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

Objective:To study the effect of Ginkgo biloba extract (EGb761) on metabolism of aflatoxin B_1(AFB_1) in Wistar rats. Methods:Seventy one Wistar rats were divided into three groups at random: group A (AFB_1 group), group B (AFB_1+EGb761 group), and group C (control group). The rats in groups A and B were given AFB_1(intraperitoneal injection, 100-200 μg/ kg body weight, 1-3 times/week). The rats in group B were fed the food containing EGb761 while the rats in groups A and C were given normal food. Blood samples were collected and liver biopsy was performed on the 14th, 28th and 42nd week. All the rats were sacrificed at the 64th week. The incidence of hepatoma was observed. The hepatic phase Ⅰ drug-metabolizing enzyme CYP450 and phase Ⅱ enzyme GST were detected by spectrometry. The serum AFB_1-lysine adduct was determined by high performance liquid chromatography (HPLC). The expression of 8-hydroxydeoxyguanosine(8-OHdG) was measured by immunohistochemistry. Results:The incidence of hepatocellular carcinoma (HCC) in group B was significantly lower than that in group A (26.92% vs 76.00%,P<0.001). No hepatocellular carcinoma developed in group C. EGb761 had no effects on the activities of CYP450 and GST in rat liver tissues. The level of AFB_1-lysine adduct reached the peak (4 356.01 pg/mg albumin) at the 14th week in group A. EGb761 significantly inhibited the formation of AFB_1-lysine adducts in serum by 13.07% at the 14th week (P＝0.033), and 73.63% at the 42nd week (P＝0.002). The expression of 8-OHdG protein in rat liver tissues in group B was significantly lower than that in group A at the 28th, 42nd, and 64th week (P<0.05). Conclusion:The main mechanism underlying the effect of EGb761 in blocking hepatogenesis induced by AFB_1 may not be fully related with its influence on the activity of liver phase Ⅰ and phase Ⅱ metabolizing enzymes. EGb761 inhibites the production of AFB_1-lysine addcuts, decreases the expression of 8-OHdG protein, and finally alleviates the DNA oxidative injury, which may be one of the mechanisms for the effects of EGb761 in inhibiting or delaying hepatogenesis induced by AFB_1.

ObjectiveTo investigate the concentration of water fluoride and the application of water defluoridation projects in Suizhou,and to provide a basis for water improvement and prevention of the disease.MethodsCross-sectional study was carried out to investigate completely the water defluoridation projects built between 1986 and 2009 in the area under Suizhou's jurisdiction; 3 source water and terminal water samples of the projects in use were collected,respectively; 1 water sample was collected,respectively in the 3 projects with out-ofcommission or discarded water sources that used to be major water supply,for detection of fluoride,chloride,total hardness,heavy metals and other indicators.ResultsA total of 21 projects were investigated,14 projects were in normal use (66.7％),7 projects were out of order and abandoned (33.3％).Of the 14 projects normally used,13projects with water fluoride concentration ≤ 1.0 mg/L(92.9％)and 1 project with water fluoride concentration higher than 1.0 mg/L(7.1％),the concentration of water fluoride in the 7 abandoned projects was higher than 1.0 mg/L(100％).Arsenic and lead levels of all the water samples were normal.There were two water samples with iron content exceeded the standard,and one total hardness of water samples exceeded the standard in the normally used 14 peripheral water sources.One chloride content exceeded the standard,one manganese content exceeded the standard,two iron content exceeded the standard and three total hardness exceeded the standard in the seven abandoned projects.ConclusionsSome defluoridation projects are stopped using and abandoned in Suizhou,and the water fluoride exceeds the standard.

Objective To provide a better cell model of closely nature infectious state for further research of hepatitis B virus(HBV). Methods Primary tupaia hepatocytes were isolated by the two-step perfusion method. The hepatocytes were then infected with purified serum from patients with hepatitis B. DNA and RNA isolated from the hepatocytes were detected with Southern blot and Northern blot. HBsAg in supernatant was tested by immunohistochemical method. Results cccDNA, pgRNA and sgRNA could be detected by Southern blot and Northem blot, and strong signals could be seen from day 7 to day 14 post-in-fection. The S/CO value of HBsAg in supernatant decreased from day 1 to day 5 and then increased after 5 day. Conclusion Primary tupaia hepatocytes are competent for infection with HBV. HBV can stably repli-cate and express in HBV-infected tupaia hepatocytes.

Objective To assess the quantitative relationship between the distribution of Himalayan marmot and its ecological environment,the terrain,the temperature and the precipitation,using remote sensing and geographic information system in Qinghai province.Methods The distribution of Himalayan marmot was located by Google Earth and ArcGIS software and by using field survey data provided by Chinese Center for Disease Control and Prevention.The corresponding ecological environment of marmot including terrain,temperature and precipitation were derived from the spatial information datasets.All results were processed according to the overlay and statistics analysis using ArcGIS software.Results Seventy-seven point twenty-seven percent(153/198) of Himalayan marmot were distributed in the area of elevation between 3000 and 4000 meters.The number of marmot reached the highest when the slope was between 0 and 17 degrees,and aspect range was between 91 and 270 degrees,180 degree was as south direction.During the period with the maximum temperature of the warmest month of 14.3-17.5 ℃,17.6-20.8 ℃ and 20.9-24.0 ℃,the distribution of marmot reached 95％(186/198) of the total area.Meanwhile,most of the marmot were presented in the area with average precipitation of 46-108 mm.Conclusions A quantitative analysis of appropriate ecological environment of Himalayan marmot in a large scope is carried uul successfully using remote sensing and geographic information system.The study indicates that spatial information technology has important applications in plague prevention and control.

AIMTo investigate whether mitochondrial calcium uniporter participates in the cardioprotection of tumor necrosis factor alpha (TNFalpha) pretreatment in isolated rat hearts subjected to ischemia/reperfusion.

METHODSIsolated perfused rat hearts were subjected to 30 min regional ischemia (occlusion of left anterior descending artery) and 120 min reperfusion. The infarct size, coronary flow (CF) and lactate dehydrogenase (LDH) release during reperfusion were measured. The mitochondria of the heart were isolated and suspended in the swelling buffer for measurement of absorbance at 520 nm.

RESULTSPretreatment with TNFa at 10 U/ml for 7 min followed by 10 min washout reduced the infarct size and LDH release, and improved the recovery of CF during reperfusion. Administration of spermine (20 micromol/L), an opener of mitochondrial calcium uniporter, for 10 min during early reperfusion attenuated the reduction of infarct size and LDH release, and improvement of CF induced by TNFalpha. In isolated mitochondria of the heart pretreated with TNFalpha, the absorbance at 520 nm decreased less than that of mitochondria without TNFalpha pretreatment. Administration of spermine (50 micromol/L) attenuated the change of the absorbance induced by TNFalpha.

CONCLUSIONThe findings indicate that TNFalpha protects myocardium against ischemia/reperfusion injury via inhibiting mitochondrial calcium uniporter opening as well as mitochondrial permeability transition pore opening.

Animals ; Calcium Channels ; drug effects ; metabolism ; Cardiotonic Agents ; pharmacology ; In Vitro Techniques ; Ischemic Preconditioning, Myocardial ; methods ; Male ; Mitochondrial Membrane Transport Proteins ; drug effects ; Myocardial Reperfusion Injury ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Spermine ; pharmacology ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo construct a recombinant plasmid vector of the RV specific fragment for expressing the specific fragment of RV E1 protein.

METHODSRNA of the RV attenuated live vaccine Wistar RA27/3 strain was extracted and reversely transcribed. The specific fragment of the E1 gene was amplified and the PCR products cloned in the vector pGEX-2T after purification. Positive clones were selected and identified by two-enzyme digestion and sequence analysis.

RESULTSA 330 bp target fragment was successfully cloned, and the sequence of the recombinant plasmid was consistent with the original sequence.

CONCLUSIONSuccessful cloning of the RV El specific fragment and the construction of the recombinant plasmid have laid a foundation for further expressing the recombinant protein.

Base Sequence ; Cloning, Molecular ; Gene Expression ; Genetic Vectors ; Molecular Sequence Data ; Plasmids ; RNA, Viral ; Reverse Transcriptase Polymerase Chain Reaction ; Rubella virus ; genetics ; immunology ; Viral Envelope Proteins ; genetics ; immunology

OBJECTIVETo clone the glycoprotein G gene and its specific fragment with high conservation and antigenicity by culturing and amplifying herpes simplex virus type 2 and extracting its whole genome.

METHODSWe obtained a great deal of suspension with HSV-2 virus after infecting the cultured Hela cells with HSV-2 virus, extracted the whole genome of the virus by the phenol-chloroform method, and amplified the US4 gene coding gG-2 by PCR. Then we selected the specific target fragment according to the amino acid sequence alignment of the gG-2 gene and cloned it with the designed primers with restricted endonuclease sites.

RESULTSWe successfully obtained a lot of suspension with HSV-2 virus, and cloned the gG-2 gene from the whole genuine and its specific target fragment. Sequencing showed that both the sequences were identical with those printed in the GenBank.

CONCLUSIONIt is feasible to obtain the virus genome and specific fragment of the gG-2 gene from virus-infected cells, especially for HSV-2 virus with relatively stable hereditary trait. It has prepared the ground for further constructing the expression plasmid of the specific fragment, expressing related proteins and identifying their antigenicity.

Antigens, Viral ; genetics ; Cloning, Molecular ; DNA, Viral ; HeLa Cells ; Herpesvirus 2, Human ; genetics ; Humans ; Viral Envelope Proteins ; genetics ; Virus Cultivation

OBJECTIVEExperimental evidence shows that microRNAs play an important role in the initiation and progression of human malignancies. The present study aimed to investigate the expressions of 6 microRNAs in prostate cancer (PCa) and their clinical significance.

METHODSWe investigated the expression profiles of 6 microRNAs (let-7g, let-7d, miR-98, miR-96, miR-182 and miR-183) using the method of locked nucleic acid (LNA)-modified oligonucleotide in situ hybridization (ISH) and the technology of tissue microarray (TMA) with the formalin-fixed paraffin-embedded (FFPE) specimens from 52 patients with PCa and 38 with benign prostatic hyperplasia (BPH). Then we analyzed the correlation among the expressions of the 6 microRNAs in PCa and their correlation with the Gleason score and clinical stages of PCa.

RESULTSCompared with BPH, the PCa patients showed decreased expressions of miR-98, let-7d and let-7g, and decreased expressions of miR-96, miR-182 and miR-183, with statistically significant differences between the two groups (P < 0.05). The positive rate of the 6 microRNAs was significantly correlated with the Gleason grades of PCa (P < 0.05), but not with the age and serum PSA concentration of the patients (P > 0.05). The expressions of miR-96 and miR-182 were correlated with the clinical stages of the tumor (P < 0.05). There was a positive correlation among the expressions of miR-96, miR-182 and miR-183 (P = 0.00, r = 0.41), as well as between the expressions of let-7d and let-7g (P = 0.00, r = 0.46) in the PCa tissues. And the expression of miR-98 was positively correlated with those of let-7d and let-7g (P = 0.00, r = 0.46).

CONCLUSIONThe expression profiles of the microRNAs let-7d, let-7g, miR-98, miR-96, miR-182 and miR-183 reflect the biological behavior of PCa to some extent, and might be important biomarkers for the early detection and prognostic assessment of prostate cancer.

Adenocarcinoma ; diagnosis ; metabolism ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; Gene Expression Profiling ; Humans ; Male ; MicroRNAs ; classification ; metabolism ; Middle Aged ; Prognosis ; Prostatic Neoplasms ; diagnosis ; metabolism

OBJECTIVEUtilizing RP-HPLC analysis of single, merging and simultaneously boiled Lonicera japonica and Forsythia suspense to find out their identity and difference.

METHODSamples were separated by Alltech-C18 column (4. 6 mm x 250 mm, 5 microm) with a mixture solution of acetonitrile-water (containing 0.2% ethanoic acid) as mobile phase, 1.0 mL x min(-1) as flow-rate, 280 nm as detected wave-length, room temperature as temperature of column, 10 microL as injected volume.

RESULTThe linear ranges of chlorogenic acid and forsythoside A were 0.980-9.800 microg (r = 0.999 9) and 0.972-9. 720 microg (r = 0.999 9), respectively. The average recoveries of chlorogenic acid and forsythoside A were 99.85% (n = 6, RSD 2.0%) and 100.87% (n = 6, RSD 1.6%), respectively. Major peaks were confirmed to the two component herbs, contents of chlorogenic acid and forsythoside A were determinated, ingredients were represented by following peaks were compared qualitatively: peaks k, m, n: L. japonica > merging sample > boiled sample; peaks g, h: Forsythia suspense > merging sample > boiled sample; peaks d, e, i: boiled sample > L. japonica, boiled sample > merging sample.

CONCLUSIONDifference and identities among 4 experimental samples of RP-HPLC separations show that Yinqiao, L. japonica and F. suspense' substances are difference and identities, which offered direction for investigating further the topic-pharmacodynamic substance of Yinqiao, provided increasing experimental evidence for clinic medicine and pharmaceutics and supplied reference for the complex prescription of traditional Chinese medicine.

Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Forsythia ; chemistry ; Glycosides ; analysis ; Hot Temperature ; Lonicera ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results