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Biomedical subjects

X D Zhu

Publications and source records attributed to X D Zhu.

At least 19 recordsLinked to original sources

Incidence-angle dependence of optical reflectivity difference from an ultrathin film on solid surface.

We studied the incidence-angle dependence of the optical reflectivity difference in response to ultrathin films on transparent and opaque substrates. We found that the classical three-layer model reproduces the experimentally obtained angular dependence for a monolayer of xenon on Nb(110) and for a monolayer of protein molecules on functionalized glass. We explore the enhancement of the optical response near the Brewster angle (or its equivalent for opaque substrates) in thin film detection.

Adsorption↗

Mechanical properties of skeletal bone in gene-mutated stöpsel(dtl28d) and wild-type zebrafish (Danio rerio) measured by atomic force microscopy-based nanoindentation.

An atomic force microscopy (AFM)-based nanoindenter was used to evaluate the mechanical properties of skeletal bones in wild-type and gene-mutated zebrafish (Danio rerio), stöpsel(dtl28d). Both skeletons were isolated from adult zebrafish and tested under a load of 5 mN. It was found that stp/stp bone has a similar nanohardness but significantly greater elastic modulus compared with that of wild-type bone. The residual indenter impressions using AFM and the fracture surfaces of both bones using scanning electron microscopy were examined and showed that the bone of zebrafish becomes more brittle after the stp mutation. This first observation of the alteration of bone mechanical behavior by gene mutation in zebrafish system is of scientific and clinical relevance to many areas of study, such as bone fracture and fragility mechanisms in human heritable disorders and bone-materials fabrication via gene engineering.

Animals↗

[Study on the molecular mechanisms involved in the increased collagen synthesis by platelet-derived wound healing factors during wound healing in alloxan-induced diabetic rat].

OBJECTIVE: To explore the molecular mechanisms involved in the increased collagen synthesis by platelet-derived wound healing factors (PDWHF) during wound healing in alloxan-induced diabetic rats. METHODS: Thirty-three male SD rats were divided into two groups, the normal (n = 9) (group A) and the diabetic group (n = 24). Two pieces of full-thickness skin with diameter of 1.8 cm were removed from the dorsal site of diabetic rats. PDWHF (100 micrograms/wound) was topically applied to one side of the diabetic wounds (group B) on the operation day and then once a day in the next successive 6 days. Meanwhile, bovine serum albumin (100 micrograms/wound) was applied to the other side of diabetic wound as control group (group C) in the same way. Levels of transforming growth factor-beta 1 (TGF-beta 1) and procollagen I mRNA in wound tissue were inspected by dot blotting. RESULTS: TGF-beta 1 mRNA levels in group B were 4 folds and 5.6 folds compared with those in group C after 5 and 7 days (P < 0.01), however, still significantly lower than those of group A (P < 0.05). There was no significance difference among three groups on the 10th day after wounding. The levels for procollagen I mRNA in group B amounted to 2.1, 1.8 and 2.3 folds of those in group C after 5, 7, and 10 days (P < 0.01), respectively. Compared with those in the group A, procollagen I mRNA levels in the group B were significantly lower after 5 and 7 days (P < 0.05), and no significant difference was observed between group B and A after 10 days. CONCLUSION: One important way for PDWHF to enhance the collagen synthesis in diabetic wound healing is to increase the gene expression of endogenous TGF-beta 1.

Animals↗

[Determination of phenazine-1-carboxylic acid in anti-fungal agent M18 by high performance liquid chromatography].

A reversed-phase HPLC method for the determination of phenazine-1-carboxylic acid (PCA) in antifungal agent M18 is established. The mobile phase was a mixture of MeOH-5 mmol/L phosphate buffer (pH 5.0) (60:40, volume ratio). The flow rate was 1.0 mL/min, and the detection wavelength was 248 nm. The linear range and detectable limit were 50 mg/L-500 mg/L and 30 mg/L respectively. The recovery was 97.53% and RSD was 1.5%. The method of PCA extraction and detection has proven to be much faster, simpler, more sensitive, accurate and reproducible than those reported already. The assay results can be used as a very important criterion for large-scale production.

Antifungal Agents↗

Formation of calcium phosphate/collagen composites through mineralization of collagen matrix.

Several types of calcium phosphate/collagen composites, including noncrystalline calcium phosphate/collagen, poorly crystalline carbonate-apatite (PCCA)/collagen, and PCCA + tetracalcium phosphate/collagen composites, were prepared through the mineralization of collagen matrix. The type I collagen was presoaked with a PO(3-)(4) containing solution and then immersed in a Ca(2+) containing solution to allow mineral deposition. The solution of 0.56 M sodium dibasic phosphate (Na(2)HPO(4)) with a pH of nearly 14 was metastable and its crystallization produced Na(2)HPO(4) and sodium tripolyphosphate hexahydrate (Na(5)P(3)O(10)). 6H(2)O), leading to a controlled release of orthophosphate ions during the subsequent mineral precipitation. The development of the composites was investigated in detail. The mineral contributed up to 60-70% of the weight of the final composites. The strength and Young's modulus of the composites in tensile tests overlapped the lower range of values reported for bone. When implanted in muscle tissue, the composite showed biodegradability that was partly through a multinucleated giant cell mediated process. In a bone explant culture model it was observed that bone-derived cells deposited mineralizing collagenous matrix on the composite.

Animals↗

Cell-cycle-regulated association of RAD50/MRE11/NBS1 with TRF2 and human telomeres.

Telomeres allow cells to distinguish natural chromosome ends from damaged DNA and protect the ends from degradation and fusion. In human cells, telomere protection depends on the TTAGGG repeat binding factor, TRF2 (refs 1-4), which has been proposed to remodel telomeres into large duplex loops (t-loops). Here we show by nanoelectrospray tandem mass spectrometry that RAD50 protein is present in TRF2 immunocomplexes. Protein blotting showed that a small fraction of RAD50, MRE11 and the third component of the MRE11 double-strand break (DSB) repair complex, the Nijmegen breakage syndrome protein (NBS1), is associated with TRF2. Indirect immunofluorescence demonstrated the presence of RAD50 and MRE11 at interphase telomeres. NBS1 was associated with TRF2 and telomeres in S phase, but not in G1 or G2. Although the MRE11 complex accumulated in irradiation-induced foci (IRIFs) in response to gamma-irradiation, TRF2 did not relocate to IRIFs and irradiation did not affect the association of TRF2 with the MRE11 complex, arguing against a role for TRF2 in DSB repair. Instead, we propose that the MRE11 complex functions at telomeres, possibly by modulating t-loop formation.

Acid Anhydride Hydrolases↗

[Genetic study of plant height of tall plant mutant Mh-1 in rice (Oryza sativa L.)].

Mh-1 is a tall plant mutant induced by irradiation from semi-dwarfed variety Guichao 2. The genetic analyses were carried out through the F1, F2, F3 progenies and test-crossed F1 on Mh-1 crossed with materials with sd-1 gene, non-allelic gene to sd-1 and common tall plant. The results indicate that the induced tall plant trait of Mh-1 is controlled by a recessive inhibiting gene, named tentatively i-sd-1(t), that can regulate the expression of sd-1 gene but not of non-allelic gene to sd-1. The valuation of i-sd-1(t) in hybrid rice breeding was discussed.

Genes, Plant↗

Three-dimensional nano-HAp/collagen matrix loading with osteogenic cells in organ culture.

Transplantation of osteogenic cells with a suitable matrix is one strategy for engineering bone tissue. Three-dimensional distribution and growth of cells within the porous scaffold are of clinical significance for the repair of large bony defects. A nano-HAp/collagen (nHAC) composite that mimics the natural bone both in composition and microstructure to some extent was employed as a matrix for the tissue engineering of bone. A porous nHAC composite was produced in sheet form and convolved to be a three-dimensional scaffold. Using organ culture techniques and the convolving method, we have developed three-dimensional osteogenic cells/nHAC constructs in vitro. Scanning electron microscopic and histological examination has demonstrated the development of the cells/material complex. Spindle-shaped cells migrating out of bone fragments continuously proliferated and migrated throughout the network of the coil. The porous nHAC scaffold provided a microenvironment resembling that seen in vivo, and cells within the composite eventually acquired a tridimensional polygonal shape. In addition, new bone matrix was synthesized at the interface of bone fragments and the composite.

Animals↗

Tissue response to nano-hydroxyapatite/collagen composite implants in marrow cavity.

The tissue response to a nano-hydroxyapatite/collagen composite implanted in a marrow cavity was investigated by histology and scanning electron microscopy. A Knoop microhardness test was performed to compare the mechanical behavior of the composite and bone. The ultrastructural features of the composite, especially the carbonate-substituted hydroxyapatite with low crystallinity and nanometer size, made it a bone-resembling material. It was bioactive, as well as biodegradable. At the interface of the implant and marrow tissue, solution-mediated dissolution and giant cell mediated resorption led to the degradation of the composite. Interfacial bone formation by osteoblasts was also evident. The process of implant degradation and bone substitution was reminiscent of bone remodeling. The composite can be incorporated into bone metabolism instead of being a permanent implant. For lack of the hierarchical organization similar to that of bone, the composite exhibited an isotropic mechanical behavior. However, the resistance of the composite to localized pressure could reach the lower limit of that of the femur compacta.

Animals↗

Selection of novel, specific single-stranded DNA sequences by Flp, a duplex-specific DNA binding protein.

Flp is a member of the integrase family of site-specific recombinases. Flp is known to be a double-stranded (ds)DNA binding protein that binds sequence specifically to the 13 bp binding elements in the FRT site (Flprecognitiontarget). We subjected a random pool of oligonucleotides to the in vitro binding site selection method and have unexpectedly recovered a series of single-stranded oligonucleotides to which Flp binds with high affinity. These single-stranded oligonucleotides differ in sequence from the duplex FRT site. The minimal length of the oligonucleotides which is active is 29 nt. This single strand-specific DNA binding activity is located in the same C-terminal 32 kDa domain of Flp in which the site-specific dsDNA binding activity resides. Competition studies suggest that the apparent affinity of Flp for single-stranded oligonucleotide is somewhat less than for a complete duplex FRT site but greater than for a single duplex 13 bp binding element. We have also shown that Cre, another member of the integrase family of site-specific recombinases, also exhibits single-stranded DNA binding similar to that of Flp.

Base Sequence↗

The role of single-stranded DNA in Flp-mediated strand exchange.

The Flp recognition target site contains two inverted 13-base pair (bp) Flp binding sequences that surround an 8-bp core region. Flp recombinase has been shown to carry out strand ligation independently of its ability to execute strand cleavage. Using a synthetic activated DNA substrate bearing a 3'-phosphotyrosine group, we have developed an assay to measure strand exchange by Flp proteins. We have shown that wild-type Flp protein was able to catalyze strand exchange using DNA substrates containing 8-bp duplex core sequences. Mutant Flp proteins that are defective in either DNA bending or DNA cleavage were also impaired in their abilities to carry out strand exchange. The inability of these mutant proteins to execute strand exchange could be overcome by providing a DNA substrate containing a single-stranded core sequence. This single-stranded core sequence could be as small as 3 nucleotides. Full activity of mutant Flp proteins in strand exchange was observed when both partner DNAs contained an 8-nucleotide single-stranded core region. Using suicide substrates, we showed that single-stranded DNA is also important for strand exchange reactions where Flp-mediated strand cleavage is required. These results suggest that the ability of Flp to induce DNA bending and strand cleavage may be crucial for strand exchange. We propose that both DNA bending and strand cleavage may be required to separate the strands of the core region and that single-stranded DNA in the core region might be an intermediate in Flp-mediated DNA recombination.

Base Sequence↗

LDL oxidation and human placental trophoblast and macrophage cytotoxicity.

We have previously shown that LDL is oxidized by placental cells in primary tissue culture and that this process causes cytotoxicity proportional to LDL oxidation in the presence of sex steroid hormones. Here we describe further experiments linking LDL oxidation to placental cell cytotoxicity. Trophoblast and macrophages were isolated from healthy elective caesarean section placentas by enzymatic digestion and separated by centrifugation on a 40% Percoll gradient and maintained in primary culture for up to 5 days. LDL was oxidized by exposure to 5 microM CuCl2, cells were incubated in the absence of albumen to favor oxidation, and cytotoxicity was measured by 51Cr release from prelabelled cells and cell protein content. Native LDL was oxidized by both cell types with a 10%-50% increase in lipid peroxides and an approximately 4-fold increase in TBARS formation. Increasing concentrations of native LDL and oxidized-LDL increased 51Cr release and diminished protein content in cells incubated in HAM's F-10 medium. Addition of 5 microM Cu2+ augmented cytotoxicity of LDL in macrophage and trophoblast culture, but more in macrophages than trophoblast. Cytotoxicity was diminished by adding 0.001-0.1 mM EDTA to the system, diminishing 51Cr release from 91 +/- 0.5 to 40.8 +/- 1.0% in macrophages and 54.2 +/- 1.2 to 33.1 +/- 1.3% in trophoblast (P < 0.001 in both instances). Similarly, the absence of transition metal ion in culture (MEM medium) blocked an increase in 51Cr release compared to its presence (HAM's F-10 medium). An antioxidant, butylated hydroxytoluene, diminished 51Cr release and LDL electrophoretic mobility in HAM's F-10 medium in placental macrophage culture. LDL oxidation injures placental macrophages and trophoblast, the former more than the latter. The process is LDL- and transition metal ion-dependent and is inhibited by antioxidant. This model of LDL oxidation and placental cell damage in vitro provides a basis for studying mechanisms of placental dysfunction and senescence in human pregnancy.

Antioxidants↗

[Genetic analysis on a new sterile locus discovered in hybrids between indica and japonica rice (Oryza sativa L.)].

Japonica line 02428, possessing wide compatibility gene S-5n, showed semi-sterility as crossed with indica variety IR24. Genetic analysis was conducted on the spikelet fertility in segregation generations from the crosses of 02428 with IR24, Pecos and Akihikara. The results indicated that the sterility in hybrids between 02428 and IR24 was controlled by a single non-allelic to S-5 gene and its inheritance fitted to one-locus sporo-gametophytic interaction model. Pecos, an American variety, has a neutral allele. The new sterility locus was preliminarily located near G24 marker on chromosome 11 detected by RFLP. The non-allelic S-locus in the study is named tentatively S-p(t). It was found that the sterility caused by the cross of Akihikara and IR24 was controlled by two S-loci, S-5 and S-p(t), and its genetical effect was additive.

Chromosome Mapping↗

Microstructural features of non-union of human humeral shaft fracture.

Microstructures of non-unions of human humeral shaft fractures were investigated by using scanning electron microscopy, transmission electron microscopy, and X-ray microdiffraction. The non-union has a trabeculae structural framework similar to woven bone. Among the trabeculae are cavities that are subdivided into small chambers by thin plates of collagen fibrils. Some chambers are filled with variously shaped mineralized particles several micrometers in size. The collagen fibrils in both the trabeculae and the thin plates were only slightly mineralized by hydroxyapatite. Vesicles loaded with noncrystalline calcium phosphate (NCP) were observed in most mineralized particles, and brushite crystals with special morphology were seen to be embedded in some particles in irregular shapes. X-ray microdiffraction results indicated that the mineral phases in the non-unions were mainly NCP in addition to small amounts of hydroxyapatite and brushite. NCP deposition and insufficient mineralization of the collagen fibrils may be two important microstructural features of the non-unions of human humeral shaft fractures different from normally repaired bone callus.

Bony Callus↗

17beta-Estradiol, progesterone, and testosterone inversely modulate low-density lipoprotein oxidation and cytotoxicity in cultured placental trophoblast and macrophages.

OBJECTIVES: We have previously shown that low-density lipoprotein oxidation is diminished by 17beta-estradiol and enhanced by progesterone and testosterone. In these experiments we wished to learn whether sex hormone effects on low-density lipoprotein oxidation alter placental cell viability in primary tissue culture. STUDY DESIGN: Primary tissue culture of human term placental cells was performed. RESULTS: Addition of 17beta-estradiol decreased low-density lipoprotein oxidation (measured as lipid peroxides, thiobarbituric acid-reacting substances, and low-density lipoprotein electrophoretic mobility) and placental cell toxicity (measured as chromium 51 release) with maximum reductions of 28% (macrophages) (p < 0.05) and 26% (trophoblasts) (p < 0.01). Conversely, progesterone and testosterone increased low-density lipoprotein oxidation and chromium 51 release, the latter a maximum of 28% and 18%, respectively, for progesterone and testosterone in macrophages (p < 0.05 in both instances) and 23% in trophoblasts (p < 0.05, testosterone only). Collectively, cytotoxicity was proportional to low-density lipoprotein oxidation and estradiol, progesterone, and testosterone concentrations. CONCLUSIONS: Estradiol inhibits placental macrophage- and trophoblast-mediated low-density lipoprotein oxidation and cytotoxicity, whereas progesterone and testosterone promote these effects. Sex steroid hormones may modulate the effects of oxidative stress on placental function in pregnancy.

Cell Death↗

Imaging objects in tissuelike media with optical tagging and the diffuse photon differential transmittance.

In a proof-of-concept experiment, we optically tagged objects embedded in an inhomogeneous and multiple-scattering medium and measured the difference between the transmitted diffuse photons at two optical wave-lengths, one at and the other off a sharp absorption peak of the exogenous contrast agent. We demonstrated that the visibility of tagged objects was significantly enhanced in comparison with that of untagged objects. From our analysis it seems possible to use dual-wavelength differential transmittance spectrometry together with monoclonal-antibody-delivered optical contrast agents to detect tumors as small as 0.1-0.3 cm in size and embedded as deeply as a few centimeters beneath a tissue surface.

Contrast Media↗

Microstructure and micromechanical properties of the mid-diaphyses of human fetal femurs.

The microstructure, composition and the micromechanical properties across the thickness of femoral mid-diaphyses from 14 to 26 week human fetuses have been investigated. Scanning electron microscopy and transmission electron microscopy were employed to examine structural changes with maturation. The fetal bones consist of layers of woven bone. From young to old fetuses and from outer to inner bone layers, the collagen fibrils become more cross-linked, densely packed and change from disordered to an ordered arrangement. The collagen fibril bundles are also more preferentially oriented and change from a chiefly circumferential to longitudinal direction. The sizes of the apatite crystals also increase with age. The Ca/P ratio remains constant around 1.55 for all the bone layers except the outmost layer which is lower than 1.2. An nano-indenter was used to evaluate the microhardness and elastic modulus of each bone layer. The increase of microhardness and elastic modulus correlates with the maturation of bone. The mechanical properties of the mid-diaphyses of human fetal femurs are anisotropic, which is due to the preferential orientation of collagen fibrils.

Collagen↗