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Biomedical subjects

C Y Zhou

Publications and source records attributed to C Y Zhou.

At least 19 recordsLinked to original sources

Runx2 overexpression enhances osteoblastic differentiation and mineralization in adipose--derived stem cells in vitro and in vivo.

Like bone marrow stromal cells, adipose tissue-derived stem cells (ADSCs) possess multilineage potential, a capacity for self-renewal and long-term viability. To confirm whether ADSCs represent a promising source of cells for gene-enhanced bone tissue-engineering, the osteogenic potential of ADSCs under the control of certain osteoinductive genes has been evaluated. Runx2, a transcription factor at the downstream end of bone morphogenetic protein (BMP) signaling pathways, is essential for osteoblast differentiation and bone formation. In this study we used adenovirus vector to deliver Runx2 to ADSCs and then examined the enhancement of osteogenic activity. Overexpression of Runx2 inhibited adipogenesis, as demonstrated by suppression of LPL and PPARgamma expression at the mRNA level and reduced lipid droplet formation. Moreover, ADSCs transduced with Ad-Runx2 underwent rapid and marked osteoblast differentiation as determined by osteoblastic gene expression, alkaline phosphatase activity and mineral deposition. Additionally, histological examination revealed that implantation of Runx2 modified ADSCs could induce mineral deposition and bone-like tissue formation in vivo. These results confirmed, firstly, the ability of Runx2 to promote osteogenesis and cell differentiation and, secondly, the competence of ADSCs as target cells for bone tissue engineering. Our work demonstrates a potential new approach for bone repair using Runx2-modified ADSCs for bone tissue engineering.

Adenoviridae↗

In vitro and in vivo induction of bone formation based on ex vivo gene therapy using rat adipose-derived adult stem cells expressing BMP-7.

BACKGROUND: Adipose-derived adult stem (ADAS) cells are multipotent cells capable of differentiating into osteoblasts, adipocytes and chondrocytes. The aim of this study was to determine whether BMP-7-expressing ADAS cells would elicit bone formation invitro and in vivo. METHODS: ADAS cells were harvested from Lewis rats and transduced with adenovirus carrying the recombinant human bone morphogenetic protein-7 (Ad-BMP-7) gene. Untransduced cells and cells transduced with adenovirus carrying the enhanced green fluorescence protein (Ad-EGFP) gene served as controls. BMP-7 expression was assessed by RT-PCR, immunofluorescence on day 1, and Western blot on days 4, 8 and 12. Alkaline phosphatase (ALP) activity was assayed on days 2, 4, 6, 8, 10 and 12. Osteocalcin production and bone nodule formation were detected by immunohistochemistry and von Kossa stain on day 12. A total of 1 x 10(6) cells mixed with type I collagen were implanted into the subcutaneous pocket in Lewis rat and subjected to histologic analysis 1, 2 and 4 weeks post-implantation. RESULTS: The Ad-BMP-7-transduced ADAS cells expressed BMP-7 at both mRNA and protein levels. ALP activity was detected in Ad-BMP-7-transduced cells from day 2 to day 12, peaking on day 8. Osteocalcin production and matrix mineralization further confirmed that these cells differentiated into osteoblasts and induced bone formation in vitro. Histologic examination revealed that implantation of BMP-7-expressing ADAS cells could induce new bone formation in vivo. DISCUSSION: ADAS cells would be a promising source of adult autologous stem cells for BMP gene therapy and tissue engineering.

Adenoviridae↗

[Effects of modified SD therapy on Bell's palsy].

OBJECTIVE: To investigate the effects of a modified SD therapy (intravenous high dose hydrocortisone and low molecular dextran) on Bell's palsy(BP), and to avoid the severe side-effects such as hepatic and renal disorders during the treatment. METHOD: Seventy-one BP patients were treated with modified SD method (modified group), thirty-two BP patients took prednisone (control group), and the curative rates of two therapies were compared, in the meanwhile recorded the side effects in detail. RESULT: Curative rate in modified group was 95.8% (House-Brakmsnn I-II) and in control group was 81.2% (P < 0.05). In modified group, the curative rate (I grade) in the patients were treated within 24 hours following onset was 75.0%, and the curative rate in other three subgroups were treated within 24-48 hours, 2-3 day and 3-5 day following onset was 43.8%, 26.7%, 18.8% respectively. Modified group had not shown hepatic and renal disorders or gastric ulcer. CONCLUSION: Modified SD method may increase recoverable rate and can avoid significant side effects for BP, and emphasizes the importance of the administration of SD therapy in the early stage of the disease.

Adult↗

Secretion of ferritin by rat hepatoma cells and its regulation by inflammatory cytokines and iron.

The possibility that serum ferritin is a secreted protein and an acute phase reactant regulated by inflammatory hormones and iron was examined in a hepatic cell line that secretes plasma proteins. Differentiated rat hepatoma cells released albumin and ferritin into the medium, as determined by rocket immunoelectrophoresis and isolation of ferritin by standard procedures plus immunoaffinity chromatography, following labeling with radioactive amino acid. Administration of interleukin-1-beta (IL-1) or tumor necrosis factor-alpha (TNF) doubled the amounts of ferritin released into the medium over 24 and 48 hours. Together, the cytokines had more than an additive effect. Albumin secretion was diminished by IL-1, but not TNF. Iron, administered as an iron dextran complex or as a 1:1 chelate with nitrilotriacetate (Fe-NTA), also enhanced ferritin release, but had no effect on albumin. Intracellular ferritin concentrations did not change significantly with cytokine treatment, but increased in response to iron. With or without treatments, release of ferritin and albumin from cells into the medium was inhibited by brefeldin A, an inhibitor of Golgi function. The effect of each of the cytokines and of iron on ferritin and albumin was also blocked by dichlorofuranosylbenzimidazole (DRB), an inhibitor of transcription. The stimulatory effect of Fe-NTA on ferritin secretion was diminished by TNF, and this was partially counteracted by IL-1, indicating additional regulatory complexity. These results show for the first time that hepatic cells secrete ferritin, that this ferritin secretion is regulated by iron and inflammatory cytokines, and that the mechanisms of regulation differ from those for intracellular ferritin. The results would explain why serum ferritin increases in inflammation or when iron flux is enhanced.

Animals↗

Urine levels of aluminum after drinking tea.

A microwave-assisted acid digestion procedure coupled with a graphite furnace atomic absorption method has been applied in the determination of aluminum (Al) in urine to verify the correlation of free forms of Al in tea infusions and urinary excretion of Al. Significant urinary Al excretion has been found in 24-h urine of four volunteers after tea drinking. However, the difference in amount of Al excretion in urine between the consumption of Oolong (black tea) and Long-Jin (green tea), each of them with unique Al contents and species, was not significant. These findings indicated that the high levels of free Al species in tea infusions did not result in significant change in urinary excretion of the metal, possibly owing to the transformation by ligands present in food and the gastrointestinal tract (GIT). However, it could not be assumed that there was no big difference in absorption of the metal in the human body if fractions of consumed Al retained in the body or excreted by bile or feces were considered.

Adult↗

Glucocorticosteroids inhibit leukotriene production.

BACKGROUND: The mode of action of corticosteroids, important drugs in the treatment of inflammatory disease, is not yet fully understood. Corticosteroids are known to inhibit phospholipase A2 in unprimed eosinophils and basophils, preventing leukotriene synthesis, but their effect on cells that are already primed is unknown. OBJECTIVE: As inflammatory cells from atopic subjects are often primed in vivo, we studied the effects of two potent corticosteroids on basophil sulfidoleukotriene production in peripheral blood mixed leukocytes (PBML) from in-season and out-of-season atopic individuals. METHODS: Cells were incubated for 24 hours with mometasone furoate or beclomethasone dipropionate, primed with IL-3, stimulated with calcium ionophore, buffer, allergen or anti-IgE, and leukotriene production was quantified. RESULTS: Peripheral blood mononuclear leukocytes from five of ten donors (in season) produced elevated sulfidoleukotrienes without a stimulus; cells from seven donors responded to anti-IgE by increased sulfidoleukotrienes. Neither steroid consistently affected sulfidoleukotriene production in anti-IgE-stimulated cells which were releasing sulfidoleukotrienes in the absence of a stimulant. In comparison, sulfidoleukotriene production was significantly reduced by 0.01 to 10 nM beclomethasone dipropionate or mometasone furoate when the cells were primed with IL-3 after exposure to the drug and stimulated with calcium ionophore or allergen, but no dose-relationship was apparent. Leukotriene production by PBML in response to anti-IgE was potently inhibited by all concentrations of mometasone furoate (0.01 nM to 1 microM) with an inhibitory concentration50 of less than 0.01 nM. Beclomethasone dipropionate inhibited sulfidoleukotriene production in this group (inhibitory concentration50 6 nM) in a dose-dependent manner. CONCLUSIONS: Sulfidoleukotriene production and, conceivably, priming may be more effectively inhibited by mometasone furoate than beclomethasone dipropionate.

Adult↗

Anti-interleukin-4 inhibits immunoglobulin E production in a murine model of atopic asthma.

Immunoglobulin E (IgE) plays an important role in allergy, acting as an initiating factor and being involved in its persistence and exacerbations. As interleukin-4 (IL-4) is critical in IgE synthesis, we propose that treatment of mice with monoclonal anti-IL-4 (11B11) prior to active sensitization with ovalbumin will inhibit IgE synthesis, therefore arresting the allergic process at an early stage. Mice treated with 11B11 and sensitized with saline or ovalbumin had significantly less serum IgE than their respective control groups which were treated with saline (p < 0.05). This study suggests that anti-IL-4 may be a prophylactic agent in asthma and allergic disease.

Animals↗

Characterization of a kinesin-related gene ATSV, within the tuberous sclerosis locus (TSC1) candidate region on chromosome 9Q34.

In the search for candidate genes for the tuberous sclerosis (TSC1) disease locus on chromosome 9q34, we have isolated an overlapping series of 22 plasmid and phage cDNA clones covering nearly 7 kb and with an open reading frame of 5070 bp encoding a protein of 1690 amino acids. The putative protein product is a member of the kinesin superfamily and is homologous to the mouse KIF1A and the Caenorhabditas elegans unc-104 genes. Both KIF1A and unc-104 function in the anterograde axonal transport of synaptic vesicles. The human homolog is therefore termed H-ATSV (axonal transporter of synaptic vesicles, HGMW-approved nomenclature ATSV) Screening of DNA from 107 tuberous sclerosis patients and 80 unaffected individuals with H-ATSV cDNA probes by pulsed-field gel electrophoresis/Southern blotting following digestion by rare-cutting methylation-sensitive restriction enzymes showed variant banding patterns in three patients with tuberous sclerosis. However, further analysis indicated that these variant fragments represent a rare polymorphism probably associated with methylation of clustered restriction sites. There is no evidence to support H-ATSV as a candidate gene for TSC1.

Amino Acid Sequence↗

A membrane cofactor protein transgenic mouse model for the study of discordant xenograft rejection.

BACKGROUND: In recent years, interest has been revived in the possibility of transplanting organs into humans from a phylogenetically disparate species such as the pig (xenotransplantation). Such discordant xenografts, however, are subject to hyperacute rejection (HAR) and activation of host complement plays a major role in this rejection. This problem may be solved through the use of transgenic technology by providing the grafted tissue with molecules that down-regulate the action of host complement. RESULTS: Transgenesis with a yeast artificial chromosome (YAC) was used to produce transgenic mice with the complete genomic gene of the human complement regulator membrane cofactor protein (MCP). Transgenic mice were obtained that exhibit full regulation of MCP as normally observed in humans. Hearts from these mice were shown to be significantly protected from HAR caused by human serum in an in vivo experimental procedure. CONCLUSIONS: We conclude that MCP can protect discordant xenografts from HAR caused by human serum and that transgenic mice can be used effectively as in vivo models for the study of the role of human complement regulatory molecules in xenotransplantation.

Animals↗

Serum ferritin: does it differ from tissue ferritin?

Serum ferritin isolated from the horse was structurally compared with horse spleen ferritin and was found to differ markedly in molecular weight, iron content, carbohydrate, subunit size and amino acid sequence. The results are summarized and initial results obtained with candidate clones of pieces of two serum ferritin subunits are described.

Animals↗

Determination of serum aluminium using an ion-pair reversed-phase high-performance liquid chromatographic-fluorimetric system with lumogallion.

An ion-pair reversed-phase high-performance liquid chromatographic method with fluorimetric detection, using lumogallion [4-chloro-3-(2,4-dihydroxyphenylazo)-2-hydroxybenzene-1-sulphonic acid] as a ligand, has been successfully applied to the determination of aluminium in human serum. The highly fluorescent aluminium-lumogallion complex (lambda ex 505 nm, lambda em 574 nm) was separated on a LiChrosorb RP-18 column with an eluent consisting of 30% acetonitrile, 70% 0.02 M potassium hydrogen phthalate and 10 microM lumogallion. The proposed system offers a simple, rapid, selective and sensitive method for the determination of aluminium in serum. The detection limit for aluminium was 0.05 microgram/l in aqueous solution and the limit of determination was 2.2 micrograms/l in serum. The recovery of the method is generally over 90%.

Aluminum↗

Physical analysis of the tuberous sclerosis region in 9q34.

We report the construction of a physical map based on cloned DNA within the candidate region for the tuberous sclerosis complex (TSC1) gene on chromosome 9q34, between the markers D9S149 and D9S66. The DNA clones form three contigs consisting of 7 YACs, bridged by P1 and cosmid clones, and cover more than 950 kb of 9q34. Despite intensive screening of all available libraries, two gaps remain. A detailed physical map of much of this region was derived, and restriction mapping of the YAC, P1, and cosmid clones reveals novel CpG islands in this region. This set of genomic clones provides a resource for characterizing candidates for the TSC1 gene, guided by the location of CpG islands.

Base Sequence↗

A set of ninety-seven overlapping yeast artificial chromosome clones spanning the human Y chromosome euchromatin.

Contiguous arrays of yeast artificial chromosomes (YACs) extending from proximal heterochromatic Yq into the pseudoautosomal portion of the Y chromosome and separated by a small interval at the centromere have been constructed. A total of 97 YACs have been aligned along the Y chromosome by STS content analysis using 222 sequence tagged sites (STSs) that detect 263 loci. Forty-five of the STSs used are novel. Their inclusion provides a significant improvement over previously available maps on the density of STS coverage along the Y chromosome, reducing the average spacing to 120 kb assuming a length of 30 Mb for the euchromatin. The average size of 61 YACs determined by pulsed-field gel electrophoresis analysis was at least 0.9 Mb. Minor differences noted between the ordering of STSs on this map compared with those previously reported may be attributed to inherent polymorphism between the Y chromosomes used to construct the YAC libraries.

Base Sequence↗

[The roles of protein kinase C and Na(+)-H+ exchange in the angiotensin II-induced hypertrophic response].

In order to study the roles of protein kinase C (PKC) and Na(+)-H+ exchange in [Sar1] ANG II-induced hypertrophic response, 3H-Leucine incorporation into serum-free cultured neonatal rat cardiomyocytes were interfered with staurosporine, phorbol 12-myristate 13-acetate (PMA) or amiloride. With [Sar1] ANG II, a dose dependent augmentation of protein synthesis was observed. Activator of PKC (PMA) could also accelerate the rate of protein synthesis to a significant extent, while pretreatment of cardiac myocytes with PKC inhibitor (staurosporine) or Na(+)-H+ exchanger inhibitor (amiloride) would block [Sar1] ANG II-induced increase 3H-Leucine incorporation. These results demonstrated that activation of PKC and Na(+)-H+ exchange might mediate the hypertrophic response initiated by [Sar1] ANG II. However, inhibition of cellular Na(+)-H+ exchange did not affect the increase in protein synthesis due to activation of PKC, a finding indicating that the two resorts to regulate cell growth are mediated by independent pathways.

Alkaloids↗

[Contraction stimulates incorporation of 3H-leucine and cell growth in cultured neonatal rat ventricular myocytes].

To determine whether contraction could influence cell growth, the rate of protein synthesis (3H-leucine incorporation) and cell diameter and volume were measured in cultured neonatal rat cardiac myocytes beating spontaneously or arrested by high potassium. In medium supplemented with 10% calf serum, the 3H-leucine incorporation for 24 h in contracting myocytes (CMC) was significantly higher by 14.2% than that in quiescent myocytes (QMC), i.e. 1,229 +/- 29 cpm/10(5) cells vs. 1,076 +/- 60 cpm/10(5) cells (P < 0.01, n = 5 for each group). The cell diameter and cell volume in QMC group were respectively 15.14 +/- 0.42 microns and 1,842 +/- 123 microns3, while in the CMC group the corresponding figures reached to 16.82 +/- 0.64 microns3 and 2,495 +/- 210 microns3, increased by 11.1% and 35.5% respectively (P < 0.01, n = 6 for each group). With prolongation of culture time, the differences in these parameters between CMC and QMC became even more significant. In all these experiments, there was no significant difference in cell number between the two groups (P > 0.05). It is concluded that contraction per se can accelerate protein synthesis and cell growth in neonatal rat ventricular myocardium.

Animals↗

[Left ventricular end-systolic pressure-dimension relation and its sensitility to the positive inotropic factor in the anesthetized open-chest dogs].

Left ventricular end-systolic pressure-dimension relation (ESPDR) was generated by the transient occlusion of the inferior vena cava (IVC) or thoracic aorta (AO) in 13 anesthetized open-chest dogs, while the left ventricular pressure was measured by a TP-200T pressure transducer and the anterior-posterior dimension was measured by a pair of ultrasonic crystal implanted in the endocardium and recorded by a Sonomicrometer. The results showed that: (1) the ESPDR was shown to be linear, unaltered by preload or afterload changes; (2) dobutamine produced a significant increase in the slope (Emax) of the ESPDR-IVC and ESPDR-AO relations, without remarkable effect on the dimension-axis intercept (Do). However, the change in Emax was more significant for ESPDR-IVC (174.43 +/- 23.48%, n = 18), as compared with that for the ESPDR-AO (71.02 +/- 12.46%, n = 12); (3) Do of the ESPDR-AO relation was smaller than that of the ESPDR-IVC both under control and during dobulamine infusion. We conclude that the linear ESPDR can be derived in the anesthetized open-chest dogs either by caval occlusion or by aortic occlusion, and the slope appears to reflect left ventricular contractility. However, techniques used to generate the ESPDR may influence results, including the Do and the sensitivity of Emax to the inotropic state.

Animals↗

[A new leptospiral serovar in the autumnalis serogroup].

Strain A6 of leptospire was isolated from blood culture of a case with leptospirosis on July 1, 1962 in Mengla county, Xishuangbanna, Yunnan. It is proved a new serovar belonging to serogroup Autumnalis by cross agglutination and cross agglutinin-absorption tests. The name, Leprospira interrogans serovar nanla with reference strain A6 is proposed.

Humans↗