Objective To optimize the processing method of Codonopsis pilozula prepared product by the factorial design and response surface methodology. Method In the two different optimizing processing, methods were optimized respectively on basis of preplanned experiment design and the percentage of extracts were also determined respectively. Drying temperature and time, bran parching temperature and time respectively as independent variable, their OD value as dependent variable, multiple regression equation was obtained and binomial equation was simulated as well. Results The r2 of Codonopsis pilozula dried product and Codonopsis pilozula parched product were respectively 0.976 4 and 0.987 9, meanwhile their optimized processing methods were drying temperature 80 ℃, drying time 2 hours and parching temperature 250 ℃, parching time 1 minute. Conclusion The effect of independent variable to dependent variable can be analyzed on three-dimensional chart with the factorial design and response surface methodology, consequently it will help to optimize processing method of prepared product.

Objective To investigate differential expressed protein in the intestinal mucosa of patients with inflammatory bowel disease (IBD) with antibody chips,and to explore the possible role of the screened proteins in pathogenesis of IBD.Methods The mucosa tissues of nine patients with ulcerative colitis (UC),nine patients with Crohn's disease (CD) and nine control individuals were collected.After total protein of each group was extracted,the differential expressed protein of each group was analyzed by Raybiotech L-series human cytokine antibody chips.The mucose tissues of other nine patients with UC,nine patients with CD and nine control individuals were collected,and were used to verify the greatly differential expressed proteins by Western blot.The t-test was performed to compare two groups.Results Compared with the control group,there was significantly difference in 263 cytokines of UC group,and 414 cytokines of CD group.And then the higher expressions of herpes virus entry mediator,leukemia inhibitory factor and platelet factor 4 in the mucosa tissues of IBD patients were confirmed by Western blot and the differences were statistically significant (UC:t=23.85,9.53,18.88; CD:t=13.54,16.65,13.67,all P＜0.01).Conclusion The screened differential expressed cytokines in the mucosa tissues of IBD patients by cytokine antibody chips could be helpful to reveal the pathogenesis of IBD and discover new molecular biomarkers.

Aim To establish a new simple, easy and economical PCR method for determining cyp2d6~*10 allele and studying its distributive frequencies in Chinese population. Methods The new PCR are established and compared with the classical PCR-RFLP. The 224 samples have been determined with the new PCR.Results the results of the two methods go on the way. The allele distribution frequencies of cyp2d6~*10 resembles the reports. Conclusion the new method is proved to be accurate and convenient. It provides a genetics basis for the drug therapy with individuals in clinics.

MTM1 gene is essential for SOD2 activity and normal mitochondrial function. MTM1 deletion results in decreased SOD2 activity, impaired mitochondrial function and growth defect on nonfermentable carbon source. A yeast genomic library was transformed into mtm1 deletion mutant to screen for suppressor genes of MTM1. The damage caused by MTM1 deletion is irreversible and even overexpression of MTM1 can not rescue the growth defect of mtm1 deletion mutant. Another screening strategy was adopted: a plasmid overexpressing MTM1 was transformed into wild type before the MTM1 gene on chromosome was deleted. The resulting strain, designated YES2MTM1, was transformed with a yeast genomic library. Transformants lost the plasmid overexpressing MTM1 after 5-FOA treatment. Yeast strains able to grow on nonfermentable carbon source with MTM1 deletion and overexpression of some DNA fragments were picked up and candidate suppressor genes were identified. Overexpression of five genes were identified to be able to rescue the growth defect on nonfermentable carbon source. The study will provide reference for MTM1 gene function and screening for suppressor of genes whose deletion result in irreversible damage.

Anti-Bacterial Agents ; therapeutic use ; Chronic Disease ; Humans ; Rhinitis ; drug therapy ; Sinusitis ; drug therapy

Objective To investigate the effect of radiation on the expressions of RANKL and OPG in osteoblasts in order to disclose the molecular mechanism of bone injury induced by ionizing radiation.Methods The osteoblasts were differentiated from MC3T3-E1 cells.After 2 or 4 Gy137 Cs γ-irradiation,the mRNA and protein expression levels of RANKL and OPG of osteoblast precursor and osteoblast were detected by real-time PCR and Western blot.Results The expressions of RANKL mRNA (t=5.41,P＜0.05)and protein(t=68.37,P＜0.01)were up-regulated after 4 Gy irradiation,while the expressions of OPG mRNA(t=5.20,7.02,P＜0.05)and protein(t=7.78,9.45,P＜0.05)were down-regulated after 2 and 4 Gy irradiation.Conclusions 2 and 4 Gy ionizing radiation alters RANKL/RANK/OPG pathway in osteoblasts,which may promote the osteoclast differentiation and maturation and hence promote bone resorption of osteoclasts.

Objective To improve the anderstanding of clinical profile of primary plasma cell leukemia.Methods Case report and literature review.Results A rare case of nervous system relapse in primary plasma cell leukemia was reported.Six patients were identified from the literature.The type of immunoglobulin included IgG(3 patients),IgD(2 patients).Clinical manifestations of nervous system were variable.The average interval from initial diagnosis to the development of nervous system relapse was 16.5 months.Plasma cells were found in cerebrospinal fluid in 4 patients.The mean surviaval time was 6.7 months after nervous system relapse.Conlusion Nervous system relapse in primary plasma cell leukemia is rare with poor prognosis.

This study discusses the effects of Shenlian extracts (SL) on M1 macrophages in atherosclerosis. The MTT assay was used to detect the growth inhibition rates of RAW264.7 cells. RAW264.7 cells were stimulated with murine interferon-gamma (IFN-gamma) plus lipopolysaccharide (LPS) to induce M1 macrophages. The different concentrations of SL extracts (high-dose 50 mg x L(-1), moderate-dose 25 mg x L(-1), low-dose 12.5 mg x L(-1)) were added. The CD86 of M1 macrophages in cell membrane was measured by flow cytometry. The mRNA expression of iNOS and TNF-alpha gene was detected by reverse transcription PCR (RT-PCR). And the supernatants were collected, the content of IL-6 and TNF-alpha were detected with ELISA kits. The results of this experiment show that the expression of the cell membrane molecule CD86, iNOS and TNF-alpha gene, the content of IL-6 and TNF-alpha was obviously increased in M1 macrophages by IFN-gamma and LPS. The different doses of SL extract could reduce the expression of the above indicators. The above experimental results demonstrate that IFN-gamma combined LPS can induce RAW264.7 cell to type into M1 macrophages, and SL extracts can inhibit M1 macrophages.
Animals ; Cell Line ; Cell Shape ; drug effects ; Cell Survival ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Interferon-gamma ; genetics ; immunology ; Interleukin-6 ; genetics ; immunology ; Macrophages ; cytology ; drug effects ; immunology ; Mice ; Tumor Necrosis Factor-alpha ; genetics ; immunology

Objective To study the influence of irradiation on the osteoblast function by the gene expression changes of RANKL and OPG.Methods Bone marrow stromal cells were induced to develop into early and mature osteoblasts in vitro.The characterization of osteoblasts was indentified by ALP staining.The RANKL and OPG mRNA levels in early and mature osteoblasts, which exposed to 0 -4 Gy radiation were determined by RT-PCR.Results Bone marrow stromal cells had been induced to early and mature osteoblasts by osteoblast differentiation medium in vitro.In early stage of osteoblast, RANKL mRNA expression levels treated with 1Gy irradiation was 2.83-fold higher than those other irradiation dosage groups.The RANKL mRNA expression levels of each group in early stage of osteoblasts were significantly higher than those in the mature counterpart ( t = 8.34 - 103.57, P ＜ 0.05 ).The ratio of RANKL/OPG mRNA was obviously greater in early osteoblast compared with the mature cells ( t = 2.84 - 20.99, P ＜0.05 ), and it was the highest in 1Gy irradiation treated early osteoblast.Conclusions Radiation exposure of the early osteoblasts promotes osteoclasts function and results in the bone loss.

ObjectiveNotch signaling is highly conservative in evolution and plays an important role in cell's proliferation and differentiation.Construction of tentiviral vector containing Notch intracellular domain (NICD) would lay the foundation for the study of Notch signaling.MethodsTotal RNA was extracted from the myeloid tissue of C57BL/6 mice.cDNA was composed via reverse transcription.NICD sequence was obtained by PCR and recombined into lentiviral vector.Lentiviral vector with NICD was infected into target HEK293T cell.Real-time PCR and Western blot were used to examine NICD expression in HEK293T cell.ResultsNICD expression increased in HEK293T cells.ConclusionThe successful construction of lentiviral vector involving mice NICD expression provides the foundation for the future study.