Ear Diseases ; etiology ; Humans ; Radiation Injuries

Nucleases is an important enzyme widely used in biotechnology. A codon optimized nuclease gene (SNU) from Northern Shrimps was inserted into pPICZα A vector, and expressed extracellularly in strain SMD1168H. On the basis of multi-copy recombinant strain, we further optimized the expression condition and characterized SNU. SNU was highly expressed and stable after 1% methanol induction for 72 h, yield reached 1.4×10⁵ U/mL. SDS-PAGE electrophoresis demonstrated that this is a N-linked glycoprotein of 50 kDa. It was purified by one step DEAE Sephadex chromatography to the purity of about 15 mg/L with a specific activity of 6.291×10⁶ U/mg. Functional analysis on the nuclease activity indicated that it was stimulated by bivalent iron, such as Ca²⁺, Mn²⁺, Co²⁺ and Mg²⁺, but inhibited by Zn²⁺, Cu²⁺ and high salt. Meanwhile, it was irreversibly inactivated at 70 ℃ for 10 min.
Animals ; Codon ; Electrophoresis, Polyacrylamide Gel ; Endonucleases ; biosynthesis ; Glycoproteins ; Hot Temperature ; Penaeidae ; enzymology ; Pichia ; metabolism ; Recombinant Proteins ; biosynthesis

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.

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.

Objective: To identify the spatial clustering and its temporal trends among newly detected female HIV/AIDS cases aged 15 years or older, in China from 2010 to 2016. Methods: Newly detected HIV/AIDS cases among aged 15 years or older women in China during 2010-2016 were collected, to describe their demographic characteristics, changing trends and spatial autocorrelation. This program was conducted at county level, using the ArcGIS 10.3. Results: The number of newly detected HIV/AIDS cases among aged 15 years or older women was increasing annually from 16 603 to 26 196 in 2010 and in 2016. As the main route proportion of heterosexual transmission increased from 84.25% (13 988/16 603) in 2010 to 96.29%(25 224/26 196) in 2016. Both the number and proportion of HIV/AIDS cases among elderly women ≥50 years of age increased significantly from 17.82%(2 959/16 603) to 38.10%(9 981/26 196) in 2016. Results from spatial analysis demonstrated a county-level clustered distribution of HIV/AIDS cases across the country with a rising global Moran's I value=0.55 over the years (Z=51.46, P<0.001), which was concentrating on western and southern China, covering 9 provinces/autonomous regions/municipalities (Yunnan, Guangxi, Sichuan, Xinjiang, Guizhou, Guangdong, Chongqing, Henan and Hunan). The temporal trends of hot spots differed by age groups, with the trend of epidemic shifting towards western border and southern coastal regions among women aged 15-49 years old, while the elderly women aged ≥50 years old were spreading northward from the southwestern regions. Conclusion: Our findings indicated that an increasing trend of clusters appeared on HIV epidemic among newly detected female HIV/AIDS cases aged 15 years or older in China, particularly in the western and southern regions. Prevention and intervention strategies should target on women according to their age distribution, particularly in regions with increasing trend of HIV epidemics.
Adolescent ; Adult ; Age Distribution ; Aged ; China/epidemiology* ; Epidemics ; Female ; HIV Infections/ethnology* ; Humans ; Middle Aged ; Spatial Analysis ; Young Adult

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

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.

Two-stage reimplantation is considered the gold standard for the management of periprosthetic joint infection. The first stage involves the removal of the prosthesis, followed by extensive debridement of all nonviable tissues, synovectomy, irrigation, and reaming of the medullary canals. Once the joint has been prepared, antibiotic-impregnated cement beads and/or spacer are inserted. Antibiotic-impregnated spacers are now more commonly used, and an increasing number of orthopaedic surgeons are using articulating spacers if indicated. Antibiotics are then prescribed to the patient based on the sensitivities of the infecting organism. The duration of systemic antibiotic use is decreasing, and short courses of antibiotic therapy have been shown to be as efficacious as prolonged therapy between the first and second stages. The second stage of the procedure involves removal of the antibiotic-impregnated cement beads and/or spacer, repeat irrigation and debridement, and final reconstruction with revision components. While two-stage reimplantation was considered the gold standard in many parts of the world, recent studies have demonstrated that it is associated with a considerable failure rate. This may be due to the lack of accurate diagnostic tools for infection eradication, and future investigation of risk factors for failure of the two-stage reimplantation should be conducted.
Anti-Bacterial Agents ; Arthroplasty ; Arthroplasty, Replacement, Knee ; Debridement ; Humans ; Joints ; Knee ; Prostheses and Implants ; Replantation ; Risk Factors ; Surgeons

Biotechnology ; China ; Cooperative Behavior ; Cuba ; Federal Government ; Translational Medical Research

Glucomannan is the key active ingredient of Dendrobium catenatum, and CSLA family is responsible for glucomannan biosynthesis. In order to systematically evaluate the CSLA family members of D. catenatum, the bioinformatics methods were performed for genome-wide identification of DcCSLA gene family members through the genomic data of D. catenatum downloaded from the NCBI database, and further analyses of their phylogenetic relationship, gene structure, protein conserved domains and motifs, promoter cis-elements and gene expression profiles in response to stresses. The results showed that D. catenatum contains 13 CSLA members, all of which contain 9-10 exons. In the evolutionary relationship, CSLA genes were clustered into 5 groups, DcCSLA genes were distributed in all branches. Among which the ancestral genes of groupI existed before the monocot-dicot divergence, and groupⅡ-Ⅴ only existed in the monocot plants, indicating that group Ⅰ represents the earliest origin group. CSLA proteins are characteristic of the signature CESA_CaSu_A2 domain. Their promoter regions contain cis elements related to stresses and hormones. Under different stress treatments, low temperature induces the expression of DcCSLA5 and inhibits the expression of DcCSLA3. Infection of Sclerotium delphinii inhibits DcCSLA3/4/6/8/9/10 expression. Under the treatment of jasmonic acid, DcCSLA11 expression was significantly up-regulated, and DcCSLA2/5/7/12/13 were significantly down-regulated. These results laid a foundation for further study on the function of DcCSLA genes in glucomannan biosynthesis and accumulation.
Basidiomycota ; Cold Temperature ; Dendrobium ; genetics ; Gene Expression Regulation, Plant ; Genome, Plant ; Multigene Family ; Phylogeny ; Plant Proteins ; genetics ; Stress, Physiological ; Transcriptome