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.

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.

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.

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.

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.

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

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

BACKGROUND: This study aimed to investigate the prevalence rate of critical illness-related corticosteroid insufficiency (CIRCI) and the effect of low-dose glucocorticoid on prognosis of CIRCI in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). METHODS: Since January 2010 to December 2012, 385 patients, who met the criteria of AECOPD, were enrolled in the Intensive Care Unit (ICU) of the First People's Hospital and Municipal Central Hospital of Xiangtan City. The AECOPD patients complicated with CIRCI screened by an adrenalcorticotrophic hormone test within 12 hours after admission to ICU were divided into a treatment group (n=32) and a control group (n=31) for a prospective, randomized and controlled clinical trial. Hydrocortisone (150 mg/d) or normal saline was injected intravenously for 7 days. The patients were followed up for 28 days after injection. The endpoint included 28-day survival time, non-shock time, ICU stay and the period of non-mechanical ventilation. The markers of inflammation C-reactive protein, tumor necrosis factor-α, interleukin 6 and procalcitonin were measured at baseline and 7 days after treatment. The variables were analyzed by Student's t test, the non-parametric statistical test, the Chi-square test or the Kaplan-Meier method with SPSS18.0 statistic software. A P value <0.05 was considered statistically significant. RESULTS: Totally 63 patients were diagnosed with CIRCI by an adrenalcorticotrophic hormone test and the prevalence rate was 16.4％. The shock rate of the AECOPD patients complicated with CIRCI was higher than that of the AECOPD patients without CIRCI (23.8％ vs. 8.7％, P<0.01). Kaplan-Meier analysis revealed that the 28-day survival time of the treatment group was obviously longer than that of the control group (P<0.05). Compared with the control group, shock-free days within 28 days was longer in the treatment group (18.2±9.5 vs. 25.8±4.1, P<0.05). Treatment with low-dose glucocorticoid obviously decreased the markers of infection and inflammation (P<0.01), such as C-reactive protein (13.2±5.5 mg/L vs. 8.3±3.1 mg/L for the control group; 13.5±5.9 mg/L vs. 5.1±2.3 mg/L for the treatment group), tumor necrosis factor-α (26.1±16.2 g/L vs. 17.5±11.7 g/L for the control group; 25.0±14.8 g/L vs. 10.4±7.8 g/L for the treatment group) and procalcitonin (3.88 g/L vs. 2.03 g/L for the control group; 3.77 g/L vs. 1.26 g/L for the treatment group). Furthermore, the markers in the treatment group decreased more obviously than those in the control group (P<0.01). CONCLUSION: The prevalence rate of CIRCI was higher in the patients with AECOPD in the department of critical medicine, and low-dose glucocorticoid treatment for one week reduced the 28-day mortality, shock time and markers of infection and inflammation.

The epitope-G1 gene of Bovine ephemeral fever virus (BEFV) glycoprotein was synthesised by PCR and cloned into expression vector pPIC9K to construct recombinant plasmid pPIC9K-G1. Then the pPIC9K-G1 was linearized and transformed into Pichia pastoris GS 115. The recombinant P. pastoris strains were selected by a G418 transformation screen and confirmed by PCR. After being induced with methanol, an expressed protein with 26 kDa molecular weight was obtained, which was much bigger than the predicted size (15.54 kDa). Deglycosylation analysis indicated the recombinant G1 was glycosylated. Western blot and ELISA tests, as well as rabbit immunization and specificity experiments indicated that the target protein had both higher reaction activity and higher immunocompetence and specificity. The recombinant G1 protein could be used as a coating antigen to develop an ELISA kit for bovine ephemeral fever diagnosis.