Adult ; Bone Neoplasms ; pathology ; Humans ; Lymphoma ; pathology ; Male ; Spine ; pathology

OBJECTIVETo investigate the effect of component I from agkistrodon acutus venomon (AAVC-I) the migration of human umbilical vein endothelial cells (HUVECs), and to elucidate the possible anti-angiogenic mechanism of AAVC-I.

METHODSThe effect of AAVC-I on the migration of HUVECs which was cultivated in vitro and treated with AAVC-1 at four concentrations: 0, 20, 40, 80 microg/ml, was observed by methods of scratch wound-healing and Transwell assay. The expression level of mRNA and protein of P-selectin and intercellular cell adhension molecule-I (ICAM-1) were examined by RT-PCR and Western blot assay.

RESULTSCompared with the blank group, the migration ability of HUVECs in each AAVE-I treated group was reduced in a dose-dependent manner, and the expression level of the mRNA and protein of P-selectin and ICAM-1 were decreased.

CONCLUSIONAAVC-I inhibits the migration of endothelial cell, which is acted by down-regulation of the expression content of mRNA and protein of P-selectin and ICAM-1.

Cell Movement ; drug effects ; Cells, Cultured ; Crotalid Venoms ; pharmacology ; Down-Regulation ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; P-Selectin ; metabolism ; RNA, Messenger

Objective To construct the recombinant plasmid pYES6-S and express and purify spike protein of severe acute respiratory syndrome(SANS)coronavirus in Saccharomyces cerevisiae. Methods DNA fragments of SANS coronavirus were obtained by reverse transeription.Four over- lapped fragments of spike protein genes were amplified by polymerase chain reaction(PCR)and ligated into an integral spike protein gene by restriction enzyme digestion.The spike protein gene recombined with pYES6 and cloned into E.coll.The recombinant plasmid pYES6-S was induced and expressed in Saccharomyces cerevisiae(INVScl)by galactose.Results The recombinant plasmid pYES6-S was confirmed that inserted fragment was right in length,direction and base matching by restriction enzyme digestion and sequencing.The purified protein encoded by the whole spike protein gene was about Mr 110?10~3 identified by electrophoresis.Conclusion The whole spike protein gene of SARS coronavirus is cloned into E.coli and the protein is expressed in Saccharomyces cerevisiae successful ly.which can be helpful in SARS vaccine research.

Objective To investigate the protective effects of CBD and Nimodipine against injuries secondary to glutamate-inuced or traumatic injury in rat cortical neurons in vitro. Methods Mix-cultured cortical neurons of SD rat were subjected to either glutamate injury or mechanical damage. The degree of injury were detected by cell count of trypan blue staining and measurement of lactate dehydrogenase (LDH) activity in the culture medium. Results When combined treatment with CBD and Nimodipine was used separately, moderate protective effects against the injury of cultured cortical neurons induced by glutamate or trauma were observed, which was obvious when compared with the injured group without treatment (P<0.01). When combined treatment with CBD and Nimodipine was administered, strong protective effect were observed than that with treatment using CBD or Nimodipine alone (P<0.01). Conclusion It is suggested that CBD combined with Nimodipine are synergetic. Combined use of different excitotoxic antagonists can be fruitful in the therapeutic intervention of secondary traumatic damage.

Objective To investigate the protective effects of CBD and Nimodipine against injuries secondary to glutamate-inuced or traumatic injury in rat cortical neurons in vitro. Methods Mix-cultured cortical neurons of SD rat were subjected to either glutamate injury or mechanical damage. The degree of injury were detected by cell count of trypan blue staining and measurement of lactate dehydrogenase (LDH) activity in the culture medium. Results When combined treatment with CBD and Nimodipine was used separately, moderate protective effects against the injury of cultured cortical neurons induced by glutamate or trauma were observed, which was obvious when compared with the injured group without treatment (P<0.01). When combined treatment with CBD and Nimodipine was administered, strong protective effect were observed than that with treatment using CBD or Nimodipine alone (P<0.01). Conclusion It is suggested that CBD combined with Nimodipine are synergetic. Combined use of different excitotoxic antagonists can be fruitful in the therapeutic intervention of secondary traumatic damage.

OBJECTIVETo investigate the chemical constituents in Bruguiera sexangula var. rhynchopetala.

METHODSilica gel flash chromatography together with Sephadex LH - 20 were performed for the isolation and purification of the petrol ether fraction of this plant, and the structures were elucidated by spectral analysis as well as the comparison of the spectral data with those reported in the literatures.

RESULTNine compounds were obtained and identified as lupeol (1), lupeone (2), trans-hydroxy-cinnamoyl ester of lupeol (3), taraxerone (4), beta-amyril-palmitate (5), squalene (6),beta-sitosterol (7), daucosterol (8) and 7alpha-hydroxy-sitosterol (9).

CONCLUSIONAll the compounds were first isolated from B. sexangula var. rhynchopetala.

Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Pentacyclic Triterpenes ; Plants, Medicinal ; chemistry ; Rhizophoraceae ; chemistry ; Sitosterols ; chemistry ; isolation & purification ; Triterpenes ; chemistry ; isolation & purification

BACKGROUNDA new brain region, the marginal division (MrD), was discovered at the caudal margin of the neostriatum. The MrD was shown to be involved in learning and memory in the rat. The aim of this study was to investigate the expression of the immediate-early genes c-fos and c-jun in the MrD of the striatum during learning and memory processes in the rat, immunocytochemical and Western blot methods were used to examine Y-maze trained rats.

METHODSThe rats were divided into three groups, namely the training, pseudotraining, and control groups. After Y-maze training, the expression of the immediate-early genes c-fos and c-jun in the MrD of the rats was investigated using immunocytochemical and Western blot methods.

RESULTSAfter one hour of Y-maze training, the expression of c-jun and c-fos proteins was significantly enhanced in the MrD; the c-jun protein, in particular, was more intensely expressed in this region than in other parts of the striatum. The expression of these two proteins in the training group was significantly higher than in the pseudotraining and control groups. In addition, positive expression was also found in the hippocampus, cingulum cortex, thalamus, and in other areas. Western blot disclosed two immunoreactive bands for the anti-c-fos antibody (47 kD and 54 kD) and two immunoreactive bands for the anti-c-jun antibody (39 kD and 54 kD).

CONCLUSIONSThese results indicate that the immediate-early genes c-fos and c-jun participate in signal transduction during the learning and memory processes associated with Y-maze training in rats.

Animals ; Male ; Maze Learning ; Memory ; Neostriatum ; metabolism ; Proto-Oncogene Proteins c-fos ; biosynthesis ; Proto-Oncogene Proteins c-jun ; biosynthesis ; Rats ; Rats, Sprague-Dawley

To investigate the common regulative effects of the Chinese drug Bailong and hexamethylen bisacetamide (HMBA) on expressions of oncogenes (c-H-ras and c-myc), and tumor suppressor genes (Rb，p53　and p21) of MGC80-3 in human cancer cell cycle. Methods: Adopting RNA Northern Blot to survey the levels of gene expressions of MGC80-3 different phases cells treated with Bailong and HMBA respectively. Results: In different phases of MGC80-3 cells treated with Bailong and differentiation inducer HMBA, expressions of oncogenes c-H-ras and c-myc were inhibited by over 50.0%, messenger kinase subspecies PKC-α gene is similar with the expression inhibition of oncogenes, except effect of Bailong on the G2 phase in cell cycle. Effect of Bailong differs greatly from HMBA in the expression of tumor suppression genes. The expression of Rb and p21 in cells treated by HMBA did not increase but were inhibited by 39.5% and 33.3% respectively in G1 phase. The level of Rb gene expression was decreased, too by 3.0% in S phase. Comparison with HMBA the expression of Rb and p21 genes were increased after treatment by Bailong in all cell cycle. But the effect of Bailong on the expression of p53 gene which was increased obviously by 125.0%－233.4% in majority phase of MGC80-3 cells is similar to HMBA. Conclusion: (1) The effect of Bailong on the regulation of oncogenes and tumor suppressor gene is similar to HMBA but the effect of Bailong is better than that of HMBA. (2) Molecular mechanism of the Bailong or HMBA on the proliferative inhibition and differentiation of MGC80-3 related to regulation of the Bailong and HMBA on the oncogenes and tumor suppressor genes in cell cycle of MGC80-3

In order to develop a rapid detection kit for novel avian influenza virus (AIV) subtype H7N9, two sets of specific primers and probes were designed based on the nucleotide sequences of hemagglutinin antigen (HA) and neuraminidase antigen (NA) of novel H7N9 virus (2013) available in GenBank to establish the method of TaqMan probe-based multiplex real-time RT-PCR for rapid detection of AIV subtype H7N9. The primer and probe of HA were for all H7 subtype AIVs, while the primer and probe of NA were only for novel N9 subtype AIVs. The results showed that this method had high sensitivity and specificity. This method was applicable to the testing of positive standard sample with a minimum concentration of 10 copies/microL; it not only distinguished H7 subtype from H1, H3, H5, H6, and H9 subtypes, but also distinguished novel N9 subtype from traditional N9 subtype. A total of 2700 samples from Zhuhai, China were tested by this method, and the results were as expected. For the advantages of sensitivity and specificity, the method holds promise for wide application.
Animals ; Birds ; virology ; Influenza A Virus, H7N9 Subtype ; genetics ; isolation & purification ; physiology ; Influenza in Birds ; prevention & control ; virology ; Real-Time Polymerase Chain Reaction ; methods ; Species Specificity ; Taq Polymerase ; metabolism ; Time Factors

AIMTo investigate whether NR2B-pERK1/2-pElk-1 signaling contributes to the Y-maze learning and memory of rat brain.

METHODS45 adult male SD rats were divided into 4 groups: (1) Ifenprodil peritoneal injection group (Ifenprodil ip, n = 14); (2) DMSO peritoneal injection group(DMSO ip, n = 15); (3) Ifenprodil cerebral ventricle injection group (Ifenprodil ic, n = 8); (4) DMSO cerebral ventricle injection group(DMSO ic, n = 8). Y-maze training and test were used as an learning and memory enhancing stimulus. Immunohistochemical and Western blotting methods were used for detecting pERK1/2 and pElk-1 expression intensity of different brain regions.

RESULTSCompared with the DMSO ip group, the ifenprodil ip group showed no change on the Y-maze learning score (P > 0.05), but its Y-maze memory score tested 24 after learning decreased (P < 0.05). Ifenprodil peritoneal injection made brain pERK1/2 and pElk-1 expression decreased generally. In hippocampus, marginal division of striatum(MrD), amygdala,these changes were more significant (P < 0.05). Compared with the DMSO ic group, the reconsolidation of Y-maze memory tested 6 hours after ifenprodil injection was impaired in ifenprodil ic group (P < 0.05). The OD value of pERK1/2 and pElk-1 positive bands in ifenprodil ic group attenuated generally. The pElk-1 positive bands of caudate putamen and MrD almost disappeared in ifenprodil ic group.

CONCLUSIONNR2B is essential for the formation of long-term memory, reconsolidation of Y-maze memory. The deactivation of NR2B by ifenprodil will impair these courses. Meanwhile, the deactivation of NR2B attenuates pERK1/2 and pElk-1 expression of learning and memory related regions after Y-maze learning and memory reconsolidation test. In MrD and caudate putamen, the pElk-1 expression are completely blocked by ifenprodil after memory reconsolidation test.

Animals ; Avoidance Learning ; physiology ; Dimethyl Sulfoxide ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Male ; Maze Learning ; physiology ; Memory ; physiology ; Piperidines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism ; ets-Domain Protein Elk-1 ; metabolism