OBJECTIVETo investigate the relationship between dentine phosphoprotein (DPP) and remineralization of demineralized dentine.

METHODS(1) Soluble DPP was extracted with 1 mol/L NaCl from demineralized dentine and was evaluated. (2) Soluble DPP was removed with 0.1 mol/L NaCl or was not removed from demineralized dentine sections in human tooth roots. Then all sections were subjected to remineralization treatment, and remineralization degrees were compared by atomic absorption spectrum, SEM and microradiography.

RESULTS(1) Soluble DPP was extracted with 1 mol/L NaCl. (2) Removal of soluble DPP resulted in significantly lower calcium concentration in remineralization solution (P < 0.01), less mean light-absorbed value in demineralized dentin sections by microradiography (P < 0.01).

CONCLUSIONSSoluble DPP may have an inhibiting effect on remineralization of demineralized dentine, this study suggests that the remove of soluble DPP from root caries lesions may enhance their remineralization potential.

Adolescent ; Child ; Dentin ; chemistry ; Humans ; In Vitro Techniques ; Phosphoproteins ; isolation & purification ; physiology ; Tooth Demineralization ; Tooth Remineralization

Recently, the gene encoding clathrin assembly protein of lymphoid myeloid leukemia (CALM), which is homologous to the AP180, was cloned from rat brain, and its expression differential to AP180 was reported (Kim and Lee, 1999). This gene product promotes the polymerization of clathrin into clathrin cage and found to be a regulator in membrane trafficking between intracellular compartments in eukaryotic cells (Kim et al., 2000). In this study, we have purified the CALM protein from clathrin-coated vesicles of rat liver using the monoclonal antibody against the recombinant N-terminal region of the CALM. The coated proteins extracted from the coated vesicle fraction was further purified by multi-step procedures involving gel-filtration and ion-exchange chromatography and SDS-PAGE. The purified protein with an apparent molecular weight of 100 kD promoted the assembly of clathrin triskelia into clathrin cage. In this respect the CALM protein bears a functional resemblance to the AP180 that has been previously described.
Animal ; Clathrin/*metabolism ; Clathrin-Coated Vesicles/*chemistry ; Liver/*chemistry ; Nerve Tissue Proteins/*isolation & purification ; Phosphoproteins/*isolation & purification ; Rats

OBJECTIVETo investigate the influence of soluble phosphoprotein removal from artificial dentin caries lesions by pretreatment of NaCl on subsequent remineralization.

METHODSHuman root dentin samples were demineralized in an acidic gel (pH = 4.4) at 37 degrees C for 48 hours. Samples were pretreated with 0.5 mol/L NaCl solution and 0.5 mol/L disodium EDTA solution respectively. The histomorphologic changes of root caries surface were examined by scannings electronic microscope. Mineral profiles were assessed by means of microradiography.

RESULTSNo histomorphologic changes were present on root caries surface after treated with 0.5 mol/L NaCl solution, while many opened dentin tubules were seen after treated with 0.5 mol/L disodium EDTA solution. The remineralization results showed that pretreatment with 0.5 mol/L NaCl solution increased more remineralization than pretreated with 0.5 mol/L disodium EDTA solution.

CONCLUSION0.5 mol/L NaCl solution has no damage on the root caries surface. The in vitro results suggested that removal of soluble phosphoprotein from dentin lesions is an interesting approach to enhance remineralization.

Dental Caries ; diagnostic imaging ; pathology ; physiopathology ; Humans ; Microscopy, Electron, Scanning ; Phosphoproteins ; isolation & purification ; Radiography ; Sodium Chloride ; pharmacology ; Tooth Calcification ; drug effects

OBJECTIVE: To express the recombinant SPLUNC1 protein in HNE1 cells and to study its function of bactericidal and binding to lipopolysaccharide (LPS). METHODS: Full length of SPLUNC1 gene was cloned into pCMV-tag4A vector and stably transfected into HNE1 cell lines, the supernatant of cell cultures was collected. After being treated with the supernatant, the Pseudomonas aeruginosa was seeded to LB soft agar plate, and the bacteria clones were counted and analyzed. For in vitro LPS binding assay, LPS was coated to 96-well plates. We incubated in the plate with SPLUNC1 protein, and detected the binded SPLUNC1 protein by ELISA. Incubating the FITC-LPS with the SPLUNC1 stably transfected or control cells, the intracellular intensity of fluorescence was observed under the fluorescence microscope. RESULTS: SPLUNC1 inhibited the bacteria clone formation obviously. Although the binding efficiency of LPS and SPLUNC1 in vitro was very low, more FITC-LPS entered into the SPLUNC1 stably transfected cells. CONCLUSION SPLUNC1 can inhibit the growth of Pseudomonas aeruginosa and bind LPS, and play an important defensive role in innate immunity of the upper airway.
Cell Line, Tumor ; Glycoproteins ; isolation & purification ; pharmacology ; Humans ; Membrane Proteins ; chemistry ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Phosphoproteins ; isolation & purification ; pharmacology ; Pseudomonas aeruginosa ; drug effects ; Respiratory Mucosa ; chemistry ; immunology ; Respiratory System ; chemistry ; immunology ; Transfection

Adolescent ; Adult ; Antigens, Viral ; blood ; Cytomegalovirus ; isolation & purification ; DNA, Viral ; analysis ; Female ; Fluorescence ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Phosphoproteins ; blood ; Polymerase Chain Reaction ; methods ; Viral Load ; Viral Matrix Proteins ; blood

Adolescent ; Adult ; Aged ; COVID-19/virology* ; COVID-19 Nucleic Acid Testing/methods* ; Child ; Coronavirus Nucleocapsid Proteins/genetics* ; Feces/virology* ; Female ; Humans ; Male ; Middle Aged ; Phosphoproteins/genetics* ; RNA, Viral/genetics* ; SARS-CoV-2/isolation & purification* ; Young Adult

Human cytomegalovirus (HCMV) infection is an ubiquitous herpesvirus disease in human populations. It is rarely pathogenic to healthy adults, yet it may cause severe outcome to organ transplant recipients, the immunocompromised individuals and pregnant women. Using DNA from HCMV AD169 strain as template, the UL32 gene encoding pp150 protein fragment and the UL57 gene encoding MDBP protein fragment were amplified by PCR technique. After the construction of cloning vector pMD18-T-UL32, pMD18-T-UL57, pMD18-T-UL32/UL57 and expression vector pET-11a-UL32/UL57, the recombinant fusion proteins pp150/MDBP were induced with IPTG in BL21 host strain. The results showed that the relative molecular weight of recombinant fusion proteins pp150/MDBP is about 27 kD, the product of fusion proteins takes 17.45% in the total proteins in host bacteria, the analytical result was positive to the fusion proteins pp150/MDBP via Western blot technique, while the purified recombinant fusion proteins have strong antigenicity detected by ELISA and protein chip compared with whole virus antigens from HCMV. It was demonstrated that when used for the detection of serum from the clinical patients it has the same detection rate compared with the whole virus antigen. It needs further research for application.
Cytomegalovirus ; DNA-Binding Proteins ; chemistry ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Genetic Vectors ; genetics ; Humans ; Peptide Fragments ; blood ; genetics ; immunology ; isolation & purification ; Phosphoproteins ; chemistry ; Plasmids ; genetics ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; blood ; genetics ; immunology ; isolation & purification ; Transcription Factors ; chemistry ; Viral Matrix Proteins ; chemistry

OBJECTIVEElevation of reactive oxygen species (ROS), especially the level of superoxide is a key event in many forms of cardiovascular diseases. To study the mechanism of tea polyphenols against cardiovascular diseases, we observed the expressions of ROS-related enzymes in endothelial cells.

METHODSTea polyphenols were co-incubated with bovine carotid artery endothelial cells (BCAECs) in vitro and intracellular NADPH oxidase subunits p22phox and p67phox, SOD-1, and catalase protein were detected using Western blot method.

RESULTSTea polyphenols of 0.4 microg/mL and 4.0 microg/mL (from either green tea or black tea) down-regulated NADPH oxidase p22phox and p67phox expressions in a dose-negative manner (P < 0.05), and up-regulated the expressions of catalase (P < 0.05).

CONCLUSIONSTea polyphenols regulate the enzymes involved in ROS production and elimination in endothelial cells, and may be beneficial to the prevention of endothelial cell dysfunction and the development of cardiovascular diseases.

Animals ; Camellia sinensis ; chemistry ; Carotid Arteries ; cytology ; Catalase ; biosynthesis ; Cattle ; Cells, Cultured ; Down-Regulation ; Endothelial Cells ; drug effects ; enzymology ; metabolism ; Flavonoids ; isolation & purification ; pharmacology ; Membrane Transport Proteins ; biosynthesis ; NADPH Dehydrogenase ; biosynthesis ; NADPH Oxidases ; Phenols ; isolation & purification ; pharmacology ; Phosphoproteins ; biosynthesis ; Polyphenols ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; biosynthesis ; Superoxide Dismutase-1 ; Up-Regulation

OBJECTIVE: To investigate the effect of purified vitexin compound 2 (VB2), a noval lignanoid from the acetoacetate extract of Vitex negundo seed on the proliferation and apoptosis as well as the expression of mTOR and 4E-BP1 mRNA signal pathway in human choriocarcinoma JEG-3 cell lines in vitro. METHODS: The inhibitory effect of different concentrations of VB2 on JEG-3 cells was examined by methyl thiazolyl tetrazolium (MTT) assay. Flow cytormetry was used to analyze the apoptosis after using different concentrations of VB2, and the expression of mTOR and 4E-BP1 mRNA was determined by RT-PCR. RESULTS: The inhibitory rate of JEG-3 cell growth which was cultured with different concentrations of VB2 (2.5, 5.0, 10.0, 20.0, 40.0, 80.0, and 160.0 μmol/L) for 24, 48, or 72 hours increased from (6.34±0.41)% to (85.89±0.81)%, and it was positively correlated with the dose and time of culture (P<0.05). VB2 at 5.0, 10.0, or 20.0 μmol/L increased the rate of JEG-3 cell apoptosis in vitro from (9.26±1.02)% to (35.55±1.24)% after 48 hour culture, which was in a dose dependent manner (P<0.05), while 5.0, 10.0, or 20.0 μmol/L of VB2 down-regulated the mRNA levels of mTOR and 4E-BP1 after 48 hour culture, which presented a significant negative correlation between VB2 and the mRNA levels of mTOR and 4E-BP1(P<0.05). CONCLUSION VB2 can restrain the proliferation of choriocarcinoma cell JEG-3 and induce its apoptosis. This effect may be related to the inhibition of VB2 on the mRNA expression of JEG-3 cell mTOR and 4E-BP1.
Adaptor Proteins, Signal Transducing ; metabolism ; Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; pharmacology ; Apigenin ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Choriocarcinoma ; pathology ; Female ; Humans ; Phosphoproteins ; metabolism ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; metabolism ; Uterine Neoplasms ; pathology ; Vitex ; chemistry

The nucleotide sequences of P gene from a field strain of peste des petits ruminants virus (PPRV) ("China/Tib/Gej/07-30") was firstly determined. The P gene is 1,655 nucleotides long with two overlapping open reading frames (ORFs). The first ORF is 1530 nucleotides long and would produce P protein of 509 amino acid residues. The second ORF is 534 nucleotides long and would produce C protein of 177 amino acid residues. The first ORF produces a second mRNA transcript of 897 nucleotides long with an extra G nucleotide at position 751. Translation from this mRNA would produce V protein of 298 amino acid residues. The nucleotide and deduced amino acid sequence were compared with the homologous region of other PPRV isolates. At the amino acid level, the "China/Tib/Gej/07-30" shares homology of 86.10%-97.3%, 84.3%-94.9%, and 82.9%-96.3% for P, C, and V proteins respectively. Several sequence motifs in the P genes were identified on the basis of conservation in the PPRVs and the morbilliviruses.
Amino Acid Sequence ; Animals ; China ; Female ; Goat Diseases ; virology ; Goats ; Molecular Sequence Data ; Peste-des-Petits-Ruminants ; veterinary ; virology ; Peste-des-petits-ruminants virus ; chemistry ; genetics ; isolation & purification ; metabolism ; Phosphoproteins ; chemistry ; genetics ; metabolism ; Sequence Analysis ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics ; metabolism