Using of clearing zones on pachyman agar medium, there are 217 plant endophytic actinomycetes, producing ?-1,3-glucanase were screened. 45.6% of the strains produced ?-1,3-glucanase, in which the strains from cucumber are up to 38. The percentages of endophytic actinomycetes from different hosts produceing ?-1,3-glucanases were different. The percentage of the strains in Rhizoma Polygonatum produced ?-1,3-glucanases is the highest, up to 88.9%. The Inhibited effects of plant endophytic actinomycetes which produced extracellular ?-1,3-glucanases on mycelium growth of Sclerotinia sclerotiorum were detected in vitro. Cucumber endophytic actinomycete gCLA4 strain was screened out from 99 isolates, which can strongly inhibit the growth of S. sclerotiorum. The optimal ?-1,3-glucanases fermentation conditions of strain gCLA4 were investigated, they were pachyman 0.2%, peptone as nitrogen, pH 7～8 for 5 days. The ?-1,3-glucanases of strain gCLA4 had some inhibiting efficiency on 13 plant pathogens, in which inhibiting efficiency to Botryosphaeria dothidea was the strongest.

Aneurysm, Dissecting ; genetics ; Aortic Aneurysm ; genetics ; Diseases in Twins ; Humans ; Male ; Middle Aged

Objective To investigate the expression of neural-nitric oxide synthase (nNOS) in renal clear cell carcinoma and its clinical significance.Methods The expression of nNOS mRNA in 533 samples of TCGA database was analyzed with Student t test,and statistical analysis was performed to assess the relationship between nNOS expression and clinical prognosis with Kapla-Meier test.Western blot analysis of nNOS protein expression in 10 cases of clear cell renal cell carcinoma(ccRCC) from department of urology of Wuhan union hospital with student t test.Results The mRNA levels of nNOS in 72 cases of ccRCC in tumor tissues and adjacent tissues and were 2.99 ± 0.28 and-1.57 ± 0.17,it is significantly lower than those in adjacent tissues (P ＜ 0.01).The mRNA levels of nNOS in 533 cases of ccRCC,in tumor tissues and adjacent tissues and were 2.99 ± 0.28 and-1.76 ± 0.05,it is significantly lower than those in adjacent tissues (P ＜ 0.01).A total of 533 sample studies showed a low correlation between nNOS expression and clinical T stage,T1-1.59 ±0.08,T2-1.96 ±0.13,T3-1.90 ±0.09,T4-2.38 ±0.28 (P =0.0029) and -1.63 ±0.06 and-2.16 ± 0.13 between non-metastasis and no-metastasis (P =0.0009),and-1.57 ± 0.08 and-2.03 ± 0.11 between non-recurrence and recurrence (P =0.008).Survival analysis showed that the overall survival time were (40.3 ± 5.6) months and (48.3 ± 5.7) months in lower and higher nNOS expression,and disease free survival time were (37.1 ± 2.1) months and (40.3 ± 5.6) months in lower and higher nNOS expression,both with shorter time in low expression of nNOS (P ＜ 0.01).nNOS proteins were 1.02 ± 0.16 and 0.61 ± 0.1 1 in tumor tissues and adjacent tissues with significantly lower expression(P＜0.05).Conclusions The mRNA and protein of nNOS are lower in ccRCC with a poor prognosis of ccRCC.

OBJECTIVEIn order to investigate the roles of metalloproteinase in inflammatory bone destruction in ankylosing spondylitis (AS), and analyze the mechanism of preventing inflammatory bone destruction of Bushen Qiangdu decoction (BSQDD) in AS cases. Comparisons were made on the expressions of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) by peripheral blood mononuclear cells (PBMC) between AS patients and healthy controls. The effect of BSQDD was investigated on the expression and of MMP-9 and TIMP-1 produced by PBMC in AS patients.

METHODSFrom March 2005 to March 2006, 30 active AS cases of Kidney-asthenia, Du-cold and blood-stasis syndrome were selected as patients group in the China-Japan Friendship Hospital. There are 27 male patients and 3 female patients. The age range is from 16 to 45, averaging (30.8 +/- 8.8) years. Disease duration is from 0.5 to 10 years. Cases received three-month BSQDD treatment were considered as the treatment group. Twenty healthy persons were included in the control group. Serum and PBMC were separated. The PBMC were stimulated by PHA and PMA, and the supernatant was collected. The mRNA expression of MMP-9 and TIMP-1 in PBMC was analyzed by RT-PCR. The content of MMP-9 and TIMP-1 in serum and culture supernatant of PBMC were detected by ELISA.

RESULTSCompared with health control group, the serum concentration of MMP-9 and TIMP-1 in patients group before treatment increased (P<0.01, P<0.05), but the level of MMP-9 and TIMP-1 in the serum of patients after treatment decreased compared with pre-treatment cases (P<0.05). Furthermore,compared with health control group, PBMC of patients group before treatment expressed higher levels of MMP-9 and TIMP-1 both on transcript level and at protein level (P<0.01, P<0.05), and the expression levels of MMP-9 and TIMP-1 in PBMC in patients after treatment both on transcript level and at protein level was down-regulated compared with pre-treatment (P<0.01, P<0.05).

CONCLUSIONPBMC of AS patients had a higher potential capacity for MMP-9 and TIMP-1. BSQDD possibly prevented inflammatory bone destruction of AS through inhibiting production of MMP-9 and TIMP-1 produced by PBMC.

Adolescent ; Adult ; Case-Control Studies ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Expression Regulation ; drug effects ; Humans ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Male ; Matrix Metalloproteinase 9 ; biosynthesis ; blood ; genetics ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Retrospective Studies ; Spondylitis, Ankylosing ; blood ; drug therapy ; genetics ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; blood ; genetics ; Young Adult

The study was aimed to explore clinical result of cyclosporin A (CsA) and androgens for treatment of myelodysplastic syndrome (MDS) with refractory anemia. Four cases of MDS-RA were treated with CsA and androgens, while the changes of blood counts, bone marrow and chromosome were observed. The results showed that substantial hematological response was observed in all four patients, that their anemia improved and all transfusion-dependent patients achieved transfusion independence. In conclusion, CsA and Adr therapy was well tolerated. CsA and Adr therapy offer an alternative treatment of MDS with RA. The mechanisms of the benifical effect from this therapy remain the subject of an ongoing study.
Adolescent ; Adult ; Aged ; Androgens ; therapeutic use ; Blood Cell Count ; Cyclosporine ; therapeutic use ; Drug Therapy, Combination ; Female ; Hemoglobins ; analysis ; Humans ; Immunosuppressive Agents ; therapeutic use ; Male ; Middle Aged ; Myelodysplastic Syndromes ; blood ; drug therapy ; Treatment Outcome

Objective To investigate the ostengenie potential of adipose-derived stem cells(AD- SCs)when exposed to adenovirns containing hBMP-2 cDNA(Adv-hBMP-2)and offer a choice of cell source for gene therapy and tissue engineering.Methods Human adipose tissues were obtained from patients who received orthopaedic surgery or liposuction.ADSCs were obtained by digesting the adipose tissues.Firstly,flowcytometric analysis was performed for the confirmation of mesenchymal stem cell ori- gin and the surface markers including CD34,CD44,CD68,CD71,CD90,and CD105.The ADSCs were transfected by Adv-hBMP-2 and the effects were tested in vitro,lmmunoprecipitation and Western blotting and ELISA were performed for confirming BMP gone transduction and its stable expression.The transform of ADSCs was assessed by extracellular ALP staining,intracellular ALP spectrophotometry,von Kossa staining and RT-PCR.In the in vivo experiment ADSC-Adv-hBMP-2 cells were injected into the hind limb of nude mice and analyzed radiographically and histologically.Results ADSCs were successfully isolated from human adipose tissues.The isolated ADSCs expressed CD44,CD71,CD90 and CD105 and CD34 and CD68 were absent.The result confirmed the mesenchymal stem cell origin of the cells.West- ern blotting and ELISA confirmed successful and persistent hBMP-2 production by ADSC-Adv-hBMP-2 cells.Extracellular ALP staining,intracellular ALP spectrophotometry,yon Kossa staining and RT-PCR revealed that ADSCs treated with Adv-hBMP-2 had a tendency of transfering into osteoblast.X-ray and H&E sections from hind limb of nude mice injected with ADSC-Adv-hBMP-2 cells confirmed bone forma- tion at 2 weeks.Conclusions Liposuction aspirates contain abundant ADSCs that can be transduced with hBMP-2 gene,and the tranduced ADSCs differentiate into the osteoblast.ADSCs may be an ideal source of mesenchyme-lineage stem cells for gone therapy and tissue engineering.

Objective To explore the clinical teaching effectiveness of LBL combination of PBL teaching mode in the intensive care unit. Methods The 2009-2010 cycle students 70 in the intensive care unit were randomly divided into control group and experiment group. Control group, the traditional LBL teaching;experimental group, LBL combined PBL teaching mode. Finally, different teaching effectiveness was evaluated by closed examinations and questionnaires. Results Examination total score, using the relevant sections of PBL teaching performance, comprehensive analysis of performance problems were significantly higher (P ＜ 0. 01 ),and training clinical thinking, theoretical understanding and promotion of memory, analytical ability,communication skills, team spirit of the experimental group students were significantly increased ( P ＜ 0. 05 or P ＜0. 01 ). Conclusions LBL combination of PBL teaching in the intensive care unit is feasible, effective and could help students improve overall quality.

AIMTo study the permeability of nerve growth factor (NGF) liposomes (NGF-L, NGF-SSL, NGF-SSL-T) on the blood-brain barrier (BBB) model and the distribution in vivo, and analyze the correlation between the results in vitro and in vivo.

METHODSThe BBB model in vitro was established by using mouse brain microvascullar endothelial cell, and the model was applied to study the permeability of NGF liposomes. The distribution of NGF of each group was studied by 125I labeled and SDS-PAGE method.

RESULTSThe highest encapsulation proportion was 34%, and the mean size of NGF liposomes was below 100 nm. The permeability of NGF liposomes on in vitro BBB model showed that the liposome could promote NGF to transport across the BBB, the permeability of NGF-SSL-T was the highest. The distribution in the brain showed in an order of NGF concentration NGF-SSL-T > NGF-SSL + RMP-7 > NGF-SSL > NGF-L. There was a close relationship between P(e) (permeability coefficient on in vitro BBB model) and BUI (brain uptake constant in vivo).

CONCLUSIONLiposomes can promote NGF to transport across the BBB, and the transporting ability BBB of NGF-SSL-T which RMP-7 incorporated into the surface of NGF liposomes is the best.

Animals ; Biological Transport ; drug effects ; Blood-Brain Barrier ; Bradykinin ; analogs & derivatives ; pharmacology ; Brain ; metabolism ; Cell Membrane Permeability ; Drug Delivery Systems ; Endothelial Cells ; cytology ; Liposomes ; Male ; Mice ; Nerve Growth Factor ; administration & dosage ; pharmacokinetics ; Particle Size ; Rats ; Tissue Distribution

OBJECTIVETo investigate the role of adhesion molecules alphavbeta3 and alphavbeta5 and their ligands Del-1 and L1 in the tumor-endothelial cell adhesion in vitro.

METHODSThe expression of alphavbeta3, alphavbeta5 and ICAM-1 in liver sinusoidal endothelial cells (LSEC) and liver cancer endothelial cells (T3A) cultured under normoxia or hypoxia were analyzed by RT-PCR and fluorescent activated cell sorter (FACS). The expression of Del-1 and L1 in six tumor cell lines under normoxia or hypoxia were analyzed by RT-PCR and Western blot, respectively. The adhesion of dye-labeled tumor cells and endothelial LSEC and T3A cells was measured by a fluorescence plate reader after their culture.

RESULTSThe expression of alphavbeta3 and alphavbeta5 were higher in T3A cells than that in LSEC cells, and were upregulated under hypoxia, while the expression of ICAM-1 was lower in T3A cells than that in LSEC cells, and was upregulated under hypoxia only in LSEC. The expression of Del-1 and L1 molecules were obviously different in various tumor cell lines and were differentially regulated under hypoxia. The adhesion of tumor cells with Del-1 or L1 expression was higher in T3A cells than that in LSEC cells, and was significantly increased under hypoxia condition. Furthermore, the adhesion of tumor cells to T3A could be inhibited by antibodies against alphavbeta3 and alphavbeta5, or SiRNAs for beta3 and beta5.

CONCLUSIONalphavbeta3 and alphavbeta5 and their ligands Del-1 and L1 may play an important role in tumor cell migration.

Antibodies ; immunology ; Cell Adhesion ; Cell Hypoxia ; Cell Line, Tumor ; Endothelial Cells ; cytology ; metabolism ; Humans ; Integrin alphaVbeta3 ; genetics ; immunology ; metabolism ; Intercellular Adhesion Molecule-1 ; immunology ; metabolism ; Ligands ; Neoplasms ; metabolism ; pathology ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; pharmacology ; Receptors, Vitronectin ; genetics ; immunology ; metabolism

OBJECTIVETo investigate the technique of periorbital defects reconstruction with island orbicularis oculi myocutaneous flap in orbital zone.

METHODSThe upper or lower eyelid island orbicularis oculi myocutaneous flap,medially based,were used for the defects of the periorbital area, according to the position,shape and size of the defects. The donor site was closed directly.

RESULTSFrom July 2003 to October 2009, 24 patients were treated in this method, the flaps survived totally. The flap proved to be flexible, safe, relatively simple, and provided good functional and aesthetic results with follow up 6 to 24 months. Complications were minimal.

CONCLUSIONSThe upper eyelid or lower eyelid island orbicularis oculi myocutaneous flap, medially based, is a satisfied method in repairing periorbital defects in one stage with good blood supply, excellent color texture matching and inconspicuous donor scar and deformity.

Adolescent ; Adult ; Aged ; Child ; Eyelids ; surgery ; Female ; Humans ; Male ; Middle Aged ; Oculomotor Muscles ; transplantation ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; Young Adult