Background: Human Heparansulphate interacting protein (hHip) has been shown to participate in biological processes of cells. Several studies indicated that hHip transcript is up regulated in several of cancer tissues including those of thyroid, colon, breast and prostate. Antibody against hHIP is necessary for methods to evaluate protein level of HIP in cancer tissues. Objectives:The aims of study was to induce anti hHIP antibody in rabbit and purify and conserve purified anti hHIP antibody. Subjects and method: The study included 9 adult and healthy rabbits with the weight 2 - 2.5kg. Immunization hHIP peptide-KLH in rabbit. Purify anti hHIP antibody using affinity chromatography. Results: The results shown synthesize hHIP peptide and conjugate it with carrier protein. Sensitive rabbit better meet with hHIP-KLH antibody. The Ig concentration obtained in sensitive rabbit was rather high and equal. Immunization hHIP-KLH successfully in rabbit. Obtainment valuable amount of anti hHIP antibody. Conclusion: Successfully induce and purify anti hHIP antibody from rabbit. Establish a standard protocol for polyclonal antibody against small peptide in rabbit.\r\n', u'\r\n', u'
Carrier Proteins/ administration & ; dosage ; chemistry ; Rabbits

OBJECTIVETo purify the arsenic-binding proteins (As-BP) in hamster plasma after a single oral administration of arsenite (iAs(III)).

METHODSArsenite was given to hamsters in a single dose. Three types of HPLC columns, size exclusion, gel filtration and anion exchange columns, combined with an inductively coupled argon plasma mass spectrometer (ICP MS) were used to purify the As-BP in hamster plasma. SDS-PAGE was used to confirm the arsenic-binding proteins at each purification step.

RESULTSThe three-step purification process successfully separated As-BP from other proteins (ie, arsenic unbound proteins) in hamster plasma. The molecular mass of purified As-BP in plasma was approximately 40-50 kD on SDS-PAGE.

CONCLUSIONThe three-step purification method is a simple and fast approach to purify the As-BP in plasma samples.

Administration, Oral ; Animals ; Arsenic ; blood ; Arsenites ; administration & dosage ; pharmacokinetics ; Carrier Proteins ; blood ; Chromatography, High Pressure Liquid ; methods ; Cricetinae

A series of receptors expressed in the surface of tumor cells, which are able to mediate internalizing effect by specially connecting with corresponding ligands. These receptors are potential targets for drugs combined with conjugates. So the drug-conjugate compounds can be targeted delivery to tumor cells. The folate receptor is a promising target because of its marrow tissue specificity, its overexpression in malignant tissues, especially in myeloid leukemic cells, and its ability to bind and internalize folic acid conjugates. It is a promising potential method to apply folate receptor in the receptor-mediated targeting therapy of leukemic cells. In this review, the biological features of folate receptor, its chromosome location and its interaction with ligands, the distribution characteristics of folate receptor in normal and tumor tissues, especially in myeloid leukemic cells, and progress of research on folate receptor mediated targeting tumor cells, especially leukemic cells were summarized.
Antineoplastic Agents ; administration & dosage ; Carrier Proteins ; metabolism ; Drug Delivery Systems ; methods ; Folate Receptors, GPI-Anchored ; Folic Acid ; metabolism ; Humans ; Leukemia ; drug therapy ; metabolism ; pathology ; Receptors, Cell Surface ; metabolism ; Tretinoin ; administration & dosage ; Tumor Cells, Cultured

Folate receptor (FR) is over-expressed in a variety of human cancers and it is seldom expressed or found in normal tissues. Therefore, folate receptor-mediated antitumor drugs can be targeted specially to the FR-positive tumor cells. This strategy improves the selectivity of drugs which may destroy the normal tissues in traditional chemotherapeutics. This review provides the delivery mechanism of FR-mediated antitumor drugs and highlights the novel folate-drug conjugates and their activities.
Animals ; Antineoplastic Agents ; administration & dosage ; chemistry ; pharmacology ; Carrier Proteins ; metabolism ; Cell Line, Tumor ; Drug Carriers ; Drug Delivery Systems ; Folate Receptors, GPI-Anchored ; Folic Acid ; administration & dosage ; analogs & derivatives ; chemistry ; pharmacology ; Humans ; Neoplasms ; metabolism ; pathology ; Receptors, Cell Surface ; metabolism

OBJECTIVETo investigate the effects of pegylated interferon a-2a (Peg-INFa-2a) treatment on expression of CD8 and CD38 surface molecules on lymphocytes from peripheral blood of inactive hepatitis B surface antigen (HBsAg) carriers.

METHODSForty-four patients with hepatitis B virus (HBV) chronic infection (CHB) received a 48-week course of Peg-INFa-2a treatment, with 30 administered 135 mug/week and 14 administered 180 mug/week. Every 12 weeks of treatment, the subjects were assessed for HBsAg titer, presence of anti-hepatitis B e (HBe) antibody, serum alanine amino transaminase (ALT) levels, and lymphocyte surface expression of CD8 and CD38 molecules. Patients were classified as responders and non-responders according to standard parameters. Dynamic differences between the two groups over time were assessed by multivariate repeated measures ANOVA with Greenhouse-Geisser correction and differences at single time points were assessed by univariate ANOVA. Linear regression analysis was performed to evaluate the relationship of two variables.

RESULTSThe responders showed a significantly higher increase in ALT at week 12 (60.75+/-24.95 U/L vs. non-responders: 37.03+/-18.45 U/L; t = 2.905, P less than 0.01) and significantly higher proportion of CD8+CD38+ cells at week 24 (71.20+/-11.70% vs. non-responders: 56.79+/-7.72%; F = 23.941, P less than 0.01). The decline in level of HBsAg at week 24 was positively correlated with the increase in ALT level at week 12 (r = 0.386, P less than 0.01) and with expression levels of CD8 and CD38 molecules on lymphocytes at week 24 (r = 0.397, P less than 0.01).

CONCLUSIONLower baseline levels of HBsAg correlated to better Peg-INFa-2a-related HBsAg clearance. Increased expression of CD8 and CD38 on lymphocytes is suggestive of intensive cellular immunity in CHB patients and may be related to HBV-induced hepatocyte damage and may promote the HBsAg clearance.

ADP-ribosyl Cyclase 1 ; metabolism ; Adult ; Aged ; Antiviral Agents ; administration & dosage ; therapeutic use ; CD8-Positive T-Lymphocytes ; Carrier State ; Hepatitis B Surface Antigens ; blood ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Interferon-alpha ; administration & dosage ; therapeutic use ; Middle Aged ; Polyethylene Glycols ; administration & dosage ; therapeutic use ; Recombinant Proteins ; administration & dosage ; therapeutic use ; T-Lymphocyte Subsets

Bovine serum albumin nanoparticles(BSANP) were prepared by desolvation method. The activated folic acid (N-hydroxysuccinimide ester of folic acid) was conjuated to the surface of BSANP via the amino groups. Then the folate-conjugated BSANPs (folate-BSANP) were purified with Sephadex G-50 column and completely separated from unreacted folic acid. After chymotryptic hydrolysis, the extent of folate conjugation on the BSANP was determined by quantitative ultraviolet(UV) spectrophotometric analysis. It was found that the spectrum of trypsin digest of folate-conjugate BSANP is basically identical with that of folate, thus indicating folate is successfully expressed on the surface of BSANP. The folate-BSANP was averagely 66 nm in diameter and was spherical in shape. Folate-conjugated BSANP was achieved, which represents a potential new drug carrier for tumor cell-selective targeting.
Animals ; Antineoplastic Agents ; administration & dosage ; Carrier Proteins ; chemistry ; Cattle ; Drug Delivery Systems ; methods ; Folate Receptors, GPI-Anchored ; Folic Acid ; chemistry ; Immunotoxins ; Microspheres ; Nanotechnology ; Receptors, Cell Surface ; chemistry ; Serum Albumin, Bovine ; chemistry

Inorganic phosphate (Pi) plays a critical role in diverse cellular functions, and regulating the Pi balance is accomplished by sodium-dependent Pi co-transporter (NPT). Pulmonary NPT has recently been identified in mammalian lungs. However, to date, many of the studies that have involved Pi have mainly focused on its effect on bone and kidney. Therefore, current study was performed to discover the potential effects of low Pi on the lung of developing transgenic mice expressing the renilla/firefly luciferase dual reporter gene. Two-weeks old male mice divided into 2 groups and these groups were fed either a low PI diet or a normal control diet (normal: 0.5% Pi, low: 0.1% Pi) for 4 weeks. After 4 weeks of the diet, all the mice were sacrificed. Their lungs were harvested and analyzed by performing luciferase assay, Western blotting, kinase assay and immunohistochemistry. Our results demonstrate that low Pi affects the lungs of developing mice by disturbing protein translation, the cell cycle and the expression of fibroblast growth factor-2. These results suggest that optimally regulating Pi consumption may be important to maintain health.
Animals ; Blotting, Western ; Carrier Proteins/metabolism ; Immunohistochemistry ; Lung/drug effects/enzymology/*growth & development/metabolism ; Male ; Mice ; Mice, Transgenic ; Phosphoproteins/metabolism ; Phosphorus, Dietary/*administration & dosage ; Phosphorylation ; Phosphotransferases (Alcohol Group Acceptor)/metabolism ; Proto-Oncogene Proteins c-akt/metabolism ; Sodium-Phosphate Cotransporter Proteins, Type IIa/*metabolism

OBJECTIVETo search the interaction between reduced folate carrier gene (RFC1 A80G) polymorphism of children with neural tube defects (NTDs) and maternal periconceptional no supplementation of folic acid. The purpose is to provide the epidemiological evidence for finding genetic marker of NTDs.

METHODSRFC1 (A80G) genotype was detected using PCR-restricted fragment length polymorphism for the blood DNA of 104 trios with NTDs-affected child, and 100 control families with non-malformed control children. The authors investigated the gene-environment interactions between the offspring RFC1 genotype and maternal periconceptional folic acid supplementation through a case-control study.

RESULTSIt was observed that the offspring with the GG genotype were associated with a 2.56-fold increased risk of NTDs when compared to those with the AA genotype (OR = 2.56; 95% CI = 1.04-6.36) in this population under investigation. The risk of mothers who did not take folic acid for having an NTDs-affected infants was 7.69 (95% CI = 2.86-21.75). Among the mothers who did not utilize folic acid supplements, the NTDs risk was 3.30 (95% CI = 1.15-9.65) for offspring with the GG genotype, compared to the reference (AA) genotype. Children who had the GG genotype and whose mothers did not take folic acid had an elevated risk for NTDs (OR = 8.80, 95% CI = 2.86 - 29.82), compared to "offspring with AA or GA genotype" and "maternal folic acid use", the interactive coefficient being 1.45.

CONCLUSIONThe above findings indicate that the RFC1 genotype (GG) is a possible susceptible gene marker for an increased NTDs risk in Chinese population, and there is a potential gene-nutrient interaction between offspring RFC1 GG genotype and maternal periconceptional intake of folic acid on the risk of NTDs. However,the sample size of this study was limited, a larger sample of population-based study is required to pursue the initial observation.

Adult ; Case-Control Studies ; Child ; Child, Preschool ; Dietary Supplements ; Female ; Folic Acid ; administration & dosage ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Male ; Membrane Transport Proteins ; genetics ; Neural Tube Defects ; genetics ; prevention & control ; Polymorphism, Genetic ; genetics ; Reduced Folate Carrier Protein ; Vitamin B Complex ; administration & dosage

Adult ; Age of Onset ; Amino Acid Metabolism, Inborn Errors ; complications ; diagnosis ; genetics ; therapy ; Betaine ; administration & dosage ; therapeutic use ; Carrier Proteins ; genetics ; metabolism ; Child ; China ; epidemiology ; DNA Mutational Analysis ; Gas Chromatography-Mass Spectrometry ; Genotype ; Homocysteine ; urine ; Humans ; Hydroxocobalamin ; administration & dosage ; therapeutic use ; Hyperhomocysteinemia ; complications ; diagnosis ; genetics ; therapy ; Infant ; Methylmalonic Acid ; blood ; urine ; Mutation ; Vitamin B 12 ; metabolism

BACKGROUND/OBJECTIVES: Endoplasmic reticulum (ER) stress is positively associated with atherosclerosis via elevating macrophage cell death and plaque formation, in which oxidative stress plays a pivotal role. Antioxidative, lipid-lowering, and anti-atherogenic effects of kimchi, a Korean fermented vegetable, have been established, wherein capsaicin, ascorbic acid, quercetin, 3-(4'-hydroxyl-3',5'-dimethoxyphenyl)propionic acid, and lactic acids were identified. In this study, mechanisms of action of kimchi methanol extracts (KME) on fatty streak formation via suppression of ER stress and apoptosis in aorta were examined in low-density lipoprotein receptor knockout mice. MATERIALS AND METHODS: Mice fed a high cholesterol diet with an oral administration of KME (KME group, 200 mg·kg-bw⁻¹·day⁻¹) or distilled water (control group) for 8 weeks (n = 20 for group). Plasma lipid and oxidative stress levels were evaluated. Protein expression was measured by western blot assay. Fatty streak lesion size and the degree of apoptosis were examined in the aorta. RESULTS: Compared to the control group, in the KME group, plasma lipids levels were decreased and oxidative stress was alleviated (P < 0.05). Protein expression levels of nuclear factor (erythroid-derived 2)-like 2-mediated antioxidants in aorta were increased whereas those for ER stress markers, glucose regulated protein 78, phospho-protein kinase RNA-like ER kinase, phospho-eukaryotic initiation factor 2 subunit α, X-box binding protein 1, and C/EBP homologous protein were decreased in the KME group (P < 0.05). Moreover, apoptosis was suppressed via downregulation of phospho-c-Jun N-terminal kinase, bcl-2-associated X protein, caspases-9, and -3 with a concomitant upregulation of anti-apoptotic protein, B-cell lymphoma 2 (P < 0.05). Fatty streak lesion size was reduced and the degree of apoptosis was less severe in the KME group (P < 0.05). CONCLUSIONS: In conclusion, antioxidant activity of KME might prevent fatty streak formation through, in part, inhibition of ER stress and apoptosis in aortic sinus where macrophages are harbored.
Administration, Oral ; Animals ; Antioxidants ; Aorta* ; Apoptosis* ; Ascorbic Acid ; Atherosclerosis ; bcl-2-Associated X Protein ; Blotting, Western ; Capsaicin ; Carrier Proteins ; Cell Death ; Cholesterol ; Diet ; Down-Regulation ; Endoplasmic Reticulum Stress* ; Endoplasmic Reticulum* ; Glucose ; Hypercholesterolemia ; Lactic Acid ; Lipoproteins* ; Lymphoma, B-Cell ; Macrophages ; Methanol ; Mice ; Mice, Knockout* ; Oxidative Stress ; Phosphotransferases ; Plasma ; Prokaryotic Initiation Factor-2 ; Quercetin ; Receptors, Lipoprotein* ; Sinus of Valsalva ; Up-Regulation ; Vegetables ; Water