Objective To explore interleukin 4 and immunoglobulin E levels indicators monitoring value in diagnosis of drug allergy reaction . Methods The way of intravenous injection and conventional intraperitoneal injection was used, with the corresponding drugs such as saline, horse serum sensitized, and in different time interleukin -4 and total immunoglobulin E index were recorded in guinea pig serum.Results Injection of saline and Chuankezhi injection, interleukin-4 and total immunoglobulin E protein content , the difference was not statistically significant, but in other drug induced sensitive D8, D14 and D21, interleukin-4 and total immunoglobulin E protein content increased significantly.Conclusion Interleukin 4 and immunoglobulin E levels index detection is the diagnosis of drug allergy is an effective and fast way .

ObjectiveTo observe the effects of mild hypothermia on post-resuscitation myocardial dysfunction in rabbits in order to elucidate the underlying mechanism of hypothermia.Methods After setting up rabbit model of cardiopulmonary resuscitation,20 rabbits were randomly ( random number)divided into two groups,namely normothermic resuscitation group (group A,n =10 ) and post-ROSC hypothermia group ( group B,n =10).In the group A,animals wore treated with standard CPR after cardiac arrest.In post-ROSC hypothermia group,the body temperature of animals was cooled to 32 ～ 34°C after successful ROSC.The left ventricular end-diastolic pressure (LVEDP),left ventricular pressure rise and fall rates ( ± dp/dtmax,serum concentrations of heart-type fatty acid-binding protein (H-FABP) and 8-isoprostaglandin F2a (8-iso-PGF2a) and Cyclooxygenase-2 (COX-2) were observed. Results Compared with the A group,the B group had significantly better hemodynamics,and lower serum H-FABP,8-isoPGF2a and COX-2 levels in the early stage of post-resuscitation ( both P ＜ 0.05 ).ConclusionsMild hypothermia attenuated post-resuscitation myocardial dysfunction during the early period of postresuscitation.The cryoprotective effect on myooardium is likely associated with the reduction of 8-iso-PGF2a and COX-2.

Objective To detect the genetic association between schizophrenia and polymorphism of phosphodiesterase 4B (PDE4B) gene. Methods Observed in a sample of 98 parent/offspring trios where the proband net the Amerecan Classification and diagnostic Criteria for Mental Disorders The Forth Revised Edition,criteria for schizophrenia using correlation analysis and haplotype relative risk analysis. The polymorphism of phosphodiesterase 4B gene was detected with PCR methods and SNP typing in all nucleus families. Results The rs2180335 al-lele was connected with schizophrenia (P = 0.02131). Allele A was protective factor (Z = -2. 184) and allele G was the hazard factor (Z = 2.184). The frequency of rs2180335 allele A was 0.452 and the allele G was 0.548. The rs1040716, rs 3767311 and rs472952 allele was independence with schizophrenia. Five kinds haplotypes of A/A in the rs2180335-rs3767311, A/A in the rs 3767311-rs472952, A/A/A in the rs2180335-rs3767311-rs472952,A/G/G/A and T/A/A/A in the rs1040716-rs2180335-rs3767311-rs472952 associated with schizophrenia (P values were 0. 028715,0. 034845,0. 024177,0. 023967,0. 010839,genotype frequencies were 0. 223, 0.223,0.127,0.081,0.073). Conclusion It shows an association between schizophrenia and the rs2180335 polymorphism at nucleotide of phosphodiesterase 4B gene in Chinese.

N-glycanase is a class of deglycosylation enzymes, widely used in the study of N-glycosylation modification of glycoprotein. In this study, an N-glycanase gene (OsPNGase A, XM_015775832) with high GC content (69.48%) was cloned from rice and then the yeast secretory expression vector pPICZ(α)A-OsPNGase A was constructed for the purpose of transformation to Pichia pastoris. After induction in Pichia pastoris SMD1168H, the target protein was purified by DEAE Sepharose and HisTrap HP chromatography, with a yield of 12.3 mg OsPNGase A from 1 L fermentation medium, showing a specific activity of 258 U/mg. SDS-PAGE revealed that the purified OsPNGase A was a single band and showed consistentcy with the expected molecular weight. OsPNGase A could act on transferrin recombinantly expressed in rice, avidin recombinantly expressed in corn and horseradish peroxidase. Furthermore, OsPNGase A showed higher activity than commercial PNGase F towards avidin. OsPNGase A displayed the highest digestion activity at pH 6.0 and 40 °C, and was also active in the neutral and alkaline environment. Despite the fact that OsPNGase A was inhibited by reducing agents and surfactants, it still maintained partial enzymatic activity in 100 mmol/L β-ME or DTT. Therefore, the successful expression of rice OsPNGase A provides a new tool for the study of plant glycoproteins and the establishment of yeast secretion expression system lays the foundation for the preparation of PNGase A.

Rutin-degrading enzymes (RDE) can degrade rutin into poorly water soluble compound, quercetin, and cause the bitter taste in tartary buckwheat. In the present study RDE from Yu 6-21 tartary buckwheat seeds was purified by ammonium sulphate precipitation, followed by hydrophobic interaction chromatography on Phenyl Sepharose CL-4B, ion exchange chromatography on CM-Cellulose and gel filtration chromatography on Sephadex G-150. Purified RDE showed single band with molecular weight of 66 kDa on SDS-PAGE. The optimum pH and temperature of RDE were 5.0 and 50 ℃ respectively. The Km was 0.27 mmol/L, and the Vmax was 39.68 U/mg. The RDE activity could be inhibited by Cu²⁺, Zn²⁺, Mn²⁺ and EDTA, and showed tolerance to 50% methanol (V/V). The N terminal sequence (TVSRSSFPDGFLFGL) was obtained by Edman degradation method and 15 internal peptide sequences were determined by MALDI-TOF-MS (matrix-assisted laser desorption ionization time of flight mass spectrometry). These results established the foundations for identification of the candidate gene of RDE via transcriptome data and further studying RDE biological function.

Transcription factor is a key trans-acting factor to mediate stress response by regulating gene expression. Plants have developed a series of mechanisms to modulate development, stress response, signaling and disease resistance at transcription level. DNA binding with one finger (DOF), containing one C₂-C₂ zinc finger domain, is a special plant transcription factor. Specifically, the conserved domain at N-terminus of DOF has multiple functions, including interacting with DNA and protein, which could be involved in plant development and stress response. Although many DOF family genes are characterized in plant stress response, it is not clear if DOF genes have functions in cereal plants. In the present paper, the role of DOF family genes on cereal plants were discussed based on a comprehensive phylogenetic relationship analysis, expression profiles in different tissues and various environmental conditions. The results obtained here will provide an important reference for further understanding the mechanism of gramineous crops in stress resistance.
DNA-Binding Proteins ; metabolism ; Gene Expression Regulation, Plant ; Phylogeny ; Plant Proteins ; metabolism ; Plants ; genetics ; Transcription Factors ; metabolism ; Zinc Fingers

The endophytic fungi HL-Y-3, which was isolated from the healthy leaves of Coptis chinensis, produced berberine when grown in the PDA culture medium. The presence of berberine was confirmed by the chromatographic and spectroscopic analyses. The yield of berberine was recorded as 9.313 μg•g⁻¹ by HPLC. The strain HL-Y-3 was identified as Alternaria sp.by morphological observation and 5.8S rDNA-ITS sequence analysis.The separation and purification of constituents were performed by PTLC. The mass spectrometry (MS) of the analyte was shown to be identical with authentic berberine.Further analysis with nuclear magnetic resonance (NMR) spectroscopy to showed that the chemical structure of the fungal berberine was identical with authentic berberine. The research provided new resources for the utilization of berberine.

By bioinformatics analysis, a putative keratinase gene gm2886 (Accession number: KY368946) was discovered in the genome of a feather-degrading strain, Streptomyces albidoflavus Fea-10. gm2886 was ligated into integrative Escherichia coli-Streptomyces shuttle vector pSET152 under the promoter PermE and added with C-terminal His-tag. The expression vector was transformed into Streptomyces pactum ACT12 by conjugal transfer and the recombinant protein GM2886-His6 was detected in fermentation broth. GM2886-His6 was purified and characterized. Its size was nearly 36 kDa. GM2886-His6 showed proteolytic activity towards a variety of substrates and could even degrade insoluble substrates, such as azure keratin and chicken feathers. The optimal pH and temperature of GM2886-His6 for proteolysis of casein was pH 10.0 and 50 ℃, respectively. The enzyme activity was inhibited by PMSF, but not EDTA, indicating that GM2886-His6 was a serine proteinase. Our results laid the foundation for the research of the molecular biological mechanism on feather-degrading and for the further utilization of Fea-10.

In order to investigate the enzyme production mechanism of yak rumen-derived anaerobic fungus Orpinomyces sp. YF3 under the induction of different carbon sources, anaerobic culture tubes were used for in vitro fermentation. 8 g/L of glucose (Glu), filter paper (Flp) and avicel (Avi) were respectively added to 10 mL of basic culture medium as the sole carbon source. The activity of fiber-degrading enzyme and the concentration of volatile fatty acid in the fermentation liquid were detected, and the enzyme producing mechanism of Orpinomyces sp. YF3 was explored by transcriptomics. It was found that, in glucose-induced fermentation solution, the activities of carboxymethyl cellulase, microcrystalline cellulase, filter paper enzyme, xylanase and the proportion of acetate were significantly increased (P < 0.05), the proportion of propionate, butyrate, isobutyrate were significantly decreased (P < 0.05). The results of transcriptome analysis showed that there were 5 949 differentially expressed genes (DEGs) between the Glu group and the Flp group, 10 970 DEGs between the Glu group and the Avi group, and 6 057 DEGs between the Flp group and the Avi group. It was found that the DEGs associated with fiber degrading enzymes were significantly up-regulated in the Glu group. Gene ontology (GO) function enrichment analysis identified that DEGs were mainly associated with the xylan catabolic process, hemicellulose metabolic process, β-glucan metabolic process, cellulase activity, endo-1,4-β-xylanase activity, cell wall polysaccharide metabolic process, carbohydrate catabolic process, glucan catabolic process and carbohydrate metabolic process. Moreover, the differentially expressed pathways associated with fiber degrading enzymes enriched by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were mainly starch and sucrose metabolic pathways and other glycan degradation pathways. In conclusion, Orpinomyces sp. YF3 with glucose as carbon source substrate significantly increased the activity of cellulose degrading enzyme and the proportion of acetate, decreased the proportion of propionate, butyrate and isobutyrate. Furthermore, the degradation ability and energy utilization efficiency of fungus in the presence of glucose were improved by means of regulating the expression of cellulose degrading enzyme gene and participating in starch and sucrose metabolism pathway, and other glycan degradation pathways, which provides a theoretical basis for the application of Orpinomyces sp. YF3 in practical production and facilitates the application of Orpinomyces sp. YF3 in the future.
Animals ; Cattle ; Neocallimastigales/metabolism* ; Anaerobiosis ; Rumen/microbiology* ; Propionates/metabolism* ; Isobutyrates/metabolism* ; Cellulose/metabolism* ; Fungi ; Starch/metabolism* ; Glucose/metabolism* ; Acetates ; Sucrose/metabolism* ; Cellulases ; Cellulase