The transplants of one-year-old Glycyrrhiza uralensis Fisch. were subjected to five concentrations of MnSO4-H2O (0, 1.81, 18.1, 36.2 and 54.3 mg·L(-1)) culturing in vermiculite. qRT-PCR and HPLC were respectively used to measure the relative expression of SQS1 gene and the content of glycyrrhizic acid of G. uralensis in different concentrations of MnSO4·H2O. This is to explore discuss the effects of the expression of SQS1 gene and the accumulation of glycyrrhizic acid by Mn treatment. The results showed both the expression of SQS1 gene and the content of glycyrrhzic acid of G. uralensis tended to rise after the fall of the first with the increase of concentration of Mn treatment. And they were of very significant positive correlation (P<0.01, r=0.737). Relative expression of SQS1 gene reached the highest 7.90 under 18.1 mg·L(-1) MnSO4·H2O treatment. It was very significantly different between 18.1 mg·L(-1) concentration of MnSO4·H2O treatment and CK (0 mg·L(-1)), 1.81, 36.2 and 54.3 mg·L(-1) (P<0.01), and 1.75, 1.37, 1.37, 2.33 times respectively. The content of glycyrrhizic acid reached the highest under 1.81 and 18.1 mg·L(-1) MnSO4·H2O treatment, and there were not significant difference (P>0.05). It was very significantly different between them and other concentrations of MnSO4·H2O treatment (P<0.01). This study suggests the appropriate concentration of Mn treatment could certain promote the expression of SQS1 gene and the accumulation of glycyrrhizic acid of G. uralensis.
Chromatography, High Pressure Liquid ; Genes, Plant ; Glycyrrhiza uralensis ; chemistry ; genetics ; Glycyrrhizic Acid ; analysis ; Manganese

The complete genomes of more cyanobacterial strains have been completed for sequence, and genetic engineering of cyanobacteria has evolved in the post-genome era. Since Kaneko and colleagues had completed the sequence for the complete genome of Anabaena sp. PCC 7120 in 2001, functions of some genes in this genome, including fructose-1,6-bisphosphate aldolase (FBA) gene, have been predicated using the method of bioinformatics. However, little information is available regarding whether this gene can encode FBA and its product characteristics of related enzyme. Here, to explore this information, the predicted II-FBA gene-encoding region in Cyanobase database was cloned by PCR method and then ligated into pET-32a to generate the expression vector, pET-FBA-II. The results of SDS-PAGE indicated that the expression level of the expected target polypeptide was approximate 23.4 percent as compared to total protein and the molecular weight is about 40 kDa as compared to the protein molecular marker. The results of enzyme activity analysis showed that the activity of II-FBA was ~11.8 U per mg protein and owned a standard activity of II-FBA. To sum up, the results not only prove the functional prediction of this II-FBA gene from the Cyanobase database, but also provide the important conditions for further studying its physiological and biochemical characteristics and functions of the gene expression product.

Objective:Osteoprotegerin (OPG) is a key molecule negatively regulating osteoclast differentiation and activation; and the conserved mycobacterial heat shock 70 (HSP70) peptide p111-125 has also been found to inhibit inflammation reactions in chronic arthritis. BaculoDirectTM baculovirus expression system was selected to express recombinant OPG-HSP70 in insect cells.Methods:The human functional fragment (p22-194) of OPG and functional fragment (p111-125) of mycobacterial HSP70 gene were cloned into the transfer vector pENTRTM/SD/D-TOPO. The recombinant plasmid was performed an LR reaction with the BaculoDirectTM Linear DNA to generate recombinant baculovirus DNA. The cultured Sf9 insect cells were directly transferred with the recombinant baculovirus DNA,and the pure recombinant baculovirus was obtained. Then recombinant baculovirus was infected Sf9 insect cells again to express the OPG-HSP70 gene.Results:The target protein was detected at the time of 48h post infection,reached at highest yield at the time of 72h post-infection. A 28kDa protein immunostaining band was detected by Western blotting from lysate of those cells.And the purified protein was obtained by using Ni-NTA system. Functional stuies on the fusion protein showed it significantly reduce osteoclast cell number[(3.10?0.640) cells under each microscope field in treatment group by comparing to (10.70?0.817)cells in the control group] in the osteoclast inhibition test,and reduce the inflammation reaction in a delayed type hypersensitivity (DTH) mice model (P

The genus Xestospongia is one of the most widespread genera of sponges, containing abundant secondary metatolites with novel structures and potent bioactivities. The main structure types of secondary metatolites found in this genus are alkaloids, quinines, terpens, steroids, lipids, polyketones, etc. These metatolites exhibit a variety of bioactivities, such as cytotoxic, antibacterial and antiviral activities. This paper reviews the progress in the chemistry and pharmacological activities of the second metabolities from sponges of Xestospongia, especially for recent five years, with the aim for further research.
Animals ; Secondary Metabolism ; Xestospongia ; chemistry

OBJECTIVETo investigate the time trends of cancer mortality among residents in Kaifeng county, Henan province.

METHODSData on cancer mortality from the vital registration system in Kaifeng county from 1988 to 2005 was analyzed. A total of 9543 death records (5974 males and 3567 females) due to malignant tumors were studied. A two-year-period age-specified standardized mortality rates were directly adjusted by the world standard population, and the annual percentage change (APC) of mortality were estimated by a linear logarithm regression.

RESULTSThe crude cancer death rate for male was 95.09/100,000 and its age-standardized death rate was 117.41/100,000. While, the crude cancer death rate for female was 59.13/100,000 and the age-standardized death rate was 57.15/100,000. There was a significant growth tread for lung cancer (APC: 6.54%), liver cancer (5.07%) in males and breast cancer (7.04%) in females in the groups aged over 18. On the contrary, the decreasing treads for esophageal cancer in both of sexes (-7.09%, -13.53%) were also observed in this study. Meanwhile, there was no other significant changes in the trend, either in the tumor sites or mortality, was observed.

CONCLUSIONIn the past two decades, there has been a significant increasing trend for cancer mortality in Kaifeng county, of Henan Province. Hence, it is necessary to enhance epidemiological survey to identify risk factors at the earlier stages.

China ; epidemiology ; Death Certificates ; Female ; Humans ; Male ; Mortality ; trends ; Neoplasms ; mortality ; Rural Population

OBJECTIVETo investigate the release model of insoluble drug carbamazepine (CBM) based on HPMC matrix tablets.

METHODSCBM release profile from matrices and HPMC erosion rate were determined.

RESULTThe mathematical model by matrix erosion rate and drug release from HPMC K15M were established for the fractional HPMC and CBM released as M(P(t))/M(P(infinity))=-[0.8095ln((t))+1.2775]Meq((-0.0622t-0.305)) and M(d(t))/M(d(infinity))=-[0.1891t-0.1294]Meq(-0.9326). In comparison with the data of HPMC K4M matrix erosion and CBM release from HPMC K4M matrices, theoretical value agreed well with experimental data.

CONCLUSIONThe two mathematical models can be satisfactorily applied to insoluble drug release, which is governed by matrix erosion.

Carbamazepine ; administration & dosage ; chemistry ; Delayed-Action Preparations ; Hypromellose Derivatives ; Methylcellulose ; administration & dosage ; analogs & derivatives ; Models, Theoretical ; Solubility ; Tablets

OBJECTIVETo investigate and compare the prevalence of erectile dysfunction, hyposexuality and ejaculation disorder among middle-aged and older males in the urban and suburban areas of Beijing.

METHODSUsing the random sampling method, we selected 1 656 men aged > or = 50 years from 15 communities in Beijing. We recorded their scores on the International Index of Erectile Function-5 (IIEF-5) and Brief Male Sexual Function Inventory for Urology (O'Leary 1995), and analyzed the obtained data using SPSS 17.0.

RESULTSTotally, 1 644 of the subjects were qualified for the study, 1 244 from the urban area and the other 400 from the suburbs. The median scores on IIEF-5, sexual desire and ejaculation were 5, 2 and 4 in the urban males as compared with 13, 2 and 5 in the suburban men, with statistically significant differences between the two groups (P < 0.01). The IIEF-5 score was significantly correlated with age, diabetes, cardio-cerebrovascular diseases, drinking and administration of 5alpha-reductase inhibitor in both the urban and suburban groups (P < 0.01).

CONCLUSIONThe sexual function of the middle-aged and older males is better in the suburbs than in the urban area of Beijing. The main reasons might be the lower incidences of diabetes and cardio-cerebrovascular diseases, moderate drinking and less use of 5alpha-reductase inhibitor among the suburban men.

Aged ; China ; epidemiology ; Erectile Dysfunction ; epidemiology ; Humans ; Libido ; Male ; Middle Aged ; Suburban Population ; Surveys and Questionnaires ; Urban Population

OBJECTIVETo investigate cell apoptosis, mitochondrial membrane potential, cytochrome C and mechanisms of mitochondria in lymphocyte apoptosis of sepsis.

METHODSIn the research, female C57BL/6 mice whose body weight ranged from 17 to 25 grams were utilized and assigned randomly to two groups: sham operated group (Control), cecal ligation and puncture group (CLP). The present study was undertaken by using the mice splenic lymphocyte to investigate cell apoptosis, mitochondrial membrane potential, cytochrome C. The apoptosis alteration was evaluated by Annexin V-FITC/PI double staining with flow cytometry. The alteration of mitochondrial membrane potential was investigated by Rhodamine-123 staining of cells. Cytochrome C of mitochondria and cytosol was investigated by Western blot methods. Statistical analysis was performed using SPSS 11.5 for Windows software. Experiment data was indicated with mean ± standard.

RESULTSThe splenic lymphocyte apoptosis was significantly accelerated in the CLP group when compared with that in control group (17.3% ± 2.2% vs. 3.5% ± 0.5%, P < 0.05). The Rhodamine-123 fluorescent intensity in splenic lymphocyte apoptosis was reduced in CLP group (76.2% ± 1.6%). Comparison between sham group (99.6% ± 0.4%) and CLP group had statistical significance (P < 0.05). Apoptosis could induce mitochondrial cytochrome C release into cytoplasm. In the CLP group, elevation of cytochrome C in cytosol was concurrently in accordance with decline in mitochondrial cytochrome C content.

CONCLUSIONThese data suggest that mitochondria and mitochondria signal pathway play an important role in lymphocyte apoptosis of sepsis.

Animals ; Apoptosis ; Cells, Cultured ; Cytochromes c ; metabolism ; Disease Models, Animal ; Female ; Lymphocytes ; metabolism ; pathology ; Mice ; Mice, Inbred C57BL ; Mitochondria ; metabolism ; physiology ; Sepsis ; metabolism ; pathology

A fused functional gene of human OPG and Mhsp65 was amplified by PCR,and cloned into the prokaryotic expression vector pET-28a.The BL21(DE3) strain of E.coli was transformed using the recombinant plasmid pET-28a-OPG-HSP65 and the expected protein was expressed by induction with IPTG.Result of SDS-PAGE indicated that the expected recombinant protein of 23 kDa was expressed with high yield as inclusion body.The fusion protein could be specifically recognized by both the anti-His antibody and anti-human OPG monoclonal antibody in Western blot analysis.The purified and refolded fusion protein could inhibit osteoclast proliferation and bone absorption in vitro.The results of mouse ear swelling assay and expressions of TNF-?,IFN-? and IL-17 mRNAs detected by real-time quantitative PCR demonstrated that the fusion protein had an anti-inflammation activity.The results suggest that the fusion protein of human OPG and Mhsp65 may act as a potential therapeutics for rheumatoid arthritis.

OBJECTIVETo study the chemical constituents of the volatile oil from the rhizome of Anemarrhena asphodeloides.

METHODThe volatile oil was steam distillation. Chemical constituents were separated and analyzed by GC-MS. The relative content of each component was determined by area nomalization.

RESULT24 volatile compounds were isolated and identified for the first time, representing 70.83% of the total oil.

CONCLUSIONThe main constituents of this oil were aldehydes (31.15%), terpene and their oxide (20.66%), alkyls (8.35%), Furan heterocyclic compounds (6.41%), non terpene alcohol (4.26%). There are 12 compounds with contents over 3%. Among them, borneol has the highest content (9.35%).

Aldehydes ; analysis ; Anemarrhena ; chemistry ; Bornanes ; analysis ; Gas Chromatography-Mass Spectrometry ; Oils, Volatile ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Terpenes ; analysis