects on the expansion of mature megakaryocytes. The effects of F-CM, E-CM or M-CM on the expansion of CFU-Meg were also investigated. The results indicated that F-CM and E-CM promoted the expansion of CFU-Meg in liquid culture system. M-CM had no positive effect on the expansion of CFU-Meg. the percentage of CFU-Meg compared with 0 h control were (168.18-/+30.24)%, (215.17-/+17.4)% and (85.0-/+7.0)%. The results of reverse transcription-polymerase chain reaction (RT-PCR ) showed that transforming growth factor -beta1 (TGF-beta1) mRNA was expressed in bone marrow endothelial cells and was not expressed in bone marrow fibroblasts. Thrombopoietin (TPO) mRNA was expressed in bone marrow fibroblasts and was not expressed in bone marrow endothelial cells. These results suggest that F-CM, >10kDa F-CM and E-CM, >10kDa E-CM significantly promoted the expansion of mature megakaryocytes and CFU-Meg in liquid culture system. The effect of F-CM on the expansion of mature megakaryocytes is better than that of E-CM.
Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cells, Cultured ; Colony-Forming Units Assay ; Culture Media, Conditioned ; pharmacology ; Female ; Male ; Megakaryocyte Progenitor Cells ; cytology ; Megakaryocytes ; cytology ; Mice ; Stromal Cells ; cytology

Objective To discuss the psychology and behavior of elderly hypoglycemia patients with symptomatic carotid artery stenosis, and compare the differences in psychology and behavior between groups.Methods Measurement and assessment with fall efficacy scale on psychology towards prevention of fall among 408 elderly hypoglycemia patients (＞ 60 years old) with symptomatic carotid artery stenosis were taken; They were divided for hypoglycemia history, carotid artery stenosis and ischemia sympton, and fall history; SPSS 17.0software was applied to do statistical analysis. Results There is significant difference on fall efficacy scale score between groups with and without hypoglycemia, carotid artery stenosis and ischemia sympton, and fall history (P ＜0. 0 1). Conclusions Subdividing symptomatic groups, applying preventive intervention procedure on prevention of fall, education on psychology and behavior towards prevention of fall are the effective psychological intervention measures on prevention of fall.

OBJECTIVETo establish a mouse model of asthmatic airway remodeling and investigate the effects of 1,25-(OH)2D3 on airway structure and T cell immunoglobulin mucin protein-4 (TIM-4) expression in asthmatic mice.

METHODSThirty female mice (BALB/c strain) were randomly divided into control, asthma and 1,25-(OH)2D3 intervention groups. An asthmatic mouse model was induced using ovalbumin. Lung tissue of the mice was collected, mRNA expression of TIM-4 was evaluated by RT-PCR and airway remodeling and protein expression of TIM-4 were observed by hematoxylineosin staining and immunohistochemistry.

RESULTSTypical airway remodeling was found in the asthma group, and TIM-4 expression in this group was significantly higher than in the control group (105±9 vs 42±5; P<0.05). Compared with the asthma group, the 1,25-(OH)2D3 intervention group showed improvement in airway remodeling and a decrease in TIM-4 expression (78±6) (P<0.05).

CONCLUSIONSTIM-4 may be involved in the airway remodeling of mice. As a new type of immunoregulator, 1,25-(OH)2D3 can downregulate expression of TIM-4 in the lungs and improve airway remodeling in asthmatic mice.

Airway Remodeling ; Animals ; Asthma ; metabolism ; Calcitriol ; pharmacology ; Female ; Gene Expression Regulation ; drug effects ; Lung ; metabolism ; Membrane Proteins ; analysis ; genetics ; physiology ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; analysis

OBJECTIVETo construct recombinant lentiviral vectors for gene delivery of the glial cell line-derived neurotrophic factor (GDNF), and evaluate the neuroprotective effect of GDNF on lactacystin-damaged PC12 cells by transfecting it into bone marrow stromal cells (BMSCs).

METHODSpLenti6/V5-GDNF plasmid was set up by double restriction enzyme digestion and ligation, and then the plasmid was transformed into Top10 cells. Purified pLenti6/V5-GDNF plasmids from the positive clones and the packaging mixture were cotransfected to the 293FT packaging cell line by Lipofectamine2000 to produce lentivirus, then the concentrated virus was transduced to BMSCs. Overexpression of GDNF in BMSCs was tested by RT-PCR, ELISA and immunocytochemistry, and its neuroprotection for lactacystin-damaged PC12 cells was evaluated by MTT assay.

RESULTSVirus stock of GDNF was harvested with the titer of 5.6 x 100,000 TU/mL. After transduction, GDNF-BMSCs successfully secreted GDNF to supernatant with higher concentration (800 pg/mL) than BMSCs did (less than 100 pg/mL). The supernatant of GDNF-BMSCs could significantly alleviate the damage of PC12 cells induced by lactacystin (10 micromol/L).

CONCLUSIONOverexpression of lentivirus-mediated GDNF in the BMSCs cells can effectively protect PC12 cells from the injury by the proteasome inhibitor.

Acetylcysteine ; analogs & derivatives ; pharmacology ; Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cells, Cultured ; Culture Media, Conditioned ; metabolism ; DNA, Recombinant ; Genetic Therapy ; methods ; Genetic Vectors ; Glial Cell Line-Derived Neurotrophic Factor ; genetics ; metabolism ; Lentivirus ; genetics ; Neurons ; drug effects ; Neuroprotective Agents ; metabolism ; PC12 Cells ; drug effects ; Plasmids ; genetics ; Protease Inhibitors ; pharmacology ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Stromal Cells ; Transduction, Genetic ; methods

Adolescent ; Adult ; Dust ; analysis ; Female ; Health Status ; Humans ; Industry ; Male ; Middle Aged ; Occupational Exposure ; analysis ; Quartz ; Young Adult

The aim of this study was to obtain the capsid protein VP2 of human bocavirus (HBoV) identified in Beijing recently and construct virus-like particles (VLPs) in insect cells for further study of this virus. The full-length VP2 gene of HBoV from BJ3722 was inserted into the baculovirus expression transfer vector (pFastBac 1) to obtain the recombinant Bacmid, and generation of recombinant baculoviruses was followed by transfection of the recombinant Bacmid into insect cells. Then the recombinant VP2 protein was recognized by SDS-PAGE using Coomassie-blue staining and Western blot using hyper-immune serum against VP2 of HBoV from rabbit. The recombinant baculoviruses were harvested and amplified to gain large amounts of viruses with high titers to infect insect cells at a multiplicity of infection (MOI) of 0. 5. After 7-10 days or 4-5 days of the infection, the supernatants of culture or the cell lysates treated with lysing solution were harvested, and ultracentrifuged twice through 40% sucrose cushion to obtain purified VLPs, which were followed by Western blot and IFA for VLPs' composition and specificity analysis, by electron microscopy for VLPs' morphologic structure. The recombinant VP2 protein with molecular weight of approximately 61 kD expressed in recombinant baculoviruses was recognized by SDS-PAGE using Coomassie-blue staining and Western blot. The presence of VP2 on VLPs was demonstrated by Western blot and IFA from samples collected during the purification of VLPs from the supernatants of culture or the cell lysates, and the expression of VP2 in insect cells led to the formation of VLPs which formed the typical icosahedral appearance of parvoviruses with a diameter of approximately 20 nm. In conclusion, the recombinant baculoviruses were constructed, the HBoV VP2 protein was expressed in insect cells with high specific antigenicity and VLPs was formed successfully.
Animals ; Blotting, Western ; Capsid Proteins ; genetics ; metabolism ; Cell Line ; Electrophoresis, Polyacrylamide Gel ; Fluorescent Antibody Technique, Indirect ; Human bocavirus ; genetics ; metabolism ; Microscopy, Electron, Transmission ; Polymerase Chain Reaction ; Spodoptera ; Virion ; genetics ; metabolism ; ultrastructure

Objective To observe the alteration of glomerular anionic sites and renal heparanase (Hpa)expression in respiratory syncytial virus(RSV) nephropathy in rats,and to investigate the role of Hpa in the pathogenesis of proteinuria in RSV nephropathy.Methods Twenty-five Sprague-Dawley (SD) rats were inoculated with 6 x 106plaque forming units(PFU) RSV and were sacrificed on days 4,8,14 and 28 post-inoculation(RSV4,RSV8,RSV14 and RSV28 group).Five normal SD rats served as normal control.The proteinuria and serum parameters were measured.Glomerular anionic sites were measured by histochemical electron microscopy.The expression of Hpa in kidney was determined by immunohistochemical staining.The relationship between the expression level of Hpa and the quantity of 24-hour urine protein was studied.Results After inoculation,the proteinuria increased,especially in RSV14 group.The 24-hour urine protein of RSV14 group,RSV8 group,RSV28 group,RSV4 group and normal group were (30.9860 ± 3.3464)mg,(15.3212 ± 1.2249) mg,(13.9193-2.1409) mg,(11.5857± 1.5705) mg,(3.9780 ± 0.6224) rag,respectively.The serum albumin of RSV14 group decreased.The anionic sites of glomerular basement membrane(GBM) decreased in RSV nephropathy.The number of anionic sites per 1000 nm GBM of RSV14 group,RSV8 group,RSV28 group,RSV4 group and normal control group were 12.0000 ± 1.5811,14.0000 ± 1.0000,14.6000 ± 1.1401,16.8000 ± 0.8366 and 21.2000 ± 1.3038,respectively.Hpa in glomeruli couldn't be detected in normal rats.Glomerular Hpa expression was up-regulated in RSV nephropathy.The expression of glomerular Hpa of RSV14 group,RSV8 group,RSV28 group and RSV4 group were 0.6622 ±0.1145,0.5511 ± 0.0257,0.3524 ± 0.0296 and 0.4521 ± 0.0087,respectively.The expression level of Hpa in RSV8 group and RSV14 group was higher than that in RSV4 group and RSV28 group.There was a linear positive correlation between the expression level of glomerular Hpa and the quantity of 24-hour urine protein (r =0.783,P ＜ 0.05).Conclusions The increased expression of glomerular Hpa in RSV nephropathy of rats leads to loss of the glomerular anionic sites and damage of the electrostatic barrier of GBM,which promote the proteinuria.