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Biomedical subjects

Qiang Zhao

Publications and source records attributed to Qiang Zhao.

At least 55 records · Page 3Linked to original sources

Autologous mesenchymal stem cell transplantation induce VEGF and neovascularization in ischemic myocardium.

Neovascularization induced by vascular endothelial growth factor (VEGF) represents an appealing approach for treating ischemic heart disease. However, VEGF therapy has been associated with transient therapeutic effects and potential risk for hemangioma growth. Adult mesenchymal stem cells (MSCs) derived from bone marrow are a promising source for tissue regeneration and repair. In order to achieve a safe and persistent angiogenic effect, we have explored the potential of autologous MSCs transplantation to enhance angiogenesis and cardiac function of ischemic hearts. One week after myocardial infarction induced by occlusion of left anterior descending artery, autologous MSCs expanded in vitro was administrated intramyocardially into the infarct area of the same donor rats. By 2 months, MSCs implantation significantly elevated VEGF expression levels, accompanied by increased vascular density and regional blood flow in the infarct zone. The neovascularization resulted in a decreased apoptosis of hypertrophied myocytes and markedly improved the left ventricular contractility (ejection fraction: 79.9+/-7.6% vs. 37.2+/-6.9% in control animals). Therefore, mechanisms underlying MSCs improvement of cardiac functions may involve neovascularization induced by differentiation of MSCs to endothelial cells and para-secretion of growth factors, in addition to the apoptosis reduction and previously reported cardiomyocytes regeneration. Two months after cell transplantation, there are significant improvement of left ventricular function. Hence, autologous MSCs transplantation may represent a promising therapeutic strategy free of ethical concerns and immune rejection, for neovascularization in ischemic heart diseases.

Angiogenesis Inducing Agents↗

In vitro chondrogenic phenotype differentiation of bone marrow-derived mesenchymal stem cells.

In order to study the chondrogenic phenotype differentiation of adult sheep bone marrow-derived mesenchymal stem cells (MSCs) in a defined medium as potential seed cells for cartilage tissue engineering. MSCs were isolated by density centrifugation with Percoll solution from bone marrow aspirated from sheep iliac crest. The third passage of MSCs were induced with H-DMEM containing TGF-beta3, IGF-I, Dexamethasone and VitC. The shape and ultrastructure of cells were observed, toluidine blue stain for GAG and immunohistochemistry for type II collagen were applied for chondrogenic phenotype identification. After 14 days of induction, MSCs changed from a spindle-like appearance to a polynal shape, a large amount of endoplasmic reticulum, Golgi complex and mitochondria were observed, and the differentiation of MSCs chondrogenic phenotype was verified by positive staining of toluidine blue and immunohistochemistry. MSCs derived from bone marrow can differentiate to chondrogenic phenotype when induced in vitro and can be used as optimal seed cells for cartilage tissue engineering.

Animals↗

Inducible defense mechanism against nitric oxide in Candida albicans.

The yeast Candida albicans is an opportunistic pathogen that threatens patients with compromised immune systems. Immune cell defenses against C. albicans are complex but typically involve the production of reactive oxygen species and nitrogen radicals such as nitric oxide (NO) that damage the yeast or inhibit its growth. Whether Candida defends itself against NO and the molecules responsible for this defense have yet to be determined. The defense against NO in various bacteria and the yeast Saccharomyces cerevisiae involves an NO-scavenging flavohemoglobin. The C. albicans genome contains three genes encoding flavohemoglobin-related proteins, CaYHB1, CaYHB4, and CaYHB5. To assess their roles in NO metabolism, we constructed strains lacking each of these genes and demonstrated that just one, CaYHB1, is responsible for NO consumption and detoxification. In C. albicans, NO metabolic activity and CaYHB1 mRNA levels are rapidly induced by NO and NO-generating agents. Loss of CaYHB1 increases the sensitivity of C. albicans to NO-mediated growth inhibition. In mice, infections with Candida strains lacking CaYHB1 still resulted in lethality, but virulence was decreased compared to that in wild-type strains. Thus, C. albicans possesses a rapid, specific, and highly inducible NO defense mechanism involving one of three putative flavohemoglobin genes.

Amino Acid Sequence↗

[Effect of programmed humidification and temperature on drug stability].

AIM: To simplify the study on the effect of relative humidity and temperature on drug stability. METHODS: The stability of penicillin potassium as a model was studied with programmed humidifying and heating. RESULTS: Results of our programmed humidifying and heating experiments are comparable to those of traditional experiment at constant humidity and temperature. CONCLUSION: Programmed humidifying and heating experiments are applicable to drug stability study.

Drug Stability↗

Identification of a conserved sequence motif that promotes Cdc37 and cyclin D1 binding to Cdk4.

Cdc37 is a molecular chaperone that is important for the stability and activity of several protein kinases, including Cdk4 and Raf1. We first determined, using in vitro assays, that Cdc37 binds to the amino-terminal lobe of Cdk4. Subsequent mutagenesis revealed that Gly-15 (G15A) and Gly-18 (G18A) were critical for Cdc37-Cdk4 complex formation. Gly-15 and Gly-18 of Cdk4 are within the conserved Gly-X-Gly-X-X-Gly motif that is required for ATP binding to the kinase. Mutation of either glycine at the equivalent positions of Raf1 (G358A and G361A) also inhibited Cdc37 binding to Raf1. Replacing another conserved residue critical for ATP binding and kinase activity, Lys-35 (K35A), reduced Cdc37-Cdk4 complex formation but to a lesser extent. The interaction of Cdk4 with Cdc37 in vitro was not sensitive to changes in ATP levels. Cell-based assays indicated that Cdk4(G15A) and Cdk4(G18A) were present at the same level as wild type Cdk4. Equivalent amounts of p16 bound to Cdk4(G15A) and Cdk4(G18A) relative to wild type Cdk4, suggesting that Cdk4(G15A) and Cdk4(G18A) adopt significant tertiary structure. However, in contrast to wild type Cdk4, Cdk4(G15A), and Cdk4(G18A) had greatly reduced binding of cyclin D1, Cdc37, and Hsp90. Importantly, overexpression of Cdc37 not only stimulated cyclin D1 binding to wild type Cdk4 but also restored its binding to Cdk4(G15A). Under the same conditions, p16 binding to wild type Cdk4 was suppressed. Our findings show that the interaction of Cdc37 with its client protein kinases requires amino acid residues within a motif that is present in many protein kinases.

Adenosine Triphosphate↗

Structural analysis of lipid complexes of GM2-activator protein.

The GM2-activator protein (GM2-AP) is a small lysosomal lipid transfer protein essential for the hydrolytic conversion of ganglioside GM2 to GM3 by beta-hexosaminidase A. The crystal structure of human apo-GM2-AP is known to consist of a novel beta-cup fold with a spacious hydrophobic interior. Here, we present two new structures of GM2-AP with bound lipids, showing two different lipid-binding modes within the apolar pocket. The 1.9A structure with GM2 bound shows the position of the ceramide tail and significant conformational differences among the three molecular copies in the asymmetric unit. The tetrasaccharide head group is not visible and is presumed to be disordered. However, its general position could be established through modeling. The structure of a low-pH crystal, determined at 2.5A resolution, has a significantly enlarged hydrophobic channel that merges with the apolar pocket. Electron density inside the pocket and channel suggests the presence of a trapped phospholipid molecule. Structure alignments among the four crystallographically unique monomers provide information on the potential role for lipid binding of flexible chain segments at the rim of the cavity opening. Two discrete orientations of the S130-T133 loop define an open and a closed configuration of the hydrophobic channel that merges with the apolar pocket. We propose: (i) that the low-pH structure represents an active membrane-binding conformation; (ii) that the mobile S130-T133 loop serves as a gate for passage of ligand into the apolar pocket; and (iii) that this loop and the adjacent apolar V59-W63 loop form a surface patch with two exposed tryptophan residues that could interface with lipid bilayers.

Apoproteins↗

Locality of quark-hadron duality and deviations from quark counting rules above the resonance region.

We show how deviations from the dimensional scaling laws for exclusive processes may be related to a breakdown in the locality of quark-hadron duality. The essential principles are illustrated in a pedagogic model of a composite system with two spinless charged constituents, for which a dual picture for the low-energy resonance phenomena and high-energy scaling behavior can be established. We introduce the concept of "restricted locality" of quark-hadron duality and show how this results in deviations from the perturbative quantum chromodynamics quark counting rules above the resonance region. In particular, it can be a possible source for oscillations about the smooth quark counting rule, as seen, e.g., in the 90-degree differential cross sections for gammap-->pi(+)n.

Journal Article↗

[Experimental study of the isolation, culture and in chondrogenic differentiation of human bone mesenchymal stem cell].

OBJECTIVE: To study the isolation of human bone marrow mesenchymal stem cells (MSCs) and in vitro differentiation into chondrocytes as potential seed cell for condyle cartilage tissue engineering. METHODS: Human MSCs were isolated by percoll solution from normal human bone marrow sample and cultured in flasks. Specific cell surface markers were identified by flow-cytometry. After the cells were treated with inductive medium containing insulin, transferrin, pyruvate, dexathemesone and TGF-beta for 7 - 14 days, microscopic, histological and immuno-histo-chemical studies were performed for chondrogenic phenotype identification. RESULTS: Primary cultures of human MSCs express CD29 and CD44 positively and meanly, but CD34, CD45 and HLA-DR negatively. After 14 days of induction, the cells were positively stained by safranin O. Immunohistochemical analysis proved strong type II collagen expression. CONCLUSIONS: Percoll helps to generate a better isolation of MSCs from human bone marrow aspirates with a purity more above 95%. The isolated MSCs can be expanded and induced in vitro to differentiate into chondrocytes by inductive medium.

Bone Marrow Cells↗

[Research in synthesis of bioactive peptide RGD and the method for its grafting on PET surface].

The aim of this experiment is to graft synthesizing Arg-Gly-Asp peptides (RGD) on the surface of polymer materials, combine endothelial cells with its special site, enhance the adhesion of endothelial cells on the surface, promote the blood compatibility of the biomaterials. Carboxy group (-COOH) was grafted on the materials surface by ultraviolet (UV) radiation, and the RGD serial obtained by liquid phase synthesis was successfully grafted on the disposed materials by chemical reaction. The endothelialization experiment was made also. The grafting results were measured by X-ray photoelectron spectroscopy (XPS), and endothelialization was observed using optical microscope and scanning electron microscope (SEM). The results indicated that the method improves the effect of materials endothelialization. The experiment has made successful use of UV grafting and chemical coupling methods to graft bioactive RGD onto PET film surface. This is a new method of grafting bioactive peptide.

Biocompatible Materials↗

[Expression of CD44v6 in gastric cancer and its correlation with Helicobacter pylori infection].

BACKGROUND & OBJECTIVE: It was considered that the expression of CD44v6 in gastric cancer was high and Helicobacter pylori (HP) infection was type I carcinogen of gastric cancer. So far, no document has been reported about the relationship between HP infection and CD44v6 expression. This study was designed to investigate the relationship between expression of CD44v6 in gastric cancer and HP infection. METHODS: The specimens were divided into three groups:gastric cancer (56 cases), cancer adjacent tissue (56 cases), and gastric mucosa dysplasia (32 cases). The expression of CD44v6 was determined by reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analysis. HP was determined with the rapid urease test and Warthin-Starry staining. Simultaneously, HP cagA antibody was assayed in the serum of 56 patients with gastric cancer. RESULTS: HP positive infection group: The positive expression rates of CD44v6 in metastasis cancer (100%, 12/12) and non-metastasis cancer (90.5%, 19/21) were higher than those in paracancer tissue (48.6%, 24/35) and in gastric mucosa dysplasia (23.6%, 6/23). No statistical difference was found between non-metastasis cancer and metastasis cancer (P >0.25). But there were significant differences between gastric mucosa dysplasia and gastric cancer or cancer adjacent tissue (P< 0.005). HP negative infection group: The positive expression rates of CD44v6 in metastasis cancer, non-metastasis cancer, cancer adjacent tissue, and gastric mucosa dysplasia were 66.7%, 51.4%, 35.1%, and 18.1%, respectively; no statistical difference was found between each group. There was a positive correlation between HP infection and CD44v6 expression (P< 0.01). Both CD44v6 expression and HP infection in intestinal type gastric cancer were higher than those in diffuse type gastric cancer (P >0.05). The positive rate of HP cagA antibody in serum was higher than that of HP infection in gastric cancer tissue (P >0.05). CONCLUSION: The positive rate of CD44v6 was high in gastric cancer and may serve as a marker for diagnosing gastric cancer. There is obvious correlationship between CD44v6 expression and HP infection.

Aged↗

Novel paromamine derivatives exploring shallow-groove recognition of ribosomal-decoding-site RNA.

Natural aminoglycoside antibiotics recognize an internal loop of bacterial ribosomal-decoding-site RNA by binding to the deep groove of the RNA structure. We have designed, synthesized, and tested RNA-targeted paromamine derivatives that exploit additional interactions on the shallow groove face of the decoding-site RNA. An in vitro transcription-translation assay of a series of 6'-derivatives showed the 6'-position to be very sensitive to substitution. This result suggests that the group at the 6'-position plays a pivotal role in RNA target recognition.

Aminoglycosides↗

Sequence and analysis of rice chromosome 4.

Rice is the principal food for over half of the population of the world. With its genome size of 430 megabase pairs (Mb), the cultivated rice species Oryza sativa is a model plant for genome research. Here we report the sequence analysis of chromosome 4 of O. sativa, one of the first two rice chromosomes to be sequenced completely. The finished sequence spans 34.6 Mb and represents 97.3% of the chromosome. In addition, we report the longest known sequence for a plant centromere, a completely sequenced contig of 1.16 Mb corresponding to the centromeric region of chromosome 4. We predict 4,658 protein coding genes and 70 transfer RNA genes. A total of 1,681 predicted genes match available unique rice expressed sequence tags. Transposable elements have a pronounced bias towards the euchromatic regions, indicating a close correlation of their distributions to genes along the chromosome. Comparative genome analysis between cultivated rice subspecies shows that there is an overall syntenic relationship between the chromosomes and divergence at the level of single-nucleotide polymorphisms and insertions and deletions. By contrast, there is little conservation in gene order between rice and Arabidopsis.

Arabidopsis↗

A fine physical map of the rice chromosome 4.

As part of an international effort to completely sequence the rice genome, we have produced a fine bacterial artificial chromosome (BAC)-based physical map of the Oryza sativa japonica Nipponbare chromosome 4 through an integration of 114 sequenced BAC clones from a taxonomically related subspecies O. sativa indica Guangluai 4 and 182 RFLP and 407 expressed sequence tag (EST) markers with the fingerprinted data of the Nipponbare genome. The map consists of 11 contigs with a total length of 34.5 Mb covering 94% of the estimated chromosome size (36.8 Mb). BAC clones corresponding to telomeres, as well as to the centromere position, were determined by BAC-pachytene chromosome fluorescence in situ hybridization (FISH). This gave rise to an estimated length ratio of 5.13 for the long arm and 2.9 for the short arm (on the basis of the physical map), which indicates that the short arm is a highly condensed one. The FISH analysis and physical mapping also showed that the short arm and the pericentromeric region of the long arm are rich in heterochromatin, which occupied 45% of the chromosome, indicating that this chromosome is likely very difficult to sequence. To our knowledge, this map provides the first example of a rapid and reliable physical mapping on the basis of the integration of the data from two taxonomically related subspecies.

Chromosomes↗

[Cartilage engineering utilizing a porous bioceramic (beta-TCP)-cell complex].

OBJECTIVE: To evaluate the feasibility of growing tissue-engineered cartilage using chondrocytes seeded onto a biodegradable porous bioceramic, the beta-tricalcium phosphate (beta-TCP). METHODS: A porous bioceramic template of beta-TCP was created in the shape of a disc. Chondrocytes isolated from rabbit articular cartilage were seeded on the beta-TCP template and then kept in rotatory cell culture system (RCCS) for 1 week prior to subcutaneous transplantation into athymic mice. The three-dimensional structure was well-maintained 16 weeks after implantation. After 4, 8, 16 weeks, the specimens were harvested and examined macroscopically, histologically and immunohistochemically. RESULTS: Gross morphological and histological analysis of the specimens from the chondrocyte-beta-TCP complex demonstrated new cartilage construction. The overall configuration of the experimental specimens closely resembled the structure of beta-TCP template. CONCLUSION: These findings suggest that porous bioceramic (beta-TCP) is a good "matrix" for chondrocyte, and can be used for cartilage engineering.

Animals↗

Repair of large articular cartilage defects with implants of autologous mesenchymal stem cells seeded into beta-tricalcium phosphate in a sheep model.

Tissue engineering has long been investigated to repair articular cartilage defects. Successful reports have usually involved the seeding of autologous chondrocytes into polymers. Problems arise because of the scarcity of cartilage tissue biopsy material, and because the in vitro expansion of chondrocytes is difficult; to some extent, these problems limit the clinical application of this promising method. Bone marrow-derived mesenchymal stem cells (MSCs) have been proved a potential cell source because of their in vitro proliferation ability and multilineage differentiation capacity. However, in vitro differentiation will lead to high cost and always results in decreased cell viability. In this study we seeded culture-expanded autologous MSCs into bioceramic scaffold-beta-tricalcium phosphate (beta-TCP) in an attempt to repair articular cartilage defects (8 mm in diameter and 4 mm in depth) in a sheep model. Twenty-four weeks later, the defects were resurfaced with hyaline-like tissue and an ideal interface between the engineered cartilage, the adjacent normal cartilage, and the underlying bone was observed. From 12 to 24 weeks postimplantation, modification of neocartilage was obvious in the rearrangement of surface cartilage and the increase in glycosaminoglycan level. These findings suggest that it is feasible to repair articular cartilage defects with implants generated by seeding autologous MSCs, without in vitro differentiation, into beta-TCP. This approach provides great potential for clinical applications.

Animals↗