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Biomedical subjects

W Zhao

Publications and source records attributed to W Zhao.

At least 307 records · Page 17Linked to original sources
PubMed1996

Experimental allergic encephalomyelitis in the rat is inhibited by aminoguanidine, an inhibitor of nitric oxide synthase.

This study assessed the role of de novo nitric oxide (NO) production in the pathogenesis of experimental allergic encephalomyelitis (EAE) by using aminoguanidine (AG), an inhibitor of nitric oxide synthase (NOS), which preferentially inhibits the cytokine- and endotoxin-inducible isoform of NOS versus the constitutive isoforms consisting of endothelial and neuronal NOS. The maximum clinical severity of EAE and the duration of illness were significantly reduced or totally inhibited by twice daily subcutaneous injection of 100 mg/kg body weight AG. Histochemical staining for NADPH diaphorase, which detects enzymatic activity of NOS, revealed positive reactivity in untreated EAE rats both in parenchymal blood vessel walls and in anterior horn cell neurons, while normal rats and rats with EAE treated with AG showed predominantly the neuronal positivity. Moreover, this NADPH staining pattern was further supported by the immunohistochemical findings that endothelial NOS (eNOS) expression was increased in blood vessels in the inflamed lesions of untreated EAE rats and that inducible NOS (iNOS) was detected in some inflammatory cells, while treatment with AG could significantly reduce both iNOS and eNOS production. These results suggest that: (i) both iNOS and eNOS are upregulated in inflamed areas of the rat central nervous system in EAE; (ii) increased NO production plays a role in the development of clinical signs in EAE; and (iii) selective inhibitors of iNOS and/or eNOS may have therapeutic potential for the treatment of certain autoimmune diseases.

Animals↗
PubMed1996

L-arginine does not improve cortical perfusion or histopathological outcome in spontaneously hypertensive rats subjected to distal middle cerebral artery photothrombotic occlusion.

The potential of nitric oxide (NO) to influence positively or negatively the outcome of mechanically induced focal cerebral ischemia is still controversial. Recent evidence suggests that NO of vascular origin, whether synthesized from exogenously administered L-arginine (L-Arg) or from NO donor compounds, is beneficial but that of neuronal origin is not. However, the therapeutic potential of NO to ameliorate stroke induced by arterial thrombosis has not been reported. We assessed the therapeutic effect of L-Arg administration in spontaneously hypertensive rats (SHR) subjected to permanent photothrombotic occlusion of the distal middle cerebral artery (dMCA). The ipsilateral carotid artery was left unligated to enhance L-Arg delivery into the putative penumbral region. Local CBF (LCBF) was assessed at 30 min by the [14C]iodoantipyrine technique (n = 9), while histological infarct volumes and index of peripheral ischemic cell change were determined at 3 days (n = 7). Rats (n = 9) given 300 mg/kg L-Arg at 18 and 3 h before photothrombotic dMCA occlusion and at 5 min afterward displayed no significant differences in LCBF compared with animals (n = 8) injected with water (the carrier vehicle) and similarly irradiated. Infarct volumes were also similar, being 37.0 +/- 9.7 mm3 (SD) in the vehicle-treated and 49.1 +/- 17.2 mm3 (SD) in the L-Arg-treated groups (both n = 7), as were assessments of ischemic neuronal density in the penumbra. In contrast, L-Arg administered intravenously in a dose of 300 mg/kg to nonischemic SHR (n = 5) increased cortical CBF by approximately 75% during a 70-min observation period. We conclude that thrombotic processes superimposed upon cerebral ischemia may facilitate tissue reactions that offset the potentially beneficial effect of L-Arg, and this caveat must be considered when proposing L-Arg for clinical treatment of focal thrombotic stroke.

Animals↗
PubMed1996

Hypericin-induced phototoxicity in cultured fibroblasts and swine erythrocytes.

Hypericin is a naturally occurring photosensitizer, whose presence in plants has been responsible for cutaneous phototoxicity in grazing animals. The photosensitizing properties of this agent have recently been exploited in models for anti-tumor and anti-viral activity. The cytotoxicity of hypericin and light was assessed in 3T3 mouse fibroblasts using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)] assay and the lactate dehydrogenase (LDH) leakage assay. Membrane damage was assessed in swine erythrocytes using hemolysis, potassium (K+) leakage and formation of lipid hydroperoxides. Concentration- and light-dependent decreases in fibroblast viability were seen starting at hypericin concentrations of 1.25 microM and light power flux levels of 24 J/cm2 using a visible light source and at 0.417 microM hypericin and a similar light dose using a solar simulator. No LDH leakage was observed at hypericin concentrations up to 30 microM and visible light up to 144 J/cm2. Light-and/or concentration-dependent increases in hemolysis, K+ leakage and formation of lipid hydroperoxides in red blood cell (RBC) membranes were observed, but at concentrations and light doses much greater than those required to induce cytotoxicity in fibroblasts. Lipid peroxidation and hemolysis occurred at 15 microM hypericin and 24 J/cm2 (visible light source). Potassium ion leakage occurred at concentrations and light levels as low as 5 microM and 12 J/cm2 or 15 microM and 4.8 J/cm2 (visible light source) but was still a less sensitive indicator than fibroblast cytotoxicity. Evidence for both type I and type II reactions was shown in RBC membranes by TLC analysis of cholesterol products. In the absence of light, hypericin appears to be relatively nontoxic in the models tested.

3T3 Cells↗
PubMed1996

Evidence for independent Cl- and HCO3- secretion and involvement of an apical Na(+)-HCO3- cotransporter in cultured rat epididymal epithelia.

Electrogenic chloride and bicarbonate secretion by cultured rat epididymal epithelia was studied using the short-circuit current (ISC) technique. When incubated in normal solution, 8-(4-chlorophenylthio)-adenosine 3',5'-cyclic monophosphate (cpt-cAMP) caused a rise in the ISC, which was attributable to Cl- and HCO3- secretion. Cl- secretion was found to contribute to the initial transient phase, whereas HCO3- secretion contributed to the sustained phase of the response. HCO3- secretion involves a basolaterally placed Na(+)-H+ exchanger and apical anion channel, most probably the cystic fibrosis transmembrane conductance regulator (CFTR). There is also evidence that an apical electrogenic Na(+)-HCO3- cotransporter is involved in HCO3- exit. CFTR accounted for 70% of HCO3- secretion, while the Na(+)-HCO3- cotransporter accounted for 30%. The possibility that the cotransporter may serve as an alternative pathway for HCO3- secretion in cystic fibrosis is discussed.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1996

Complex alternative RNA processing generates an unexpected diversity of poly(A) polymerase isoforms.

Multiple forms of poly(A) polymerase (PAPs I, II, and III) cDNA have previously been isolated from bovine, human, and/or frog cDNA libraries. PAPs I and II are long forms of the enzyme that contain four functional domains: an apparent ribonucleoprotein-type RNA-binding domain, a catalytic region that may be related to the polymerase module, two nuclear localization signals (NLSs I and 2), and a C-terminal Ser/Thr-rich region. PAP III would encode a truncated protein that lacks the NLSs and the S/T-rich region. To investigate further the structure and expression of these forms, we isolated the mouse PAP gene and an intronless pseudogene from a mouse liver genomic library. The structure of the gene indicates that different forms of PAP are produced by alternative splicing (PAPs I and II) or by competition between polyadenylation and splicing (PAP III). The pseudogene appears to reflect yet another form of long PAP, which we call PAP IV. Mouse PAP III and two additional truncated forms, PAPs V and VI, which would be produced by use of poly(A) sites in adjacent introns, were also isolated from a mouse brain cDNA library. RNase protection and reverse transcription-PCR analyses showed that PAP II, V, and VI are expressed in all tissues tested but that PAP I and/or IV and III are tissue specific. However, immunoblot analysis detected only the long forms, raising the possibility that the short-form RNAs are not translated. Purified recombinant baculovirus-expressed PAPs were tested in several in vitro assays, and the short forms were found to be inactive. We discuss the possible significance of this complex expression pattern.

Alternative Splicing↗
PubMed1996

Depiction of infarct frequency distribution by computer-assisted image mapping in rat brains with middle cerebral artery occlusion. Comparison of photothrombotic and intraluminal suture models.

BACKGROUND AND PURPOSE: Histopathologic analysis of experimental brain damage has traditionally been performed by measuring areas of infarction and/or selective neuronal alterations on a section-by-section basis in individual animals. For series containing multiple replicate animals, quantitation of tissue injury is typically performed at similar coronal levels throughout an experimental group. A means of facilitating pictorial group comparisons of these histopathologic alterations between different series of replicate studies is highly desirable. METHODS: We introduce a newly designed approach to achieve this goal, based on a linear affine transformation that is used to map corresponding sections at the same anatomic level into a common template to yield a frequency distribution map depicting the aggregate data set. We have applied this approach to compare the histopathologic features of two models of middle cerebral artery (MCA) occlusion in rats: (1) photothrombotically induced permanent distal MCA occlusion in spontaneously hypertensive rats (SHR) and (2) temporary MCA occlusion by intraluminal suture in Wistar rats. RESULTS: The brains of SHR rats with permanent distal MCA occlusion showed a high frequency of infarction involving the dorsolateral and lateral portions of the ipsilateral neocortex, whereas Wistar rats with 90-minute MCA suture occlusion showed a zone of infarction largely concentrated in the dorsolateral portion of the ipsilateral caudoputamen. Infarct frequency distributions for the two animal groups were compared statistically at three corresponding anatomic levels by Fisher's exact test; the resulting statistical parametric maps are shown. CONCLUSIONS: With the use of frequency distribution maps, the pattern of trends within a group can be observed coronally or three-dimensionally. One can directly access data as to numbers of rats with infarction for any point on the map. Studies performed under different experimental conditions can also be compared with one another by means of the generated data sets.

Animals↗
PubMed1996

Middle cerebral artery occlusion in the rat by intraluminal suture. Neurological and pathological evaluation of an improved model.

BACKGROUND AND PURPOSE: The purpose of the present study was to evaluate a modified method of intraluminal suture occlusion of the middle cerebral artery (MCA) on the volume of brain infarction and on neurobehavioral function in rats subjected to a temporary focal ischemic insult. METHODS: Male Sprague-Dawley rats were anesthetized with halothane and subjected to 60 minutes or 2 hours of temporary MCA occlusion (MCAo) by an intraluminal thread. In one group of rats, the suture was coated with poly-L-lysine, while in a second group, a conventional uncoated suture was used. Behavioral function was evaluated at 50 to 60 minutes after occlusion and during a 3-day period after MCAo. Three days after MCAo brains were perfusion-fixed and infarct volumes were measured. RESULTS: In rats with 60-minute MCAo, only 3 of 7 animals with uncoated sutures had infarcts, whereas in the group with poly-L-lysine-coated sutures, all rats (n = 7) exhibited infarction (P = .009, Fisher's exact test). With 2 hours of MCAo, total infarct volume (corrected for brain edema) was significantly larger in rats with poly-L-lysine-coated sutures than in the group with uncoated sutures (mean +/- SEM, 122.1 +/- 4.8 versus 67.0 +/- 18.2 mm3, respectively; P = .03; n = 4 in each group). In the 2-hour MCAo study, infarct volumes in the uncoated-suture group tended to be variable and inconsistent (coefficient of variation, 54%) compared with the group in which sutures were coated with poly-L-lysine, in which a highly consistent infarct was produced (coefficient of variation of infarct volume, 8%). CONCLUSIONS: Reversible MCAo in which a poly-L-lysine-coated intraluminal suture was used proved to be a reliable and effective modification of this technique, yielding consistently larger infarcts and greatly reduced interanimal variability.

Animals↗
PubMed1996

Localization and distribution of angiotensin II in the rat epididymis.

The localization and distribution of angiotensin II (Ang II) in the rat epididymis was studied using immunohistochemical and RIA techniques. The immunohistochemical results showed that Ang II-like immunoreactivity progressively increased along the length of the rat epididymis (cauda > corpus >> caput) and was predominately localized in the basal region of the epididymal epithelium. Occasionally, immunostaining of lighter intensity was also found in the apical region. The concentration of Ang II in cultured rat cauda epididymal epithelial cells was further measured by RIA. In addition to that found in cultured epithelial cells, Ang II activity was also detected in the culture medium, suggesting a secretory role of the epithelium. These findings suggest that Ang II could be derived locally from epididymal epithelium and that it could play a role in local regulation of epithelial transport and, possibly, in the maintenance of sperm function as well, by exerting its paracrine and/or autocrine effect in various regions of the epididymis.

Angiotensin II↗
PubMed1996

[Anti-embryonation immunity and cultivation of Schistosoma japonicum eggs in vitro].

OBJECTIVE: To study anti-SIEA super-immune sera against the influence on schistosoma japonicum in vitro cultivation of newly laid eggs by adult worms, their hatches and development. METHODS: The adult worm pairs of Schistosoma japonicum were collected from test animals, portae vena which infected with the worms for 38-45 days. The worm pairs were injected divided into control group with normal rabbit sera and experimental group with anti-SIEA super-immune sera. The output of newly laid eggs and the embryonic development were observed with immunofluorescence staining. RESULTS: Female worms cultured in vitro with normal rabbit serum not only discharged eggs already contained in its uterus, but also produced newly formed ones. They however, did not occur in female worms cultured with anti-SIEA super-immune sera and the output of eggs by the latter was much lower than that of the former. There was also a marked decrease in the proportion of developed eggs in the experimental group compared with that in the control group. In addition, immunofluorescence staining of immature eggs and female worms by anti-SIEA super-immune sera showed that a lightened fluorescence was not only seen in the embryonic cells of immature eggs, but also in the vitelaria and in the internal membrane tissue of gut cavity of the female worms. CONCLUSION: SIEA was possibly involved in the immunity against egg development and fecundity of schistosoma japonicum.

Animals↗
PubMed1996

Effects of alveolar macrophage conditioned media from interstitial lung disease patients on the procollagen mRNA expression in human lung fibroblasts.

Progressive inflammation and fibrosis are the central processes in the pathogenesis of pulmonary fibrosis. It is believed that macrophages in areas of chronically inflamed lung play a key role in fibrotic response. Therefore, we investigated the effects of alveolar macrophage (Am phi) conditioned media from interstitial lung disease (ILD) patients on lung fibroblast proliferation and procollagen mRNA expression. After stimulating with Am phi conditioned media from ILD patients, the fibroblast proliferation increased 71.4% compared with the control, but for media from bronchial carcinoma (BC) patients, it just increased 14.3%. There is a significant difference between the two groups (P < 0.05). The procollagen alpha, (I) mRNA in fibroblasts stimulated with Am phi conditioned media from ILD patients was increased 21.3%, and alpha 1 (III) was 37.2% higher than control (P < 0.05). It increased 6.8% and 12.8% for media from BC patients respectively, but there was no difference when compared to the control. We considered that Am phi from ILD patients might be in an activated state and could release some growth factors to stimulate fibroblast proliferation and promote collagen DNA expression.

Carcinoma, Bronchogenic↗
PubMed1996

[Factors influencing long-term survival in patients with nonoperable lung cancer: an analysis by Cox model].

This paper reports a prospective survey of 173 patients with nonoperable lung cancer between January. 1, 1983 to March. 1, 1985. The follow-up rate was 97.7% over five years. Fourteen factors including sex, age, course of disease before treatment, clinical stage, performance status, size of mass, metastatic status, hemoglobin before treatment, short-term response to treatment and so on which might influence long term survival were studied by univariate analysis (Kruskal-Wallis test for Kaplan-Meier survival curve) and by multivariate analysis (Cox's proportional hazad model and audio-visual chart test for goodness of fit). Multivariate analysis using Cox's model revealed 6 significant prognostic factors: performance status, short-term response to treatment, clinical stage, hemoglobin before treatment, smoking index and method of treatment. The survival prediction equation was chi 2 = 72.14, nu = 6, P < 0.0001. The results indicate that the performance status and the CR rate of the initial treatment, among other things, is the major factors affecting prognosis.

Adenocarcinoma↗
PubMed1995

Protein kinase C regulates pleckstrin by phosphorylation of sites adjacent to the N-terminal pleckstrin homology domain.

Pleckstrin is a substrate for protein kinase C in activated platelets that contains at its N and C termini two of the pleckstrin homology (PH) domains that have been proposed to mediate protein-protein and protein-lipid interactions. We have recently shown that pleckstrin can inhibit agonist-induced phosphoinositide hydrolysis and that this inhibition requires an intact N-terminal PH domain (residues 6 to 99). In the present studies, we have identified the sites of phosphorylation in pleckstrin and examined their contribution to pleckstrin function. In human platelets activated with thrombin or phorbol esters, and in COS-1 cells expressing pleckstrin, a combination of phosphopeptide analysis and site-directed mutagenesis shows that three residues in the intervening sequence between the two pleckstrin PH domains become phosphorylated: Ser113, Thr114, and Ser117. Replacing all three of these sites with glycine decreased phosphorylation by > 90% and reduced pleckstrin's ability to inhibit phosphoinositide hydrolysis by as much as 80%. Replacing the phosphorylation sites with alanine residues had a similar effect, while substitution with aspartate, glutamate, or lysine residues produced pleckstrin variants that were fully active even in the absence of phosphorylation. These results suggest that phosphorylation enhances pleckstrin's activity by introducing a cluster of charges into a region adjacent to, but not within, the N-terminal PH domain. This may have an allosteric effect on the N-terminal PH domain, regulating its interaction with other molecules necessary for the inhibition of phosphoinositide hydrolysis.

Amino Acid Sequence↗
PubMed1995

Phosphatidylinositol (3,4,5)-trisphosphate stimulates phosphorylation of pleckstrin in human platelets.

We have reported that platelets exposed to thrombin or thrombin receptor-directed ligand activate phospholipase C and rapidly accumulate phosphatidylinositol (3,4,5)-trisphosphate (PtdIns(3,4,5)P3) and phosphatidylinositol (3,4)-bisphosphate (PtdIns(3,4)P2) as a function of the activation of phosphoinositide (PI) 3-kinases in a GTP-binding protein-dependent manner. In such platelets, serine- and threonine-directed phosphorylation of pleckstrin also occurs and has been attributed to protein kinase C activation. We now report that the phosphorylation of pleckstrin is partially dependent upon PI 3-kinase. Pleckstrin phosphorylation in response to thrombin receptor stimulation is progressively susceptible to inhibition by wortmannin, a potent and specific inhibitor of platelet PI 3-kinases. PI 3-kinase thus seems to play a gradually increasing role in promoting pleckstrin phosphorylation. The IC50 for wortmannin in inhibiting SFLLRN-stimulated 3-phosphorylated phosphoinositide accumulation is 10 nM, and that (i.e. 50% of maximum inhibition) for inhibiting pleckstrin phosphorylation is 15 nM. Synthetic PtdIns(3,4,5)P3, when added to saponin-permeabilized (but not intact) platelets, causes wortmannin-insensitive phosphorylation of pleckstrin. PtdIns(3,4,5)P3 also overcomes the inhibition by wortmannin of thrombin- or guanosine 5'-3-O-(thio)trisphosphate-stimulated pleckstrin phosphorylation. In contrast, PtdIns(4,5)P2 or inositol (1,3,4,5)-tetrakisphosphate are ineffective in these respects. The pattern of phosphorylation of pleckstrin activated by PtdIns(3,4,5)P3 is not distinguishable from that of pleckstrin phosphorylated in intact platelets exposed to protein kinase C-activating beta-phorbol myristate acetate, mimicking diacylglycerol. Activation of protein kinase(s) by PtdIns(3,4,5)P3 thus offers a route for pleckstrin phosphorylation in vivo that is an alternative to activation of phospholipase C-->diacylglycerol-->protein kinase C.

Androstadienes↗
PubMed1995

Local hemodynamic changes during transient middle cerebral artery occlusion and recirculation in the rat: a [14C]iodoantipyrine autoradiographic study.

We evaluated acute alterations of local cerebral perfusion following 30 min of transient right proximal middle cerebral artery (MCA) clip-occlusion in the rat and following two intervals of postischemic reperfusion. Local cerebral blood flow (lCBF) was assessed by [14C]iodoantipyrine autoradiography. Brain temperature was controlled at 35.5-36.5 degrees C throughout the experiment. We measured lCBF in four groups of rats: (a) sham-operated controls (n = 5), (b) following 30 min MCA occlusion (n = 5), (c) following 30 min of MCA occlusion with 15-min reperfusion (n = 6) and (d) following 30 min of MCA with 120-min reperfusion (n = 6). lCBF was measured in seven regions of the ischemic and non-ischemic hemispheres. MCA occlusion induced an ipsilateral reduction of lCBF, which was most severe in the parietal cortex (8.4 +/- 4.0% of control, mean +/- S.D.), and dorsolateral caudoputamen (20.0 +/- 13.4% of control). lCBF in the non-ischemic hemisphere and in ipsilateral regions lying outside the MCA territory also decreased significantly. lCBF recovery was incomplete when assessed following only 15 min of reperfusion. Reperfusion of 120 min led to return of cortical CBF to control levels, but lCBF in the caudoputamen remained depressed (50-55% of control values). Caudoputaminal CBF and cortical CBF values were highly correlated with one another under normal and ischemic conditions, but this correlation was disrupted following reperfusion. On the basis of these results, we speculate that, if a means were found to enhance the early recovery of lCBF following transient ischemia, this might expand the therapeutic window of opportunity for the institution of other neuroprotective strategies.

Acute Disease↗