Objective To investigate the effect of enamel matrix proteins (EMPs) on gene expression in human bone marrow stromal cells (hBMSCs) by cDNA microarray. Methods hBMSCs were cultured in vitro by whole bone marrow technique. Those stimulated with EMPs at a concentration of 200 ?g/mL in vitro were classified as test group, and those normally cultured were served as control. Afer culturing for 5 days, a cDNA microarray containing 4 096 genes was used to detect and analyze the gene expression. Four differentially-expressed genes in cDNA microarray were determined by real-time PCR for validation of the microarray data. Results Twenty-seven genes of hBMSCs were differentially expressed in the presence of EMPs, among which 18 were up-regulated and 9 down-regulated. The result of real-time PCR was in accordance with that of the cDNA microarray. Conclusion cDNA microarray technique can screen the differentially-expressed genes in hBMSCs treated with EMPs, which lays a foundation for the research of molecular mechanism of EMPs in inducing the regeneration of periodontal tissue.

American Heart Association ; Anti-Bacterial Agents ; administration & dosage ; therapeutic use ; Cardiac Surgical Procedures ; Child ; Drug Resistance, Bacterial ; Drug Therapy, Combination ; Endocarditis ; drug therapy ; surgery ; Endocarditis, Bacterial ; drug therapy ; surgery ; Humans ; Practice Guidelines as Topic ; United States

Child ; Congresses as Topic ; Coronary Disease ; diagnosis ; etiology ; therapy ; Humans ; Immunoglobulins, Intravenous ; administration & dosage ; therapeutic use ; Incidence ; Injections, Intravenous ; International Cooperation ; Mucocutaneous Lymph Node Syndrome ; epidemiology ; etiology ; therapy

Antigens, CD34 ; metabolism ; Craniotomy ; Diagnosis, Differential ; Follow-Up Studies ; Hemangiosarcoma ; metabolism ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Skull ; pathology ; surgery ; Skull Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

Anthracyclines ; administration & dosage ; adverse effects ; Antineoplastic Agents ; administration & dosage ; adverse effects ; Biomarkers ; blood ; Cardiomyopathies ; chemically induced ; diagnosis ; prevention & control ; Cardiotonic Agents ; therapeutic use ; Child ; Child, Preschool ; Echocardiography ; Heart ; drug effects ; Heart Diseases ; chemically induced ; diagnosis ; prevention & control ; Humans ; Risk Factors ; Survival Analysis ; Troponin I ; analysis

Objective: To establish the methods for extraction and isolation of punicalagin from pomegranate peel, and further study the purification and quantification of punicalagin. Methods: Using an ultrasonic-assisted extraction method, punicalagin in pomegranate peel was extracted at room temperature by 50% ethanol with 20-fold volume of raw material. The content of punicalagin in the crude extract was determined by HPLC. To optimize the purification process of punicalagin, static adsorption and desorption experi-ments were employed to study five kinds of macroporous adsorbent resins (D101, A8-8, NKA-9, HPD-100 and HPD-500) for the one with the highest purification efficacy of punicalagin. In addition, the technical parameters of the macroporous adsorbent resin were opti-mized to obtain punicalagin with higher purity. Punicalagin was further separated and purified by using a reverse phase MCI GEL CHP20P column. Results:HPD500 resin showed the best ability to absorb and separate punicalagin in among five kinds of macro-porous adsorbent resins. The best technical parameters were as follows:the mass concentration of sample solution was 15 mg·ml-1 , the loading amount was 2BV, the pH was 2 and the eluting solvent was 8BV of 30% ethanol. With the best process as described a-bove, the content of punicalagin extracted from pomegranate peel increased from 10. 3% to 30. 7%. The obtained punicalagin could be further purified to 61. 3% from 30% in ethanol eluate by the reverse phase MCI GEL CHP20P column. Conclusion:HPD500 resin is the most effective in the purification of punicalagin from pomegranate peel, and the content of punicalagin can be dramatically increased after the purification by a reverse phase MCI GEL CHP20P column. The optimized process shows good reproducibility and stability.

OBJECTIVETo determine the compound Daqiqi decoction( CDQD) combined cisplatin plays on the cell apoptosis of subcutaneous transplanted ovarian cancer in nude mice, to provide theory evidence for clinical treatment.

METHODMaking the models of subcutaneous transplanted ovarian cancer in nude mice, and divide the 40 mice with tumor into 5 groups (n = 8), the model control group, CDQD low dosed group, CDQD high dosed group, cisplatin group, cisplatin combined CDQD group, killing all the mice after 3 weeks' treatment and stripping tumors. Measure the volume and weight of the tumor and calculate tumor growth curve damps, the inhibition rate. Examining the expression of the Bcl-2(B-cell leukemia /lymphoma 2) and Bax (Bcl-2 associated x protein) mRNA and protein by the RT-PCR and the Western blot.

RESULT(O)The tumor weight shows that there was certain lighter effect in each CDQD group, and compared with cisplatin groups has no statistical significance, the cisplatin combined CDQD group is obviously lighter than that of the other group(P <0.01). The tumor growth curve damps shows that compared with model control group, the treatment group tumors had some extent narrowing (P < 0.01). (2) RT-PCR results: Bax (Bcl-2 associated x protein) mRNA expression shows that compared with model control group, the treatment group has increased (P < 0.01), and the cisplatin combined CDQD group compared with the other group is the highest, there was significant difference with the rest of the treatment group (P < 0.01). In the Bcl-2 mRNA express lowest in cisplatin combined CDQD group (P < 0.01), there has no difference between CDQD high dose group and cisplatin group. (Western blot shows: compared with model control group, the Bax protein of treatment group has increased expression (P < 0.01), the expression of Bax protein in cisplatin combined CDQD group is the highest(P < 0.01). There has no difference between CDQD high dose group and cisplatin group. The Bcl-2 protein expression of the cisplatin combined CDQD group is the lowest (P < 0.01), there has no difference between CDQD high dose group and cisplatin group.

CONCLUSIONThe effect of CDQD on subcutaneous transplantation ovarian tumor has promoting apoptosis function, its mechanism may be related to downgrade the Bcl-2 expression, higher expression of bax, stimulation on the apoptosis of tumor cells; cisplatin combined CDQD have synergistic effect.

Animals ; Cell Line, Tumor ; Cisplatin ; administration & dosage ; Disease Models, Animal ; Drug Therapy, Combination ; Female ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Ovarian Neoplasms ; drug therapy ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

By a orthogonal experiment, the influence of different ratio of phosphorus and potassium fertilizers on imperatorin, isoimperatorin and psoralen contents and yield of Glehnia littoralis were studied. The results showed that root dry weight and the yield of G. littoralis increased when reasonably applied phosphorus fertilizer combined with potassium fertilizer within a certain range. And the influence of phosphorus fertilizer was greater than that of potassium fertilizer. The optimal value of root dry weight and yield achieved at both P2O5 360 kg x hm(-2), K2O 270 kg x hm(-2) and P2O5 360 kg x hm(-2), K2O 180 kg x hm(-2). The effects of different phosphorus and potassium treatments on the content of imperatorin, isoimperatorin and psoralen in G. littoralis were determined, which shows that the content increased with the moderate increase of phosphorus and potassium. And the effects of phosphorus fertilizer were more significantly. The isoimperatorin content achieved the largest value at P2O5 360 kg x hm(-2), K2O 270 kg x hm(-2), also a larger content of imperatorin and psoralen. The imperatorin content is the largest when applied P2O5 360 kg x hm(-2), K2O 180 kg x hm(-2), and the isoimperatorin content was higher as well. So that the treatment of P2O5 360 kg x hm(-2), K2O 180 kg x hm(-2) are suitable for promote to the agricultural production, which could improve the quality and yield of G. littoralis.
Agriculture ; Apiaceae ; chemistry ; growth & development ; metabolism ; Coumarins ; analysis ; metabolism ; Drugs, Chinese Herbal ; analysis ; metabolism ; Fertilizers ; analysis ; Phosphorus ; analysis ; metabolism ; Potassium ; analysis ; metabolism

OBJECTIVETo observe features of Icariin in promoting osteogenic differentiation of SD rat bone marrow mesenchymal stem cells (BMSCs) in vitro.

METHODS(1) SD rats' BMSCs were isolated and purified by mechanically isolated and cultured by whole bone marrow adherent method. Effects of various concentrations Icariin on serum activities of alkaline phosphatase (ALP) were detected using amino antipyrine phenol determination method at day 3, 6, 9, 12, 15, 18, and 21. Calcium nodes of each groups were detected using alizarin red staining. Roles of various concentrations Icariin in promoting osteogenic differentiation of BMSCs were observed. (2) BMSCs were divided into the blank control group, the osteogenic induced group, and the Icariin group (0.5 microg/mL). ALP activities were detected at day 7, 14, and 21 of culture. Meanwhile, ALP positive staining rate and calcium nodes were detected at day 14 and 21 respectively. Additionally, mRNA expressions of Runt-related transcription factor-2 (Runx2) and Osteocalcin were detected at day 7, 14, and 21 by real-time fluorescent quantitative PCR.

RESULTS(1) 0.05-5.0 microg/mL Icariin could significantly elevate serum ALP activities. Of them, 0.2-2.0 microg/mL Icariin significantly increased calcium nodes numbers (P < 0.01). (2) When Icariin promoted osteogenic differentiation of BMSCs, Runx2 mRNA expression levels and ALP activities increased earlier and then decreased, while osteocalcin mRNA expression levels continued to increase (P < 0.01). Compared with the osteogenic induced group, ALP activities and ALP positive staining rate were both elevated after 14 days of Icariin treatment in the Icariin group (P < 0.05, P < 0.01).

CONCLUSIONSIcariin could promote the differentiation of BMSCs to osteoblasts by up-regulating Runx2 mRNA expression levels. It also could promote the mineralization by increasing ALP secretion and Osteocalcin mRNA expression levels, thereby promoting mature of newly generated osteoblasts.

Animals ; Bone Marrow Cells ; Cell Differentiation ; drug effects ; Flavonoids ; pharmacology ; Hematopoietic Stem Cells ; Mesenchymal Stromal Cells ; physiology ; Osteoblasts ; Osteocalcin ; Osteogenesis ; Rats ; Rats, Sprague-Dawley

Objective To establish an ultraviolet-irradiation damage model in cultured fibroblasts derived from human skin and to explore the potential protective effects and mechanisms of amyloid precursor protein 17-met peptide (APP17-mer peptide) on the oxidative damage and collagen metabolism in cultured fibroblasts after ultraviolet irradiation. Methods Human dermal fibroblast cultures were established by outgrowth from foreskin biopsies of a healthy donor and were irradiated by a single exposure to ultraviolet rays and cultured in a series of concentrations of APP17-mer peptide (0, 20, 40, 80 μmol/L).The activity of fibroblasts was detected by the assay of MTT. The intracellular ROS level was measured with a confocal microscope. The expression of MMP-1 mRNA was analyzed real-time quantitatively following RT-PCR. Results Primary cultures of human skin fibroblasts were established from human foreskin in DMEM supplemented with 10 % fetal bovine serum. UV irradiation depressed cellular activity and increased intracellular level of ROS (P<0.05). 40μmol/L and 80μmol/L APP17-mer peptide increased the cellular activity in both UV irradiated fibroblasts and unirradiated fibroblasts (P<0.05), however,20 μmol/L did not show such protective effects (P>0. 05). 40μmol/L APP17-mer peptide could depress the level of ROS in irradiated libroblasts. A single exposure of fibroblasts to UV irradiation resulted in 1.78 foldup-regulation of MMP-1 mRNA compared with unirradiated sample, 40μmol/L and 80μmol/L APP17-mer peptide decreased the expression of MMP-1 mRNA (P<0.05 and P<0.01, respectively).Conclusion APP17-mer peptide can enhance cellular activity under UV-induced oxidative stress and in-hibit collagen degradation in fibroblasts irradiated with ultraviolet rays. Inhibition of ROS production may be involved in the protective mechanism of APP17 peptide.