Staphylococcus aureus is the most important Gram-positive colonizer of human skin and nasal passage, causing high morbidity and mortality. SD-repeat containing protein D (SdrD), an MSCRAMM (Microbial Surface Components Recognizing Adhesive Matrix Molecules) family surface protein, plays an important role in S. aureus adhesion and pathogenesis, while its binding target and molecular mechanism remain largely unknown. Here we solved the crystal structures of SdrD N2-N3 domain and N2-N3-B1 domain. Through structural analysis and comparisons, we characterized the ligand binding site of SdrD, and proposed a featured sequence motif of its potential ligands. In addition, the structures revealed for the first time the interactions between B1 domain and N2-N3 domain among B domain-containing MSCRAMMs. Our results may help in understanding the roles SdrD plays in S. aureus adhesion and shed light on the development of novel antibiotics.
Amino Acid Sequence ; Bacterial Proteins ; chemistry ; genetics ; metabolism ; Binding Sites ; Calcium ; chemistry ; metabolism ; Calcium-Binding Proteins ; chemistry ; genetics ; metabolism ; Hydrogen Bonding ; Ligands ; Molecular Sequence Data ; Protein Binding ; Protein Structure, Tertiary ; Receptors, Cell Surface ; chemistry ; metabolism ; Recombinant Proteins ; biosynthesis ; chemistry ; genetics ; Sequence Alignment ; Staphylococcus aureus ; metabolism

Steroid cell tumor, not otherwise specified (NOS), are rare ovarian tumor, in addition, it is more rare in children. The majority of these tumors produce several steroid hormones, particularly testosterone. Estrogen also secreted by steroid cell tumor, NOS, but it is uncommon. Furthermore, hypertension is an infrequent sign in steroid cell tumor, NOS. An 8.5-yr-old girl with hypertension and frequent vaginal spotting visited at our clinic. On laboratory evaluation, secondary hypertension due to an elevated plasma renin level and isosexual pseudoprecocious puberty was diagnosed. Right solid ovarian mass was detected in radiologic tests. She underwent a right ooporectomy and it revealed renin and progesterone receptor positive steroid cell tumor, NOS. After operation, her blood pressure returned to normal level and vaginal bleeding disappeared. Even though this case is very rare, when hypertension coincides with virilization or feminization, a renin-secreting ovarian steroid cell tumor, NOS, should be considered.
Child ; Female ; Humans ; Hypertension/*etiology ; Ovarian Neoplasms/complications/*diagnosis/pathology ; Puberty, Precocious/enzymology/*etiology ; Receptors, Cell Surface/metabolism ; Receptors, Progesterone/metabolism ; Renin/blood ; Sex Cord-Gonadal Stromal Tumors/complications/*diagnosis/pathology ; Steroids/biosynthesis ; Tomography, X-Ray Computed ; Vacuolar Proton-Translocating ATPases/metabolism

OBJECTIVE: To investigate the expression and the relativity of RECK,RAGE and MMP-9 in nasopharyngeal carcinoma (NPC) tissues. METHOD: RECK, RAGE and MMP-9 were detected with immunohistochemical methods in 64 NPC specimens(30 cases with cervical lymph nodes metastasis, while the other 34 cases are not) and 30 specimens with chronic nasopharyngitis. RESULT: RECK hardly expressed in chronic nasopharyngitis and NPC tissues, However, strong expression of RECK could be seen in the surrounding inflammatory cells and matrix of cancer nests. The positive rates of RECK in NPC with and without cervical lymph nodes metastasis were 3.3% (1/30) and 11.8% (4/34), separately, There was no statistical significance between the groups (P > 0.05). The positive rates of RAGE in chronic nasopharyngitis and NPC tissues were 100% (30/30) and 70.3% (45/64) respectively, while MMP-9 were 46.7% (14/30) and 78.1% (50/64). The positive rates of RAGE in NPC with and without cervical lymph nodes metastasis were 86.7% (26/30) and 55.9% (19/34) separately, whereas MMP-9 were 90.0% (27/30) and 67.7% (23/34) respectively. They all showed statistical significance between the groups. MMP-9 had a negative correlation with RECK (r = -0.369, P < 0.05) and a positive correlation with RAGE in NPC (r = 0.471, P < 0.05). CONCLUSION The expression of RECK and RAGE in NPC were down-regulated, the expression of MMP-9 was up-regulated. While the expression of RECK in NPC with cervical lymph nodes metastasis was down-regulated, while RAGE and MMP-9 were up-regulated. The abnormal expression of the three genes may be related to the progression of NPC. RECK and RAGE may be involved in the invasion and metastasis of NPC by regulating the expression of MMP-9.
Adolescent ; Adult ; Aged ; Female ; GPI-Linked Proteins ; metabolism ; Glycation End Products, Advanced ; metabolism ; Humans ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; RNA, Messenger ; genetics ; Receptor for Advanced Glycation End Products ; metabolism ; Receptors, Cell Surface ; biosynthesis ; Young Adult

Mammalian zona pellucida 3(ZP3) plays an important role in the induction of capacitating sperm acrosome reaction. In this study, we obtained the soluble mZP3 fusion protein and identified its immunoreactivity. mZP3 cDNA was cloned into plasmid pMAL-p2x, and the recombinant plasmid was transformed into Escherichia coli BL21. To get the soluble mZP3 fusion protein, we tried to optimize the expression conditions, including additives, IPTG concentrations, temperatures and induction duration. Then, Western blotting and ELISA were used to identify the immunoreactivity of the purified protein. Based on the optimization experiments, we concluded that the best soluble expression conditions for the mZP3 fusion protein involved incubation to an A600 of 0.6, addition of glucose to a final concentration of 0.02 mol/L, addition of IPTG to a final concentration of 0.6 mmol/L and then further incubation for 4 h at 25 degrees C. Western blotting and ELISA showed that the mZP3 fusion protein retained immunoreactivity. The fusion protein can be used as solubility antigens for developing the immunocontraception vaccines of mZP3 and detecting the immune effects of the vaccine.
Animals ; Egg Proteins ; biosynthesis ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Female ; Membrane Glycoproteins ; biosynthesis ; genetics ; immunology ; Mice ; Receptors, Cell Surface ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Solubility ; Vaccines, Contraceptive ; immunology ; Zona Pellucida Glycoproteins

GPR43 (G protein-coupled receptor 43) is a recently discovered short-chain free fatty acid receptor which plays important role in adipogenesis. Here we explored the transcriptional expression rule of GPR43 in porcine adipose tissue and primary cultured adipocytes. Partial cDNA of GPR43 was successfully cloned from swine by RT-PCR and the expression profile of GPR43 mRNA was studied from different types, different growing stages, and different sites of porcine adipose tissue as well as porcine primary cultured adipocytes. The results showed that porcine GPR43 shared high homology with human (89%), mouse (84%) and rat (83%). The expression level of GPR43 mRNA was significantly higher in adipose tissue of obese pigs than that of lean pigs, and also the expression level gradually increased with age. Further, the abundance of GPR43 mRNA level was higher in subcutaneous fat than in visceral fat. In addition, during the adipocytes differentiation, the expression of GPR43 mRNA increased in a time-dependent manner. These data indicated that GPR43 gene expression was relate to the site of adipose tissue, economic type, and age of pig as well as differentiating state of adipocytes, implying that GPR43 can be a potential factor to regulate adipogenesis.
Adipocytes ; cytology ; metabolism ; Adipose Tissue ; metabolism ; Animals ; Cells, Cultured ; DNA, Complementary ; biosynthesis ; genetics ; Gene Expression Profiling ; Humans ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Cell Surface ; biosynthesis ; genetics ; Receptors, G-Protein-Coupled ; biosynthesis ; genetics ; Swine ; Transcription, Genetic

In order to study how to induce tolerogenic dendritic cells in vitro and its mechanism, the K562 cells transduced with HLA-G construct were used to co-culture with DC. Then their related immunological changes, such as membrane molecules CD80, CD86, ILT3 and ILT4 expression levels were detected by flow cytometry. Allogeneic proliferation of peripheral blood mononuclear cells (PBMNC) was detected by mixed lymphocyte reaction. The results showed that CD80 and CD86 expressions on DC were downregulated, while ILT3 and ILT4 expressions were upregulated after co-culturing with K562-HLA-G cells. The DCs were less able to stimulate the allogenic PBMNC. It is concluded that the membrane-bound HLA-G can upregulate expression of inhibitory receptors ILT3 and ILT4, inducing tolerogenic DC in vitro, which may provide a novel strategy for transplant tolerance induction.
B7-1 Antigen ; biosynthesis ; B7-2 Antigen ; biosynthesis ; Coculture Techniques ; Dendritic Cells ; cytology ; immunology ; HLA Antigens ; immunology ; HLA-G Antigens ; Histocompatibility Antigens Class I ; immunology ; Humans ; Immune Tolerance ; immunology ; K562 Cells ; Lymphocyte Culture Test, Mixed ; Membrane Glycoproteins ; biosynthesis ; Receptors, Cell Surface ; biosynthesis ; Receptors, Immunologic ; biosynthesis ; T-Lymphocytes ; immunology ; Transfection

OBJECTIVETo investigate whether the sonic hedgehog signaling pathway is involved in the development of human fetal prostate, and to evaluate the changing staining patterns of its molecules, sonic hedgehog (SHH), patchedl (PTC1), smoothened (SMO), and GLI1, in the human fetal prostate at various gestation stages.

METHODSFifteen human fetal prostate specimens at various developmental stages (10 - 39 weeks) were included in this study. SHH, PTC1, SMO and GLI1 were detected in all the specimens by immunohistochemical technique. All the slides were observed and assessed under the light microscope.

RESULTSSHH, PTC1, SMO and GLI1 could be detected in human fetal prostate tissues, and their expression formed two surges, the former at week 16, and the latter at week 28. The staining of SHH and SMO was distributed only in the ductal epithelium but not in the stroma. The expression of PTC1 and GLI1 could be found mainly in the epithelium, with minimal staining in the stroma.

CONCLUSIONThe sonic hedgehog signaling pathway is involved in the development of the human fetal prostate. The high expression of its molecules at early gestation stages might be associated with the induction of prostatic buds, while their abundant expression at later gestation stages might be related to the prostate ductal branching, growth, differentiation and morphogenesis.

Gene Expression Regulation, Developmental ; physiology ; Hedgehog Proteins ; biosynthesis ; Humans ; Male ; Oncogene Proteins ; biosynthesis ; Patched Receptors ; Prostate ; embryology ; metabolism ; Receptors, Cell Surface ; biosynthesis ; Receptors, G-Protein-Coupled ; biosynthesis ; Signal Transduction ; physiology ; Smoothened Receptor ; Trans-Activators ; biosynthesis ; Zinc Finger Protein GLI1

Adolescent ; Adult ; Aged ; Antigens, CD ; biosynthesis ; Brain Neoplasms ; blood supply ; metabolism ; pathology ; Child ; Endoglin ; Female ; Glioma ; blood supply ; metabolism ; pathology ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; biosynthesis ; Immunohistochemistry ; Male ; Middle Aged ; Neovascularization, Pathologic ; metabolism ; pathology ; Receptors, Cell Surface ; biosynthesis ; Telomerase ; biosynthesis ; Young Adult

OBJECTIVESTo study the fibrinolytic activity and the expression of uPAR and Annexin II in leukemic cell lines and their alterations on all-trans retinoic acid ( ATRA) treatment.

METHODSThe fibrinolytic activity was measured by chromogenic assay in NB4, SHI-1, K562, Jurkat and Raji cell lines. The protein expression of uPAR and Annexin II on cells surface and the mRNA expression of uPAR and Annexin II in cells of these cell lines were detected using flow cytometry and RT-PCR method respectively.

RESULTSThe plasmin activity in supernatant was increased significantly after incubation of SHI-1 and NB4 cells with plasminogen. The plasmin activity of NB4 cells was obviously decreased by ATRA. The plasmin activity of NB4 and SHI-1 cells was significantly decreased by uPAR monoclonal antibodies. The expressions of uPAR and Annexin II and their mRNA in SHI-1 and NB4 cells were higher than that in other cell lines. ATRA could remarkably decrease the expressions of Annexin II and uPAR and their mRNA in NB4 cells.

CONCLUSIONIn leukemia cell lines, NB4 and SHI-1 cells have stronger fibrinolytic activity. Both Annexin II and uPAR on the leukemic cell membranes might contribute to this activity. The high fibrinolytic activity can be corrected by ATRA by down-regulating Annexin I and uPAR mRNA and protein expression in NB4 cells.

Annexin A2 ; biosynthesis ; genetics ; Cell Line, Tumor ; Fibrinolysis ; drug effects ; Humans ; Leukemia ; metabolism ; pathology ; RNA, Messenger ; genetics ; Receptors, Cell Surface ; biosynthesis ; genetics ; Receptors, Urokinase Plasminogen Activator ; Reverse Transcriptase Polymerase Chain Reaction ; Tretinoin ; pharmacology

OBJECTIVETo investigate the protective mechanisms of Radix Salviae miltiorrhizae (RSM) on chronic alcoholic liver injury in mice.

METHODSThe chronic alcoholic liver injury mouse model was established. The morphologic change of hepatic tissue was observed with hematoxylin-eosin (HE) staining; the levels of toll-like receptor-4 (TLR-4) mRNA in hepatic tissue and hemeoxygenase-1 (HO-1) mRNA were determined using reverse transcription polymerase chain reaction (RT-PCR) technique; and the expression of TLR-4 protein was determined by immunohistochemistry method.

RESULTSRSM could alleviate the fatty degeneration and adiponecrosis of hepatic cells induced by alcohol, down-regulate the expressions of TLR-4 mRNA and HO-1 mRNA, and significantly decrease the number of TLR-4 positive cells.

CONCLUSIONRSM could prevent liver injury from alcohol by way of influencing TLR-4 signal transcription.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Heme Oxygenase (Decyclizing) ; biosynthesis ; genetics ; Heme Oxygenase-1 ; Hepatitis, Alcoholic ; drug therapy ; pathology ; Liver ; metabolism ; Male ; Membrane Glycoproteins ; biosynthesis ; genetics ; Membrane Proteins ; Mice ; Phytotherapy ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Cell Surface ; biosynthesis ; genetics ; Salvia miltiorrhiza ; Toll-Like Receptor 4 ; Toll-Like Receptors