Adolescent ; Adult ; Female ; Fenestration, Labyrinth ; methods ; Humans ; Male ; Middle Aged ; Neuroma, Acoustic ; surgery ; Young Adult

Miniature inverted-repeat transposable elements transposon is a special transposon that could transpose by "cut-paste" mechanism, which is one of characteristics of DNA transposons. Otherwise, the copy number of MITEs is very high, which is one of characteristics of RNA transposons. Many MITE families have been reported, but little about active MITEs. We summarize recent advances in studying active MITEs. Most the MITEs belong to the Tourist-like family, such as mPing, mGing, PhTourist1, Tmi1 and PhTst-3. Additionally, DTstu1 and MITE-39 belong to Stowaway-like family, and AhMITEs1 belongs to Mutator-like family. Moreover, we summarize the structure (terminal inverse repeats and target site duplications), copy number, evolution pattern and transposition characteristics of these active MITEs, to provide the foundation for the identification of other active MITEs and subsequent research on MITE transposition and amplification mechanism.

Juvenile dermatomyositis (JDM) is an autoimmune disease manifesting as proximal muscle weakness and skin rash and can involve multiple systems and visceral organs. Myositis-specific autoantibodies (MSAs) are highly associated with various complications and prognosis in JDM. Patients with anti-Mi-2 antibodies tend to have good prognosis and typical clinical symptoms. Patients with anti-MDA5 antibodies often have diffuse interstitial lung disease and skin ulcer, with mild symptoms of myositis. Patients with anti-NXP2 antibodies often have calcinosis, and such antibodies are associated with gastrointestinal bleeding and perforation. Patients with anti-TIF1-γ antibodies have diffuse and refractory skin lesions. Anti-SAE antibodies are rarely detected in children, with few reports of such cases. This article reviews the features of clinical phenotypes in JDM children with these five types of MSAs, so as to provide a basis for the clinical treatment and follow-up management of children with JDM.
Autoantibodies ; Dermatomyositis ; Humans ; Lung Diseases, Interstitial/etiology* ; Myositis ; Prognosis

MCL-1 is encoded by myeloid cell leukemia-1 gene (mcl-1), which is one of the anti-apoptotic members of bcl-2 cell apoptotic gene superfamily. ChanSu is made of dorsal secretions of several Bufo species, commonly used in the prescriptions of traditional Chinese medicine for treating many diseases including cancer. To clarify if mcl-1 is expressed in the dorsal skin of B. gargarizans, the PCR (polymerase chain reaction) was performed with its dorsal skin first strand cDNA as the template and a pair of specific primers of mcl-1, and PCR products were cloned into the pGM-T vector. DNA sequencing indicated that the ORF length was 639 bp encoding 212 amino acid residues, and the homology of 44%-95% with the MCL-1 of several other animals. For the further studies on MCL-1 biological functions during the oncogenesis and preparation of its antibody, the prokaryotic expression construct of pET-28b-mcl-1 was prepared which was confirmed by DNA sequencing, and its recombinant protein expression (0.02% wet weight) in E. coli BL21 (DE3) strain was confirmed by SDS-PAGE and Western blotting.
Amino Acid Sequence ; Animals ; Bufonidae ; classification ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; metabolism ; Myeloid Cell Leukemia Sequence 1 Protein ; genetics ; metabolism ; Open Reading Frames ; genetics ; Phylogeny ; Recombinant Proteins ; genetics ; metabolism ; Sequence Alignment ; Sequence Homology ; Skin ; metabolism

Headspace-solid phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS) were used to analyze the interaction between the β-lactoglobulin (β-LG) and the botany volatile organic compounds (BVOCs) from pomelo peel to screen out the pharmacodynamic active BVOCs substance group. The selective binding effect between β-LG and BVOCs was analyzed by quantitative recovery of BVOCs, and the binding parameters were calculated. Then, the molecular model of BVOCs binding with β-LG was established by molecular docking and spectroscopic method, and the molecular mechanism of interaction between pharmacodynamic active BVOCs and β-LG was discussed from the perspective of omics. The results showed that dipentene (Dt), linalylacetate (La) and nootkatone (Nt) of BVOCs were selected by HS-SPME/GC-MS by the interaction of β-LG and BVOCs substance group. Parameter calculation showed that β-LG had the strongest affinity with Nt, but the binding force was not strong, and the affinity for La was the weakest. The affinity of β-LG to Dt was weak, but the binding force was the strongest, with a binding rate of 54. 66%, indicating that the selective binding strength of β-LG with the pharmacodynamic active BVOCs depended on the chemical structure of BVOCs molecules. The β-LG preferred to bind to the aldehyde and ketone BVOCs molecules containing carbonyl oxygen structure. The molecular model of β-LG and BVOCs group (Dt, La, Nt) was established to evaluate the binding position of BVOCs group (Dt, La, Nt) on β-LG. The loosening, extension and conformational change of β-LG secondary structure caused by the introduction of BVOCs are the result of van der Waals force, hydro-phobicity and hydrogen bonding. This study provides a new method for screening pharmacodynamic active BVOCs from the perspective of whole substance group of BVOCs, and provides a useful reference for investigating the binding mechanism between pharmacodynamic active BVOCs and functional protein molecules from the perspective of omics.

Molecular mechanisms underlying the effects of Fyn on cell morphology, pseudopodium movement, and cell migration were investigated. The Fyn gene was subcloned into pEGFP-N1 to produce pEGFP-N1-Fyn. Chinese hamster ovary (CHO) cells were transfected with pEGFP-N1-Fyn. The expression of Fyn mRNA and proteins was monitored by reverse transcription-PCR and Western blotting. Additionally, transfected cells were stained with 4',6-diamidino-2-phenylindole and a series of time-lapse images was taken. Sequences of the recombinant plasmids pMD18-T-Fyn and pEGFP-N1-Fyn were confirmed by sequence identification using National Center for Biotechnology Information in USA, and Fyn expression was detected by RT-PCR and Western blotting. The morphology of CHO cells transfected with the recombinant vector was significantly altered. Fyn expression induced filopodia and lamellipodia formation. Based on these results, we concluded that overexpression of mouse Fyn induces the formation of filopodia and lamellipodia in CHO cells, and promotes cell movement.
Animals ; Blotting, Western ; CHO Cells ; Cricetinae ; Cricetulus ; Genetic Vectors ; Green Fluorescent Proteins/genetics ; Mice ; Proto-Oncogene Proteins c-fyn/genetics/*metabolism ; Pseudopodia/*metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Time-Lapse Imaging ; Transfection

OBJECTIVEMcAbs against rabies nucleocapsid were used to detect rabies street viruses in animal brain specimens with indirect immunofluorescent assay to evaluate the sensitivity and specificity of this assay.

METHODS62 specimen from rabid animal brains including genotype 1 to 7 and 271 specimens from different normal animal brains collected in Pasteur Institute in 2003 were tested and compared, using indirect immunofluorescent assay. All these specimens were identified and compared using rapid rabies enzyme immunodiagnosis, fluorescent antibody test and rabies virus isolation assay in neuroblastoma cell culture which were all provided by Pasteur Institute.

RESULTSBoth sensitivity and the specificity of the indirect immunofluorescent assay were 100%.

CONCLUSIONThe results showed a positive of rabies virus detection with these methods.

Animals ; Antibodies, Monoclonal ; immunology ; Antibodies, Viral ; immunology ; Brain ; virology ; Dogs ; Fluorescent Antibody Technique ; methods ; Genotype ; Nucleocapsid ; immunology ; Rabies virus ; isolation & purification ; Sensitivity and Specificity

OBJECTIVETo observe the activity of repeated extracts of bone matrix and the production of purified bone morphogenetic proteins (BMPs).

METHODSBMPs were extracted 1- 4 times from fresh bovine cortical bone by the modified Urist's method, with each collected precipitate separated and lyophilized as partially purified BMPs. Another fresh bovine bone was extracted three times and the precipitates were mixed and lyophilized. Meanwhile,the alkaline phosphatase (ALP) activity was measured by an in vitro assay employing cultured C2C12 mouse myoblast cells through the osteoinductivity of bovine BMPs extracted four times at days 1, 4, 7, and 14, and the correlation between BMPs quantities and costing during extraction processes was analyzed.

RESULTSThe BMPs purified and the cost showed a positive correlation (r=0.969). To separate and lyophilize each collected precipitate as partially purified BMPs raised the cost, and mixed precipitates also cost much. ALP activities of the 1st and mixed extractions of BMPs were shown to be highly osteoinductive and keep a significantly high level (P<0.05-0.01) 4 days after culturing, compared with the 2nd, 3rd and 4th extractions, especially the control group. However, the more times the extraction was done, the less activity of BMPs was shown and more costing was. The x-ray and histological analysis also showed that the 1st extraction of BMPs induced more ossicles and new bone formation.

CONCLUSIONSThe results indicated that BMPs enhanced the abilities of osteoinductivity in C2C12 culture in vitro. The first extraction of BMPs from bone is fitfull, the second extraction should be enough, while, the 3rd and 4th extractions are unnecessary for they cost more and waste more time, say nothing of mixed extractions.

Alkaline Phosphatase ; analysis ; Analysis of Variance ; Animals ; Biopsy, Needle ; Bone Matrix ; pathology ; physiology ; Bone Morphogenetic Proteins ; analysis ; metabolism ; Cattle ; Immunohistochemistry ; Mice ; Osteogenesis ; physiology ; Probability ; Sensitivity and Specificity ; Time Factors ; Tissue Culture Techniques

Field experiments were conducted in Shangluo pharmaceutical base in Shaanxi province to study the effect of nitrogen, phosphorus and potassium in different fertilization levels on Platycodon grandiflorum soil microorganism and activities of soil enzyme, using three-factor D-saturation optimal design with random block design. The results showed that N0P2K2, N2P2K0, N3P1K3 and N3P3K1 increased the amount of bacteria in 0-20 cm of soil compared with N0P0K0 by 144.34%, 39.25%, 37.17%, 53.58%, respectively. The amount of bacteria in 2040 cm of soil of N3P1K3 increased by 163.77%, N0P0K3 increased the amount of soil actinomycetes significantly by 192.11%, while other treatments had no significant effect. N2P0K2 and N3P1K3 increased the amounts of fungus significantly in 0-20 cm of soil compared with N0P0K0, increased by 35.27% and 92.21%, respectively. N3P0K0 increased the amounts of fungus significantly in 20-40 cm of soil by 165.35%, while other treatments had no significant effect. All treatments decrease soil catalase activity significantly in 0-20 cm of soil except for N2P0K2, and while N2P2K0 and NPK increased catalase activity significantly in 2040 cm of soil. Fertilization regime increased invertase activity significantly in 2040 cm of soil, and decreased phosphatase activity inordinately in 0-20 cm of soil, while increased phosphatase activity in 2040 cm of soil other than N1P3K3. N3P0K0, N0P0K3, N2P0K2, N2P2K0 and NPK increased soil urease activity significantly in 0-20 cm of soil compared with N0P0K0 by 18.22%, 14.87%,17.84%, 27.88%, 24.54%, respectively. Fertilization regime increased soil urease activity significantly in 2040 cm of soil other than N0P2K2.
Bacteria ; enzymology ; growth & development ; isolation & purification ; metabolism ; Bacterial Proteins ; analysis ; metabolism ; Catalase ; analysis ; metabolism ; Fertilizers ; analysis ; Fungal Proteins ; analysis ; metabolism ; Fungi ; enzymology ; growth & development ; isolation & purification ; metabolism ; Nitrogen ; metabolism ; Phosphoric Monoester Hydrolases ; analysis ; metabolism ; Phosphorus ; metabolism ; Potassium ; metabolism ; Soil ; chemistry ; Soil Microbiology ; Urease ; analysis ; metabolism

OBJECTIVETo study the expression of bone matrix protein (BMP) induced by bovine bone morphogenetic proteins (BMPs) in vitro.

METHODSType I collagen, osteopontin (OPN), osteonectin (ON), osteocalcin (OC), and bone sialoprotein (BSP) were detected by immunohistochemistry in C2C12 cultured from day 1 to day 28.

RESULTSThe signaling of bone matrix protein expression became weaker except for type I collagen, OC and BSP after 5 days. Fourteen days after culture, the positive signaling of type I collagen, OPN, ON, OC, and BSP was gradually declined, and could be detected significantly as compared with that of the negative control on day 28. BMP assay showed that the alkaline phosphatase (ALP) activity was higher in C2C12 culture than in the control during the 14-day culture. Also, total protein and DNA significantly increased during the 14-day culture. High levels of ALP were seen in preosteoblasts and osteoblasts in vivo and in differentiating osteoblasts in vitro. ALP was well recognized as a marker reflecting osteoblastic activity.

CONCLUSIONNative bovine BMP induces conversion of myoblasts into osteoblasts, produces type I collagen, and plays significantly role in osteoinduction and bone matrix mineralization of C2C12 in vitro.

Alkaline Phosphatase ; metabolism ; Animals ; Bone Matrix ; metabolism ; Bone Morphogenetic Proteins ; pharmacology ; Cattle ; Cell Line ; DNA ; metabolism ; Gene Expression Regulation ; physiology ; Mice ; Osteoblasts ; drug effects ; metabolism