A cDNA sequence of Arnebia euchroma AP2/ERF named AeAP2/ERF was cloned by in silico cloning in this study, using ACX71873 sequence from Lithospermum erythrorhizon as the probe sequence. Some characters of the AP2/ERF gene and encoded protein sequences were predicted and analyzed by the bioinformatics methods, including general physical and chemical properties, hydrophobieity, signal peptide, secondary structure, localization sites in cells. Results showed that the 876 bp long gene included a 1 077 bp ORF and encoding 205 amino acid. The AeAP2/ERF protein had no signal peptide, it was a hydrophilic proteins located in nucleus. The function of the AP2/ERF protein was mainly involved with metabolism controlling and signal transduction.
Amino Acid Sequence ; Base Sequence ; Boraginaceae ; classification ; genetics ; Cloning, Molecular ; Computational Biology ; Computer Simulation ; Hydrophobic and Hydrophilic Interactions ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Protein Structure, Secondary ; Protein Structure, Tertiary ; Transcription Factors ; chemistry ; genetics

Arnebia euchroma is the main source for medicinal herb Zicao. and its most important component shikonin compounds have high medicinal and industrial value. This research is aimed to build overexpression vectors and RNAi vectors for key secondary metabolism genes of A. euchroma, and bulid platform for constructions of related transgenic lines using GATEWAY technology. To build genetic material based genetic research platform is to provide a great convenience for digging and functional verification of the genes on secondary metabolic pathway, and also to fill the gaps in transgenic research of A. euchroma. This study is also important for the cultivation of shikonin high-yielding strains of A. euchroma.
Boraginaceae ; genetics ; metabolism ; Genetic Vectors ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; RNA Interference ; Secondary Metabolism

Twenty-seven ERF transcription factor family genes were isolated from Arnebia euchroma, with an average size of 1,010 bp, each gene encoded a 212 amino acids on average. The gene structure and expression of physicochemical properties, subcellular localization, signal peptides, senior structural domains and conservative forecasting, and analysis of A. euchroma were studied comparing with ERF gene gi261363612 of Lithospermum erythrorhizon, and phylogenetic analysis of A. euchroma ERF family was carried out. The results showed the existence of three conserved domains in this family, the senior structure based on random coil and it clustered into CBF/DREB and ERF subfamilies.
Amino Acid Motifs ; Amino Acid Sequence ; Boraginaceae ; genetics ; Cloning, Molecular ; Computational Biology ; Genome, Plant ; genetics ; High-Throughput Nucleotide Sequencing ; Medicine, Chinese Traditional ; Molecular Sequence Data ; Multigene Family ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; Plants, Medicinal ; Protein Structure, Secondary ; Protein Structure, Tertiary ; Sequence Analysis, DNA ; Transcription Factors ; chemistry ; genetics ; Transcriptome

In the late 20th century, the rapid development of life sciences resulted masses of biological data, in which laws of life sciences urgently needed to be excavated. Together with the progress of computer science and technology, bioinformatics was born. Chinese medicinal resources, especially the plant resources, are the basis for the development of the Chinese pharmaceutical industry. As a new discipline and important life science research tools, bioinformatics applied rapidly in the study of Chinese medicine resources and involved in different aspects of genomics in recent years. Bioinformatics provides a new research approach to reveal the biological principles, methods and techniques and powerful tools of excavation, at the same time it is speeding researches in the field of Chinese medicine resources and the development of the industry. This article summarizes a comprehensive collection of bioinformatics application data in the study of Chinese medicine resources, prospects the application in Chinese medicine resources, and provides a basis bioinformatics information for the study of Chinese medicine resources.
Biotechnology ; methods ; Computational Biology ; methods ; Drug Industry ; methods ; Genomics ; methods ; Medicine, Chinese Traditional ; methods ; Metabolomics ; methods ; Plants, Medicinal ; genetics ; growth & development ; metabolism ; Proteomics ; methods

OBJECTIVETo identify the relationship between amino acid mutations in Neisseria gonorrhoeae isolates and their antibiotic resistance.

METHODSPI gene fragments of Neisseria gonorrhoeae from 17 clinical isolates were obtained with PCR amplification. They were cloned into the PCR cloning vector pBS-T to form pBS-T-PI and sequenced. The sequences of PI genes were analyzed. At the same time, minimum inhibitory concentration (MIC) of penicillin and tetracycline to these 17 isolates were measured and contrasted with the corresponding PI sequence.

RESULTSThe recombinants of PI gene from 17 clinical isolates of Neisseria gonorrhoeae were successfully constructed and sequenced. They were divided into PIA and PIB subtypes according to the results from blastn software by comparing the sequences with the GenBank. Mutations were found at the sites of 120 and 121. There were only some of the sequences having an aspartic acid (D) mutation on 120 and 121 sites, which was not the same as reported. On the other hand,there were two PI sequences,5-9 and 6-1, whose mutations on No. 120 were lysine, similar to those documented.

CONCLUSIONSome relationship between PI amino acids mutations at sites 120 and 121 in Neisseria gonorrhoeae isolates from Chengdu, China and their resistance to penicillin and tetracycline were found. However,further studies need to be done in the future to confirm this hypothesis.

Amino Acid Sequence ; Anti-Bacterial Agents ; pharmacology ; DNA Mutational Analysis ; DNA, Bacterial ; Drug Resistance, Bacterial ; Mutation ; Neisseria gonorrhoeae ; drug effects ; genetics ; isolation & purification ; Polymerase Chain Reaction

Hydrogel nanoparticles (NPs) were synthesized by precipitation polymerization method with N-isopropyl acrylamide (NIPAm), acrylic acid (Aac), N-tert butyl acrylamide (tBAM) and N,N'-methylene bisacrylamide(Bis) as thermosensitive monomer,negative monomer,hydrophobic monomer and crosslinker, respectively. The morphology of the resulting NPs was characterized by scanning electron microscopy(SEM),and the size and the particle size distribution were investigated by dynamic light scattering (DLS). The dynamics test was also carried out to investigate adsorption property of NPs. The results showed that NPs was spherical with uniform particle size and narrow distribution. NPs had the best adsorption performance to lysozyme when the monomer molar ratio was optimized to Aac 20%,tBAM 40%,NIPAm 38% and Bis 2%. Meanwhile, when the particle size of NPs decreased from 386. 20 nm to 77. 25 nm, the adsorption capacity increased gradually. The adsorption rate could reach up to 67.8% within 5 minutes. The thermosensitive of NPs provided a new candidate for the adsorption and separation of lysozyme with good reusability.

Objective To observe the anti-inflammatory and immune-modulating effects of Dilmang Heji(DHHJ),a compound traditional Chinese medicinal preparation,in patients with multiple sclerosis(MS)and explore the possible mechanisms underlying these effects. Methods Forty MS patients were randomized into prednisone treatment and prednisone+DHHJ group for the corresponding treatments.Another 20 surgical patients without immune or inflammatory diseases undergoing lumbar anesthesia served as the control group.Glial fibrillary acidic protein(GFAP)and S100B levels in the cerebrospinal fluid(CSF)and the peripheral blood of these subjects were detected using ennzynle-linked immtmosorbent assay(ELISA), and the numbers 0f CD4+ and CD8+ cells were detected by flow cytometry.The ambulation index(AD,expanded disability status scale(EDSS)and 9-hole PEG test (9HPT)were used to assess the patients'clinical symptoms.All the patients were followed up for3 years to record the number of times of MS relapse. Results GFAP and S100B levels in the CSF were significantly higher in the MS patients than in the healthy subjects,but lower in MS patients treatedwith prednisone plus DHHJ thanin those with prednisone treatment only(P<0.05).In the MS patients,AI and 9HPT scores were correlated to the GFAP and S100B levels in the CSF.Irednisone plus DHHJ treatment was associated with significantly reduced MS relapse in comparison with prednisone treatment alone(P<0.05).Before the treatment,the MS patients showed increased CD4+ cell number and decreased CD8+ cell number especially in the CSF;atter the treaRnents,the CD4+ ceils decreased and CD8+ cells increased,and this effect was stronger with prednisone plus DHHJ treatment(P<0.05).Conclusions DHHJ produces anti-inflammatory effect by inhibiting glial cell activation and modulating immune balance in MS,thus alleviating the symptoms of MS and reducing MS relapse.DHHJ may provide anideal adjuvanttherapy for MS.

OBJECTIVEVentricular resynchronization might be achieved via minimally invasive left ventricular epicardial lead placement.

METHODSix patients with congestive heart failure underwent minimally invasive left ventricular epicardial lead placement after failed coronary sinus cannulation were followed up for 1 year, cardiac function and LV lead threshold were evaluated.

RESULTSThere were no in-hospital deaths, intraoperative complications and diaphragm stimulation. Correct lead positioning was achieved in all 6 patients. LV lead thresholds remained unchanged [(1.2 ± 0.5) V vs (1.1 ± 0.4) V, P = 0.68] at 12 months follow-up. Improvements on 6 min walking test [(327 ± 77) m vs (267 ± 68) m, P = 0.001], LVEF [(26.1 ± 6.0)% vs (38.2 ± 4.7)%, P = 0.004], and NYHA functional class were evidenced at 12 months follow-up.

CONCLUSIONMinimally invasive left ventricular epicardial lead placement is a safe and reliable technique and should be considered as an alternative option in case of difficult coronary venous anatomy and inability to position the lead for resynchronization therapy.

Adult ; Cardiac Pacing, Artificial ; methods ; Cardiac Resynchronization Therapy ; Female ; Heart Failure ; surgery ; Heart Ventricles ; surgery ; Humans ; Male ; Middle Aged ; Pericardium ; surgery

OBJECTIVETo compare differences of angiogenesis among collagen- chitosan, collagen-sulfonated carboxymethyl chitosan porous scaffolds and acellular dermal matrix after these three different scaffolds with silicone membrane were transplanted on the wounds of full thickness burn, and the wound repair of different scaffolds with epidermis grafting on.

METHODSAngiogenesis in different dermal scaffolds, the wound surface and epidermis survival were observed in 1, 2, and 3 weeks after the three different scaffolds were respectively transplanted on wounds of full thickness burn with debridement in 6 Bama miniature pigs (total 18 pigs in 3 groups). At the same time, CD34 positive signals (neo-forming microvessels) were detected by immunohistochemical staining. The wounds without any scaffold transplantation were studied as the control.

RESULTSAngiogenesis had been fundamentally finished in 2 weeks after implantation of collagen- sulfonated carboxymethyl chitosan porous scaffold. And fundamental angiogenesis in collagen- chitosan porous scaffolds and acellular dermal matrix needed at least 3 weeks. Neo-forming micro-vessels perpendicular to wound beds with these three different scaffolds were more than those in the control wounds without scaffold. CD34 positive signals (neo-forming micro-vessels) were significantly higher in wounds at the second week than those in wounds at the first week. And those in wounds at the third week were significantly higher than those in wounds at the second week in all wounds with different scaffold transplantations and the control wounds. CD34 positive signals in the group of sulfonated carboxymethyl chitosan porous scaffold on the 1st, 2nd and 3rd week after the scaffold transplantation were significantly higher than those corresponding signals in the other three groups. Epidermis on the sulfonated carboxymethyl chitosan porous scaffold which had been transplanted on burn wound for 1 week could survive perfectly, however, epidermis on the collagen- chitosan porous scaffold or acellular dermal matrix could not survive until these two scaffolds had been transplanted on the burn wounds for at least 2 weeks.

CONCLUSIONSThese three different scaffolds could repair the full thickness skin defects caused by burn, and angiogenesis of sulfonated carboxymethyl chitosan porous scaffold is the best.

Animals ; Burns ; surgery ; Chitosan ; analogs & derivatives ; Collagen ; Disease Models, Animal ; Female ; Silicones ; Skin Transplantation ; Skin, Artificial ; Swine ; Swine, Miniature ; Tissue Scaffolds

Via studying the phenotype, growth curve and secondary metabolites of two kinds of suspension culture cell of Arnebia euchroma, the kinetics parameters of growth and accumulation of shikonin compounds in cell suspension culture of A. euchroma was obtained through simulating and modeling. This Study found that the red high-yielding one was a fine cell line for producing shikonin compounds, and the white low-yielding one may be a mutant. The first-order and second-order derivative of the fitting function were obtained by fitting the Logistic model of growth curve to get the growth rate and growth acceleration curve of the suspended cells. It is found that the best period to subculture was the 15th day cultured in fresh medium, and the best period of the induction process was the 13th-14th day. When compared the growth rate of the red line and the shikonin compounds accumulation curve, it is found that the rapid growth of the biomass of cells was not conducive to the synthesis and accumulation of shikonin compounds.
Boraginaceae ; chemistry ; cytology ; metabolism ; Cell Culture Techniques ; Cell Proliferation ; Naphthoquinones ; metabolism ; Plant Cells