Biomedical subjects

Y Zhang

Publications and source records attributed to Y Zhang.

At least 505 records · Page 28Linked to original sources

PubMed2001

Murine high-fat diet and laser photochemical model of basal deposits in Bruch membrane.

OBJECTIVE: To examine the histologic, histochemical, and ultrastructural changes in Bruch membrane in mice on a high-fat diet with and without laser photochemical injury. METHODS: Five groups of C57BL/6 mice were studied. Group 1 included 2-month-old mice on a normal diet; group 2 included 8-month-old mice on a normal diet; group 3 included 8-month-old mice on a high-fat diet; groups 4 and 5 included 8-month-old mice on a normal diet or high-fat diet, respectively, that underwent laser application of one eye with argon blue laser (488 nm). The mice were killed and plasma lipid levels were measured. The eyes were examined by standard electron microscopy, filipin histochemistry for unesterified cholesterol (UC) and esterified cholesterol (EC), and the osmium-tannic acid-phenylenediamine method for preserving extracellular lipid particles. RESULTS: The plasma cholesterol level was significantly higher in the mice on the high-fat diet than the controls (P<.001). Bruch membrane was thicker in group 2 than group 1 (P =.04) and group 3 had a thicker Bruch membrane than group 2 (P =.003). All eyes in group 3 exhibited accumulation of electron-lucent debris. There was no histochemical and ultrastructural evidence that this material represented accumulated UC or EC. Seven of 9 laser-injured eyes in group 5 accumulated basal laminar deposit (BlamD)-like material in Bruch membrane (P =.02). CONCLUSIONS: Electron-lucent debris accumulates in murine Bruch membrane, and the amount correlates with age and high-fat diet. This debris has some similarities with basal linear deposits, although the debris does not form a discrete layer external to the basement membrane of the retinal pigment epithelium as occurs in basal linear deposits. These deposits do not appear to be UC or EC. Laser photochemical injury of the retinal pigment epithelium may result in the appearance of BlamD-like deposits in eyes with electron-lucent debris. The BlamD-like deposits in this model are similar to the basal laminar deposits that occur in age-related macular degeneration. CLINICAL RELEVANCE: This is an animal model of ultrastructural BlamD-like material in Bruch membrane that is very similar to the deposits that occur in age-related macular degeneration.

Animals↗

PubMed2001

Experimental study on facial nerve regeneration with or without geniculate ganglionectomy.

OBJECTIVE: To investigate regeneration of the distal facial nerve following nerve grafting within the tympanic segment with geniculate ganglion preservation or dissection. DESIGN: Randomized controlled trial. SUBJECTS: Twenty-three adult New Zealand albino rabbits were used in this study. INTERVENTIONS: A 2-mm tympanic segment of the facial nerve was removed, and the greater auricular nerve was harvested for grafting in all animals. In group 1 (10 rabbits), the geniculate ganglion was preserved. In group 2 (13 rabbits), the geniculate ganglion was dissected. Mastoidal and extratemporal segments of the facial nerve were harvested 3 months postoperatively for histological examination by electron microscopy. RESULTS: The number of myelinated axons in normal facial nerves was 1819.6 +/- 535.6. In group 1, the number of myelinated axons was 123.6 +/- 31.1, and, compared with normal facial nerves, the diameter of the regenerative axons was decreased and the sheath thickness in the regenerative fiber was diminished. In group 2, the number of myelinated axons was 515.1 +/- 103.1, while the myelin sheath thickness was proportionate to axon diameter. (Data are given as mean +/- SD.) CONCLUSION: Geniculate ganglionectomy may improve motor axon regeneration.

Animals↗

PubMed2001

Biodegradable dextran-polylactide hydrogel network and its controlled release of albumin.

The objective of this paper was to study the release of bovine serum albumin (BSA) from a series of biodegradable hydrogels having a wide range of hydrophilicity to hydrophobicity, swelling, and biodegradation properties. BSA was incorporated into a series of biodegradable hydrogels made from a dextran derivative of allyl isocyanate (dex-AI, as the hydrophilic constituent) and poly(DL-lactic acid) diacrylate macromer (PDLLAM, as the hydrophobic constituent). The release kinetics of BSA from these dex-AI/PDLLAM hydrogels was studied. Laser confocal scanning microscopy was used to investigate the morphological change of the hydrogels, as well as BSA distribution in the hydrogels, as a function of dex-AI to PDLLAM composition ratio and incubation time. We found that the incorporation of PDLLAM into dex-AI reduced the initial burst release of BSA due to its more homogeneous distribution in the hydrogels. As the PDLLAM component increased, the rate of formation of a loose three-dimensional (3D) network structure increased; consequently, the sustained rate and extent of BSA release increased. Both release index and diffusion coefficient (from release kinetics data) increased as the PDLLAM component increased in the hydrogels. The data suggest that the release of BSA was controlled by both diffusion of BSA through swelling of the hydrophilic phase during an early stage, and degradation of the hydrophobic phase during a late stage, and also that the magnitude of diffusion versus degradation controlled release is dependent on composition ratio and immersion time.

Biocompatible Materials↗

PubMed2001

High-throughput polymerase chain reaction analysis of clinical samples by capillary electrophoresis with UV detection.

Routine genetic analysis of large numbers of individuals by polymerase chain reaction (PCR) using capillary electrophoresis is often restricted by the low throughput of standard protocols and the tedious sample preparation process. Here, we demonstrate that capillary electrophoresis with UV detection can be used in PCR-based DNA analysis starting from clinical samples without purification or complicated sample manipulation. After PCR reaction using cheek cells, blood, or HIV-1 gag DNA, the reaction mixtures were injected into a capillary array either on-line or off-line by base stacking. The use of multiplexed absorption detection and the elimination of any purification steps both before and after PCR reaction can potentially provide significant benefits compared to current methods for DNA analysis with regard to time, cost, and labor.

DNA↗

PubMed2001

Comparison of the prevalence of knee osteoarthritis between the elderly Chinese population in Beijing and whites in the United States: The Beijing Osteoarthritis Study.

OBJECTIVE: To estimate the prevalence of radiographic and symptomatic knee osteoarthritis (OA) in a population-based sample of elderly subjects in Beijing, China and compare it with that reported in the Framingham (Massachusetts) OA Study. METHODS: We recruited a sample of persons age > or = 60, using door-to-door enumeration in randomly selected neighborhoods in Beijing. Subjects completed a home interview including questions on knee symptoms and a hospital examination including knee radiographs obtained during weight bearing. The protocol was identical to that used in the Framingham OA Study. A reader read intermingled Beijing and Framingham Study films to ensure high reliability. We defined a subject as having radiographic knee OA when the Kellgren/Lawrence grade was > or = 2 in at least 1 knee. Symptomatic knee OA was recorded as present when knee pain was reported and the symptomatic knee had radiographic OA. We estimated the prevalence of these entities in elderly subjects in Beijing and compared it with OA prevalence in Framingham, using an age-standardized prevalence ratio. RESULTS: Of 2,180 age-eligible Beijing subjects contacted, knee radiographs were obtained in 1,787 (82.0%). The prevalence of radiographic knee OA was 42.8% in women and 21.5% in men. Symptomatic knee OA occurred in 15.0% of women and 5.6% of men. Compared with women of the same age in Framingham, women in Beijing had a higher prevalence of radiographic knee OA (prevalence ratio 1.45, 95% confidence interval 1.31-1.60) and of symptomatic knee OA (prevalence ratio 1.43, 95% confidence interval 1.16-1.75). The prevalence of knee OA in Chinese men was similar to that in their white US counterparts (for radiographic OA, prevalence ratio 0.90; for symptomatic OA, prevalence ratio 1.02). CONCLUSION: Using identical methods and definitions to evaluate the prevalence of OA across populations, we found, surprisingly, that older Chinese women have a higher prevalence of knee OA than women in Framingham, Massachusetts. The prevalence in men was comparable. Possible explanations for these differences range from genetic differences to heavy physical activity among Chinese.

Age Distribution↗

PubMed2001

Regulation of systemic acquired resistance by NPR1 and its partners.

The NPR1 protein of Arabidopsis thaliana has been shown to be an important regulatory component of systemic acquired resistance (SAR). Mutations in the NPR1 gene block the induction of SAR by the signal molecule salicylic acid (SA). NPR1 contains an ankyrin repeats and a BTB domain which are involved in interaction with other protein(s). To further study the function of NPR1 and the regulatory mechanism of SAR, we used both molecular and genetic approaches to identify additional SAR regulatory components. Through a yeast two-hybrid screen we found that NPR1 interacts specifically with bZIP transcription factors. The involvement of bZIP transcription factors in controlling the SA-induced genes had been suggested by a number of promoter studies performed on these genes. It was found that as1 element, which is a binding site for bZIP transcription factors, is essential for SA-induced gene expression. In a genetic screen for suppressors of npr1, we found a mutant, sni1, that restored the responsiveness to SAR induction in npr1. The genetic characteristics of the sni1 mutant and the sequence of SNI1 suggest that the wild-type SNI1 protein is a negative regulator of SAR. We believe that SAR is controlled by both positive regulators and negative regulators.

Arabidopsis↗

PubMed2001

Morphologic predictors of papillary carcinoma on fine-needle aspiration of thyroid with ThinPrep preparations.

Although the cytologic features of papillary carcinoma of the thyroid are well-known, none is entirely specific. We conducted this study to determine the minimal criteria necessary to achieve 100% specificity for the diagnosis of papillary carcinoma on fine-needle aspiration (FNA). Forty patients with histologically confirmed papillary carcinoma and 17 patients with other thyroid lesions who underwent preoperative FNA at Beth Israel Deaconess Medical Center during a 4-yr period were included in the study. All cytology slides were prepared with the ThinPrep processing technique. Various architectural and nuclear features were evaluated, with a score assigned to each feature, and correlated with the histologic diagnosis of papillary carcinoma. Intranuclear inclusions, papillary and/or sheet arrangements, nuclear grooves, powdery chromatin, nuclear molding, high cellularity, and small nucleoli were significantly associated with papillary carcinoma (P < 0.05). The requirement of any intranuclear inclusions and many nuclear grooves, or a minimum of sum of scores (of the above eight features) of 10, yields 100% specificity and approximately 70% sensitivity. Cases with fewer features can be reported as suspicious or indeterminate for papillary carcinoma. A quantitative/probabilistic approach in the reporting of thyroid FNA provides a practical guide for management of patients with thyroid nodules.

Adult↗

PubMed2001

Erythrocytapheresis for Plasmodium falciparum infection complicated by cerebral malaria and hyperparasitemia.

In malaria due to Plasmodium falciparum, life-threatening complications are in part related to the degree of parasitemia. Whole blood exchange and red blood cell exchange (RCE) have been used for the rapid removal of parasites from the circulation of patients with a high parasite load complicated by cerebral, pulmonary, and renal dysfunction. We have treated three 5-45-year-old patients with hyperparasitemia and end-organ dysfunction with red cell exchange by automated apheresis as an adjunct to specific anti-malarial chemotherapy. Parasitemia dropped more than 80% in all three patients immediately after the exchange, and all patients had an uneventful and full recovery. In combination with effective anti-malarial chemotherapy, apheresis RCE is a safe and rapid approach to treat complicated malaria due to P. falciparum.

Adult↗

PubMed2001

Heat shock factor-4 (HSF-4a) is a repressor of HSF-1 mediated transcription.

Heat shock transcription factors (HSFs) regulate the expression of heat shock proteins and other molecular chaperones that are involved in cellular processes from higher order assembly to protein degradation and apoptosis. Among the human HSFs, HSF-4 is expressed as at least two splice variants. One isoform (HSF-4b) possesses a transcriptional activation domain, but this region is absent in the other isoform (HSF-4a). We have recently shown that the HSF-4a isoform represses basal transcription from heterologous promoters both in vitro and in vivo. Here we show that HSF-4a and HSF-4b have dramatically different effects on HSF-1-containing nuclear bodies, which form after heat shock. While the expression of HSF-4b colocalizes with nuclear granules, the expression of HSF-4a prevents their formation. In addition, there is a concurrent reduction of HSF-1 in the nucleus, and there is reduction in its DNA binding activity and in HSE-dependent transcription of a reporter gene. To better understand the mechanism by which HSF-4a represses transcription, we inducibly expressed HSF-4a in cells and found that HSF-4a binds to the heat shock element (HSE) during attenuation of the heat shock response. Thus HSF-4a is an active repressor of HSF-1-mediated transcription. This repressor function makes the HSF-4a isoform unique within the HSF family.

Cell Nucleus↗

PubMed2001

Ebp1, an ErbB-3 binding protein, interacts with Rb and affects Rb transcriptional regulation.

Ebp1, an ErbB-3 binding protein, inhibits the proliferation and induces the differentiation of human breast cancer cells. The mechanisms of these effects are unknown. Rb, the product of the retinoblastoma gene, is an important modulator of cell cycle progression and cellular differentiation. We report that Rb is a binding target for Ebp1. Ebp1 was localized to both the nucleus and the cytoplasm of logarithmically growing AU565 breast cancer cells and HeLa cells as determined by confocal immunofluorescent microscopy. Ebp1 was present in Rb immunoprecipitates derived from AU565 breast cancer cells. GST-Rb also bound endogenous Ebp1. Using GST-Ebp1 constructs, we determined that the 72 C-terminal amino acids of Ebp1 were sufficient to bind Rb. Dephosphorylation of Ebp1 enhanced the interaction of Ebp1 with Rb. The overexpression of Ebp1 in MCF-7 and AU565 (Rb(+)) cells inhibited the activity of the E2F1 regulated cyclin-E promoter. Ebp1 bound E2F1 indirectly via Rb in lysates of MCF-7 cells. The interaction of Ebp1 with Rb may prove to be an important mechanism of Ebp1 induced changes in cell proliferation and differentiation.

Adaptor Proteins, Signal Transducing↗

PubMed2001

Studies of apoptosis of malignant lymphoma cells induced by arsenic trioxide.

The present study investigated the effect of As(2)O(3)on malignant lymphoma cells. Cell apoptosis was detected by cell staining and TdT-mediated dUTP Nick-end Labelling (TUNEL). Cellular DNA and protein expression content were determined by immunohistochemistry and flow cytometry. It was found that 0.5-2.0 microm /l As(2)O(3)could inhibit cell growth, including Raji cells and lymphoma cells from patients, and induce apoptosis, such as condensed chromatin and nuclear fragmentation with intact cell membrane, i.e. apoptotic body. It was also found that the cells of the sub-G(1)phase increased significantly and bcl-2 gene expression was greatly downregulated. However, this effect was not observed for Jurkat cells under the same conditions. We concluded that As(2)O(3)at a range of 0.5-2.0 microm /l can inhibit the growth and induce apoptosis in malignant lymphoma cells, which may have therapeutic potential.

Antineoplastic Agents↗

PubMed2001

Correlation between putative inhibitory molecules at the dorsal root entry zone and failure of dorsal root axonal regeneration.

The molecular mechanisms involved in preventing regenerating dorsal root axons from entering the spinal cord at the dorsal root entry zone (DREZ) are obscure. We used immunohistochemistry, in situ hybridization, and electron microscopy to study axonal regeneration after dorsal rhizotomy in adult rats and its relationship to cellular changes and the distribution of putative growth inhibitory molecules in this region. Astrocyte processes, ending as bulb-shaped expansions, grew up to 700 microm into the basal lamina tubes of injured roots, where regenerating axons were also present. Some of these axons approached or reached the DREZ but grew no further; others turned back toward the ganglion, suggesting the presence of repulsive cues in or near the DREZ. Tenascin-C mRNA and protein and CSPG stub immunoreactivity were strongly upregulated in the roots after rhizotomy, but were only weakly expressed in the DREZ. Tenascin-R immunoreactivity was confined to CNS tissue, and unaffected by rhizotomy. Large, rounded GFAP-negative, NG2-immunoreactive cells, a few of which were OX42 positive, were found in the DREZ following rhizotomy. Astrocyte processes projecting into the roots were tenascin-R and NG2 negative. Hence, only NG2-expressing cells and tenascin-R were appropriately situated to inhibit regeneration through the DREZ.

Animals↗

PubMed2001

Escherichia coli K1 purA and sorC are preferentially expressed upon association with human brain microvascular endothelial cells.

In order to better understand the events that allow Escherichia coli K1 to cross the blood-brain barrier we used differential fluorescence induction to identify bacterial genes that are preferentially expressed when associated with human brain microvascular endothelial cells (HBMEC), which comprise the blood-brain barrier. Random gene fusions of E. coli K1 DNA were created in a promoterless gfp vector and gene fusion libraries were incubated with and without HBMEC. The cells were subjected to a series of fluorescence-activated cell sorting screens to identify promoter fusions which lead to fluorescence when bacteria were associated with HBMEC, yet not fluorescent when grown in media alone. Two genes were identified, purA (encodes adenylosuccinate synthetase) and a sorC homologue (encodes a member of the sorC family of transcriptional regulators), whose expression were preferentially induced when bacteria were associated with eukaryotic cells. Individual gene disruption mutants of E. coli K1 purA and sorC demonstrated significantly decreased HBMEC invasion phenotype in vitro, when compared to the wild-type strain, and could be complemented when the respective wild-type sequences were supplied in trans. The purA and sorC mutants were deficient in their ability to grow in defined minimal media, without adenine, and with sorbose as sole carbon source, respectively, yet capable of normal growth in complex media. We have identified novel phenotypes associated with E. coli K1 purA and sorC, which provide evidence that these genes contribute to the invasion of HBMEC.

Artificial Gene Fusion↗

PubMed2001

Molecular systematics of Xenocyprinae (teleostei: cyprinidae): taxonomy, biogeography, and coevolution of a special group restricted in East Asia.

We surveyed mitochondrial DNA (mtDNA) sequence variation in the subfamily Xenocyprinae from China and used these data to estimate intraspecific, interspecific, and intergeneric phylogeny and assess biogeographic scenarios underlying the geographic structure of lineages. We sequenced 1140 bp of cytochrome b from 30 individuals of Xenocyprinae and one putative outgroup (Myxocypris asiaticus) and also sequenced 297 bp of ND4L, 1380 bp of ND4, 68 bp of tRNA(His), and 69 bp of tRNA(Ser) from 17 individuals of Xenocyprinae and the outgroup (M. asiaticus). We detected high levels of nucleotide variation among populations, species, and genera. The phylogenetic analysis suggested that Distoechodon hupeinensis might be transferred to the genus Xenocypris, the taxonomic status of the genus Plagiognathops might be preserved, and species of Xenocypris and Plagiognathops form a monophyletic group that is sister to the genus Distoechodon and Pseudobrama. The introgressive hybridization might occur among the populations of X. argentea and X. davidi, causing the two species to not be separated by mtDNA patterns according to their species identification, and the process and direction of hybridization are discussed. The spatial distributions of mtDNA lineages among populations of Xenocypris were compatible with the major geographic region, which indicated that the relationship between Hubei + Hunan and Fujian is closer than that between Hubei + Hunan and Sichuan. From a perspective of parasite investigation, our data suggested that the fauna of Hexamita in Xenocyprinae could be used to infer the phylogeny of their hosts.

Animals↗

PubMed2001

Acute cytokine response to systemic adenoviral vectors in mice is mediated by dendritic cells and macrophages.

We investigated the cellular basis for secretion of inflammatory cytokines in mice following intravenous administration of adenoviral vectors (Ad). Serum inflammatory cytokines including interleukin-6 (IL-6), IL-12, and tumor necrosis factor-alpha (TNF-alpha) were detected as early as 6 h following intravenous injection of Ad-expressing Escherichia coli beta-galactosidase (Ad-lacZ). Ad-lacZ readily accumulated in the splenic marginal zone 1 h after intravenous infusion, where both dendritic cells (DCs) and macrophages were transduced and activated within 6 h. Flow cytometric analyses showed that the expression of Ia and CD86 antigens was markedly enhanced on splenic DCs indicating their activation in vivo by Ad-lacZ. Upon ex vivo culture, these early-activated splenic DCs spontaneously produced high levels of IL-6 and IL-12. By contrast, activated splenic macrophages spontaneously secreted only IL-6. Elimination of tissue macrophages and splenic DCs in vivo considerably reduced the early release of IL-12, IL-6, and TNF-alpha and significantly blocked the specific cellular immune response to Ad and the transgene product in vivo. Our findings indicate that preferential activation of DCs and macrophages may account for Ad-triggered acute inflammatory response in vivo in mice. Moreover, DCs and macrophages may play different roles in this process in terms of their abilities to produce distinct patterns of inflammatory cytokines.

Adenoviridae↗

PubMed2001

Activation of innate immunity in nonhuman primates following intraportal administration of adenoviral vectors.

The innate immune response to intraportally infused adenoviral vector was evaluated in rhesus monkeys. A first-generation adenovirus-expressing lacZ (Ad-lacZ) was administered at a dose just below that which causes severe morbidity. The response to vector was evaluated for the initial 24 h following infusion. Clinical findings during this time were primarily limited to petechiae, consistent with the development of thrombocytopenia and biochemical evidence of disseminated intravascular coagulation. Serum transaminases were elevated and a lymphopenia developed. Tracking of fluorescent-labeled vector demonstrated distribution to macrophages and dendritic cells of the spleen and Kupffer cells of the liver. A systemic release of the cytokine IL-6 occurred soon after vector infusion. Analysis of splenic cells revealed acute activation of macrophages and dendritic cells followed by massive apoptosis. Bone marrow cultures demonstrated normal erythroid and primitive progenitors with a significant decrease in myeloid progenitors. Similar findings, except the abnormality in bone marrow cultures, were observed in monkeys who received an identical dose of Ad-lacZ in which vector genes were inactivated with psoralen and UV irradiation. These data suggest that inadvertent targeting of antigen-presenting cells following intraportal infusion of vector leads to a systemic cytokine syndrome which may be triggered by the viral capsid proteins.

Adenoviridae↗

PubMed2001

Analysis of the first heart sound using the matching pursuit method.

It is acknowledged that the first heart sound S1 consists of two major, high-frequency components M1 and T1, corresponding, respectively, to the vibrations of the mitral and tricuspid valves and their surrounding tissues following valve closure in early systole. In this study, the matching pursuit (MP) method was used to decompose S1 into a series of time-frequency atoms. M1 and T1 were separated from the parameterised atoms of S1. The first two dominant frequencies of M1 were identified and used as features of a linear classifier to diagnose mitral valve abnormality. This method was applied to two sets of S1 data recorded from 15 patients with normal, and 15 patients with abnormal, bioprosthetic mitral valves, respectively. It was found that the two features exhibit significant differences between the normal and abnormal sets (p< 0.001). Using these two features, a correct classification of 93% was obtained. In addition, when the Wigner distribution of S1 was calculated from the decomposed atoms and compared with a spectrogram, the MP method provided better results. The study demonstrates that the MP method may be a promising technique for heart sound analysis.

Heart Murmurs↗

PubMed2001

Effect of antisense oligodeoxynucleotide directed to NF-kappa B-RelA on Bcl-XL mRNA in extended drug resistance leukemia cell line HL-60/E6.

To explore the effect of NF-kappa B on bcl-x gene transcription in extended drug resistance leukemia cell line HL-60/E6, drug-resistant subline HL-60/E6 was derived by intermittently exposing HL-60 cells to 6 ng/ml epirubicin. Indirect immunofluorescence was used to demonstrate the location of NF-kappa B-RelA in HL-60/E6 cells. FCM analysis and RT-PCR were used to detect the efficiency of liposome-mediated ODN transfection and the change of bcl-XL mRNA levels after 5 mumol/L phosphorothioate (PS)-derivatized antisense (AS) oligodeoxynucleotide (ODN) directed to RelA was transferred into HL-60/E6 cells. The results showed that RelA remained persistently active and located at the nuclei of HL-60/E6 cells, but in the cytoplasm of HL-60 cells, the efficiency of liposome-mediated ODN transfection was significantly higher than that of null ODN (P < 0.01 in 4 h, 6 h, 12 h, 24 h). Exposure of HL-60/E6 cells to 5 mumol/L AS-PS-ODN directed to RelA led to a maximal 40% decline of bcl-XL mRNA levels within 8 h. The inhibition rate of bcl-XL mRNA was (15 +/- 1.79)%, (28 +/- 2.34)%, (40 +/- 3.47)%, (20 +/- 1.54)%, in 4 h, 6 h, 8 h, 15 h, respectively, but it was less than 15% in control group. It was concluded that NF-kappa B was involved in regulating bcl-x transcription. It was suggested that NF-kappa B was an important factor for drug resistance in leukemia cells.

Apoptosis↗