Adolescent ; Adult ; Dysgerminoma ; etiology ; genetics ; Female ; Gonadal Dysgenesis ; genetics ; Gonadal Dysgenesis, 46,XY ; genetics ; Humans ; Ovarian Neoplasms ; genetics

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.

Humans ; Osteogenesis ; Mesentery/surgery* ; Ossification, Heterotopic/surgery*

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 importance of vaccine on public health is related to the herd protection related to the levels of vaccine coverage, which directly influences the vaccinated individuals as well as the unvaccinated community. Reaching the level of herd protection by increasing vaccine coverage is the basic strategy to eradicate related infectious diseases. Again, herd protection has played an important role in public health practices. With the increasing interests in estimating the vaccine herd protection, we however, have seen only few relevant papers including observational population-based and cluster-randomized clinical trials reported in China. We hope to discuss the study designs for evaluating the vaccine herd protection in order to generate evidence-based related research in this field.
China/epidemiology* ; Communicable Disease Control ; Communicable Diseases/immunology* ; Humans ; Immunity, Herd/immunology* ; Research Design ; Vaccination/trends* ; Vaccines/immunology*

Petite integration frequency 1 (PIF1) helicases are ubiquitous enzymes which play vital roles in nearly all DNA metabolic processes. In recent years, the biochemical activity and three-dimensional structure of several PIF1 helicases have been reported, but there are few reports on the PIF1 helicase of bacteria living in extreme environments. In this paper, a series of biochemical and biophysical techniques were used to study the Thermodesulfovibrio yellowstonii PIF1 (Ty.PIF1) helicase in many aspects. Ty. PIF1 was obtained with a purity of over 90% and good uniformity using the prokaryotic expression and purification system. Ty.PIF1 is a monomer with a calculated molecular weight of 60 kD in solution. Ty. PIF1 has high thermal stability. The secondary structure remains stable when the temperature is below 65 ℃, and the secondary structure changes only when the temperature is above 70 ℃. The optimal unwinding temperature of Ty.PIF1 in vitro is 45 ℃, which is not the optimal temperature for the survival of thermodesulfovibrio yellowstonii. It indicates that when Ty.PIF1 exerts its enzymatic activity in vivo, it may require the participation of other cofactors. Ty.PIF1 can exert unwinding activity in a wide temperature range (20-55 ℃), and the presence of enzyme activity at 55 ℃ indicates that Ty.PIF1 has heat-resistant properties. Ty.PIF1 prefers to bind to substrates containing ssDNA, but there is certain requirement for the length of the ssDNA, which is at least 4 nt in length. Ty.PIF1 can also bind to the G

OBJECTIVETo study the effects of fertilizers with the different proportional of nitrogen, phosphorus and potassium on growth and active ingredient of Astragalus membranaceus.

METHODField experiment was conducted based on the D-saturation optimal design with three factors of nitrogen, phosphorus and potassium. The effects on growth and active ingredient of A. membranaceus were analyzed.

RESULTFertilization promoted the seedling growth and provided abundant supply of nutrition for growth of root, yield and accumulation of active ingredient at the later growth stage, and increased the accumulation of dry matter of stem-leaf and root system. The effect of nitrogen, phosphorus and potassium application on the total dry matter accumulation of A. membranaceus was as following: nitrogen > potassium > phosphorus; the effect on the stem-leaf dry matter accumulation was as following: nitrogen > phosphorus > potassium; the effect on the root dry matter accumulation was as following: nitrogen > potassium > phosphorus. Nitrogen, phosphorus and potassium fertilizer increased the root yield of A. membranaceus. Obviously, the effect on the root yield was as following: nitrogen > potassium > phosphorus. The application of different proportional with nitrogen, phosphorus and potassium increased the content of polysaccharide and astragaloside, but had no distinct effect on the content of total flavonoids. The effect on the content of polysaccharide was as following: potassium > phosphorus > nitrogen, but the effect on the conten,t of astragaloside was as following: nitrogen > potassium > phosphorus.

CONCLUSIONNitrogen and potassium fertilizer application had more important effect on growth, yield and the contents of polysaccharide and astragaloside in A. membranaceus. During medicinal plants cultivation process, it should pay attention to the application of nitrogen fertilizer and potassium fertilizer and make balance application of nitrogen, phosphorus and potassium fertilizer.

Astragalus membranaceus ; drug effects ; growth & development ; metabolism ; Fertilizers ; Nitrogen ; pharmacology ; Phosphorus ; pharmacology ; Potassium ; pharmacology

Abstract. Species-diagnostic anatomical characters of fleshflies are not known for most immature stages or even adults, and an existing key may be incomplete or difûcult for nonspecialists to use. The use of sarcophagids for PMI estimations has been greatly hampered by their highly similar morphological characters. DNA-based method can be used as a supplemental means of morphological method in identification of forensically important sarcophagid flies. However, relying solely on single DNA fragment for delimiting species is considered to be unreliable, especially when the fragment was small. Sequence data of selected regions of the cytochrome oxidase subunit two (COII) and 16S ribosomal RNA (16SrRNA) genes of the most important Chinese fleshfly taxa associated with cadavers are presented, which can be instrumental for implementation of the Chinese Sarcophagidae database. Phylogenetic analysis of the sequenced segments showed that all sarcophagid specimens were properly assigned into five species, which indicated the possibility of separation congeneric species with the short fragments.

BACKGROUND: Aerococcus urinae is a rare pathogen of endocarditis with high rates of embolic events, valvular damage and mortality.
CASE 1:A 24 y/o male, with mitral valve prolapse, presented with recurrent fever and body malaise for four months. ?-hemolytic streptococci was isolated in his blood 3 months prior, antibiotics for 10 days temporarily relieved his symptoms. He denied illicit drug use, recent dental, genitourinary manipulations. On admission, he was febrile with a 4/6 holosystolic murmur at the apex. He had leukocytosis and elevated acute phase reactants. Blood cultures: Aerococcus urinae. A TEE revealed: ruptured chordae and vegetation at the posterior mitral valve leaflet. Gentamicin for 14 days and Ceftriaxone for 28 days was completed. Mitral valve replacement was done and LV dimension returned to normal.
CASE 2:A 51 y/o male presented with 9 days of fever, chills, and malaise. He was treated with norfloxacin with no relief of symptoms. On admission, he was febrile, with a grade 2/6 holosystolic murmur at the apex and left parasternal area. He had leukocytosis and blood culture grew Aerococcus urinae. Echocardiogram showed mitral stenosis, aortic stenosis and vegetations at the mitral valve and non coronary cusp. Pen-G plus Gentamicin for 14 days and upon discharge, amoxicillin for 2 weeks was completed.
DISCUSSION: Risk factors associated with A. urinae endocarditis are >65 year s of age, male, urologic abnormalities, malignancy and diabetes. Diagnosis isusually made by culture as our cases and both were found to have vegetations by echocardiography. B-lactam and amino glycoside treatment is effective and although mortality is high, both patients improved and were discharged.
CONCLUSION: A. urinae endocarditis does occur in a young population and to those without urologic abnormality.

Human ; Male ; Middle Aged ; Young Adult ; Aerococcus ; Amoxicillin ; Anti-bacterial Agents ; Aortic Valve Stenosis ; Diabetes Mellitus ; Endocarditis ; Mitral Valve ; Risk Factors ;