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Biomedical subjects

Z Liu

Publications and source records attributed to Z Liu.

At least 397 records · Page 22Linked to original sources

Effect of raloxifene-analog (LY 117018-Hcl) on the bone marrow of ovariectomized mice.

The effects of LY117018-Hcl (Ralox-A) on body metabolism and differentiation of bone marrow cells were studied in ovariectomized (OVX) mice. We used a mouse model in which estrogen depletion was established for a period of three months before treatment. After that period the animals were divided into three experimental groups consisting of sham-operated, OVX, and OVX-Ralox-A-treated mice. The OVX animals received daily treatment of Ralox-A during two time periods (35 and 65 days). After the treatment we measured the serum levels of protein, ion(s), lipid content, liver, and kidney functions. Our findings indicated that a change in hormonal state did not affect basic body metabolism except for causing an increase in triglycerides (TG) in the OVX mice, which was lowered by the Ralox-A. A higher alkaline phosphatase (ALK-P) level was observed in serum of the OVX-Ralox-A-treated mice than in serum of the OVX mice. We investigated the effects of estrogen depletion on the differentiation of hematopoietic and stromal cells that directly affect bone resorption and formation. OVX and OVX-treated mice were compared with the sham group and assessed for the alteration of these cells' differentiation. The proliferation of stromal stem cells was measured by CFU-F assay in vitro. A decrease in CFU-F colonies derived from OVX mice was observed and after the Ralox-A treatment the number of CFU-F reached sham levels. On the contrary, an upregulation of myeloid cells was observed when analyzed by FACS and by granulocyte/macrophage-colony forming unit (G/M-CFU) assay in selective culture conditions. The G/M-CFUs were increased in the OVX mice and were reduced to sham levels after Ralox-A treatment. In this study, we demonstrated cellular changes of stromal and hemopoietic cells in OVX mice and a beneficial Ralox-A effect that protected such cellular changes.

Alkaline Phosphatase↗

Synthesis and characterization of surface-hydrophobic ion-exchange microspheres and the effect of coating on drug release rate.

Biodegradable, dextran-based ion-exchange microspheres (IE-MS) have been used for localized delivery of anticancer drugs and chemosensitizers. Because of their hydrophilic nature, the IE-MS release their payload quickly. The purpose of this work was to develop an IE-MS system that could provide a broad range of release rates for in vitro and in vivo applications. Sulfopropylated dextran microspheres (SP C25 MS) were surface-modified by acylation. These hydrophobically modified sulfopropylated dextran microspheres (HM-MS) were further coated with the cationic acrylic polymer Eudragit RL100 (EU-MS). The changes in chemical composition after the surface modification and coating were characterized by X-ray photoelectron spectroscopy. The effects of the modification and coating on the surface hydrophobicity, equilibrium swelling, surface morphology, and drug loading capacity were investigated. The HM-MS showed little change in swelling and functionality, despite significantly increased affinity to oil and carbon content on the surface. The coated microspheres (EU-MS) exhibited a profoundly decreased swelling ratio, an even higher affinity to oil, a higher loading capacity, and a lower drug release rate. Through further coating of the EU-MS with different amounts of corn oil, the rate of drug release could be tailored to cover a relatively wide range. These results suggest that a versatile delivery system with various release profiles can be prepared by a combination of hydrophobic modification, polymer coating, and oil coating.

Ion Exchange Resins↗

Skeletal-specific expression of Fgd1 during bone formation and skeletal defects in faciogenital dysplasia (FGDY; Aarskog syndrome).

FGD1 encodes a guanine nucleotide exchange factor (GEF) that specifically activates the Rho GTPase Cdc42; FGD1 mutations result in Faciogenital Dysplasia (FGDY, Aarskog syndrome), an X-linked developmental disorder that adversely affects the formation of multiple skeletal structures. To further define the role of FGD1 in skeletal development, we examined its expression in developing mouse embryos and correlated this pattern with FGDY skeletal defects. In this study, we show that Fgd1, the mouse FGD1 ortholog, is initially expressed during the onset of ossification during embryogenesis. Fgd1 is expressed in regions of active bone formation in the trabeculae and diaphyseal cortices of developing long bones. The onset of Fgd1 expression correlates with the expression of bone sialo-protein, a protein specifically expressed in osteoblasts at the onset of matrix mineralization; an analysis of serial sections shows that Fgd1 is expressed in tissues containing calcified and mineralized extracellular matrix. Fgd1 protein is specifically expressed in cultured osteoblast and osteoblast-like cells including MC3T3-E1 cells and human osteosarcoma cells but not in other mesodermal cells; immunohistochemical studies confirm the presence of Fgd1 protein in mouse calvarial cells. Postnatally, Fgd1 is expressed more broadly in skeletal tissue with expression in the perichondrium, resting chondrocytes, and joint capsule fibroblasts. The data indicate that Fgd1 is expressed in a variety of regions of incipient and active endochondral and intramembranous ossification including the craniofacial bones, vertebrae, ribs, long bones and phalanges. The observed pattern of Fgd1 expression correlates with FGDY skeletal manifestations and provides an embryologic basis for the prevalence of observed skeletal defects. The observation that the induction of Fgd1 expression coincides with the initiation of ossification strongly suggests that FGD1 signaling plays a role in ossification and bone formation; it also suggests that FGD1 signaling does not play a role in the earlier phases of skeletogenesis. With the observation that FGD1 mutations result in the skeletal dysplasia FGDY, accumulated data indicate that FGD1 signaling plays a critical role in ossification and skeletal development.

3T3 Cells↗

Rapid identification of saponins in plant extracts by electrospray ionization multi-stage tandem mass spectrometry and liquid chromatography/tandem mass spectrometry.

Electrospray ionization multi-stage tandem mass spectrometry (ESI-MS(n)) and liquid chromatography coupled with on-line mass spectrometry (LC/MS/MS) were applied to characterize saponins in crude extracts from Panax ginseng. The MS(n) data of the [M - H](-) ions of saponins can provide structural information on the sugar sequences of the saccharide chains and on the sapogins of saponins. By ESI-MS(n), non-isomeric saponins and isomeric saponins with different aglycones can be determined rapidly in plant extracts. LC/MS/MS is a good complementary analytical tool for determination of isomeric saponins. These approaches constitute powerful analytical tools for rapid screening and structural assignment of saponins in plant extracts.

Chromatography, Liquid↗

Construction of protein binding sites in scaffold structures.

We have developed a strategy for grafting a protein-protein interface based on the known crystal structure of a native ligand and receptor proteins in a complex. The key interaction residues at the ligand protein binding interface are transferred onto a scaffold protein so that the mutated scaffold protein will bind the receptor protein in the same manner as the ligand protein. First, our method identifies key residues and atoms in the ligand protein, which strongly interact with the receptor protein. Second, this method searches the scaffold protein for combinations of candidate residues, among which the distance between any two candidate residues is similar to that between relevant key interaction residues in the ligand protein. These candidate residues are mutated to key interaction residues in the ligand protein respectively. The scaffold protein is superposed onto the ligand protein based upon the coordinates of corresponding atoms, which are assumed to strongly interact with the receptor protein. Complementarity between scaffold and receptor proteins is evaluated. Scaffold proteins with a low superposing rms difference and high complementary score are accepted for further analysis. Then, the relative position of the scaffold protein is adjusted so that the interfaces between the scaffold and receptor proteins have a reasonable packing density. Other mutations are also considered to reduce the desolvation energy or bad steric contacts. Finally, the scaffold protein is cominimized with the receptor protein and evaluated. To test the method, the binding interface of barstar, the inhibitor of barnase, was grafted onto small proteins. Four scaffold proteins with high complementary scores are accepted.

Bacillus↗

Pulse Pyrolysis Infrared Laser Jet Spectroscopy of Chloroketene.

The short-lived molecule chloroketene (ClHCCO) was generated by the pyrolysis of chloroacetyl chloride in a high-temperature nozzle and the infrared laser-absorption spectrum of its nu(2) fundamental band was measured between 2153 and 2161 cm(-1). The spectrum was greatly simplified by comparison with a room-temperature spectrum enabling 230 lines of the (35)ClHCCO isotopomer to be assigned and measured. A least-squares fit of these transitions yielded accurate molecular parameters for the v(2) = 1 state using ground state constants from microwave spectroscopy. The nu(2) band origin of (35)ClHCCO is 2157.19238(16) cm(-1). A comparison of simulated and experimental spectra showed that the rotational temperature of the jet spectrum was around 35 K. Copyright 2000 Academic Press.

Journal Article↗

Pre-clinical experimental studies of indomethacin-releasing copper intrauterine device.

Effects of two types of intrauterine device (IUD) on the prostaglandins and endothelin (ET) in uterus and on the endometrial morphology in rats and rabbits, and Cu2+ releasing amounts of both IUDs in vitro were observed. The results showed that the inhibiting action of the indomethacin-releasing copper IUD (FICu-IUD) on the PGI2 was stronger than that on the TXA2, the ratio of 6-keto-PGF1 alpha/TXB2 was reduced with the increase of the doses. There were significant differences between the groups. The FICu-IUD could inhibit the rising of the ET level and lighten the endometrial impairment caused by the FCu-IUD, and promote copper ion release. It was suggested that indomethacin released by FICu-IUD could effectively reduce abnormal uterine bleeding.

Animals↗

Platelet fibrinogen in the patients with idiopathic bleeding platelet disorder.

The changes of platelet fibrinogen in patients with obscure cutaneous mucosal bleeding and their relation with disease were evaluated. By using flow cytometry and Western blot, the platelet binding site of fibrinogen and the expression of platelet fibrinogen in 8 cases of obscure bleeding were detected. The results showed that platelet surface binding sites of fibrinogen were 65.38 +/- 3.62 in the resting state and 65.25 +/- 5.78 after activated by adenosine diphosphate, showing no significant difference as compared with control group (P > 0.05). The expression of platelet fibrinogen was obviously decreased in 4 patients and increased in one case as compared with control group. It was concluded that the binding sites of fibrinogen on the platelet surface were normal in the patients with idiopathic bleeding platelet disorder. The abnormal of platelet fibrinogen might be one of hemorrhagic reasons in partial patients.

Adult↗

Study on the effects of losartan on cardiomyocyte apoptosis and gene expression after ischemia and reperfusion in vivo in rats.

In order to study the effects of losartan on cardiomyocyte apoptosis following ischemia (0.5 h) and reperfusion (48 h) in vivo and bcl-2 and bax gene expression, TUNEL staining method, immunohistochemistry and in situ hybridization histochemistry (ISHH) were used to monitor the apoptotic cells, mRNA and protein of gene expression, respectively. Image processing system was used to quantitatively dispose the positive metric substance of both immunohistochemistry and ISHH through the average optical density (OD) value. The number of the apoptotic cells were 38 +/- 9 (control group), 0-1 (sham operation group) and 9 +/- 4 (losartan-treated group) in each visual field respectively with the difference among the groups being significant (P < 0.001). OD values of bcl-2 (ISHH) were 0.07425 +/- 0.02029 (control group), 0.05961 +/- 0.009932 (sham operation group) and 0.07619 +/- 0.01445 (losartan-treated group) respectively, while OD values of bcl-2 (immunohistochemistry) were 0.1374 +/- 0.01367 (control group), 0.08510 +/- 0.01862 (sham operation group) and 0.1252 +/- 0.02064 (losartan-treated group). bcl-2 gene expression was increased significantly in the control group and losartan-treated group as compared with sham operation group (P < 0.05). OD value of bax (immunohistochemistry) was 09727 +/- 0.02230 (control group), 0.06182 +/- 0.01430 (sham operation group) and 0.06213 +/- 0.01420 (losartan-treated group). bax gene expression was decreased very significantly in losartan-treated group and sham operation group as compared with control group (P < 0.001). Bcl-2/bax ratio was 1.413 (control group), 1.376 (sham operation group) and 2.016 (losartan-treated group) respectively. The results indicated that losartan might inhibit cardiomyocyte apoptosis following ischemia and reperfusion. The mechanism might be that bax gene expression was inhibited to increase bcl-2/bax ratio.

Animals↗

Identification of a hybrid-specific expressed gene encoding novel RNA-binding protein in wheat seedling leaves using differential display of mRNA.

A hybrid-specific expressed cDNA fragment, designated as AG5, has been identified in wheat seedling leaves using differential mRNA display. AG5 contains an open reading frame (ORF) encoding 183 amino acid residues. Comparison with amino acid sequences in GenBank revealed that the AG5 protein is homologous to a group of Gly-rich proteins with consensus sequence-type RNA-binding domains (CS-RBD). Structural analysis showed that AG5 protein contains five motifs, including a consensus sequence-type RNA-binding domain near its N-terminus, arginine/aspartic acid repeats and a Gly-rich region in its center, a Cys-X2-Cys-X4-His-X4-Cys (CCHC) zinc finger motif in the Gly-rich region, and TrySer2ArgAsp2Arg repeats towards its C-terminus. Of all previously described RNA-binding proteins, only RZ-1 from tobacco has a similar structure to the AG5 protein, but RZ-1 lacks a TrySer2ArgAsp2Arg repeat motif, indicating that the two proteins may belong to a family of closely related proteins in plants. The possible role of AG5 and its relation to wheat heterosis are discussed.

Amino Acid Sequence↗

An amperometric biosensor based on the coimmobilization of horseradish peroxidase and methylene blue on a beta-type zeolite modified electrode.

A new biosensor for the amperometric detection of hydrogen peroxide was developed based on the coimmobilization of horseradish peroxidase (HRP) and methylene blue on a beta-type zeolite modified glassy carbon electrode without the commonly used bovine serum albumin-glutaraldehyde. The intermolecular interaction between enzyme and zeolite matrix was investigated using FT-IR. The cyclic voltammetry and amperometric measurement demonstrated that methylene blue co-immobilized with HRP in this way displayed good stability and could efficiently transfer electrons between immobilized HRP and the electrode. The sensor responded rapidly to H2O2 in the linear range from 2.5 x 10(-6) to 4.0 x 10(-3) M with a detection limit of 0.3 microM. The sensor was stable in continuous operation.

Biosensing Techniques↗

Resonance Rayleigh scattering method for the determination of trace amounts of cadmium with iodide-basic triphenylmethane dye systems.

In dilute phosphoric acid solution, cadmium (II) reacts with a large excess of I- to form [CdI4]2- which reacts further with basic triphenylmethane dyes such as crystal violet (CV), ethyl violet (EV), methyl violet (MV), brilliant green (BG) or malachite green (MG) to form an ion-association complex. This results in a significant enhancement of resonance Rayleigh scattering (RRS) intensity and the appearance of new RRS spectra. The characteristics of RRS spectra of the ion-association complexes, the influencing factors and the optimum conditions of these reactions have been investigated. The intensity of RRS is directly proportional to the concentration of cadmium from 0 to 60 ng mL(-1) for EV and MV systems, 0 to 80 ng mL(-1) for CV system, and 0 to 100 ng mL(-1) for BG and MG systems. The methods exhibit high sensitivities and the detection limits for cadmium are between 0.35 and 2.00 ng mL(-1) depending on the different reaction systems. The new RRS method was applied to the direct determination of traces of cadmium in pure zinc and synthetic water samples.

Journal Article↗

Sequence, organization, and evolution of Rh50 glycoprotein genes in nonhuman primates.

The human RHAG locus encodes Rh50 glycoprotein, a polytopic protein that modulates expression of Rh antigens carried by Rh30 polypeptides. Rh50 is almost invariant, whereas Rh30 shows high polymorphism. To assess the relative conservation and phylogenetic relationship of RHAG genes, we characterized their protein expression, transcript structure, genomic organization, and noncoding regions (promoter and introns) in seven nonhuman primate species. Western blot showed that only ape Rh50 glycoproteins are recognized by the antibody 2D10 specific for the human counterpart. Analysis of RHAG gene and its transcript showed a high degree of sequence identity and features of interspecific diversity. The nonhuman primate RHAG genes are highly similar in promoter region and identical in exon-intron organization. Genomic sequencing identified one retro-transposon-like element in intron 2 and three types of Alu elements in intron 4 and 9, with varying copies of minisatellites. Reconstruction of coding and noncoding sequence trees revealed concordances and discordances with regard to the branching of RHAG-like genes in higher primates. A joined tree of Rh50 glycoproteins and Rh30 polypeptides shows that the former evolved at a rate about two times slower than the latter. Statistical tests demonstrated that at least a portion of the RHAG gene was subjected to a positive selection during evolution of anthropoids.

Amino Acid Sequence↗

Structure and mapping of the mouse matrilin-3 gene (Matn3), a member of a gene family containing a U12-type AT-AC intron.

The gene for murine matrilin-3, an extracellular matrix protein present in cartilage, was isolated and further characterized. The gene spans 23.4 kb and comprises 8 exons; with one exception, this reflects the modular structure of the protein. The major and a minor transcription start site were determined by RNase protection assays to positions approximately 72 nt and 87 nt upstream of the ATG codon, respectively. The promoter contains a TATA-like box 32 bp upstream of the main transcription start as well as several potential binding sites for eukaryotic transcription factors. As in all known matrilin genes, the last intron, separating the exons coding for the coiled-coil domain, does not follow the GT-AG rule and belongs to the subgroup of introns having AT-AC at the ends that are spliced by the U12-type spliceosome. The mouse matrilin-3 gene does not contain hidden exon sequences coding for the second vWFA-like domain present in all other matrilins. The intron that could possibly contain such sequences instead shows 75% repetitive sequences, indicating an evolutionary process that has led to the loss of sequences coding for vWFA2. Single-strand conformation polymorphism analysis was used to map the Matn3 gene to the proximal end of Chr 12, linked to the genes Synd1, Apob, Dntb, and Kif3c.

Amino Acid Sequence↗

Increased expression of the type 1 growth factor receptor family in the conjunctival epithelium of patients with keratoconjunctivitis sicca.

PURPOSE: To investigate the expression of type 1 growth factor receptors (epidermal growth factor receptor, ErbB2, and ErbB3) in the conjunctival epithelium of patients with keratoconjunctivitis sicca. METHODS: Immunofluorescent staining and Western blotting were performed to grade the level of expression of the epidermal growth factor receptor ErbB2, and ErbB3 in conjunctival epithelial impression cytologies taken from both eyes of seven normal subjects and 22 patients with keratoconjunctivitis sicca. RESULTS: Epidermal growth factor receptor staining was observed in a greater percentage of keratoconjunctivitis sicca than normal samples (P <.05). ErbB2 and ErB3 staining in the apical conjunctival epithelium was observed in both groups, but stronger ErbB2 and ErbB3 staining was noted in keratoconjunctivitis sicca conjunctival samples (P <.05). The relative levels of expression of these receptor proteins on immunoblots were consistent with immunofluorescent staining. On immunoblots, epidermal growth factor receptor protein was detected in 50% of keratoconjunctivitis sicca samples, but none of the normal samples (P <.025). The expression of ErbB2 and ErbB3 on immunoblots was also greater in the keratoconjunctivitis sicca samples (P <.05). Immunofluorescent staining scores for these receptors were correlated with conjunctival lissamine green staining scores (r =. 574, P <.01 for epidermal growth factor receptor; r =.620, P <.0025 for ErbB2; r =.502, P <.025 for ErbB3) and with corneal fluorescein staining (r =.409, P <.05 for ErbB2; r =.588, P <.005 for ErbB3). CONCLUSION: The expression of the type 1 growth factor receptors is significantly greater in the conjunctival epithelium of eyes with keratoconjunctivitis sicca than normal eyes. The increased expression of these receptors was positively correlated with ocular surface dye staining. The increased expression of these receptors may contribute to the abnormal growth and differentiation of the conjunctival epithelium that occurs in keratoconjunctivitis sicca.

Adult↗