Integrative Medicine ; education ; Internal Medicine ; education

Adult ; Cranial Fossa, Middle ; anatomy & histology ; surgery ; Ear, Inner ; anatomy & histology ; surgery ; Facial Nerve ; anatomy & histology ; surgery ; Humans ; Male

Child ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell

Altitude ; Animals ; Arvicolinae ; Gene Expression ; Hypoxia ; Rats ; Tumor Suppressor Protein p53 ; genetics

OBJECTIVETo explore the effect of total flavonoids of Herba Epimedium (FHE) on BMP-2/RunX2/OSX signaling pathway in promoting osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs).

METHODSPassage 3 BMSCs were randomly divided into the control group, the experimental group, and the inhibitor group. BMSCs in the control group were cultured in 0.2% dimethyl sulfoxide + Osteogenuxic Supplement (OS) fluid + DMEM/F12 culture media. BMSCs in the experimental group were intervened by 20 microg/mL FHE. BMSCs in the inhibitor group were intervened by 20 microg/mL FHE and 1 microg/mL NOGGIN recombinant protein. At day 9 alkaline phosphatase (ALP) activity was measured. Calcium nodules were stained by alizarin red staining and the density was observed. The transcription expression of osteogenic differentiation-related proteins (type I collagen, osteocalcin, and osteopontin) and related factors of BMP-2/RunX2/OSX signaling pathway was assayed by RT-PCR.

RESULTSCompared with the control group, ALP activities were enhanced and the density of calcium nodules significantly increased; type I collagen, osteocalcin, and osteopontin expression levels were increased in the experimental group. The expression of osteogenesis-related transcription factor was also increased in the experimental group. Noggin recombinant protein inhibited FHE promoting BMSCs osteogenesis in the inhibitor group. Compared with the experimental group, ALP activity decreased (P < 0.05), the density of calcium nodules was lowered, expression levels of type I collagen, osteocalcin, osteopontin significantly decreased (P < 0.05) in the inhibitor group.

CONCLUSION20 microg/mL FHE promoted osteogenic differentiation process of BMSCs by BMP-2/RunX2/OSX signaling pathway.

Bone Morphogenetic Protein 2 ; metabolism ; Cell Differentiation ; drug effects ; Cells, Cultured ; Collagen Type I ; metabolism ; Core Binding Factor Alpha 1 Subunit ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Epimedium ; chemistry ; Flavonoids ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Osteocalcin ; metabolism ; Osteogenesis ; drug effects ; Osteopontin ; metabolism ; Signal Transduction ; Sp7 Transcription Factor ; Transcription Factors ; metabolism

To examine the effects of heterologous expression of ZrGPD1 (encoding glycerol 3-phosphate dehydrogenase ) cloned from osmotolerant yeast Zygosacharomyces rouxii on glycerol production in wild Pichia farinosa,the URA3 gene was amplified from P. farinosa as the homology integrative region. A recombinant plasmid (pUR-ZG) was constructed then transformed into P. farinosa by electroporation. The transformant pfa-gu was obtained by the selectable marker Zeocin TM . Primary results showed that the biomass of pfa-gu was higher than the wild type in the flask and after 72h fermentation the concentration of glycerol of pfa-gu was 37g/L enhanced 30% in comparison with the wild type. It is concluded that heterologous expression of ZrGPD1 is useful for increasing glycerol production and the ability of osmoregulation in P. farinosa.

Objective To induce mouse embryonic stem(ES)cells to differentiate into insulin-secreting cells by means of a 5-step model system.Methods E14.1 mouse ES cells were cultured in the presence of leukemia inhibitory factor(LIF)for 2 days(step 1),then the cells were cultured in hanging drops to form embryonic bodies(EBs)and the resulting EBs were cultured in suspension for 6 days in the presence of basic fibroblast growth factor bFGF(step 2).Subsequently the EBs were cultured in the medium containing glucagon- like peptide 1(GLP-1),hepatocyte growth factor(HGF),nerve growth factor(NGF)and nicotinamide for 10 days(step 3).After that,the EBs were dissociated into single cells,and the cells were cultured in monolayer in the presence of GLP-1,betacellulin,activin A,bFGF and nicotinamide for 10 days(step 4).Finally,the cells were cultured in low-glucose medium containing nicotinamide for 4 days(step 5).Insulin and some other islet- related genes expressions were investigated using RT-PCR and insulin expression was also investigated by DTZ- staining and immunohistochemistry.The percentage of insulin-secreting cells was evaluated by flowcytometry and insulin concentrations were measured by RIA.Results mRNA expression of insulin became visible at step 3 and more evident at step 5.Additionally,at step 5,mRNAs of glucagon,somatostatin,pancreatic polypeptide(PP), pancreatic duodenal homeobox 1(PDX-1),beta-cell E box transactivator 2(Beta2)and neurogenin 3(Ngn3) were detected.DTZ-staining positive cells and insulin immunohistochemical staining positive cells were observed. The percentage of insulin-positive cells was(24.0?2.5)%(n=6).In the presence of 5.6 mmol/L and 25 mmol/L glucose,insulin concentrations were(0.05?0.01)?g/L and(0.13?0.02)?g/L respectively(n= 6).Conclusion E14.1 mouse ES cells can be induced to differentiate into insulin-secreting cells by the 5-step model system.Insulin-secreting cells can release insulin into culture medium when treated with glucose,and insulin concentrations increase with rising concentration of glucose.

Objective To determine whether Fudenine is a novel membrane protein. Methods Green fluorescence protein(GFP)was used to localize Fudenine in vivo. GFP, as a control, was targeted to cytoplasm.Epithelial cell, CBRH7919, and non-epithelial cell, L-6TG, were cultured and transiently transfected by using the lipofectamine reagent. After 48 h, intact cells were examined with fluorescence microscope for Fudenine. Results Reporter plasmid pEGFP-N1, as a control , was expressed and localized to cytoplasm. But Fudenine, driven by the cytomegalovirus promoterenhancer contained in the pEGFP-N1 vector, was overexpressed and targeted to cellular membrane. Conclusions Fudenine is a novel membrane protein. It may play the similar role with its homologues AC133 antigen and prominin in human and mouse.respectively. It might be involved in signaling transduction and regulate blood glucose metabolism in vivo.

OBJECTIVETo evaluate the efficacy and safety of Fulu Baoxinping (FLBXP) oral liquid in the treatment of coronary heart disease patients with premature ventricular beat (PVB), differentiated as qi-yin deficiency with phlegm-stasis syndrome type.

METHODSAdopting randomized, double-blinded, double-simulated, positive drug parallel controlled and multi-centered clinical research method, 240 patients enrolled were randomly assigned equally to the treatment group treated with FLBXP 10 mL (containing 13.33 g of crude drug) thrice a day and the control group treated with Wenxin Granule 9 g thrice a day. Meanwhile, simulator of the test or positive control drug was given to them all correspondingly. The therapeutic course for them all was 28 days. Efficacy on PVB and TCM syndrome was observed.

RESULTSThe markedly effective rate and total effective rate on PVB were 55.0% and 78.4% in the treatment group, and 37.2% and 53.1% in the control group, significant difference between groups was shown in comparison of both indexes (P < 0.05). Dynamic ECG showed the total number of PVB decreased for 3460.59 +/- 6516.56 beats/24 h in the treatment group, and for 2148.36 +/- 5129.47 beats/24 h in the control group, difference between them showed no statistical significance (P > 0.05). The TCM syndrome score in both groups was markedly decreased after treatment when compared with before treatment (P < 0.01); the differences of the treated and the control groups were -9.34 +/- 4.21 and -8.08 +/- 4.33 respectively, showing sigificant difference (P < 0.05).

CONCLUSIONFLBXP oral liquid has certain effect on PVB in CHD patients of qi-yin deficiency with phlegm-stasis syndrome type, no obvious adverse reaction was found in the clinical trial.

Administration, Oral ; Adult ; Aged ; Coronary Disease ; complications ; drug therapy ; physiopathology ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Electrocardiography ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Ventricular Premature Complexes ; complications ; drug therapy ; physiopathology

OBJECTIVETo explore the roles of granulocyte colony-stimulating factor in the pathogenesis of moyamoya disease.

METHODSSerum G-CSF concentrations were measured using enzyme linked immunosorbent assay (ELISA) in 20 children with moyamoya disease and 20 healthy children.

RESULTSSerum G-CSF concentration (35.7+/-10.3 pg/mL) in children with moyamoya disease was significantly higher than that in healthy controls (23.5+/-3.8 pg/mL) (p<0.01).

CONCLUSIONSThe elevated serum G-CSF concentration in children with moyamoya disease suggests that G-CSF may play an important role in the pathogenesis of moyamoya disease.

Child ; Child, Preschool ; Female ; Granulocyte Colony-Stimulating Factor ; blood ; physiology ; Humans ; Male ; Moyamoya Disease ; blood ; etiology ; Vascular Endothelial Growth Factor A ; analysis ; physiology