OBJECTIVETo observe the effect of total flavones of Epimedium leptorrhizum (YYH-C) on osteoporosis in ovariectomized rats.

METHODOvariectomized female rats were randomly divided into the model group, YYH-C lower, middle and high dose (0.7, 1.4, 2.8 g x kg(-1)) groups, the positive drug Bujiale (0.15 mg x kg(-1)) group, and the sham group. The rats were orally ad-ministrated with drugs for three months. Parathyroid hormone (PTH), procollagen I N-terminal peptide (PINP), alkaline phosphatase (ALP), calcium (Ca) and phosphrous (P) in serum were detected. Femur bones and vertebrae bones of left side were collected to determined bone metrological indexes, including bone mineral density (BMD), bone Ca, and bone ash weight/dry weight percentage. Femur bones of right side were collected to for a morphological observation of bone.

RESULTCompared with the sham group, the model group showed significantly higher PTH and ALP content but obviously lower PINP and Ca content. The three YYH-C 3 groups could resist the decrease of PINP. Specifically, low and middle dose groups could remarkably inhibit the increase of PTH, and the high dose group could increase the Ca content in serum, but without significant effect on the rise in ALP. There was no significant difference in P content in serum in each group. BMD, ash weight/dry weight percentage, Ca and P content of the model group were significantly lower than those in the sham group. The high dose YYH-C group could significantly increase BMD. All of the three YYH-C groups could notably increase ash weight/dry weight percentage and Ca, P content in femur bones and vertebrae bones. YYH-C could significantly increase average thickness, area, area percentage of bony trabeculae, cortical bone area percentage of femoral shaft and the number of osteoblasts on the surface of bony trabeculae, and decrease the number of osteoclasts.

CONCLUSIONYYH-C can effectively control the bone mass loss of rats with ovariectomy-induced osteoporosis, prevent the changes in bone microstructure, and inhibit bone absorption, so as to resist high turn-over osteoporosis after ovariectomy. [Key words] total flavones of Epimedium leptorrhizum; ovariectomized rat; osteoporosis

Alkaline Phosphatase ; metabolism ; Animals ; Bone Density ; drug effects ; Calcium ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Epimedium ; chemistry ; Female ; Flavones ; administration & dosage ; Humans ; Osteoporosis, Postmenopausal ; drug therapy ; metabolism ; physiopathology ; Ovariectomy ; Parathyroid Hormone ; metabolism ; Rats ; Rats, Sprague-Dawley

Cadherins ; metabolism ; Female ; Humans ; MCF-7 Cells ; Neoplastic Stem Cells ; metabolism ; Receptor, Notch1 ; metabolism ; Receptors, CXCR4 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor C ; metabolism

By studying the literatures on the adverse reaction of Yuxingcao injection, the clinical features of ADR were summed up, and the causes of ADR were analyzed through raw material, technology, chemical composition, compatibility and the clinical usage. The causes of ADR induced by Yuxingcao injection are complicated, maybe both due to the medicine itself and the incorrect clinical usage. Multi-measures including manufacture, techniques, clinical usage and some other cases should be taken to prevent the occurrence of adverse reaction of Yuxingcao injection.
Dosage Forms ; Drug Administration Schedule ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Humans ; Injections

OBJECTIVETo obtain the normal values of four-phase rhinomanometry specific parameters of normal adult Chinese and analyze the correlation between four-phase rhinomanometry and acoustic rhinometry measurement results.

METHODSEighty-five normal adults were recruited. The HRR2 four-phase rhinomanometry was used to acquire the effective resistances in inspiration, expiration and total breathing process (Reffin, Reffex, Refft) and vertex resistance in the process of inspiration and expiration (Vrin and Vrex). The Eccovision acoustic rhinometry was used to measure the minimum cross-sectional area (MCA) and the nasal volume of 0-5 cm nasal cavity (V5).

RESULTSReffin (x(-) +/- s) was (1.28 +/- 1.02) Pa/(cm(3).s) for male, (1.55 +/- 1.03) Pa/(cm(3).s) for female; Reflex (x(-) +/- s) was (1.43 +/- 1.07) Pa/(cm(3).s) for male, (1.75 +/- 1.14) Pa/(cm(3).s) for female; Refft (x(-) +/- s) was (1.34 +/- 0.99) Pa/(cm(3).s) for male, (1.62 +/- 1.03) Pa/(cm(3).s) for female; Vrin (x(-) +/- s) was (1.31 +/- 1.03) Pa/(cm(3).s) for male; (1.60 +/- 1.03) Pa/(cm(3).s) for female, Vrex (x(-) +/- s) was (1.46 +/- 1.04) Pa/(cm(3).s) for male, (1.82 +/- 1.17) Pa/(cm(3).s) for female. No statistically significant difference was found between men and women (r = 0.661, -0.397, 0.127, 0.649, -0.684, P > 0.05, respectively). There was no significant correlation between Reffin, Reflex, Refft, Vrin, Vrex and age, height, weight, head circumference, body surface area, body mass index (P > 0.05, respectively). However, there was significant correlation between Reffin, Reflex, Refft, Vrin, Vrex and MCA, V5 (P < 0.05, respectively).

CONCLUSIONSThe results of four-phase rhinomanometry show significant correlation to acoustic rhinometry.

Adolescent ; Adult ; Airway Resistance ; physiology ; Female ; Humans ; Male ; Middle Aged ; Nasal Cavity ; physiology ; Reference Values ; Respiration ; Rhinomanometry ; methods ; Rhinometry, Acoustic ; methods ; Young Adult

OBJECTIVETo investigate the effect of cytochrome P450 isozymes on aristolochic acid induced cytotoxicity on renal proximal tubular epithelial cell (cell line HK-2).

METHODHuman renal tubular cells (cell line HK-2), were treated with aristolochic acid (AA) alone or in combination with cytochrome P450 isozymes inhibitors, including alpha-naphthoflavone (CYP450 1A1 and 1 A2 inhibitors), ketoconazole (CYP450 3A4 inhibitor), sodium diethyldithiocarbamate (CYP450 2A6 and 2E1 inhibitors), quinidine (CYP450 2D inhibitor), alpha-lipoic acid (NADPH: P450 reductase inhibitor), sulfaphenazole (CYP450 2C inhibitor) in the presence or absence of liver microsome(S9). The inhibition of cell proliferation rate was studied by MTT assay and the lactate dehydrogenase release rate was determined with continuous monitoring method.

RESULTAA inhibits cell proliferation and promotes the release of LDH over the range of 12.5-100 mg x L(-1), in a dose-dependent manner. Addition of S9 into the culture system reduced AA cytotoxicity, with the cell proliferation inhibition reducing and the release of LDH decreasing (AA + S9 group vs the same concentration of AA alone group, P < 0.05). In the absence of S9, ketoconazole or alpha-naphthoflavone has no obvious effect on AA cytotoxicity, however,under the conditions of adding S9, ketoconazole or alpha-naphthoflavone enhances AA cytotoxicity. Other inhibitors of CYP450 isozymes has no distinct effect on AA cytotoxicity.

CONCLUSIONThe microsomal enzyme of Liver can reduce the AA cytotoxicity, and CYP450 3A, CYP450 1A may be the major cytochrome P450 isozymes which impact AA cytotoxicity.

Animals ; Aristolochic Acids ; toxicity ; Cell Line ; Cell Proliferation ; drug effects ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; metabolism ; Cytotoxicity, Immunologic ; drug effects ; Enzyme Inhibitors ; pharmacology ; Humans ; Kidney Tubules ; drug effects ; enzymology ; immunology ; Male ; Rats ; Rats, Wistar

OBJECTIVETo study the clinical and molecular epidemiology characteristics of human Bocavirus 1-3 (HBoV1-3) in children for acute respiratory infection in Lanzhou area.

METHODSNasopharyngeal aspiration samples and throat swabs were collected from 524 children with ARTI at the First Hospital of Lanzhou University, Gansu Province, China, between December 2009 and November 2010. Nested PCR was employed to screening HBoV1-3, which amplified a 518-bp fragment of the partial NS1 gene. Furthermore, a standard reverse transcription-PCR was used to screen for other common respiratory viruses.

RESULTSThe overall frequency of HBoV was 8.2% (43/524), lining up behind human rhinovirus, RSV, parainfluenza virus 3. Thirty of the HBoV-postive children(69.8%) were co-infected with other respiratory viruses. The prevalence of HBoV1 in ALRTI was obviously higher than that in AURI. The 2 HBoV2 NS1 sequences shared 99% and 100% nucleotide sequence identity with HBoV2 strain CU47TH respectively. Two cases of HBoV2 postive children appears gastrointestinal symptoms. The one HBoV3 NS1 sequences shared 99% nucleotide sequence identity with HBoV3 isolate 46-BJ07.

CONCLUSIONThe HBoV3 was detected at the first time in lanzhou area. HBoV1-3 infection exists in children with acute respiratory tract infections in Lanzhou region, HBoV1 were dominant. The mixed infection rate was higher.

Acute Disease ; China ; Female ; Human bocavirus ; classification ; genetics ; isolation & purification ; Humans ; Infant ; Male ; Phylogeny ; Respiratory Tract Infections ; virology

A recombinant plasmid pET28a-HBcAg-delta n was constructed, in which three mimic B-epitopes of HER family were inserted into the truncated HBc vector. The fusion protein expressed was purified and used to immunize BALB/c mice to induce antibody against the epitopes. Three mimic epitope genes were inserted into the sequences of amino acid residues 78 and 79 of HBcAg by overlap PCR. The PCR product was then cloned into pET28a to construct recombinant expression plasmid which was transformed to E. coli BL21 (DE3) and induced by IPTG. After purification, the fused protein designed HBHE was used to immunize BALB/c mice to detect humoral immunoresponse. The recombinant plasmid was successfully constructed by DNA sequencing analysis. A fusion protein with correct molecular mass was expressed and confirmed by SDS-PAGE. High titre antibody was elicited in the mice immunized with HBHE by indirect ELISA and Western blotting. The HBc particle vector containing three B-epitopes of HER family had been successfully prepared, purified and high titre antibody against HBHE was detected. All these data are helpful in further research of the broad-spectrum anti-tumour effect of combine polypeptide epi-position vaccine of EGFR and HER2.
Animals ; Cancer Vaccines ; immunology ; Cell Line, Tumor ; Epitopes ; immunology ; Genetic Vectors ; Hepatitis B Core Antigens ; genetics ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Plasmids ; Random Allocation ; Receptor, Epidermal Growth Factor ; genetics ; immunology ; Receptor, ErbB-2 ; genetics ; immunology ; Recombinant Fusion Proteins ; genetics ; immunology ; Vaccination ; methods ; Vaccines, Combined ; immunology

OBJECTIVEAlthough chondroprotective activities have been documented for polysaccharides, the potential target of different polysaccharide may differ. The study was aimed to explore the effect of glucan HBP-A in chondrocyte monolayer culture and chondrocytes-alginate hydrogel constructs in vivo, especially on the expression of type II collagen.

METHODSChondrocytes isolated from rabbit articular cartilage were cultured and verified by immunocytochemical staining of type II collagen. Chondrocyte viability was assessed after being treated with HBP-A in different concentrations. Morphological status of chondrocytes-alginate hydrogel constructs in vitro was observed by scanning electron microscope (SEM). The constructs were treated with HBP-A and then injected to nude mice subcutaneously. Six weeks after transplantation, the specimens were observed through transmission electron microscopy (TEM). The mRNA expressions of disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTs-5), aggrecan and type II collagen in both monolayer culture and constructs were determined by real time polymerase chain reaction (PCR). The expression of type II collagen and matrix metalloproteinases-3 (MMP-3) in chondrocyte monolayer culture was also tested through Western blot and enzyme linked immunosorbent assay (ELISA), respectively.

RESULTSMMP-3 secretion and ADAMTs-5 mRNA expression in vitro were inhibited by HBP-A at 0.3 mg/mL concentration. In morphological study, there were significant appearance of collagen in those constructs treated by HBP-A. Accordingly, in both chondrocyte monolayer culture and chondrocytes-alginate hydrogel constructs, the expression of type II collagen was increased significantly in HBP-A group when compared with control group (P<0.001).

CONCLUSIONSThe study documented that the potential pharmacological target of glucan HBP-A in chondrocytes monolayer culture and tissue engineered cartilage in vivo may be concerned with the inhibition of catabolic enzymes MMP-3, ADAMTs-5, and increasing of type II collagen expression.

ADAM Proteins ; genetics ; metabolism ; Aggrecans ; genetics ; metabolism ; Alginates ; pharmacology ; Animals ; Cartilage, Articular ; drug effects ; physiology ; Cell Proliferation ; drug effects ; Cell Shape ; drug effects ; Cell Survival ; drug effects ; Chondrocytes ; cytology ; drug effects ; metabolism ; ultrastructure ; Collagen Type II ; genetics ; metabolism ; Female ; Glucans ; pharmacology ; Glucuronic Acid ; pharmacology ; Hexuronic Acids ; pharmacology ; Hydrogel, Polyethylene Glycol Dimethacrylate ; pharmacology ; Immunohistochemistry ; Matrix Metalloproteinase 3 ; metabolism ; Mice, Nude ; RNA, Messenger ; genetics ; metabolism ; Rabbits ; Tissue Engineering ; methods

OBJECTIVETo explore a simple, rapid and efficient way to generate dendritic cells from leukemic cells.

METHODSK562 cells were cultured with calcium ionosphere A23187 alone, A23187 plus GM-CSF, or a DC differentiation cocktail consisting of GM-CSF, IL-4 and TNF-alpha, respectively. The expression of surface markers of induced DCs was analyzed by flow cytometry. The K562-DCs stimulating the proliferation of allo-genetic naive T cells and inducing cytotoxicity of T cells were determined by MTT assay.

RESULTSMicroscopic examination revealed that under all the three culture conditions, K562 cells became displaying DC morphology. At 72 hours in the two culture systems containing A23187, there were higher proportions of cells with dendritic morphology [(69.5 +/- 17.2)% and (73.1 +/- 13.9)%, respectively] than that in the cocktail system [(28.5 +/- 12.3)%] (P < 0.05). And the same did when cultured for 7 days [(69.5 +/- 17.2)%, (73.1 +/- 13.9)% respectively vs (51.2 +/- 10.7)%, P < 0.05]. In the 7-day cultures, the percentage of CD1a expressing cells was lower [(8.2 +/- 2.3)% and (10.3 +/- 5.1)% vs (17.2 +/- 1.6)%, respectively] while the CD83 expressing cells was higher [(85.6 +/- 8.8)% and (82.4 +/- 9.1)% vs (77.4 +/- 12.9)%, respectively] compared with that in the cocktail system (P < 0.05). No significant difference was found in the allogeneic T cell proliferation response and induced T cell cytotoxicity between A23187 containing and cocktail groups (P > 0.05).

CONCLUSIONSA23187 treatment is a simple, rapid and efficient in vitro strategy for inducing dendritic cell from leukemic cells.

Calcimycin ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Coculture Techniques ; Dendritic Cells ; cytology ; immunology ; metabolism ; Humans ; K562 Cells ; cytology ; drug effects

OBJECTIVETo investigate the effects of liver X receptor (LXR) agonist on adipose-derived mesenchymal stem cells (AD-MSCs) implantation into infarcted hearts of mice.

METHODAD-MSC(Fluc+) which stably expressed firefly luciferase (Fluc) were isolated from β-actin-Fluc transgenic mice and characterized by flow cytometry. Male FVB mice were randomly allocated into the following four groups (n = 10 each): (1) sham group; (2) MI + PBS group; (3) MI + AD-MSC(Fluc+) group; (4) MI + AD-MSC(Fluc+) + LXR agonist (T0901317) group. AD-MSC(Fluc+) or PBS were injected intramyocardial into peri-infarcted region of mice heart after permanent left anterior descending (LAD) artery ligation. Bioluminescence imaging (BLI) was performed for quantification of injected cells retention and survival. Cardiac function was evaluated by echocardiography.

RESULTSThe AD-MSC(Fluc+) were positive for CD44 and CD90 by flow cytometry. BLI evidenced the firefly luciferase expression of AD-MSC(Fluc+) which was positively correlated with cell numbers (r(2) = 0.98). The results of BLI in vivo revealed that LXR agonist could improve the survival of AD-MSC(Fluc+) at day 7, 14 and 21 after transplantation compared with AD-MSC(Fluc+) alone group. Cardiac function was further improved in combination therapy group compared with AD-MSC(Fluc+) alone group (P < 0.05).

CONCLUSIONSLXR agonist T0901317 can improve the retention and survival of intramyocardial injected AD-MSC(Fluc+) post-MI, and the combination therapy of T0901317 and AD-MSC(Fluc+) has a synergetic effect on improving cardiac function in this model.

Animals ; Hydrocarbons, Fluorinated ; therapeutic use ; Liver X Receptors ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Mice ; Mice, Transgenic ; Myocardial Infarction ; mortality ; surgery ; Orphan Nuclear Receptors ; agonists ; Sulfonamides ; therapeutic use ; Treatment Outcome