Biomedical subjects

H Zhao

Publications and source records attributed to H Zhao.

At least 379 records · Page 21Linked to original sources

PubMed1998

[Study on the contents of HBV DNA in sera of patients with hepatic diseases by QPCR technique].

In order to test the content of serum HBV DNA for evaluating the effectiveness of clinical antiviral treatment and the relation of serum HBV DNA level to state of illness and prognosis, a QPCR method of energy transfer in amplification by semi-nested fluorescein labeled primer was established. Serum HBV DNA contents of 63 cases of hepatic diseases with abnormal liver function were detected by QPCR, PCR and ELISA. The positive detectable rate of the three methods were compared. The results showed that the positive detectable rate of QPCR was 82.54%, PCR 71.43% and ELISA 63.50%. The detectable rate of QPCR was compared with PCR, and uniformity rate of the both methods was 88.89%, P < 0.05; the positive results of QPCR were compared with ELISA, and their uniformity rate was 80.95%, P < 0.05; when serum average content (mean +/- s) of HBV DNA by QPCR method was compared with serum average titer of HBV by ELISA method, serum HBV-DNA average content of the group in which both QPCR and ELISA were positive was much higher than that of the group of QPCR(+) and ELISA(-), P < 0.05; serum QPCR-HBV-DNA of 30 cases of healthy persons were all negative. The results indicated that QPCR method is of highest positive detectable rate and more specific in detecting serum HBV DNA and had no nonspecific amplifying reaction. It has the superiority of detecting low level serum HBV DNA.

Adult↗

PubMed1998

[The construction of poliovirus chimera and the analysis of its antigenic structure].

The construction of poliovirus chimera is an useful way to investigate the antigenic structure conformation of poliovirus. In this research, the poliovirus chimera containing a piece of antigenic fragment of hepatitis A virus was constructed for analysis of poliovirus antigenic structure conformation. Depending on the characterization of this chimera, the conformation of poliovirus neutralization antigenic site I in VP1 was analyzed, and that the possible influence of hepatitis A virus antigenic fragment inserted to the structure conformation of poliovirus antigen was also investigated.

Animals↗

PubMed1998

[FTIR microscope study on the change of membrane protein and lipids of erythrocyte damaged by free radicals].

The FTIR Microscope and Computer Aided Analysis were used to study not only the change of secondary structure of erythrocyte membrane protein attacked by the free radicals, but also the effect of oxygen centered free radicals on the membrane lipids in this paper. The results indicate that the content of alpha-helix in the secondary structure of protein clearly changed because of the attack of the free radicals. Damaged by free radicals in 30 minutes, the protein structure could not be restored after 3 hours. The number of the P=O (vp=0), C=O and C=C in the lipids of the membrane was changed. These changes were caused by the peroxidation of the oxygen centered free radicals.

Adult↗

PubMed1998

Experimental study on the scavenging effects of ginsenosides on oxygen free radicals using model of heterotopic heart transplantation in rats.

Oxygen-derived free radicals play an important role in myocardial injury associated ischemia and reperfusion. To investigate whether ginsenosides, as an additive agent of cardioplegic solution, can decrease toxicity of oxygen free radicals in myocardial injury, a heterotopic heart transplantation model in Wistar rats was employed. St. Thomas II cold cardioplegia containing ginsenosides 80 mg/l was used in the experimental group. St. Thomas II cold cardioplegia alone was used in the control group. After global ischemia for 60 minutes and reperfusion for 30 minutes of a transplanted heart, SOD activity in the myocardium treated with ginsenosides was significantly higher than that in the control group (N=10, p< 0.01), whereas the MDA in the myocardium treated with ginsenosides were markedly lower than that of the control group (p< 0.01). The amounts of oxygen free radicals in the myocardium treated with ginsenosides were significantly lower than that of the control group (p <0.001). This study demonstrates that ginsenosides, as a proper additive agent of cardioplegic solution, can decrease toxicity of oxygen free radicals, suggesting one of the mechanisms for its protective effects against myocardial ischemia and reperfusion injury.

Animals↗

PubMed1998

Haemorheological consequences of hyperglycaemic spike in healthy volunteers and insulin-dependent diabetics.

The present study has been designed to examine the role of a hyperglycaemic spike of short duration as a factor possibly involved in haemorheological microcirculatory and (or) haemostatic dysfunctions in ten insulin-dependent diabetes mellitus patients (IDDM) and five healthy volunteers. The hyperglycaemic spike was induced by glucose infusion via GCIIS Biostator. Blood viscosity, erythrocyte aggregation, erythrocyte deformability, plasma viscosity and proteins' levels, cell counts, and transcutaneous oxygen pressure were investigated during normoglycaemic period and at the end of a 1 h hyperglycaemic spike. Hyperglycaemia induced in IDDM patients significant decreases in erythrocyte aggregation, in blood and plasma viscosities and in both fibrinogen and albumin levels. The number of platelets was significantly decreased as a result of the hyperglycaemic spike. There was no significant change induced by the hyperglycaemic spike in healthy volunteers. These findings show that a hyperglycaemic spike of 280 mg/dl (15.6 mmol/l) of short duration in IDDM patients causes numerous significant changes in both blood components and rheological behaviour. These changes were accompanied by a significant decrease in transcutaneous oxygen tension indicating an impaired tissue oxygenation during the hyperglycaemia.

Adult↗

PubMed1997

Immunochemical evidence for increased formation of advanced glycation end products and inhibition by aminoguanidine in diabetic rat lenses.

Water-soluble and urea-soluble protein fractions from control and streptozotocin-diabetic rats were analyzed for AGEs with a CML-specific monoclonal anti-AGE antibody and a polyclonal anti-AGE antibody. AGEs, CML in particular, were significantly increased in the diabetic rats whereas aminoguandine treatment resulted in significant decrease in AGEs. The data also confirm that CML, a glycoxidation product, is a major epitope of AGE structures in lenses.

Animals↗

PubMed1997

Direct projections from the lumbosacral spinal cord to Barrington's nucleus in the rat: a special reference to micturition reflex.

In the present study, direct projections from the lumbosacral cord to Barrington's nucleus in the rat were investigated by using retrograde and anterograde tracing techniques. After injection of cholera toxin B subunit (CTb) into Barrington's nucleus, a number of moderately CTb-labeled neurons were observed in the lumbosacral cord, with a slight ipsilateral dominance; most were located in the spinal parasympathetic and dorsal commissural nuclei of the lumbosacral cord. In addition, some retrogradely labeled neurons were found in the periaqueductal gray (PAG). These findings were confirmed by an anterograde labeling experiment. After biotinylated dextran amine (BDA) was injected into the lumbosacral cord, dense BDA-labeled axon terminals were found in Barrington's nucleus as well as in the PAG. Injection of BDA into the PAG resulted in many BDA-labeled terminals in Barrington's nucleus. The present results provided clear evidence for a direct projection from the spinal parasympathetic and dorsal commissural nuclei to Barrington's nucleus that could subserve conveying bladder-filling information from the lumbosacral cord to Barrington's nucleus in the micturition reflex of the rat.

Animals↗

PubMed1997

OlP-1, a novel protein that distinguishes early oligodendrocyte precursors.

Oligodendrocyte development may be divided into three distinct stages: I) commitment of neuroectoderm cells to the oligodendrocyte lineage, II) migration of precursors into the surrounding parenchyma concomitant with increased proliferation, and III) cessation of migration and proliferation and initiation of myelination. Stage II of development has remained enigmatic because of the paucity of known molecules that distinguish these immature migratory cells. We describe a novel surface protein, termed OlP-1, which is restricted in expression to this developmental stage in the mouse. Cytofluorographic comparisons with known developmental markers showed OlP-1 to be expressed primarily by stage II precursors in vitro. Histologic analyses supported this conclusion by showing co-localization of OlP-1 with stage II molecules in vivo. Two conclusions were drawn from these results. First, OlP-1 was a novel protein expressed by murine oligodendrocyte precursors at a point in development that suggested a role in migration or proliferation. Second, dispersal of OlP-1-positive cells throughout the developing brain did not correlate with the location of myelination which, observed days later, progressed in a caudal to rostral manner. These data supported the concept that the final steps of maturation and myelin gene expression may be dependent upon extrinsic factors located predominantly within white matter tracts.

Animals↗

PubMed1997

High levels of expression and nuclear localization of interleukin-1 beta converting enzyme (ICE) and CPP32 in favorable human neuroblastomas.

Neuroblastomas frequently show spontaneous regression and differentiation, which may at least partly be regulated by signaling through nerve growth factor and its receptors, TRK-A and p75LNTR. We studied 52 neuroblastic tumors to test whether the cell death-related proteases, interleukin-1 beta converting enzyme (ICE), CPP32, and Ich-1, were involved in the regression of the tumors. High levels of expression of ICE and CPP32 were significantly correlated with a high level of TRK-A expression, single copy of N-myc, younger age, lower stages, and better prognosis. The immunohistochemical studies and Western analyses as well as the terminal dUTP-biotin nick end labeling (TUNEL) method revealed that both ICE and CPP32 were translocated from the cytoplasm into the nuclei in regressing, apoptotic tumor cells. Our results suggest that ICE and CPP32 cysteine proteases may play an important role in regulating the apoptotic process of the favorable neuroblastomas.

Age Factors↗

PubMed1997

T cell receptor-induced phosphorylation of Sos requires activity of CD45, Lck, and protein kinase C, but not ERK.

Stimulation of the T cell antigen receptor (TCR) activates signaling pathways involving protein kinases, phospholipase Cgamma1, and Ras. How these second messengers interact to initiate distal activation events is an area of intense scrutiny. In this report, we confirm that TCR ligation results in phosphorylation of Sos, a guanine nucleotide exchange factor for Ras. This requires expression of both the CD45 tyrosine phosphatase and the Lck protein tyrosine kinase and depends upon signaling via protein kinase C. In contrast to previous studies examining requirements for Sos phosphorylation following insulin and epidermal growth factor receptor engagement, we show that TCR-induced phosphorylation of Sos does not require activation of the mitogen-activated protein kinase/extracellular-signal regulated kinase (MEK/ERK) pathway. However, the basal phosphorylation of Sos in T cells is affected by either MEK or MEK-dependent kinases. Although Sos phosphorylation results in its dissociation from Grb2 following insulin stimulation in Chinese hamster ovary cells, TCR engagement on the Jurkat T cell line fails to elicit a similar effect. These data demonstrate that the kinases responsible for Sos phosphorylation differ following ligation of various cell surface receptors and that the consequences of Sos phosphorylation relies, at least in part, on sites of its phosphorylation.

Adaptor Proteins, Signal Transducing↗

PubMed1997

Functional and nonfunctional mutations distinguished by random recombination of homologous genes.

We describe a convenient method for distinguishing functional from nonfunctional or deleterious mutations in homologous genes. High fidelity in vitro gene recombination ("DNA shuffling") coupled with sequence analysis of a small sampling of the shuffled library exhibiting the evolved behavior allows identification of those mutations responsible for the behavior in a background of neutral and deleterious mutations. Functional mutations are expected to occur in 100% of the sequenced screened sample; neutral mutations are found in 50% on average, and deleterious mutations do not appear at all. When used to analyze 10 mutations in a laboratory-evolved gene encoding a thermostable subtilisin E, this method rapidly identified the two responsible for the observed protease thermostability; the remaining eight were neutral with respect to thermostability, within the precision of the screening assay. A similar approach, coupled with selection for growth and survival of the host organism, could be used to distinguish adaptive from neutral mutations.

Calorimetry, Differential Scanning↗

PubMed1997

Adjuvant chemotherapy for the treatment of intracranial ependymoma of childhood.

BACKGROUND: Current treatment for childhood intracranial ependymomas with surgery and radiation therapy (RT) yields 5-year survival rates ranging from 50-70% after complete resection to 0-30% after incomplete surgical resection. The role of chemotherapy in the treatment of ependymoma has not been established. In this pilot study, children with newly diagnosed intracranial ependymoma were treated with RT and chemotherapy using agents comparable to those found to be active in the treatment of intracranial ependymoma in infants. METHODS: Nineteen children age 3-14 years (median, 7.5 years) were treated with postoperative RT and chemotherapy. Chemotherapy was comprised of carboplatin, 560 mg/m2, with vincristine, 1.5 mg/m2, weekly for 3 weeks, alternating at 4-week intervals with ifosfamide, 1.8 g/m2, and etoposide, 100 mg/m2, for 5 consecutive days for a total of 4 cycles. RESULTS: The 5-year progression free survival (PFS) estimate was 74%. The extent of surgical resection was not a significant prognostic factor in this study. By contrast, ependymomas located in the posterior fossa were associated with a higher rate of progression (P = 0.036). Toxicity, limited predominantly to myelosuppression, was manageable. CONCLUSIONS: The PFS for children with postoperative residual ependymoma treated with RT and chemotherapy in this study was higher than published survival results for RT alone. These results suggest a role for multialkylator chemotherapy in incompletely resected intracranial ependymoma and provide the rationale for a randomized trial comparing this strategy with conventional postoperative RT.

Adolescent↗

PubMed1997

The 25-kDa soluble CD23 activates type III constitutive nitric oxide-synthase activity via CD11b and CD11c expressed by human monocytes.

CD23, a low-affinity receptor for IgE, was recently shown to bind to CD11b and CD11c molecules on human monocytes. The 25-kDa soluble fragment of CD23 (sCD23), was tested for its capacity to elicit various signaling pathways in human monocytes. sCD23 was found to trigger an early increase in cGMP accumulation, through the generation of nitric oxide. This was a result of activating the L-arginine pathway, since the sCD23-mediated effect was inhibited in the presence of substituted nonmetabolizable L-arginine analogues. In addition, the increase in cGMP was suppressed by calcium chelators and inhibitors of the calcium/calmodulin complex, suggesting the involvement of a constitutive, calcium-dependent nitric oxide synthase (NOS). Indeed, the presence of an endothelial constitutive type III NOS mRNA was detected in nonactivated human monocytes, and the corresponding protein has been detected by flow cytometry. Moreover, sCD23 was shown to induce a calcium influx in monocytes, in accordance with an activation of a constitutive NOS through a transient increase in [Ca2+]i. As expected, these events were mimicked by mAbs against CD11b and CD11c, the macrophage receptors for CD23. In addition to these early events, sCD23 and the agonistic anti-CD11b and CD11c mAbs, which all trigger the release of proinflammatory mediators such as TNF-alpha, were shown to act through an NO-dependent process.

Cell Line↗

PubMed1997

Real-time monitoring of the effects of normothermia and hypothermia on extracellular glutamate re-uptake in the rat following global brain ischemia.

Brain hypothermia during ischemia may have a neuroprotective effect on pathological and functional outcomes in vivo. Although a microdialysis study demonstrated that hypothermia decreases glutamate release into the extracellular space, the issue of whether this suppression of the glutamate elevation normally accompanying ischemia is attributable to inhibition of intra-ischemic release or acceleration of post-ischemic re-uptake was not addressed. Recently, we established a real-time method for monitoring glutamate levels in extracellular space, utilizing a dialysis electrode. This method allows detailed analysis of the in vivo dynamics of biphasic glutamate elevation in the extracellular space during the intra-ischemic period and post-ischemic re-uptake. The present results show that post-ischemic hypothermia has little effect on the initial glutamate release, but remarkably enhances post-ischemic glutamate re-uptake.

Animals↗

PubMed1997

Arylamide inhibitors of HIV-1 integrase.

Based on data derived from a large number of HIV-1 integrase inhibitors, similar structural features can be observed, which consist of two aryl units separated by a central linker. For many inhibitors fitting this pattern, at least one aryl ring also requires ortho bis-hydroxylation for significant inhibitory potency. The ability of such catechol species to undergo in situ oxidation to reactive quinones presents one potential limitation to their utility. In an effort to address this problem, a series of inhibitors were prepared which did not contain ortho bishydroxyls. None of these analogues exhibited significant inhibition. Therefore an alternate approach was taken, whose aim was to increase potency rather than eliminate catechol substructures. In this latter study, naphthyl nuclei were utilized as aryl components, since a previous report had indicated that fused bicyclic rings may afford higher affinity relative to monocyclic phenyl-based systems. In preliminary work with monomeric units, it was found that the 6,7-dihydroxy-2-naphthoic acid (17) (IC50 = 4.7 microM) was approximately 10-fold more potent than its 5,6-dihydroxy isomer 19 (IC50 = 62.4 microM). Of particular note was the dramatic difference in potency between free acid 17 and its methyl ester 21 (IC50 > 200 microM). The nearly total loss of activity induced by esterification strongly indicates that the free carboxylic -OH is important for high potency of this compound. This contrasts with the isomeric 5,6-dihydroxy species 19, where esterification had no effect on inhibitory potency (23, IC50 = 52.7 microM). These data provide evidence that the monomeric 6,7- and 5,6-dihydroxynaphthalenes may be interacting with the enzyme in markedly different fashions. However, when these naphthyl nuclei were incorporated into dimeric structures, significant enhancements in potencies each relative to the monomeric acids were observed, with bis-6,7-dihydroxy analogue 49 and bis-5,6-dihydroxy analogue 51 both exhibiting approximately equal potencies (IC50 values of 0.81 and 0.11 microM, respectively).

Caffeic Acids↗

PubMed1997

Attenuation of insulin secretion by insulin-like growth factor 1 is mediated through activation of phosphodiesterase 3B.

Both insulin and insulin-like growth factor 1 (IGF-1) are known to reduce glucose-dependent insulin secretion from the beta cells of pancreatic islets. In this paper we show that the mechanism by which IGF-1 mediates this effect is in large part through activation of a specific cyclic nucleotide phosphodiesterase, phosphodiesterase 3B (PDE3B). More specifically, in both isolated pancreatic islets and insulin-secreting HIT-T15 cells, IGF-1 inhibits insulin secretion that has been increased by glucose and glucagonlike peptide 1 (GLP-1). Moreover, IGF-1 decreases cAMP levels in parallel to the reduction of insulin secretion. Insulin secretion stimulated by cAMP analogs that activate protein kinase A and also are substrates for PDE3B is also inhibited by IGF-1. However, IGF-1 does not inhibit insulin secretion stimulated by nonhydrolyzable cAMP analogs. In addition, selective inhibitors of PDE3B completely block the ability of IGF-1 to inhibit insulin secretion. Finally, PDE3B activity measured in vitro after immunoprecipitation from cells treated with IGF-1 is higher than the activity from control cells. Taken together with the fact that pancreatic beta cells express little or no insulin receptor but large amounts of IGF-1 receptor, these data strongly suggest a new regulatory feedback loop model for the control of insulin secretion. In this model, increased insulin secretion in vivo will stimulate IGF-1 synthesis by the liver, and the secreted IGF-1 in turn feedback inhibits insulin secretion from the beta cells through an IGF-1 receptor-mediated pathway. This pathway is likely to be particularly important when levels of both glucose and secretagogues such as GLP-1 are elevated.

Animals↗

PubMed1997

Optimization of DNA shuffling for high fidelity recombination.

A convenient 'DNA shuffling' protocol for random recombination of homologous genes in vitro with a very low rate of associated point mutagenesis (0.05%) is described. In addition, the mutagenesis rate can be controlled over a wide range by the inclusion of Mn2+or Mg2+during DNase I digestion, by choice of DNA polymerase used during gene reassembly as well as how the genes are prepared for shuffling (PCR amplification versus restriction enzyme digestion of plasmid DNA). These protocols should be useful for in vitro protein evolution, for DNA based computing and for structure-function studies of evolutionarily related genes.

Biological Evolution↗

PubMed1997

Discovery of HIV-1 integrase inhibitors by pharmacophore searching.

Based upon a class of known HIV-1 integrase inhibitors, several pharmacophore models were proposed from molecular modeling studies and validated using a 3D database of 152, compounds for which integrase assay data are known. Using the most probable pharmacophore model as the query, the NCI 3D database of 206,876 compounds was searched, and 340 compounds that contain the pharmacophore query were identified. Twenty-nine of these compounds were selected and tested in the HIV-1 integrase assay. This led to the discovery of 10 novel, structurally diverse HIV-1 integrase inhibitors, four of which have an IC50 value less than 30 microM and are promising lead compounds for further HIV-1 integrase inhibitor development.

4-Hydroxycoumarins↗