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Biomedical subjects

W Zhao

Publications and source records attributed to W Zhao.

At least 109 records · Page 6Linked to original sources
PubMed2001

[Characters of uncoupling protein and its relation with obesity].

Uncoupling proteins(UCP) are carrier proteins in mitochondria. In eukaryotic cells, ATP is generated by oxidative phosphorylation, an energetic coupling at mitochondria level. The oxidative reactions occurring in the respiratory chain generate an electrochemical proton gradient at both sides of the inner membrane of mitochondria. This gradient is used by the ATP synthase to phosphorylate ADP into ATP. The coupling of cell respiration with ADP phosphorylation is only partial in brown adipose tissue (BAT) mitochondria, where UCP causes a reentry of protons into the matrix and abolishes the electrochemical proton gradient. The liberated energy is then dissipated as heat and the synthesis of ATP is reduced. Recently, the cloning of new UCPs expressed in other tissues revealed the importance of this kind of regulation of respiratory control in metabolism and energy expenditure. The newly characterized UCPs are potential target drugs for obesity treatment, which could be favor of energy expenditure and diminish the metabolic efficiency.

Energy Metabolism↗
PubMed2001

[Analysis of the mitochondria-related protein of Schistosoma japonicum and its antigen epitopes].

OBJECTIVE: To sequence the cloned gene Sj338 and to identify the encoded protein and its antigen epitopes. METHODS: The Sj338 gene fragment obtained from adult S. japonicum cDNA library amplified by PCR method was subcloned into pGEM-T vector for sequencing. The sequence of nucleotides and the characteristics of the encoded protein were analyzed by DNASIS Program and Goldkey DNA and Protein Analytical Program, and then the homology of the amino acid sequence was searched on the BLAST net. RESULTS: The cloned rSj338 gene was demonstrated to be 487 bp containing one 459 bp ORF, encoding a protein consisted of 153 amino acids with a molecular weight of 17.6 kDa. The amino acid sequence of the recombinant protein rSj338 shared 46% identity with that of the corresponding part of human mitochondrial import receptor and 44% identity with that of the Rattus sp. mitochondrial precursor receptor. The possible antigen epitopes were predicted within the peptide fragments of 26-32 aa, 37-46 aa and 147-151 aa. CONCLUSION: The protein encoded by rSj338 gene fragment might be the mitochondria-related protein of Schistosoma japonicum.

Amino Acid Sequence↗
PubMed2001

[IR studies on hydrogen bonding in hard segment domains of segmented polyester polyurethaneurea].

Hydrogen bonding in hard segment domains of segmented polyester polyurethaneurea (PUU) based on 4,4'-diphenylmethane diisocyanate (MDI), ethylene diamine (ED) and polybutylene adipate (PBA) (M.W.1,975 and M.W.1,228) was studied by means of IR spectroscopy. It was found that the double peak absorption of urea C=O groups occurred in the studied PUU with the increase of hard segment content. Two hydrogen bonding models of urea C=O groups in different hard segment sequence lengths of PUU were proposed. Three dimensional hydrogen bonding in hard segment domains can be formed by further bonding of the above planar hydrogen bondings.

Hydrogen Bonding↗
PubMed2001

[IR studies on segmented polyester polyurethaneurea elastomers].

The studies segmented polyester polyurethaneurea (PUU) elastomers based on 4,4'-diphenylmethane diisocyanate (MDI), ethylene diamine (ED) and polybutylene adipate(PBA) (M.W.1,975 and M.W.1,228) were studied by means of IR spectrascopy. The major IR bands of PUU were first assigned and the views were proposed for the bands at 1,600, 1,317, 1,520 and 1,230 cm-1 assigned by other authors. The reliable theoretical basis for the assignments of IR bands and the studies of hydrogen bonding in these elastomers has been proposed by use of deuteration and varying temperature. The results indicated that substantial hydrogen bonding mainly occurred in the aggregate region of hard segments, a small amount of hydrogen bonding can be formed at the interface between the soft and hard segments. It can be concluded from this work that the studied PUU materials were phase segregation.

Elastomers↗
PubMed2001

[Effects of aspiration of airway dead space on blood gases and respiratory mechanics in acute hypercapnia model dogs].

OBJECTIVE: To establish the method of aspiration of airway dead space (ASPIDS) and evaluate the effects of ASPIDS on elimination of CO2 and improvement of ventilation efficiency. METHODS: Using self-designed ASPIDS device, physiological parameters such as PaCO2, P(peak), P(pause), end expiratory carbon dioxide pressure (PaCO2), lung statistic compliance (Cst), respiratory resistance (Re) and hemodynamics in two animal groups of conventional mechanical ventilation (CMV) and hypercapnia (HC) were observed by applying tracheal gas insufflation (TGI 4.0 L/min) and ASPIDS (4.0 L/min). RESULTS: ASPIDS significantly decreased PaCO2 level, enhanced CO2 elimination and maintained PaCO in normal range while decreased VT by 30%, the elimination of CO2 of ASPIDS was superior to that of TGI. ASPIDS significantly decreased airway pressure in both animal groups . P(et)CO2 and Re levels significantly decreased after ASPIDS. ASPIDS showed no influence on Cst, hemodynamics and oxygenation. CONCLUSIONS: The ASPIDS device is a simple and practical adjunct method to mechanical ventilation, it can more effectively decrease PaCO2 than TGI, and maintain lower airway pressure.

Animals↗
PubMed2000

Nonreceptor tyrosine protein kinase pp60c-src in spatial learning: synapse-specific changes in its gene expression, tyrosine phosphorylation, and protein-protein interactions.

c-src is a nonreceptor tyrosine protein kinase that is highly concentrated in synaptic regions, including synaptic vesicles and growth cones. Here, we report that the mRNA signal of pp60c-src is widely distributed in the rat brain with particularly high concentrations in the hippocampus. After spatial maze learning, up-regulation of c-src mRNA was observed in the CA3 region of the hippocampus, which was accompanied by increases in pp60c-src protein in hippocampal synaptosomal preparations. Training also triggered an increase in c-src protein tyrosine kinase activity that was correlated with its tyrosine dephosphorylation in the synaptic membrane fraction. After training, pp60c-src from hippocampus showed enhanced interactions with synaptic proteins such as synapsin I, synaptophysin, and the type 2 N-methyl-d-aspartate receptor, as well as the cytoskeletal protein actin. The association of pp60c-src with insulin receptor in the synaptic membrane fraction, however, was temporally decreased after training. Furthermore, in vitro results showed that Ca(2+) and protein kinase C might be involved in the regulation of protein-protein interactions of pp60c-src. These results suggest, therefore, that pp60c-src participates in the regulation of hippocampal synaptic activity during learning and memory.

Animals↗
PubMed2000

Differential changes of bax, caspase-3 and p21 mRNA expression after transient focal brain ischemia in the rat.

Recent studies of transient focal ischemia have focused interest on apoptotic mechanisms of neuronal cell death involving constitutive pro-apoptotic proteins. The finding of specific patterns of novel gene expression might indicate the activation of pro-apoptotic genes in previously ischemic areas. Thus, we investigated gene expression for the pro-apoptotic regulators, Bax and caspase-3, after transient focal brain ischemia, together with the p53-regulated cell cycle inhibitor, p21/WAF1/CIP1. Reversible occlusion of the middle cerebral artery for 2 h was carried out in halothane-anesthetized rats using the poly-L-lysine coated filament method. In situ hybridization was performed at 0, 1, 3, 6 h and 1, 3 and 7 d of recirculation and in sham controls. Radioactive antisense probes served for detection of bax, p21 and caspase-3 mRNAs on brain sections, and quantitative film autoradiography was combined with image-averaging techniques. Bax mRNA tended to decline after focal brain ischemia within 1 d. p21 mRNA was upregulated with a perifocal pattern at 3 h and 1 d after ischemia whereas the ischemic regions themselves failed to show significant upregulation. Caspase-3 mRNA was elevated in the resistant dorsomedial cortex at 1 d. A pro-apoptotic pattern of novel gene expression, involving Bax and caspase-3, was not observed after transient focal brain ischemia. Rather, the perifocal expression of p21 and caspase-3 mRNAs observed at 1 d after ischemia points to reactive changes in resistant brain areas.

Analysis of Variance↗
PubMed2000

Newly isolated archaerhodopsin from a strain of Chinese halobacteria and its proton pumping behavior.

A strain of extremely salt-loving halobacteria Halobacterium species xz515 from a salt lake in Tibet was isolated. SDS-polyacrylamide gel electrophoresis shows that there is only one protein on claret membrane, which is the same membrane fraction as purple membrane from Halobacterium salinarum, with a molecular weight close to bacteriorhodopsin (br). The purified retinal containing protein from xz515 has an absorption peak at around 550 nm. These facts indicate that it is a br-like protein. The partial sequence determination [H. Wang et al., Chin. Sci. Bull., 45 (2000)] shows that this br-like protein belongs to the archaerhodopsin family. The measurements of light-induced medium pH change in intact cells and cell envelope vesicles of xz515 suggest that this type of archaerhodopsin has a proton pumping function. However, the study about the dynamics of pumped protons across the membrane reveal that the proton release and proton uptake is in reverse order compared to br. The probable reason, attributing to regulating the rate of proton release is discussed.

Archaeal Proteins↗
PubMed2000

alpha-Amylase production by Bacillus subtilis with dregs in an external-loop airlift bioreactor.

An external-loop airlift bioreactor, with a low ratio 2.9 of height-to-diameter of the riser and a ratio 6.6 of riser-to-downcomer diameter, was used to produce alpha-amylase from fermentation with dregs by Bacillus subtilis. The effects of gas flow rate and liquid volume on alpha-amylase production were investigated. After a 36-h fermentation time, an average of 432.3U/ml alpha-amylase activity was obtained under the conditions of liquid volume 8.5l and gas flow rate 1.2vvm for the first 12h of fermentation, 1.4vvm from 12 to 27h, and 1.2vvm from 27h to the end. The activity was higher than that obtained in shaking flasks (409.0U/ml) and in a mechanically stirred tank bioreactor (397.2U/ml) under optimized operating conditions. The fermentation cycle of the airlift bioreactor was shorter than the 48h required for the shaking flasks and close to the 36h of the mechanically stirred tank bioreactor. It was demonstrated that the external-loop airlift bioreactor could substitute for the traditional mechanically stirred tank bioreactor to produce alpha-amylase from fermentation by Bacillus subtilis with dregs.

Journal Article↗
PubMed2000

MRZ 2/579, a novel uncompetitive N-methyl-D-aspartate antagonist, reduces infarct volume and brain swelling and improves neurological deficit after focal cerebral ischemia in rats.

The purpose of this study was to evaluate the effects of MRZ 2/579, an uncompetitive N-methyl-D-aspartate antagonist, on infarct size, extent of swelling and neurological deficit in a model of transient middle cerebral artery occlusion in rats. Physiologically controlled Sprague-Dawley rats received 2 h MCAo by retrograde insertion of an intraluminal suture coated with poly-L-lysine. The agent (MRZ 2/579) or vehicle (sodium chloride 0.9%) was administered i.v. immediately after suture removal following a 2-h period of MCAo. Two experimental groups were studied: group A was treated by vehicle (bolus infusion:1 ml/kg for 10 min followed by infusion of 6 ml/kg/h over 6 h). Group B was treated by MRZ 2/579 (bolus infusion:10 mg/kg for 10 min followed by infusion of 6 mg/kg/h over 6 h). The neurological status was evaluated during occlusion (at 60 min) and daily for 3 days after MCAo. Brains were then perfusion-fixed, and infarct volumes and brain swelling were determined. MRZ 2/579 significantly improved the neurological score compared to vehicle-treated rats at 48 h (6.2+/-0.6 and 8.7+/-0.5, respectively; P<0.004) and 72 h after MCAo (5.2+/-0.6 and 8.4+/-0.5, respectively; P<0.001). Treatment with MRZ 2/579 also significantly reduced total infarct volume (29.3+/-11.1 and 83.2+/-16.5 mm(3), respectively; P<0. 01), cortical infarct volume (24.8+/-11.2 and 70.0+/-18.0 mm(3), respectively; P<0.04) and subcortical infarction (21.2+/-4.1 and 49. 6+/-4.5 mm(3), respectively; P<0.0002). Brain swelling was also markedly reduced compared with vehicle-treated rats (4.7+/-1.3 and 10.8+/-2.1%, respectively; P<0.02). These results demonstrate that treatment with MRZ 2/579, when administered promptly after reperfusion, confers neuroprotective effects on infarct volume, brain swelling, and neurological score compared to the vehicle group.

Animals↗
PubMed2000

Role of a bacillus Calmette-Guérin fibronectin attachment protein in BCG-induced antitumor activity.

Intravesical Mycobacterium bovis bacillus Calmette-Gu*erin (BCG) is the treatment of choice for superficial bladder cancer. Previous studies showed that attachment of BCG to fibronectin within the bladder was necessary for mediation of the antitumor response. Further studies identified a bacterial receptor, fibronectin attachment protein (FAP), as an important mediator of BCG attachment to fibronectin. In vitro studies showed that a stable BCG/fibronectin interaction was dependent on FAP binding to fibronectin; however, no role for FAP in the attachment of BCG in vivo has been characterized. We now report the cloning of the M. bovis BCG FAP (FAP-B) and demonstrate an important role for FAP in the in vivo attachment of BCG to the bladder wall and in the induction of BCG-mediated antitumor activity. The predicted amino acid sequence for FAP-B shows 61% and 71% homology, respectively, with Mycobacterium avium FAP (FAP-A) and Mycobacterium leprae FAP (FAP-L). Rabbit polyclonal antibodies against Mycobacterium vaccae FAP (FAP-V) reacted with all 3 recombinant FAP proteins on Western blots. Functional studies show FAP-B to bind fibronectin via the highly conserved attachment regions previously identified for FAP-A and FAP-L and also to competitively inhibit attachment of BCG to matrix fibronectin. In vivo studies show FAP to be a necessary protein for the stable attachment of BCG to the bladder wall. Moreover, stable binding of BCG via FAP was shown to be necessary for the expression of BCG-induced antitumor activity. Our results demonstrate a biological role for FAP in the mediation of BCG-induced antitumor activity.

Adhesins, Bacterial↗
PubMed2000

Human vascular smooth muscle cells possess functional CCR5.

CC chemokine receptors are important modulators of inflammation. Although CC chemokine receptors have been found predominantly on leukocytes, recent studies have suggested that vascular smooth muscle cells respond to CC chemokines. We now report that human smooth muscle cells express CCR5, a co-receptor for human immunodeficiency virus. CCR5 mRNA was detectable by RNA blot hybridization in human aortic and coronary artery smooth muscle cells. The cDNA generated by reverse transcription-polymerase chain reaction from aortic smooth muscle cells had 100% identity throughout the entire coding region with the CCR5 cloned from THP-1 cells. By immunohistochemistry, CCR5 and the CCR5 ligand, macrophage inflammatory protein-1beta (MIP-1beta), were detected in smooth muscle cells and macrophages of the atherosclerotic plaque. In smooth muscle cell culture, MIP-1beta induced a significant increase in intracellular calcium concentrations, which was blocked by an antibody to CCR5. In addition, MIP-1beta caused a calcium-dependent increase in tissue factor activity. Tissue factor is the initiator of coagulation and is thought to play a key role in arterial thrombosis. These data suggest that human arterial smooth muscle cells express functional CCR5 receptors and MIP-1beta is an agonist for these cells.

Aorta↗
PubMed2000

Cloning and distribution of the rat parkin mRNA.

We have isolated by RT-PCR and sequenced a partial cDNA coding for the rat homolog of parkin, a gene mutated in autosomal recessive juvenile parkinsonism. The 1.46 kb rat cDNA clone contains a 1376 bp coding sequence that shares strong similarity with the human parkin cDNA. RT-PCR and in situ hybridization revealed widespread expression of parkin in the rat brain and the periphery. The availability of the rat parkin cDNA and the initial elucidation of its distribution should facilitate further research on the pathophysiological role of parkin in the nervous system.

Amino Acid Sequence↗
PubMed2000

Fructose-mediated damage to lens alpha-crystallin: prevention by pyruvate.

Post-translational modifications in lens crystallins due to glycation and oxidation have been suggested to play a significant role in the development of cataracts associated with aging and diabetes. We have previously shown that alpha-keto acids, like pyruvate, can protect the lens against oxidation. We hypothesize that they can also prevent the glycation of proteins competitively by forming a Schiff base between their free keto groups and the free -NH(2) groups of protein as well as subsequently inhibit the oxidative conversion of the initial glycation product to advanced glycation end products (AGE). The purpose of this study was to investigate these possibilities using purified crystallins. The crystallins isolated from bovine lenses were incubated with fructose in the absence and presence of pyruvate. The post-incubation mixtures were analyzed for fructose binding to the crystallins, AGE formation, and the generation of high molecular weight (HMW) proteins. In parallel experiments, the keto acid was replaced by catalase, superoxide dismutase (SOD), or diethylene triaminepentaacetic acid (DTPA). This was done to ascertain oxidative mode of pyruvate effects. Interestingly, the glycation and consequent formation of AGE from alpha-crystallin was more pronounced than from beta-, and gamma-crystallins. The changes in the crystallins brought about by incubation with fructose were prevented by pyruvate. Catalase, SOD, and DTPA were also effective. The results suggest that pyruvate prevents against fructose-mediated changes by inhibiting the initial glycation reaction as well as the conversion of the initial glycated product to AGE. Hence it is effective in early as well as late phases of the reactions associated with the formation of HMW crystallin aggregates.

Animals↗
PubMed2000

Doubly vibrationally enhanced four wave mixing: the optical analog to 2D NMR.

We report the development of the four wave mixing vibrational analog to 2D NMR and demonstrate its spectral selectivity, sensitivity to the interactions causing mode coupling, and ability to spectrally resolve isotopic mixtures. The method discriminates against uncoupled vibrational modes and isolates the features that are associated with intra- or intermolecular interactions.

Absorption↗
PubMed2000

Neuroprotection by LY341122, a novel inhibitor of lipid peroxidation, against focal ischemic brain damage in rats.

LY341122 (2-(3, 5-di-t-butyl-4-hydroxyphenyl)-4-(2-(4-methylethylaminomethyl-ph enylox y)ethyl)oxazole) is a potent inhibitor of lipid peroxidation which has been shown to protect against global ischemia and traumatic brain injury in rats. The purpose of this study was to examine the effect of LY341122 on ischemic injury in a highly reproducible model of focal cerebral ischemia in rats. Male Sprague-Dawley rats were anesthetized with halothane and subjected to 120 min of temporary middle cerebral artery occlusion by retrograde insertion of an intraluminal nylon suture coated with poly-L-lysine. The drug (LY341122, n=19) or vehicle (phosphate-buffered saline (PBS), n=10) was administered i.v. (as a 5 or 10 mg/kg bolus followed by a 5 or 10 mg/kg/h infusion for 20 h, respectively, starting 1 or 2 h after the onset of middle cerebral artery occlusion). Neurological status was evaluated during middle cerebral artery occlusion (60 min) and daily for 3 days thereafter. Three days after ischemia, brains were perfusion-fixed and infarct volumes and brain edema were determined. LY341122 significantly improved the neurological score compared to vehicle at 24, 48 and 72 h after middle cerebral artery occlusion. Treatment with LY341122 significantly reduced total infarct volume in all treated groups compared to vehicle rats. Cortical infarct volume was significantly reduced by LY341122 treatment in the 10 mg/kg (1 h) and LY341122 10 mg/kg (2 h) groups compared to vehicle rats (14.7+/-9.5 vs. 106.8+/-20.9 mm(3), and 36.9+/-20.1 vs. 106. 8+/-20.9 mm(3), respectively (mean+/-S.E.M.)). Striatal infarct volume was also significantly reduced by treatment with LY341122 in the 10 mg/kg (1 h) group compared to vehicle (23.7+/-3.4 vs. 68. 2+/-6.7 mm(3)). These results demonstrate the neuroprotective efficacy of LY341122 in focal cerebral ischemia.

Animals↗
PubMed2000

TNF-alpha induced over-expression of GFAP is associated with MAPKs.

Increased levels of tumor necrosis factor-alpha (TNF-alpha), a pluripotent cytokine that is reportedly mitogenic to astrocytes, are associated with the expression of glial fibrillary acidic protein (GFAP), the most specific marker for astrocytes, in many neuropathological conditions, including brain injury, CNS infection, Creutzfeldt-Jakob disease and Alzheimer's disease. Here, we show that treatment of cultured astrocytes with TNF-alpha resulted in dramatic over-expression of GFAP, associated with a substantial activation of the mitogen activated protein kinase (MAPK) Erk2 (extracellular signal-regulated protein kinase). We also demonstrate that TNF-alpha-induced over-expression of GFAP was significantly attenuated by the MAPK inhibitor PD98059. We conclude that TNF-alpha may upregulate GFAP through the MAPK signaling pathway. Because increased GFAP is a hallmark of reactive gliosis, understanding the mechanisms that regulate GFAP expression may facilitate development of strategies to minimize the gliosis associated with many brain diseases.

Animals↗