OBJECTIVETo inspect the quality of commercially available Chaihu (Bupluri Radix, a common traditional Chinese herbal drug) in Guangzhou by determining the content of saikosaponin A (SSa) using enzyme-linked immunosorbent assay (ELISA).

METHODSA competitive ELISA system using mouse anti-saikosaponin A monoclonal antibody was established for determining SSa content. Commercial samples of Chaihu were obtained from 10 drug stores in Guangzhou, and SSa contents in the methanol extracts of these samples were determined using the ELISA system.

RESULTSThe detection range of this competitive assay was 0.16-2.5 µg/ml for determining SSa contents. In the 10 commercial Chaihu samples, SSa contents in the methanol extract determined by this method ranged from 0.32 µg/mg to 6.87 µg/mg, and 3 samples showed a SSa content lower than the minimum requirement documented in the Chinese Pharmacopeia.

CONCLUSIONThis competitive ELISA is sensitive, rapid, economic and environment-friendly for SSa determination, especially suitable for batch determination. The results of SSa detection for the commercial Chaihu samples demonstrate an uneven quality of Chaihu in Guangzhou market, suggesting the necessity of more rigorous quality control measures for this drug.

Antibodies, Monoclonal ; Bupleurum ; chemistry ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Oleanolic Acid ; analogs & derivatives ; analysis ; Quality Control ; Saponins ; analysis

AIM: To investigate the status of myopia in students from different kinds of schools in Shantou city, China.
METHODS: In this cross-sectional study, 2 418 students from 4 middle and high schools in Shantou were surveyed through randomized cluster sampling. Ophthalmic examination and auto-refraction were performed for all subjects.
RESULTS: Among all 2 394 students examined. The myopia rates for students from middle and high school were 48. 07% and 68. 28%. In normal middle school, students from rural area had lower rate of myopia than students from the urban area (χ2 = 13. 879, P<0. 01 ). Students from key rural high schools had lower rate of myopia than students from key urban high school (χ2 =175. 231, P<0. 01).
CONCLUSION:The prevalence of myopia in the middle school of urban area in Shantou was higher than rural area. The increasing rate of myopia in youngsters is correlated with study stress. In order to decrease the myopia rate, it is extremely important to improve study environment in students.

BACKGROUNDOxidative stress and inflammation are important steps in the pathogenesis of atherosclerosis. We postulated that therapeutic concentrations of aspirin and pravastatin, especially in combination, may suppress oxidative stress and inflammation in endothelial cells, and this concept was examined in human coronary artery endothelial cells (HCAECs).

METHODSHuman coronary artery endothelial cells were cultured and treated with oxidized-low density lipoprotein (ox-LDL, 60 microg/ml for 24 hours) alone, or pre-treated with aspirin (1, 2 or 5 mmol/L), pravastatin (1, 5 or 10 micromol/L) or their combination (1 mmol/L aspirin and 5 micromol/L pravastatin), followed by ox-LDL treatment. After respective treatment, superoxide anion production, p38 mitogen activated protein kinase and transcription factor NF-kappaB activation, protein expression of lectin-like ox-LDL receptor-1 (LOX-1) and adhesion molecules, and monocyte adhesion were measured.

RESULTSOx-LDL treatment greatly elicited its receptor LOX-1 expression, superoxide anion production and inflammatory response, which were minimally affected by low concentration of aspirin (1 mmol/L) or pravastatin (5 micromol/L), but were markedly decreased by their combination. Activation of p38 mitogen activated protein kinase and NF-kappaB, the expression of intercellular adhesion molecule-1 and monocyte chemotactic protein-1, which were only mildly affected by aspirin or pravastatin alone, were significantly attenuated by their combination. As a consequence, monocyte adhesion to endothelial cells was markedly attenuated by the combination of the two agents. Well-known anti-oxidants alpha-tocopherol and gamma-tocopherol had similar inhibitory effects on ox-LDL-mediated oxidative stress and LOX-1 expression as well as monocyte adhesion as did the combination of aspirin and pravastatin.

CONCLUSIONSThese studies point to a positive interaction between aspirin and pravastatin with regard to endothelial biology. Anti-oxidant and subsequent anti-inflammatory effect may be one of the potential underling mechanisms.

Anticholesteremic Agents ; pharmacology ; Aspirin ; pharmacology ; Blotting, Western ; Cell Adhesion Molecules ; metabolism ; Cells, Cultured ; Coronary Vessels ; cytology ; Cyclooxygenase Inhibitors ; pharmacology ; Electrophoretic Mobility Shift Assay ; Endothelial Cells ; drug effects ; metabolism ; Humans ; Oxidative Stress ; drug effects ; Pravastatin ; pharmacology ; Scavenger Receptors, Class E ; metabolism ; Superoxides ; metabolism

Objective To establish an isolated rat pancreas perfusion technique,a method for the precise measurement of insulin secretion in vitro.Methods An isolated rat pancreas perfusion technique was applied in the study of insulin secretion from?-cells in 10 high-fat diet-induced obese Wistar rats.Results For the assessment of the functional integrity of the perfused pancreas,the isolated pancreas of 6 rats met all the criteria: (1)The constancy of perfusion pressure was kept over the whole experiment time[(70?5)mm Hg,1 mm Hg= 0.133 kPa].(2)The duodenal peristaltic activity of isolated pancreas and duodenum block was present after perfusion experiment.(3)Total insulin response to arginine stimulation was significantly increased as compared with glucose stimulation[maximum insulin secretion rate:(987?100)?U/min vs(545?50)?U/min,P

OBJECTIVETo study the correlation between magnetic resonance imaging (MRI) findings and proliferating cell nuclear antigen (PCNA) expression in peripheral lung cancer.

METHODSThe expression of PCNA was detected by means of SABC immunohistochemistry in 45 cases of surgically and pathologically confirmed peripheral lung cancer. The correlation between PCNA expression in the tumors and the MRI findings was analyzed.

RESULTSPCNA expression was correlated to the differentiation, tumor size, lobulation, and mediastinal lymph node metastasis of the tumors (P<0.05), but not to the histological type, clinical stage, pleural retraction, spiculation, or signal feature.

CONCLUSIONCorrelations are found between MRI findings of lung cancer and abnormal expression of PCNA.

Adenocarcinoma ; pathology ; Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; pathology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Proliferating Cell Nuclear Antigen ; metabolism

Objective: To observe the effect of emodin on the biological behavior of human fibroblasts (FB) in culture of kidney in patients with lupus nephritis (LN). Methods: FB were isolated from kidney culture of LN patients, and the effect of emodin on 3　H-TdR incorporated rate of FB was observed. The apoptosis and c-myc gene expression were detected in the same way by flow cytometry. Results: Emodin could markedly inhibit the proliferation of human kidney FB, and inducing cell apoptosis through up-regulating c-myc gene expression in human renal FB. Conclusion: Emodin can inhibit proliferation and promote apoptosis of FB, which may be important in ameliorating interstitial fibrosis, and thus improve prognosis of LN.

OBJECTIVETo explore an effective method to culture oral mucosa epithelial cells (OMECs) of canine in vitro, and to observe the biological characteristics of OMECs growing on small intestinal submucosa (SIS) in order to provide the experimental basis for epithelium tissue engineering.

METHODSThe primary OMECs were cultivated with DKSFM (defined keratinocyte serum free medium) containing 6% fetal bovine serum (FBS). The morphological characteristics and the growth curve of OMECs were observed. The expressions of OMECs marker (CK19) were examined by immunocytochemistry. The 2nd passage of OMECs were seeded on SIS, OMECs co-cultured with SIS were observed by hematoxylin-eosin staining, immunohistochemical staining, and scanning electron microscope (SEM).

RESULTSOMECs were grown well in DKSFM. Immunohistochemical staining of the 2nd passage cultured canine OMECs with broadly reacting anti-cytokeratin anyibodies (CK19) was positive. OMECs formed a single layer on the surface of SIS, and eight days later the cells were polygong and arranged like slabstone.

CONCLUSIONCulture of canine OMECs in DKSFM containing 6% FBS is a simple and feasible method. SIS has good biocompatibility, it is a kind of good bioscafold in the tissue-engineered epithelium.

Animals ; Cattle ; Cell Culture Techniques ; Cells, Cultured ; Coculture Techniques ; Epithelial Cells ; In Vitro Techniques ; Intestine, Small ; Mouth Mucosa ; Tissue Engineering

BACKGROUNDThis study was designed to investigate the potential pathogenicity of Mycoplasma penetrans (M. penetrans) and its molecular mechanisms responsible for the induction of iNOS gene expression in mouse macrophages stimulated by lipid-associated membrane proteins (LAMPs) prepared from M. penetrans.

METHODSMouse macrophages were stimulated with M. penetrans LAMPs to assay the production of nitric oxide (NO). The expression of inducible nitric oxide synthase (iNOS) was detected by RT-PCR and Western blotting. The activity of nuclear factor kappaB (NF-kappaB) and the effects of pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-kappaB, on the production of nitric oxide and the expression of iNOS were also assessed in mouse macrophages treated with M. penetrans LAMPs by indirect immunofluorescence and Western blotting.

RESULTSM. penetrans LAMPs stimulated mouse macrophages to produce nitric oxide in a dose- and time-dependent manner. The mRNA and protein levels of iNOS were also upregulated in response to LAMP stimulation and inhibited by PDTC treatment. M. penetrans LAMPs were found to trigger NF-kappaB activation, a possible mechanism for the induction of iNOS expression.

CONCLUSIONThis study demonstrated that M. penetrans may be an important etiological factor of certain diseases due to the ability of M. penetrans LAMPs to stimulate the expression of iNOS, which is probably mediated through the activation of NF-kappaB.

Animals ; Bacterial Proteins ; pharmacology ; Cells, Cultured ; Enzyme Induction ; Lipoproteins ; pharmacology ; Membrane Proteins ; pharmacology ; Mice ; Mycoplasma penetrans ; chemistry ; NF-kappa B ; metabolism ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase ; biosynthesis ; Nitric Oxide Synthase Type II ; RNA, Messenger ; analysis

OBJECTIVETo observe the hair development after subcutaneous implantation of embryonic skin cells in nude mice, and construct a model of hair development.

METHODSDermal and epidermal cells isolated from embryonic mouse skin were mixed at a given ratio and injected subcutaneously in nude mice, and hair formation after the implantation was observed under a microscope.

RESULTSFormation of the hair follicles and fibers under the skin of the recipient nude mice and emergence of the hair shaft were observed microscopically.

CONCLUSIONEmbryonic skin cells be used to construct a complete model of hair development after implantation in vivo.

Animals ; Cell Transplantation ; Cells, Cultured ; Dermis ; cytology ; transplantation ; Embryo, Mammalian ; Epidermis ; cytology ; transplantation ; Hair ; growth & development ; Mice ; Mice, Inbred C57BL ; Mice, Nude ; Models, Animal ; Skin ; cytology