OBJECTIVETo investigate the distribution, changes and a possible role for retinal dopamine transporter (DAT) in experimental myopia in chickens.

METHODSTwo-day-old chickens were divided into four groups. Chicken eyes were fitted with lenses of -10D,-20D and translucent goggles unilaterally. Normal eyes were used as controls. After 3 wk, all chickens were given an intramuscular injection of (125)I-beta-CIT 2beta-carbomethoxy-3beta-(4-iodophenyl)tropane and sacrificed two hours post injection. Retinal pigment epithelium (RPE) and the neural retina were obtained together or RPE was dissected out from the neural retina. Radioactive DAT from each specimen was assayed by gamma-counter.

RESULTSRetinal DAT was detected in RPE specimens rather than in the neural retina in all eyes. Radioactive DAT in myopic eyes was higher, compared with control eyes.

CONCLUSIONSRetinal DAT is mainly located in the RPE and may be involved in the formation of lens induced myopia (LIM) and form deprivation myopia (FDM). These methods may provide a new approach for further studying the role of the dopamine system in experimental myopia.

Animals ; Chickens ; Dopamine Plasma Membrane Transport Proteins ; Eye ; growth & development ; Membrane Glycoproteins ; Membrane Transport Proteins ; analysis ; physiology ; Myopia ; metabolism ; Nerve Tissue Proteins ; Retina ; chemistry

BACKGROUNDThe diagnosis of Parkinson's disease is presently based on non-specific symptoms. However, radionuclide dopamine transporters imaging can provide specific diagnostic tool for Parkinson's disease. This study was designed to investigate the effects of imaging of dopamine transporters with 99mTc-TRODAT-1 in early diagnosis or differential diagnosis of Parkinson's disease.

METHODSNine normal monkeys were used to establish N-methyl-4-phenyl-1, 2, 3, 6-tetra-hydropyridine (MPTP) hemi-Parkinsonian animal models, and they were subjected to imaging. Twenty-nine patients with Parkinson's disease, 12 age-matched healthy volunteers, and 18 age-matched patients with Parkinson's syndrome were investigated. Single photon emission computer tomography (SPECT) was performed 3 hours after intravenous injection of 740 MBq 99mTc-TRODAT-1. Striatum specific uptake of 99mTc-TRODAT-1 was calculated according to the ratio of striatum (ST) to cerebellum (CB) in dopamine transporters uptake.

RESULTSIn normal monkeys, bilateral ratio of ST/CB was 2.34 +/- 0.41. After the injection of MPTP, uptake rate of 99mTc-TRODAT-1 at damaged region was much lower than that at the contralateral region, resulting in a significant difference in the ratio of ST/CB (right: ST/CB = 1.73 +/- 0.35; left: ST/CB = 1.90 +/- 0.30), especially in hemi-Parkinsonian model monkeys (right: ST/CB = 1.29 +/- 0.17; left: ST/CB = 1.80 +/- 0.33). The ratios of ST/CB were 1.57 +/- 0.17 and 1.61 +/- 0.14 for the right and left respectively in the healthy volunteers, 1.04 +/- 0.29 and 1.06 +/- 0.30 in the age-matched patients with Parkinson's disease, and 1.56 +/- 0.17 and 1.59 +/- 0.18 in the age-matched patients with Parkinson's disease syndrome. A significant difference was noted between group of Parkinson's disease, normal controls and Parkinson's disease syndrome.

CONCLUSIONThe results suggest that 99mTc-TRODAT-1 dopamine transporters SPECT has clinical application value in early diagnosis or differential diagnosis of Parkinson's disease.

Adult ; Aged ; Animals ; Dopamine Plasma Membrane Transport Proteins ; Female ; Haplorhini ; Humans ; Male ; Membrane Glycoproteins ; analysis ; Membrane Transport Proteins ; analysis ; Middle Aged ; Nerve Tissue Proteins ; analysis ; Organotechnetium Compounds ; Parkinson Disease ; diagnosis ; Radiopharmaceuticals ; Tomography, Emission-Computed, Single-Photon ; Tropanes

OBJECTIVESTo study the interaction of beta-2-glycoprotein I (beta 2GPI) with the membrane of hepatocytes and determine whether beta 2GPI participates in HBV infection.

METHODSLigand blotting, fluorescence microscopy, and fluorescence activated cell sorter (FACS) analysis were used to detect the specific interaction of beta 2GPI with the hepatoma cell line smmc7721, the gastric carcinoma cell line SGC7901, and the lymphoma cell line HL-60.

RESULTSA specific 40 kDa beta 2GPI band was observed by ligand blotting in the case of smmc7721 cells. No such band was observed in SGC7901 or HL-60 cells. Fluorescence microscopy also revealed specific binding of FITC-beta 2GPI to smmc7721 cells, but neither to SGC7901 nor HL-60 cells. FACS analysis demonstrated that the binding rate of FITC-beta 2GPI to smmc7721 cells was significantly higher than these in SGC7901 and HL-60 cells (P < 0.01). The binding rate to smmc7721 cells did not increase with increasing amounts of FITC-beta 2GPI.

CONCLUSIONSThere is a specific beta 2GPI-binding protein on the membrane of hepatoma cells in cell line smmc7721 which as the beta 2GPI receptor may participate in HBV infection of hepatocytes.

Animals ; Carcinoma, Hepatocellular ; Cell Membrane ; metabolism ; Flow Cytometry ; Glycoproteins ; metabolism ; Hepatocytes ; metabolism ; Liver Neoplasms, Experimental ; metabolism ; Platelet Glycoprotein GPIb-IX Complex ; analysis ; Platelet Membrane Glycoproteins ; Tumor Cells, Cultured ; beta 2-Glycoprotein I

OBJECTIVETo investigate the effect of triggering receptor expressed on myeloid cells 2 (TREM-2) overexpression on airway inflammation and remodeling in mice with asthma.

METHODSA total of 40 BALB/c mice were randomly divided into normal control, asthma, empty vector, and TREM-2 overexpression groups (n=10 each). Ovalbumin (OVA) sensitization and challenge were performed to establish the model of asthma. The mice in the control group were given normal saline, and those in the empty vector and TREM-2 overexpression groups were transfected with adenovirus vector and TREM-2 adenovirus, respectively. RT-PCR and Western blot were used to measure the expression of TREM-2, MMP-2, MMP-9, ADAM33, and ADAM8. Bronchoalveolar lavage fluid (BALF) was collected to perform cell counting and classification. ELISA was used to measure the total serum level of IgE and the levels of cytokines in BALF.

RESULTSCompared with the control group, the asthma group showed significant reductions in the mRNA and protein expression of TREM-2 (P<0.05), a significantly increased level of Th2 cytokine (P<0.05), and significantly increased numbers of total cells and classified cells. Compared with the asthma group, the TREM-2 overexpression group showed a significantly reduced level of Th2 cytokine (P<0.05), a significantly reduced level of IgE (P<0.05), and significantly reduced numbers of total cells and classified cells (P<0.05), as well as significantly downregulated expression of the inflammatory factors and growth factors MMP-2, MMP-9, TGF-β1, ADAM8, and ADAM33 (P<0.05).

CONCLUSIONSTREM-2 overexpression significantly alleviates airway inflammation and airway remodeling in mice with asthma and may become a potential target for the prevention and treatment of childhood asthma.

Airway Remodeling ; Animals ; Asthma ; etiology ; immunology ; Cytokines ; analysis ; Female ; Membrane Glycoproteins ; genetics ; physiology ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; immunology ; RNA, Messenger ; analysis ; Receptors, Immunologic ; genetics ; physiology

OBJECTIVETo study the expression of glucose-regulated protin 78 (GRP78) and glucose-regulated protin 94 (GRP94) in the liver tissues from children with hepatoblastoma (HB) and to investigate the possible clinicopathological values of GRP78 and GRP94 in HB.

METHODSLiver tissue specimens from 15 children with HB and 10 specimens of normal liver tissues were obtained. EnVison immunohistochemistry was used to detect the expression of GRP78 and GRP94 in the conventional paraffin-embedded liver sections.

RESULTSThe positive rates of GRP78 expression (53% vs 10%; P<0.05) and GRP94 expression (60% vs 10%; P<0.05) in HB liver tissues were significantly higher than those in the normal liver tissues. The positive rates of GRP78 expression in the cases without lymphnode metastasis or in clinical stage I-II were significantly lower than those in the cases with lymphnode metastasis or in clinical stage III-IV (P<0.05). GRP94 showed a decreased tendency of positive expression in the cases without lymphnode metastasis or in clinical stage I-II when compared with the cases with lymphnode metastasis or in clinical stage III-IV, although there were no statistical differences between them.

CONCLUSIONSGRP78 and GRP94 expression might play important roles in the pathogenesis and progression of pediatric HB.

Child ; Child, Preschool ; Female ; Heat-Shock Proteins ; analysis ; Hepatoblastoma ; chemistry ; pathology ; Humans ; Immunohistochemistry ; Infant ; Liver ; chemistry ; Liver Neoplasms ; chemistry ; pathology ; Male ; Membrane Glycoproteins ; analysis ; Neoplasm Staging

Prostate stem cell antigen (PSCA) is a cell surface antigen expressed in normal prostate and overexpressed in cancers associated with prostate, bladder and pancreas. The sensitivity of PSCA labeling is higher than PSA in prostate cancer. PSCA can be used in the preparation of protein vaccine and nucleic acid vaccine. Further studies are required to confirm its safety and efficacy as a diagnostic means.
Antigens, Neoplasm ; GPI-Linked Proteins ; Humans ; Immunotherapy ; Male ; Membrane Glycoproteins ; analysis ; genetics ; immunology ; Neoplasm Proteins ; analysis ; genetics ; immunology ; Pancreatic Neoplasms ; diagnosis ; Prostatic Neoplasms ; diagnosis ; Urinary Bladder Neoplasms ; diagnosis

Adult ; Antigens, CD ; analysis ; B7-1 Antigen ; analysis ; B7-2 Antigen ; Dendritic Cells ; immunology ; Female ; HLA-DR Antigens ; analysis ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Immunophenotyping ; Intercellular Adhesion Molecule-1 ; analysis ; Interferon-alpha ; therapeutic use ; Male ; Membrane Glycoproteins ; analysis

The epidermal growth factor receptor (EGFR) is a transmembrane glycoprotein whose expression is a possible cause of increased tumor cell proliferation and has recently been proposed as a prognostic parameter in some tumors. Expression of EGFR was studied immunohistochemically in 62 cases of human renal cell carcinomas to evaluate their possible prognostic roles. We also examined the correlation between EGFR expression and cell proliferation by immunohistochemical staining for proliferating cell nuclear antigen (PCNA). Fifty-six cases (90.3%) expressed EGFR, with staining largely confined to the cell membrane and cytoplasm. Staining intensity of EGFR was directly correlated with nuclear grade (p=0.000) and TNM stage (p=0.015). PCNA index was significantly higher in EGFR-positive tumors than in EGFR- negative tumors. There was a statistically significant positive correlation between PCNA index and increasing staining intensity of EGFR (p=0.000). In univariate survival analysis, EGFR expression was significantly associated with shortened survival. However, EGFR expression was not an independent prognostic factor by multivariate analysis. These findings suggest that EGFR expression may be an important cause of tumor cell proliferation in renal cell carcinoma and further studies are needed to evaluate whether EGFR expression analysis provides independent prognostic information.
Carcinoma, Renal Cell* ; Cell Membrane ; Cell Proliferation* ; Cytoplasm ; Epidermal Growth Factor* ; Glycoproteins ; Humans ; Multivariate Analysis ; Proliferating Cell Nuclear Antigen ; Receptor, Epidermal Growth Factor*

The possibility of immunotherapy for lymphoma by single FasL-Fas way was investigated. After pBillneo-mFasL was transformed into competent E. coli DH5alpha and amplified, the plasmid DNA was prepared and purified from the DH5alpha. To determine the primary structure and inserting direction of mFasL cDNA gene in pBillneo-mFasL, the plasmid DNA was cleaved by restriction enzyme, and the mFasL cDNA of pBillneo-mFasL was amplified by polymerase chain reaction (PCR), the DNA sequence of the PCR product was analysed by automatic DNA sequencing. After pBillneo-mFasL was transfected into COS-7 cells by liposome, the COS-7 cells were selected with G418 selective medium, and the expressing levels of mFasL cDNA on the COS-7 cell membrane was assayed by Western Blot. After the COS-7 cells higher expressing mFasL protein and mouse lymphoma cell line Yac-1 expressing Fas were cocultured for 5 hours, the suspending Yac-1 cells were collected and labeled by annexin V/PI kit. The apoptosis rate of the Yac-1 cells was tested by flow cytometry. The EcoRI cleaving products of pBillneo-mF asL included 920 bp and 7227 bp fragments. Its Hind III cleaving products included 1293 bp and 6807 bp fragments. These results showed: (1) the length of DNA sequence containing mFasL cDNA within pBillneo-mFasL is the same as theoretical length; (2) the inserting of mFasL cDNA in pBillneo-mFasL was in positive orientation. The expected 890 bp DNA fragments of mFasL cDNA (from ATG to +36 bp following TAA) emerged in PCR product with pBillneo-mFasL as a template. The sequencing result of the PCR product equaled the known mFasL cDNA sequence in the gene bank. The COS-7 cells transfected by pBillneo-mFasL and selected with G418 culture medium expressed more mFasL membrane protein assayed by Western Blot. After the COS-7 cells were cocultured with Fas(+) Yac-1 cells in different E:T ratios (1:1, 5:1 and 10:1) for 5 hours, the apoptosis rates of Yac-1 cells were (22 +/- 4.8)%, (32.18 +/- 7.8)%, and (51.8 +/- 5.4)%, respectively. These were obviously different from the control group (P < 0.01), in which the COS-7 cell was transfected by pBillneo (not carrying mFasL gene). It was concluded that lymphoma cells highly expressing Fas can be effectively killed through single Fas-FasL way in vitro.
Animals ; Apoptosis ; COS Cells ; Fas Ligand Protein ; Immunotherapy ; Lymphoma ; pathology ; therapy ; Membrane Glycoproteins ; genetics ; physiology ; Mice ; Transfection ; Tumor Cells, Cultured ; fas Receptor ; analysis

As one important member of B7/CD28/CTLA-4 costimulatory signal pathway, B7-2 molecule plays a critical role in regulating T-cell response. In order to further explore its effects on regulation of T cell activation, proliferation and associated signal pathways, the cDNA encoding extracellular region of human B7-2 was amplified via PCR and subcloned into some prokaryotic expression vectors to express target protein in host strains. The expressed protein was identified with Western blot and MTT. Results showed that after screening, the expression level of the protein of interest attained the yield of over 20% total bacterial protein by using pGEX-4T-2 vector and E. coli BL21 (DE3)-CodonPlus-RIL host cells. The recombinant protein could specially react with B7-2 McAb and could stimulate T-cell proliferation combined with anti-CD3 antibody. In conclusion, the recombinant protein was bioactive, therefore the study will make it possible for the research of relationship between B7-2 structure and its function.
Antigens, CD ; biosynthesis ; genetics ; pharmacology ; B7-2 Antigen ; Blotting, Western ; Cloning, Molecular ; DNA, Complementary ; analysis ; Humans ; Membrane Glycoproteins ; biosynthesis ; genetics ; pharmacology ; Recombinant Proteins ; biosynthesis ; isolation & purification ; pharmacology