Objective To construct mouse granulocyte-macrophage colony stimulating factor (mGM-CSF) gene eukaryotic expressing plasmid pcDNA3-GM-CSF, to transfect the recombinant into erythroleukemia cell line FBL-3, and identify their biological activity.Methods GM-CSF gene eukaryotic expressing plasmid was constructed by subclone and recombinant was transfected into FBL-3 cells by electroporation. After screening by G418 and cloning by limiting dilution,we obtained positive cell clones(FBL-3-GM-CSF). PCR and RT-PCR were used to identify the integration and stable expression of GM-SF gene in FBL-3-GM-CSF cells. The biological activity was confirmed by the hematopoietic progenitor cell proliferative assay and hematopoietic progenitor cell colony formation assay. Results Mouse GM-CSF cDNA was amplified from the prokaryotic expressing plasmid PET-30a(+)-GM-CSF by PCR firstly and BamH Ⅰ and EcoRⅠrestriction sites were introduced. The inserted fragment was cut by BamH Ⅰ and EcoR Ⅰ digestion and ligated into pcDNA3 vector. The pcDNA3-GM-CSF eukaryotic expressing plasmid was constructed. The recombinant was cleared with appropriate endoneucleases and sequenced. The findings showed that the orientation of the insert was correct, while no rearrangement or mutation was found. PCR and RT-PCR assay showed that GM-CSF gene had integrated into FBL-3-GM-CSF cells and stably expressed. The hematopoietic progenitor cell proliferative assay and hematopoietic progenitor cell colony formation assay demonstrated that the cultured supernatant of FBL-3-GM-CSF cells of expressing GM-CSF should obviously stimulate proliferation of murine marrow mononuclear cells, and could stimulate hematopoietic progenitor cell colony formation. The number of colony formation was 54.67?4.83. The rate of colony formation was 0.547 %.Conclusions GM-CSF gene eukaryotic expressing plasmid is constructed successfully. A cell clone, which can express stably GM-CSF gene and possess biological activity,is obtained. Our studies have founded the base for the preparation of GM-CSF gene-modified vaccine of tumor cell and the study of feasibility of immune therapy of leukemia.

Objective To investigate the regional cerebral blood flow (rCBF) in amnestic mild cognitive impairment (aMCI) and vascular cognitive impairment-no dementia (VCI-ND) subjects. Methods Sixteen normal elders, 10 patients of aMCI, 12 patients of VCI-ND who were aged from 50 to 80 years old and received an education of middle school or higher. All participants finished cranial CT or MRI. Xe-CT was used to evaluate rCBF of different cerebral regions of all participants. Results The distribution of rCBF of basal ganglia, the cortex and white matter was (76. 4 ± 8. 6), (48.0 ± 7. 1) and (20. 5 ± 1.7) ml· 100 g-1 ·min-1, respectively. When compared in 3 groups, the temporal and parietal lobe rCBF had a decreasing tendency in aMCI group, while in VCI-ND group, the most dominant decreasing parts were mainly concentrated in white matter region ((17. 7±2. 3) ml·100 g-1·min-1, F = 5. 740, P = 0. 002). Whatever the depth or the width was, beth periventricular and subeortical deep white matter, anterior and posterior ventricular regions were all involved. There are no dominant difference of rCBF in caudate nucleus, lentiform nucleus and thalamus. Conclusion The difference in rCBF reflects the pathological difference between aMCI and VCI-ND.

OBJECTIVETo analyze the relationship between symptoms and signs of temporomandibular disorders (TMD) and the patients' quality of life (QOL).

METHODSA total of 492 TMD patients were included in this study. The clinical examination results were recorded using Fricton index of temporomandibular joint function. "Visual analog scale (VAS) evaluation of QOL disturbance" was designed to quantitate patients' QOL, to evaluate the degree that the patients QOL was affected.

RESULTSChewing, daily life and emotion among all 8 items of QOL were frequently affected by TMD, and joint clicking had the least influence on QOL. Intermittent closed lock had more severe interference with QOL than joint clicking only. Severe and moderate pain or limited mouth opening affected the QOL more severely than mild pain or mild limited mouth opening. The simple linear relationship between Fricton index and patients' QOL was poor (r < 0.4).

CONCLUSIONSPain is the most frequently seen symptom in TMD. TMD could affect patients' QOL, including both physical and social-psychological functions. The results suggest that the patients' QOL as well as TMD symptoms and signs should be considered in the management of TMD.

As the major commercial source of natural rubber,Hevea brasiliensis attracts much attention.However,the heterozygous nature,long breeding cycle are strong limitations for conventional breeding.While genetic engineering,which can be used to widen the germplasm base and produce desirable agronomic traits quickly and efficiently,offers a viable alternative approach to complement traditional breeding.Comprehensive analysis indicates that in the past two decades,with calli derived from immature anther or integumental tissues of immature fruit as receptors,both biolistic and Agrobacterium-mediated transformation methods were employed for developping rubber genotypes with improved latex yield,tolerance to tapping panel dryness syndrome,producing high-value recombinant proteins,etc.Being recalcitrant to tissue culture,the transformation efficiency of Hevea is comparatively low,and the procedures are still needed to optimize.Finally,breeding objectives and strategies to improve transformation efficiency were also proposed in the review.

Objective: To study the quality and transdermal properties of matrine microemulsion-based hydrogel (MBH) to provide basis for the development of the preparation.Methods: The stability of MBH was observed at 4 ℃ for 3 months and the changes of particle appearance, viscosity, pH and matrine content were observed.The transdermal permeation of MBH was investigated by a dual chamber permeation and diffusion device with excised mouse skin as the barrier.Taking rabbits as the experimental subjects, the irritation of MBH to the normal skin and damaged skin was investigated.Results: The appearance, viscosity, pH and matrine content of MBH at 4 ℃ in 3 months did not change significantly.In vitro transdermal test showed that MBH had a good penetration rate on mouse skin, and no skin irritation occurred after single or multiple administrations.Conclusion: MBH has good stability and high rate of transdermal penetration without skin irritation, which is a promising drug delivery system of matrine with good application prospects.

Objective To set up and verify a new scoring system of clock drawing test(CDT).Methods CDT and other neuropsychological tests were applied to 180 normal individuals.170 subjects with mild cognitive impairment(MCI),and 31 patients with mild Alzheimer's disease(AD).The first step of the 30-score system of CDT was to anchor the 4 points of"12-3-6-9".totaling a score of 4 which was the "30-score system A(Anchoring)part";and the other 13 items reflecting the drawing results and totaling a score of 26 made up the so called"30-score system C(Clockfaee)part".the two together had a total score of 30.Thirty-eight subjects underwent Xe-enhanced computed tomography(Xe-CT)scanning for quantitively measuring regional cerebral blood flow(rCBF)of 26 regions of brain.Results Correlations coefficients of the scores of 14 items with the total score in 30-score system of CDT were in the range of 0.48to 0.71(P＜0.01).30-score system C part was closely correlated with indicators of visuospatial and executive function,while 30-score system A part with memory indicators.The sensitivity and specificity of the 30-score system A part score≤2 for detecting MCl were 70.6%and 73.9%.respectively.The sensitivity the specificity of 30-score system C part score≤17 for mild AD were 75.3%and 75.9%.respectively.In the regression equation of 30-score system A and C part.the combined independent variables were the CBF of left frontal cortex and right white matter,respectively.Conclusion The 30-score system A and C part have difierent meanings:30-score system C part could be used to identify AD while 30-score system A part is helpful for identiring MCI.

Objective To analyze the security and effectiveness of the accurate resection and reconstruction of pelvic tumor with computer navigation and to evaluate the feasibility of pelvic ring structure and mechanics reconstruction using finite element analysis.Methors From December 2008 to June 2010,tumor resections assisted by computer navigation technology and pelvic reconstructions with assembly prosthesis designed by finite element analysis were performed in 12 patients with internal hemi-pelvic malignant tumors.There were 8 male and 4 female patients,with the average age of 39.7 years(range,25-53).The malignant tumors involved chondrosarcoma(4 cases),osteosarcoma(2),Ewing sarcoma(2),malignant fibrohistiocytoma(1),and metastatic tumor(3,in which including renal cell carcinoma 1 case,thyroid carcinoma 1,and breast carcinoma 1).According to Enneking staging classification,there were 1 case in Ⅰ A,2 cases in ⅠB,2 cases in Ⅱ A.and 4 cases in Ⅱ B.The local recurrence of tumor and functional results were followed up postoperatively.Results There were no nerves,vessels and pelvic organs injuries intra-operatively.The postoperative X-rays showed that the excised regions were proper to the preoperative schedule and en bloc resections were performed in all patients.All assembly prostheses were in satisfactory position.and the screw fixation was accordance with the finite element biomechanical analysis.All the cases were followed up for average 18.2 months (range,8-26).There were 1 case with local recurrence and pulmonary metastasis,1 case with deep infection,and 1 case with deep vein thrombosis.No prosthesis dislocation,loosening,breaking,and limb length discrepancy occurred in all patients.The average MSTS93 functional score was 70%in all patients at 6 months,and 68 % in 6 patients at 20 months follow.up.Conclusion The resection and reconstruction of pelvic tumor assisted by computer navigation is a safe and effective procedure.The finite element analysis can provide accurate biomechanical guidance to the reconstruction of pelvic ring.

There are 250 species of Zanthoxylum (Rutaceae) in the world. This genus distributed in tropical and subtropical regions. Alkaloids are the major and representative ingredients in these plants including quinolines, isoquinolines, and amide alkaloids, with such biological activities as anti-tumor, anti-inflammatory, analgesic, anti-virus, anti-platelet aggregation, anti-bacteria and anti- oxidant. These species have been used for a long time to treat toothache, urinary and venereal diseases, lumbago and rheumatism. This review summarizes the chemical constituents and pharmacological activities from the Z. sppplants, in an effort to the systematic research and application of the alkaloids of this genus.
Alkaloids ; chemistry ; pharmacology ; Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Molecular Structure ; Zanthoxylum ; chemistry

Isolated alkaloids from Coptis chinensis Franch. The compounds were identified as berberine, columbamine, groenlandicine, jatrorrhizine, magnoflorine, corydaldine and ferulic acid methylester. Then measured their bitter degree based on the electronic tongue and evaluated the antibacterial. The results based on the Electronic Tongue showed that berberine, columbamine, groenlandicine and jatrorrhizine have higher bitter degree than magnoflorine and corydaldine. And they also appeared better antibacterial activity on E. coli and S. aureus. The correlation coefficients between bitter degree and the two bacteria antibacterial activity were 0.983 and 0.911. So there was close relationship between the bitter degree and antibacterial activity of bitter components. Thus, it is confirmed further that bitter components are the material foundation of medicinal effectiveness of bitter herbs.
Aporphines ; analysis ; Berberine ; analogs & derivatives ; analysis ; Berberine Alkaloids ; analysis ; Biomedical Research ; instrumentation ; methods ; Coptis ; chemistry ; Drugs, Chinese Herbal ; analysis ; chemistry ; pharmacology ; Electronics ; instrumentation ; methods ; Escherichia coli ; drug effects ; growth & development ; Microbial Sensitivity Tests ; Reproducibility of Results ; Staphylococcus aureus ; drug effects ; growth & development ; Taste

To study the role of anti- C. albicans IgY and serum in protection of C. albicans infection of several animal models. To develop three animal models of C. albicans infection: a burned rat model of C. albicans infection, a mouse model of vaginal candiasis and a immunosuppression mouse model of C. albicans infection. And we compared the contribution of anti- C. albicans IgY and serum to the clearance of the C. albicans in three animal models of C. albicans infection. Anti- C. albicans IgY can protect against C. albicans infection in a burned rat model of C. albicans infection and a mouse model of vaginal candidiasis. The serum can effectively protect the mice from disseminated candidiasis in a immunosuppression mouse model. Humoral immunity component involving anti- C. albicans IgY and serum protect against C. albicans infection in a burned rat model of C. albicans infection ,a mouse model of vaginal candidiasis and a immunosuppression mouse model of C. albicans infection.