ABSTRACT
BACKGROUND: LincRNAs have been revealed to be tightly associated with various tumorigeneses and cancer development, but the roles of specific lincRNA on tumor-related angiogenesis was hardly studied. Here, we aimed to investigate whether linc-OIP5 in breast cancer cells affects the angiogenesis of HUVECs and whether the linc-OIP5 regulations are involved in angiogenesis-related Notch and Hippo signaling pathways. METHODS: A trans-well system co-cultured HUVECs with linc-OIP5 knockdown breast cancer cell MDA-MB-231 was utilized to study the proliferation, migration and tube formation abilities of HUVECs and alterations of related signaling indicators in breast cancer cells and their conditioned medium through a series of cell and molecular experiments. RESULTS: Overexpressed linc-OIP5, YAP1, and JAG1 were found in breast cancer cell lines MCF7 and MDA-MB-231 and the expression levels of YAP1 and JAG1 were proportional to the breast cancer tissue grades. MDA-MB-231 cells with linc-OIP5 knockdown led to weakened proliferation, migration, and tube formation capacity of co-cultured HUVECs. Besides, linc-OIP5 knockdown in co-cultured MDA-MB-231 cells showed downregulated YAP1 and JAG1 expression, combined with a reduced JAG1 level in conditioned medium. Furthermore, a disrupted DLL4/Notch/NRP1 signaling in co-cultured HUVECs were also discovered under this condition. CONCLUSION: Hence, linc-OIP5 in MDA-MB-231 breast cancer cells may act on the upstream of the YAP1/Notch/NRP1 signaling circuit to affect proliferation, migration, and tube formation of co-cultured HUVECs in a non-cellular direct contact way through JAG1 in conditioned medium. These findings at least partially provide a new angiogenic signaling circuit in breast cancers and suggest linc-OIP5 could be considered as a therapeutic target in angiogenesis of breast cancers.
Subject(s)
Humans , Female , Transcription Factors/metabolism , Breast Neoplasms/pathology , Neuropilin-1/metabolism , Adaptor Proteins, Signal Transducing/metabolism , Receptors, Notch/metabolism , Tumor Microenvironment , Human Umbilical Vein Endothelial Cells/cytology , Breast Neoplasms/metabolism , Immunohistochemistry , Signal Transduction , Blotting, Western , Reverse Transcriptase Polymerase Chain Reaction , Cell Line, Tumor , Real-Time Polymerase Chain ReactionABSTRACT
Aseptic loosening is the most common chronic complication of total joint arthroplasty,which significantly shortens the life span of prosthesis and leads to mental stress and heavy economic burden of patients.Hence,how to overcome aseptic loosening is a focus in the research of joint arthroplasty.A series of mechanisms participates the pathogenesis of aseptic loosening,including oste-olysis induced by wear particles,micromotion,stress shielding,high fluid pressure,endotoxin and genetic factors.Osteolysis induced by wear particles is the most recognized mechanism.The latest study suggested that autophagy,epigenetic regulation and immunologic derangement all participate the development of osteolysis induced by wear particles.However,the prevention,treatment and even the concept of aseptic loosening is still controverial.We hope that the future study will provide a definite mechanism for aseptic loosening and provide new theoretic evidence for the prevention and treatment of aseptic loosening.
ABSTRACT
Background: Glycine oxidase (GO), a type of D-amino acid oxidase, is of biotechnological interest for its potential in several fields. In our previous study, we have characterized a new glycine oxidase (BceGO) from Bacillus cereus HYC-7. Here, a variant of N336K with increased the affinity against all the tested substrate was obtained by screening a random mutant library of BceGO. It is observed that the residue N336 is invariable between its homogeneous enzymes. This work was aimed to explore the role of the residue N336 in glycine oxidase by site-directed mutagenesis, kinetic assay, structure modeling and substrate docking. Results: The results showed that the affinity of N336H, N336K and N336R increased gradually toward all the substrates, with increase in positive charge on side chain, while N336A and N336G have not shown a little significant effect on substrate affinity. The structure modeling studies indicated that the residue Asn336 is located in a random coil between -J-18 and a-10. Also, far-UV CD spectra-analysis showed that the mutations at Asn336 do not affect the secondary structure of enzyme. Conclusion: Asn336 site was located in a conserved GHYRNG loop which adjoining to substrate and the isoalloxazine ring of FAD, and involved in the substrate affinity of glycine oxidase. This might provide new insight into the structure-function relationship of GO, and valuable clue to redesign its substrate specificity for some biotechnological application.
Subject(s)
Bacillus cereus/metabolism , Amino Acid Oxidoreductases/metabolism , Glycine/analogs & derivatives , Substrate Specificity , Kinetics , Polymerase Chain Reaction/methods , Mutagenesis, Site-Directed , Amino Acid Oxidoreductases/geneticsABSTRACT
<p><b>BACKGROUND</b>Gingerol is the generic term for pungent constituents in ginger, which has been reported to be effective for inhibiting vomiting. We attempted to investigate the antiemetic effect of gingerol and its effective mechanism on substance P and NK(1) receptors in minks.</p><p><b>METHODS</b>The antiemetic effect of gingerol was investigated during a 6-hour observation on a vomiting model in minks induced by cisplatin, (7.5 mg/kg, intraperitoneal). The distribution of substance P and NK(1) receptors in the area postrema and ileum were measured by immunohistochemistry, and the expression of NK(1) receptor in the area postrema and ileum were measured by Western blotting.</p><p><b>RESULTS</b>The frequency of cisplatin induced retching and vomiting was significantly reduced by pretreatment with gingerol in a dose-dependent manner (P < 0.05). Substance P-immunoreactive was mainly situated in the mucosa and submucosa of the ileum as well as in the neurons of the area postrema. The immunoreactive production of NK(1) receptor was mainly situated in the muscular and submucosa of ileum and the neurons of area postrema, gingerol markedly suppressed the increased immunoreactivity of substance P and NK(1)1 receptor induced by cisplatin in a dose-dependent manner (P < 0.05), and exhibited effective inhibition on the increased expression levels of NK(1) receptor in both the ileum and area postrema dose-dependently (P < 0.05).</p><p><b>CONCLUSIONS</b>Gingerol has good activity against cisplatin-induced emesis in minks possibly by inhibiting central or peripheral increase of substance P and NK(1) receptors.</p>
Subject(s)
Animals , Male , Area Postrema , Metabolism , Blotting, Western , Catechols , Therapeutic Uses , Disease Models, Animal , Fatty Alcohols , Therapeutic Uses , Ileum , Metabolism , Immunohistochemistry , Mink , Receptors, Neurokinin-1 , Metabolism , Substance P , Metabolism , Vomiting , Drug TherapyABSTRACT
<p><b>OBJECTIVE</b>To screen and identify the peptides that specifically bind to CD13 on monocytes.</p><p><b>METHODS</b>The phages capable of specific binding to CD13 were screened in the phage-displayed 12-peptide library. The affinity of the selected phages with CD13 was verified with enzyme-linked immunosorbent assay (ELISA). The sequences of the peptides bound to the phages were deduced according to the phage DNA sequences, and the functional peptides aligned using the BLASTP on the Website NCBI were synthesized. To analyze the biological function of the screened peptides, the location of the peptides bound to THP-1 cells was detected using immunofluorescence assay. The blocking effect of WM15 on the peptide binding to THP-1 cells was assessed by immunofluorescence assay.</p><p><b>RESULTS</b>The phages that specifically bound to CD13 were effectively enriched to approach saturation after 4 rounds of panning. The recovery rate in the fourth round was 30 times that in the first round. Twenty selected phages were verified by ELISA, and the signals of 10 phages were higher than the control. The sequences of the peptides P9 and P7 showed 83% and 100% identity with those of human cytomegalovirus (HCMV) UL38 and UL105, respectively. The peptides bound to the cell membrane of THP-1 cells as shown by immunofluorescence assay. The binding of the peptides P9 and P7 to THP-1 cells was blocked by CD13-specific monoclonal antibody WM15 at different levels.</p><p><b>CONCLUSION</b>Two peptides (P7 and P9) that can specifically bind to CD13 have been screened successfully, and these two peptides show specific binding to CD13 on the membrane of THP-1 cells.</p>
Subject(s)
Humans , Amino Acid Sequence , Binding, Competitive , CD13 Antigens , Metabolism , Cell Line , Molecular Sequence Data , Peptide Library , Peptides , Metabolism , Protein BindingABSTRACT
This article describes how to construct a integrated wireless clinical information system based on the original information system. It is constructed on the basis of the hospital HIS through the middle ware which provides a data-interaction platform for all the existing data and the data which would be added in future for various information management systems. The software system adopts the Web Service technology in information services. The SOAP protocol is applied to the data interaction between the foreground and the background. The mode of the network structure is "Wireless Switchboard + Intelligence antenna". Relying on the wireless network, the handheld data terminal and the barcode, the network information can be extended to the patient bedsides and the mobile medical personnels.
Subject(s)
Hospital Information Systems , Local Area Networks , Medical Records Systems, Computerized , Software , Systems Integration , User-Computer Interface , Wireless TechnologyABSTRACT
The complete nucleotide sequence of a cDNA clone encoding weever ribosomal protein L8, WeevL8, is described here. The WeevL8 cDNA has 848 nucleotides in full length and encodes a 257 amino-acid protein with a calculated molecular mass of 28.02 kDa. Two conserved domains have been identified in WEEVL8. It was concluded fromsequence alignment that WeevL8 gene was quite conservative, consistent with its role as a house-keeping gene. A phylogenetic analysis made by L8 protein showed the similar phylogenetic relationship to that with 18s rDNA. The high similarity supports the notion that ribosomal protein L8 can also be used as phylogenetic criterion.
ABSTRACT
The complete nucleotide sequence of a cDNA clone encoding weever ribosomal protein L8, WeevL8, is described here. The WeevL8 cDNA has 848 nucleotides in full length and encodes a 257 amino-acid protein with a calculated molecular mass of 28.02 kDa. Two conserved domains have been identified in WEEVL8. It was concluded fromsequence alignment that WeevL8 gene was quite conservative, consistent with its role as a house-keeping gene. A phylogenetic analysis made by L8 protein showed the similar phylogenetic relationship to that with 18s rDNA. The high similarity supports the notion that ribosomal protein L8 can also be used as phylogenetic criterion.