With homology cloning approaches coupling with RACE (rapid-amplification of cDNA ends) techniques, the full-length coding sequence of pathogenesis-related protein PR10-1 with differential expression was cloned from the total RNA of the root of Panax notoginseng, and its function was explored furtherly. As a result, the longest 465 bp ORF (named as PnPR10-1 with the Accession No. KJ741402 in GenBank) was detected from the cloned sequence with full-length of cDNA of 863 bp. The corresponding peptide encoded consisted of 155 amino acids, contained some domains such as Bet-v-I, and showed high similarity with that from Panax ginseng by analysis of phylogenetic trees created from the alignments. Real-time quantitative PCR showed that the expression of PnPR10-1 gene was constitutive in different tissues of 1-3 year old plant, suggesting that it might be involved in growth, development, and secondary metabolism; yet it was up-regulated significantly with the infection of Fusarium oxysporum in root, suggesting that it might be involved in defense against many diseases including root rot in P. notoginseng.
Amino Acid Sequence ; Cloning, Molecular ; DNA, Complementary ; Genes, Plant ; Glycoside Hydrolases ; genetics ; Molecular Sequence Data ; Open Reading Frames ; Panax notoginseng ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Plant Roots

BACKGROUND:The process of oxidative stress that impacts the curative effect exists in the region which accepts cel transplantation. However, there are few reports about the effects of oxidative stress on human dental pulp stem cels and relevant mechanism.
OBJECTIVE:To understand the effect of hydrogen peroxide on the senescence of human dental pulp stem cels.
METHODS:Human dental pulp stem cels were isolated and cultured in PBS, 100 and 200 μmol/L hydrogen peroxide for 2 hours, respectively. Cel morphology was observed under inverted microscope, degree of cel senescence monitored by β-galactosidase staining, cel proliferation ability detected by BrdU kit and cel counting method, cytoskeleton of dental pulp stem cels and expression of sirt1 tested using immunofluorescence method, and expression of sirt1 and p16 proteins measured by western blot assay.
RESULTS AND CONCLUSION:Dental pulp stem cels exhibited a fibroblast-like morphology with spindle-shaped appearance. After stimulated by hydrogen peroxide, the cel volume was enlarged, theβ-galactosidase staining deepened and the proliferation of dental pulp stem cels reduced. The enhancement of senescence of dental pulp stem cels was accompanied with the increasing concentration of hydrogen peroxide, and in this process, the expression of p16 was raised while the expression of sirt1 was decreased. In conclusion, the senescence of human dental pulp stem cels can be promoted by the stimulation of hydrogen peroxide, and sirt1 and p16 are involved in this process. Our findings may provide a theoretical and experimental foundation for autologous transplantation of dental pulp stem cels.

To analyze the composition regularity of prescriptions containing Scutellaria baicalensis in Drug Standard of Ministry of Public Health of the Peoples Republic of China--Chinese Patent Medicines and Preparations on the basis of the traditional Chinese medicine inheritance support system (TCMISS), in order to provide reference for new drug R&D. the platform's software V2.0 was applied to establish a database of prescriptions containing S. baicalensis. The software's statistical statement module, association rules and improved mutual information method and other data mining technologies were adopted to analyze commonly used drugs, combination rules and core combination of S. baicalensis prescriptions. Having analyzed 477 prescriptions containing S. baicalensis, the researchers summarized 45 most commonly used drug combinations, whose ingredients mostly had functions of heat-clearing and damp-drying, purging fire for removing toxin and hemostasis. Drugs adopted in core combinations were relatively concentrated and selected according to definite composition methods. There were 23 diseases that S. baicalensis were most frequently applied in the treatment. Having compared three highly frequent diseases--cold, cough and dizziness, the researchers concluded that S. baicalensis could show different therapeutic effects through different combination ratios. Therefore, TCMISS (V2.0) is an important tool in analyzing the composition regularity of traditional Chinese medicines. The longitudinal and parallel comparison method is an effective method for studying the clinical composition regularity of S. baicalensis, while providing reference for new drug R&D.
China ; Drug Compounding ; statistics & numerical data ; Drug Therapy ; statistics & numerical data ; Humans ; Plant Extracts ; analysis ; therapeutic use ; Scutellaria baicalensis ; chemistry

Objective To investigate the effect of adenovirus-bone morphogenic protein 9 ( Ad-BMP9 ) on osteogenic differentiation of immortalized calvarial mesenchymal progenitor cells ( iCALs ) .Methods iCALs were infected with adenoviral vectors encoding BMP-9 or green fluorescent protein ( GFP) and the early osteogenic differentiation was assessed by detecting alkaline phosphatase (ALP) activity after being cultured for 3, 5 and 7 days.14 days after infection, alizarin red S staining was performed to study the formation of osteogenic calcium nodules .The expression of osteogenic marker genes Runx2 and OCN was assessed by quantitative real-time ( RT )-PCR and Western blotting .Results Significant increases in ALP activity and in the expressions of Runx 2 and OCN were detected in BMP-9 treated iCALs compared with GFP-treated cells(P<0.05).Meanwhile, alizarin red S staining showed that more mineralized nodules were found in the BMP-9 induced group .Conclusion BMP-9 can promote the osteogenic differentiation of iCALs .

The effects of active antiendotoxin chemical fraction isolated from Radix Isatidis (fraction D) on TNF-α and IL-8 secretion in HL-60 cells induced by lipopolysaccharide (LPS) were studied. The appropriate densities of cell suspension and fraction D solution were determined by MTT colorimetric method. Fraction D and LPS were added to HL-60 cell suspension with three different methods respectively. The contents of TNF-α and IL-8 in the cultured supernatant induced by LPS were detected by using ELISA method. The results showed that the absorbance (A) was directly proportional to the number of cells and the linearity was good in the range from 0.25 × 105 to 2 ×105 cell/mL cell suspension. The fraction D significantly inhibited the oversecretion of TNF-α and IL-8 in HL-60 cells induced by LPS at the concentration of 7. 812 mg/mL which had no cytotoxicity. It was indicated that the antiendotoxin mechanism of the active fraction from Radix Isatidis was contributed to the inhibition of the oversecretion of cytokines induced by LPS.

By using enrichment media NPC and CVP, 792 strains of Pseudomonas and 515 strains of Erwinia were isolated from the rhizosphere of Digitaria adscendens (H. B. K.) Hem and Setaria vindis (L.) Beanv. Following which, experiments of antimetabolic test with E. coli, seed emergence controlling of S. viridis, herbicidal activity and security with green grass were carried out to select the desired bacteria. As the result, the selected strain, S7, could wholly control the seed emergence of S. viridis without any harm to the two tested green grass. And more, S7 promoted the seed emergence of Festuca arundinacea slightly. In spite of the comparatively low corrected mortality (56. 7%) of S7 after emergence of S. viridis, Selecting of microbial herbicides from weed DRB is thought to be more prospective.

Transgenic tobacco plants expressing a stress responsive gene BoRS1, isolated from Brassica oleracea var. acephala, under the control of the 35S promoter of the Cauliflower mosaic virus were produced. Some plants were further used to test the effect of high level BoRS1 expression on drought stress resistance. The presence of transgene in putative transgenic plants was confirmed by PCR analysis. Thirty-six among 130 transformants showed amplification of predicted fragment of BoRS1 while no amplification was observed in the control. Some transgenic lines confirmed by PCR analysis were analyzed through semi-quantitative one-step RT-PCR for the expression of BoRS1 gene. Amplification of 1.4 kb cDNA product revealed transcription of BoRS1 gene. Meanwhile, differential intensity of the cDNA band indicated variable expression levels of the transgene among different transformed lines. The water loss of detached leaves from the transgenic plants was slower than that of the control. Transgenic tobaccos and the non-transgenic controls were used for further drought stress experiments by using different concentration of mannitol. The transformants showed higher tolerance to drought stress than non-transgenic plants and different transgenic lines exhibited different tolerance during drought stress. These results showed that the BoRS1 gene probably play role in enhancing the ability to drought stress.
Adaptation, Physiological ; genetics ; Agrobacterium tumefaciens ; genetics ; Brassica ; genetics ; Dehydration ; Genes, Plant ; Plant Proteins ; genetics ; metabolism ; Plants, Genetically Modified ; genetics ; growth & development ; physiology ; Stress, Physiological ; Tobacco ; genetics ; growth & development ; physiology ; Transformation, Genetic ; Transgenes

Objective To construct the protein-protein interaction network of Parkinson's disease (PD) associated proteins. Methods PD-related proteins and their interaction proteins were collected by bioinformatics method from HGNC, OPHID and UniHI database. A protein-protein interaction network of PD associated proteins was designed by ProteoLens software; each protein was evaluated by its contribution to the network and the relevance scores were calculated, thus the coefficient of association of each protein in the network was noted. Results A protein-protein interaction network containing 463 PD associated proteins and 767 their interaction proteins was obtained with its index aggregation reaching 90.5% (P=0.008). The relevance scores of SNCA, PARK2, DRD2, HTRA2,NDUFV2, DJ1, DRD1, DRD3, TRAP1 and ND3 were much higher than that of the others, which indicated their important roles in the pathogenesis of PD. The relevance scores of APP, UBE2I, CLIC6 and UBB were a little higher among all the preteins, indicating that they also participated in the pathogenesis of PD. Some novel proteins, such as FLNA, FREQ, BIRC7, EPB41, EPB41L1, GIPC1,GNAZ, GRB2, KCNJ9, MAPK1, BAG5 and CYC, may also involved in the pathogenesis of PD.Conclusion A scientific and practical protein-protein interaction network of PD associated proteins is successfully constructed, which further confirms the role of some proteins in the pathogenesis of PD.Some novel proteins that might involve in PD are obtained too.

In the present study, an attempt was made to prove the question whether endothelial cell precursors exist in blood circulation during postnatal period. CD34(+) cells were harvested from G-CSF mobilized adult blood and umbilical cord blood and incubated onto fibronectin/gelatin-coated Petric dishes in the presence of recombinant human vascular endothelial cell growth factor(rhVEGF) and basic fibroblast growth factor(rhbFGF). Endothelial cell lineage was identified by von Willebrand factor(vWF) expression and Ulex europous agglutinin I(UEA-I) binding capacity. The results showed that a firmly adherent cell monolayer formed when CD34(+) cells, but not CD34(-) cells, were cultured for 5 - 6 weeks as described before. Immunocytochemistry and flow cytometry analysis showed that almost all of the adherent cells were vWF-positive and around 90% were able to bind UEA-I specifically. These findings demonstrate that angioblasts exist in the circulation during postnatal life and therefore, vasculogenesis might occur in adults.

OBJECTIVETo explore the imaging diagnosis of pelvic eosinophilic granuloma and improve diagnosis accuracy.

METHODSA retrospective analysis of the plain film, CT and MRI was conducted in 24 cases of pathologically confirmed eosinophilic granuloma. All the patients received radiography, CT scan was performed in 17 cases and MRI was done 9 cases.

RESULTSIn the 24 cases of eosinophilic granuloma, 6 lesions were located in the pubis, 5 in the ischium and 13 in the iliac region. The lesion showed bony destruction or irregular bony destruction, with periosteal reaction and soft tissue mass.

CONCLUSIONPelvic eosinophilic granuloma has characteristic imaging features, and combination of the imaging findings can improve the diagnosing accuracy.

Adolescent ; Adult ; Bone Diseases ; diagnosis ; diagnostic imaging ; Child ; Child, Preschool ; Diagnostic Errors ; Eosinophilic Granuloma ; diagnosis ; diagnostic imaging ; Female ; Humans ; Infant ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Pelvis ; Retrospective Studies ; Tomography, X-Ray Computed ; Young Adult