Humans ; Neoplasms ; therapy ; Receptor, Epidermal Growth Factor

Objective To observe the expression of c-fos mRNA and protein in fluoride treated mouse fibroblast (FB) and osteoblast (OB) and to further explore the effects of c-fos in the osteogenic action of FB. Methods Mouse FB and OB were divided into control group and six fluoride groups (0, 0.0001, 0.0010, 0.1000, 1.0000, 10.0000,20.0000 mg/L F-), and the levels of c-fos protein at 2,4,24,48,72 h and c-fos mRNA at 48 h were measured by using ELISA and RT-PCR methods. Results Compared with the control group, fluoride increased the content of c-fos protein obviously in all FB group(P<0.01); and it is increased in 0.0001,0.0010 mg/L groups at 48 h (0.73±0.04, 0.64±0.14) and 0.0001 mg/L group at 72 h(0.70±0.17) in OB compared with the control group (0.32±0.04,0.27±0.05, P<0.01). Compared with the control group (0.95±0.11), RT-PCR revealed an increasing tendency of the expression of c-fos mRNA at 48 h in FB (1.06±0.16, 1.06±0.12,1.12±0.16,1.04±0.15,1.04±0.10,1.15±0.29), but the difference was not statistically significant(P>0.05); however, a statistically significant difference(P<0.01) of c-fos mRNA in 20.0000 mg/L group(1.40±0.17) in O B was found compared with the control group (1.06±0.06). Conclusion The higher expression of c-fos mRNA and protein in FB induced by fluoride may play an important role in the transformation of osteoblastic phenotype as well as increase the osteogenesis ability in FB.

Drugs, Chinese Herbal ; Medicine, Chinese Traditional ; Terminology as Topic ; Translations

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Eye ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Paralysis ; physiopathology ; therapy ; Trochlear Nerve Diseases ; physiopathology ; therapy ; Young Adult

Objective: To study the chemical constituents from Sinopodophyllum hexandrum and their antitumor activities. Methods: The constituents were separated by chromatography of silica gel, ODS, Sephadex LH20 and pre-TLC. Their structures were elucidated by spectroscopic means. The in vitro cytotoxic activities of the isolated compounds were studied by MTT method. Results: Nine compounds were isolated and identified as 8,2'-diprenylquercetin 3-methyl ether-4'-O-β-D-glucoside (1), 8,2'-diprenyl quercetin-3- methylether (2), 5,7,4'-trihydroxy-3'-(3-methylbut-2-enyl)-3-methoxy flavone (3), 8-prenylkaempferol (4), sophoflavescenol (5), podoverine A (6), sinoflavonoid K (7), diosmetin (8) and acacetin (9). Conclusion: Compound 1 is a new compound named sinoflavonoid glycosides A, and compounds 5-9 are isolated from S. hexandrum for the first time. Compounds 1-5 show cytotoxicities against HeLa cells with IC50 of 42.6, 46.9, 26.9, 16.1 and 31.2 μmol/L, respectively.

Objective To study the relationship of urine RBC morphology between UF-100 urine sediment analytic instrument andphase contrast microscope.Methods The UF-100 urine sediment analytic instrument to analyze 500 urine specimens and study the relation-ship of urine RBC morphology between urine sediment analytic instrument and phase contrast microscope.Results The according perceptionof Normocytic,Microcytic and Non-classified RBC between phase contrast microscope and UF-100 urine sediment analytic instrument RBC-info are 91.4%,94.4%,83.3% respectively,the according perception between phase contrast microscope and RBC-P70Fsc are 94.9%,95.7%,94.7% respectively,and the according perception between phase contrast microscope and RBC Fsc-DW are 84.4%,86.8%,90.5% respectively,the specificity of UF-100 and phase contrast microscope in glomerular hematuria and non-glomerular hematuria are84.3%,88.1% and 83.3%,87.9% respectively.Conclusion The results show that the UF-100 urine sediment analytic instrument issimply operating,fast and high accurate,and which can instruct clinical dignose,therapy and prognosis judgement.

A HPLC method of analysis of Monascus citrinin was established. More than 30 strains of Monascus spp. were cultured in steamed rice at solid state or in MSG liquid medium composed of monosodium glutamate as sole nitrogen source and glucose as sole carbon to investigate their ability of producing citrinin. The results indicated that most of the Monascus strains are able to produce citrinin. MSG medium can be used as a specific culture medium to qualitatively identify if the strain is the potential citrinin producer. But to confirm whether the Monascus strains are potential citrinin producers, these strains should be cultured in several cultivation methods, as the culture states and culture conditions influence the citrinin production greatly.

OTX1 gene is one of the pivotal transcriptional factors involved in the neurogenesis.In order to overexpress the OTX1 gene in distinct cell types and find out its contribution to the proliferation and differentiation of stem cells in vitro,OTX1 cDNA was subcloned into lentiviral vectors.The resulting constructions pDUETOTX1,pDUETGFPOTX1 and pDUETGFP were packaged in 293 cells producing viral particles to transduce 293T cells,SY5Y cells,mouse embryonic stem cells and E15 neural stem cells.It was proved that the transferred OTX1 gene was located in the nuclei of the transduced cells in stead of plasma.Lentivirus is an ideal vector delivering gene to different cells.The overexpression of OTX1 in transduced 293T cells were validated by Western blot and immunofluorescence.

Objective:To remove murine embryonic stem cells(mESC)from the differentiating cell culture and purify the differentiated cells by Magnetic Activated Cell Sorting(MACS).Methods:Neural differentiation of mESC was induced by a 5-stage method.The specific cell surface marker,SSEA-1,was used to identify ES cells in the differentiating cells.The optimal dilutions of mouse anti mouse SSEA-1 IgM primary antibody and FITC conjugated goat anti mouse secondary antibody were determined before the flow cytometry test.The incubation time and incubation temperature of primary antibody were all optimized to make the cytometry test accurate.After the optimization,stage 4 cells were dissociated into single cell suspension,incubated with antibody of SSEA-1 and microbeads conjugated goat anti mouse IgM,and then sorted through the magnetic field.The rate of SSEA-1 positive cells in pre-and post-separation groups was assessed by flow cytometry,and the viability of cells was evaluated by trypan blue staining counting under light microscopy.Results:The proportion of SSEA-1 positive cells in the separated cells can be reduced from(7.19?1.36)% to(1.34?0.80)%.The survival rate of sorted cells was more than 92%,similar to that of pre-separation cells.Conclusions:The MACS system we used can effectively remove mESC from the differentiated cells.The sorted cells will be well provided for the subsequent studies about transplantation therapy.

AIM To compare the contents of benzoyl mesaconitine,benzoyl hypaconitine,benzoyl aconitine,aconitine,hypaconitine and mesaconitine in Zingiberis Rhizoma Recens boiled juice-processed,Zingiberis Rhizoma boiled juice-processed,Zingiberis Rhizoma Recens juicing-processed,Zingiberis Rhizoma Recens slice malaxation steaming,and Zingiberis Rhizoma slice malaxation steaming Aconiti lateralis Radix Praeparata.METHODS Acid-base titration method was adopted in the content determination of total alkaloids.HPLC was applied to determining the contents of six alkaloids,the analysis was performed on a 35 ℃ thermostatic Waters Symmetry(C)C1scolumn (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.04 mol/L ammonium acetate flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 235 nm.RESULTS The contents of total alkaloids and diester-type alkaloids (mesaconitine,aconitine and hypaconitine)in various processed products,especially for Zingiberis Rhizoma Recens slice,Zingiberis Rhizoma slice malaxation steaming products,were obviously lower than those in the raw product.The contents of benzoyl hypaconitine and benzoyl aconitine in Zingiberis Rhizoma slice malaxation steaming product were the lowest,while that of benzoyl mesaconitine in the raw product was the lowest.CONCLUSION Compared with other Zingiber-processing methods,both Zingiberis Rhizoma Recens slice and Zingiberis Rhizoma slice malaxation steaming can reduce the toxicity of Aconiti lateralis Radix Praeparata more effectively.