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

OBJECTIVETo study the transfer of paralytic shellfish toxins (PST) using four simulated marine food chains: dinoflagellate Alexandrium tamarense --> Artemia Artemia salina --> Mysid shrimp Neomysis awatschensis; A. tamarense --> N. awatschensis; A. tamarense --> A. salina --> Perch Lateolabrax japonicus; and A. tamarense --> L. japonicus.

METHODSThe ingestion of A. tamarense, a producer of PST, by L. japonicus, N. awatschensis, and A. salina was first confirmed by microscopic observation of A. tamarense cells in the intestine samples of the three different organisms, and by the analysis of Chl.a levels in the samples. Toxin accumulation in L. japonicus and N. awatschensis directly from the feeding on A. tamarense or indirectly through the vector of A. salina was then studied. The toxicity of samples was measured using the AOAC mouse bioassay method, and the toxin content and profile of A. tamarense were analyzed by the HPLC method.

RESULTSBoth A. salina and N. awatschensis could ingest A. tamarense cells. However, the ingestion capability of A. salina exceeded that of N. awatschensis. After the exposure to the culture of A. tamarense (2000 cells x mL(-1)) for 70 minutes, the content of Chl.a in A. salina and N. awatschensis reached 0.87 and 0.024 microg x mg(-1), respectively. Besides, A. tamarense cells existed in the intestines of L. japonicus, N. awatschensis and A. salina by microscopic observation. Therefore, the three organisms could ingest A. tamarense cells directly. A. salina could accumulate high content of PST, and the toxicity of A. salina in samples collected on days 1, 4, and 5 of the experiment was 2.18, 2.6, and 2.1 MU x g(-1), respectively. All extracts from the samples could lead to death of tested mice within 7 minutes, and the toxin content in artemia sample collected on the 1st day was estimated to be 1.65 x 10(-5) microg STX equal/individual. Toxin accumulation in L. japonicus and N. awatschensis directly from the feeding on A. tamarense or indirectly from the vector of A. salina was also studied. The mice injected with extracts from L. japonicus and N. awatschensis samples that accumulated PST either directly or indirectly showed PST intoxication symptoms, indicating that low levels of PST existed in these samples.

CONCLUSIONParalytic shellfish toxins can be transferred to L. japonicus, N. awatschensis, and A. salina from A. tamarense directly or indirectly via the food chains.

Animals ; Artemia ; drug effects ; Cell Count ; Chlorophyll ; metabolism ; Eukaryota ; drug effects ; Feeding Behavior ; drug effects ; Fishes ; Food Chain ; Hydrolysis ; Marine Toxins ; analysis ; metabolism ; toxicity ; Mice ; Models, Biological ; Mollusca ; chemistry ; Paralysis ; chemically induced

OBJECTIVESTo study the relationship between quasispecies of hepatitis B virus and the clinical manifestations of their infection, and to find the answer of why different quasispecies of HBV with the same genotype can induce different clinical situations.

METHODSSixty serum samples, in all of which HBVs of genotype B exist, taken from 32 chronic asymptomatic carriers and 28 severe chronic hepatitis patients, were collected to detect quasispecies of HBV DNA by melt curve approach. Then the relationship between quasispecies of HBV of the same genotype and the clinical situation of their infection was studied by comparing the wave crests of the two sample groups.

RESULTSThe data of the 60 serum samples of HBV of genotype B detected by melt curve showed that HBV DNA in severe chronic hepatitis patients had more wave crests than that in chronic asymptomatic carriers (P < 0.05), suggesting that HBV in severe chronic hepatitis patients had more quasispecies than in the chronic asymptomatic carriers.

CONCLUSIONThe numbers of quasispecies of HBV correlate with the clinical situations of their infection. In the patients infected by HBV of the same genotype, those who have more HBV quasispecies would have more severe clinical manifestations.

Adolescent ; Adult ; Child ; Female ; Genotype ; Hepatitis B virus ; classification ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Male ; Middle Aged