To explore a new method to identify Moutan Cortex to guarantee its safe use, internal transcribed spacer 2 (ITS2) sequence was used to identify Moutan Cortex and its adulterants. DNA was extracted and target fragments were amplified. Sequences were analyzed and assembled by CodonCode Aligner V3.7.1. Genetic distances were computed and phylogenetic tree was constructed based on kimura 2-parameter (K2P) model by MEGA 5.0. The length of the 20 ITS2 sequences of Moutan Cortex from nine different places is 227 bp, and no variation site was detected. The maximum inter-specificK2P distance of Moutan Cortex is 0, the minimum intra-specific K2P distance is 0.041, the average intra-specific K2P distance is 0.222. According to NJ analysis, Moutan Cortex from different places can get together as one branch with bootstrap support values 99%, which indicates Moutan Cortex can be easily distinguished from its adulterants. Using ITS2 sequence can accurately identify Moutan Cortex and its adulterants, it is an effective supplementary to traditional identification methods.
Base Sequence ; China ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Paeonia ; classification ; genetics ; Phylogeny ; Quality Control

Increasing studies have demonstrated that interferon gamma (IFN-γ),which serves as a critical inflammatory cytokine,is essential to induce the immunosuppressive effects of mesenchymal stem cells (MSCs).However,the mechanisms underlying the enhanced immunosuppressive effects of IFN-γ-stimulated MSCs (γMSCs) are not fully understood.MSC-derived rnicrovesicles (MSC-MVs) have been viewed as potential pivotal mediators of the immunosuppressive effects of MSCs.Moreover,microRNAs (miRNAs) are important regulators of immunological processes and can be shuttled from cell to cell by MVs.The aim of our study was to analyze the the miRNA expression signature of MVs derived from γMSCs (γMSC-MVs),which may provide better understanding of the immunosuppressive property of their parent cells.Through miRNA microarray and bioinformatics analysis,we found 62 significantly differentially expressed miRNAs (DEMs) in γMSC-MVs compared with MSC-MVs.And the potential target genes and signaling pathways regulated by DEMs were predicted and analyzed.Interestingly,many DEMs and predicted signaling pathways had been.demonstrated to be involved in immunoregulation.Furthermore,the network between immunoregulation-related pathways and relevant DEMs was constructed.Collectively,our research on the miRNA repertoires of γMSC-MVs not only provides new perspectives into the mechanisms underlying the enhanced immunosuppressive property of γMSCs,but also paves the way to clinical application of these potent organelles in the future.

DNA barcoding method was conducted for the authentication of pollen materials due to difficulty of discriminating pollen materials bearing morphological similarity. In this study, a specific focus was to identify cattail pollen (Puhuang) and pine pollen (Songhuafen) samples from their adulterants which are frequently mixed-together. Regions of the internal transcribed spacer (ITS2) from 60 samples were sequenced, and new primers for cattail pollen were designed according to the sequence information. The results from the NJ trees showed that the species of pine pollen, Puhuang and their adulterants can be classified as obvious monophyly. Therefore, we propose to adapt DNA barcoding methodology to accurately distinguish cattail pollen, pine pollen and their adulterant materials. It is a great help for drug regulatory agency to supervise the quality of medicinal materials.
China ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Pinus ; classification ; genetics ; Pollen ; classification ; genetics ; Quality Control ; Typhaceae ; classification ; genetics

The aim of the study was to look for the chemical constituents of the herb of Lysimachia foenum-graecum. The herb of Lysimachia foenum-graecum was extracted with 70% EtOH. The isolation and purification was performed with a combination of multi-column chromatography and the structure was determined by spectral analysis. The flavonoid compound was obtained and elucidated as kaempferol-7-O(4"-(E)-p-coumaroyl-)-alpha-L-rhmanopyranosyl)-3-O-beta-D-glucopyranosyl (1-->4)-alpha-L-rhmanopyranosyl (1-->2)-beta-D-glucopyranoside. It is a new flavonoid compound.
Flavonoids ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Primulaceae ; chemistry

Objective To study the mlationship between dietary soy isoflavones and blood lipids among residents of 40-65 years old,in Guangzhou.Methods Dietary soy isoflavones and other nutrients intakes were assessed with quantitative food frequency questionnaire(FFQ).Total cholesterol(TC),triglycerides(TG),HDL cholesterol(HDL-C)and LDL cholesterol(LDL-C)in plasma were measured with colorimetry.Results Ranges of dietary soy isoflavones intake among 134 males and 261 females were fxom 0 mg/day to 61.96 mg/day and 0 mg/day to 82.52 mg/day,with means of 11.95 mg/day,14.90 mg/day,respectively.After adjusted for total energy intake and fat percent energy,difiefences of TC,LDL-C in total population and TC in women were statistically significant between groups(P value was 0.002,0.008,0.004,respectively) and dose-effect relationships(P value was <0.001.0.012.0.001,respectively)were observed between dietary soy isoflavones intake and the upper mentioned three indices.Compared with the low-intake group,tbese three indices lowered 7.06%,10.13%and 7.48%,respectively in high-intake group.Critical significance of LDL-C was observed both in women and men between groups.Further controlled for age,BMI and WHR,no obvious change of the results was observed.Conclusion Moderate intakes of soy isoflavone as part of a regular diet seemed to be associated with favorable blood lipid levels.

The cytoplasm of E. coli is a reducing environment where cysteines do not engage in disulfide bonds. Any disulfide bonds that do appear are rapidly reduced through the action of disulfide reducing enzymes such as thioredoxin and glutaredoxin. To study the influence of E. coli cytoplasm on the solubility of recombinant proteins produced in it, bovine fibroblast growth factor (BbFGF), with single disulfide bond, and anti-HBsAg single-chain Fv (HBscFv), with two disulfide bonds, were selected as the pattern molecules of simple protein and complex protein, respectively. pJN98-BbFGF, a BbFGF expressing plasmid based on the vector pET3c, was constructed and transformed into normal host BL21(DE3) and a reductase deficient strain, E. coli Origami(DE3). At the same time, pQE-HBscFv, a HBscFv expressing plasmid was constructed and transformed into M15 [pREP4] and Origami(DE3). The recombinant BbFGF and HBscFv were produced in 2 types of bacteria and their solubilities and bioactivities were determined, respectively. It was found that the majority of BbFGF had formed inclusion body in the cytoplasm of BL21 (DE3) and all of them turned into soluble protein in Origami(DE3). It was also found the productivity of BbFGF in Origami (DE3) was 5% - 10% of the total protein and the value was 15% - 23% in BL21(DE3). BbFGFs produced in 2 recombinant bacteria were purified by cation exchange and heparin affinity chromatography. MTT assay revealed that the bioactivity of BbFGF purified from Origami(DE3) was higher than its counterpart from BL21(DE3). The ED50 of BbFGFs from different bacteria was 1.6ng/mL and 2.2ng/mL, respectively. As far as HBscFvs, both of them formed inclusion body in the cytoplasm of M15 [pQE-HBscFv] and Origami [pQE-HBscFv]. The inclusion body was solubilized in 6mol/L GuHCl, purified with a His-Trap column and then refolded by dialysis step-by-step against buffers containing downtrend concentration of GuHCl. Indirect ELISA was applied to determine the HBsAg binding activity of HBscFvs. It was found there was no obvious difference between the bioactivity of refolded HBscFvs produced from 2 recombinant bacteria. On the other hand, the supernatant of Origami [pQE-HBscFv] lysate displayed weak bioactivity and its counterpart from M15 [pQE-HBscFv] displayed without any bioactivity. The soluble HBsFv in the cytoplasm of Origami [pQE-HBscFv] was purified by cation exchange and immobilized metal affinity chromatography (IMAC) and the yield was 1 - 2mg/L. Those results suggested that modification of the redox environment of E. coli cytoplasm greatly improved the solubility of recombinant disulfide-bonded proteins produced in it. In the next step, we had like to co-express of molecular chaperones or refoldase to raise the yield of soluble recombinant proteins, as well as optimizing the culture condition of the "oxidizing" E. coli.
Animals ; Antibodies ; genetics ; immunology ; metabolism ; Cattle ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; enzymology ; genetics ; metabolism ; Escherichia coli Proteins ; genetics ; Fibroblast Growth Factors ; genetics ; metabolism ; Genetic Vectors ; genetics ; Hepatitis B Surface Antigens ; immunology ; Inclusion Bodies ; chemistry ; metabolism ; Oxidoreductases ; genetics ; Plasmids ; genetics ; Protein Engineering ; Recombinant Proteins ; chemistry ; genetics ; metabolism ; Solubility

OBJECTIVESTo investigate the mechanical properties of bilateral atlantoaxial trans-articular screws and atlas laminar hooks instrumentation with finite element method.

METHODSThere was a volunteer with age of 28 years old, body height 172 cm, body weight of 60 kg and without cervical deformity by X rays. The ligamentous, nonlinear, three-dimensional finite element models of normal upper cervical spine (C0-3) was developed and validated. The destabilized model with bilateral atlantoaxial trans-articular screws and atlas laminar hooks was evaluated for quasistatic loading.

RESULTSThe finite element model of upper cervical spine consists of 229,047 nodes and 152,475 elements, and correlated well with experimental data for all load cases and could be used for experiment. The finite model with bilateral atlantoaxial trans-articular screws and atlas-laminar hooks predicted that the maximum Von Mises Stress was in the region in which screws penetrated the atlantoaxial articular facet. The novel instrumentation resulted in sufficient stability.

CONCLUSIONThe bilateral atlantoaxial trans-articular screws and atlas laminar hooks instrumentation is useful and effective for atlantoaxial arthrodesis.

Atlanto-Axial Joint ; surgery ; Biomechanical Phenomena ; Bone Screws ; Cervical Vertebrae ; diagnostic imaging ; Finite Element Analysis ; Humans ; Imaging, Three-Dimensional ; Internal Fixators ; Joint Instability ; surgery ; Male ; Radiography ; Spinal Fusion ; instrumentation ; methods

Since the first outbreak of avian influenza A(H7N9) virus in humans was identified in 2013, there have been five seasonal epidemics observed in China. An earlier start and a steep increase in the number of humans infected with H7N9 virus was observed between September and December 2016, raising great public concern in domestic and international societies. The epidemiological characteristics of the recently reported confirmed H7N9 cases were analysed. The results suggested that although more cases were reported recently, most cases in the fifth epidemic were still highly sporadically distributed without any epidemiology links; the main characteristics remained unchanged and the genetic characteristics of virus strains that were isolated in this epidemic remained similar to earlier epidemics. Interventions included live poultry market closures in several cities that reported more H7N9 cases recently.

OBJECTIVETo establish rabbit model of scoliosis induced with stable asymmetric lumbar loads.

METHODSScoliosis was induced in 10 two-month-old New Zealand rabbits using 316L stainless steel springs placed between the unilateral transverse processes of L2 and L5. Serial radiographs were documented before and at 1, 4, 8, 9 and 12 weeks after the operation. At weeks, the rabbits were randomly divided into SR group (n=5) with the spring removed and SK group (n=5) without spring removal.

RESULTSAll the rabbits survived the experiment with Cobb angle all greater than 10 degree at the end of the experiment. Significant changes were found in the Cobb angles and kyphotic angles at 1, 4 and 8 weeks after the operation (P<0.05). At 8 weeks, the Cobb angle, the kyphotic angle and the length of the spring were similar between SR and SK groups (P>0.05), and in the 4 weeks following spring removal in SR group, the Cobb angle and the kyphosis decreased significantly compared with those in SK group (P<0.05). Micro-CT showed that the BV/TV of the concave side was greater than that of the convex side. The length of the spring did not show obvious changes during the experiment (P>0.05).

CONCLUSIONSAsymmetric lumbar loading is a convenient, time-saving, and highly reproducible approach for establishing rabbit models of scoliosis.

Animals ; Disease Models, Animal ; Rabbits ; Scoliosis ; physiopathology ; Spine ; pathology

Adult ; Hemophilia A ; complications ; Humans ; Leukemia, Promyelocytic, Acute ; complications ; Male