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Biomedical subjects

J Zhu

Publications and source records attributed to J Zhu.

At least 559 records · Page 31Linked to original sources
PubMed1997

[Construction of recombinant vaccinia virus expressing HSV-2 gD gene as live recombinant vaccine strain].

We had reported that the recombinant vaccinia virus expressing glycoprotein D of herpes simplex virus type 2 (HSV-2 gD) protected mice against lethal HSV-2 challenge. Following the succeed in animal model, we continue the research to construct the recombinant vaccinia virus expressing HSV-2 gD gene as live recombinant vaccine strain in strict accordance with the guideline for human vaccine research. A PCR-modified HSV-2 gD gene was inserted into the plasmid pJSB1175, under the control of P7.5K early/late promoter of vaccinia virus. The recombinant plasmid was used, with lipofectin reagent, to transfect 2BS cells which had been infected by wild type of TK+ vaccinia virus (Tian Tan 761 strain). The recombinant vaccinia virus harboring HSV-2 gD gene was selected out by using in situ hybridization employing 32P-1a-belled HSV-2 gD fragment as probe, together with three cycles of plaque purification. Dot and Southern blot confirmed HSV-2 gD gene had been integrated into the TK region of vaccinia virus genome, as expected. Indirect immunofluorescent assay using anti-HSV-2 gD monoclonal antibody showed HSV-2 gD was expressed effectively in the recombinant virus infected cells.

Animals↗
PubMed1997

[Study of dominant negative effect of the PLZF-RARalpha against the wild-type RARalpha in acute promyelocytic leukemia].

OBJECTIVE: To explore the role of promyelocytic leukemia zinc finger (PLZF) gene on 11q23 and RARalpha gene on 17q21. METHODS: Eight PLZF/RARalpha expression plasmids with deletions of different PLZF motifs were constructed by using PCR based-method. RESULTS AND CONCLUSION: By transient co-transfection, it was confirmed that PLZF/RARalpha had a dominant negative effect against the wild-type RARalpha, the POZ domain of PLZF/RARalpha was responsible for the dominant negative effect and the PLZF/RARalpha may play certain role through a spatial interaction with POZ.

Animals↗
PubMed1997

[Preliminary study on the arsenic trioxide-induced NB4 cell apoptosis and its molecular mechanisms].

OBJECTIVE: To illustrate the possible cellular and molecular mechanisms of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia (APL). METHODS: APL cell line NB4 was used for in vitro studies. The effect of As2O3 on APL was studied by using flow cytometry, DNA electrophoresis, Narthern blotting and Western blotting. RESULTS: As2O3 induced NB4 cell apoptosis, while not inhibiting the growth and survival of two other leukemic cell lines (HL-60 and U937). Furthermore, As2O3 effectively down-regulated the expression of bcl-2 gene without changing the mRNA levels of other apoptosis-associated genes (including p53, c-myc, bax and bcl-XL). CONCLUSION: These might be one of the molecular mechanisms of As2O3 induced NB4 cell apoptosis.

Animals↗
PubMed1997

[Effects of arsenic trioxide on the subcellular localization of PML/PML-RARalpha protein in leukemic cells].

OBJECTIVE: In order to illustrate the possible roles of PML-RARalpha protein in arsenic trioxide (AsO3)-induced NB4 cell apoptosis. METHODS: Effects of As2O3 on the subcellular localization of PML-RARalpha in NB4 cells were studied. RESULTS: (1) Anti-PML serum staining was reduced and PML granules emerged in the perinuclear cytoplasm in a diffuse pattern in HL-60 cells under As2O3 treatment; (2) abnormal PML/PML-RARalpha granules were decreased; (3) NB4 cells accumulated anti-PML serum staining granules in the cytoplasms were increased and similar accumulation also found in apoptotic cells; and (4) pretreatment with all-trans retinoic acid (ATRA) for 24 or 48 hours did not alter the As2O3 effects. CONCLUSION: As2O3-induced apoptosis was independent of the retinoic acid signal pathway, and it might be regulated by PML/PML-RARalpha and/or other related genes.

Antineoplastic Agents↗
PubMed1997

[Biological characteristics of residual leukemic cells and their drug-resistant regrowth].

OBJECTIVE: To elucidate the mechanisms of residual leukemia and its drug-resistant regrowth. METHODS: Bone marrow cells from transplantable mouse leukemia models L615 and L7811 were cultivated in long-term culture system respectively. Daunorubicin (DNR) was added to the systems to mimick in vivo chemotherapy. RESULTS: Residual leukemic cells on stromal layers escaped DNR killing. They derived from the adherent leukemic cell population, which was heterogeneous with many undifferentiated and dormant cells. They were drug-resistant and expressed bcl-2 gene. CONCLUSION: Bone marrow stromal layer was the vital microenvironment for the residual leukemic cells to survive and regrow, and might increase the drug-resistance by up-regulating bcl-2 gene expression.

Animals↗
PubMed1997

High IL-6 and low IL-10 in the central nervous system are associated with protracted relapsing EAE in DA rats.

Experimental autoimmune encephalomyelitis (EAE) is a CD4+ T cell-mediated, inflammatory demyelinating disease of the central nervous system (CNS) that serves as a model for multiple sclerosis (MS). The mechanisms behind differences in clinical course of EAE in different rat strains have not been defined. We induced acute EAE in Lewis rats and protracted relapsing EAE (PR-EAE) in DA rats and examined mRNA expression of IL-1 beta, IL-6, IL-10, IL-12, and TNF-beta in brain tissue sections, cerebrospinal fluid (CSF) cells, and lymph node cells. IL-1 beta, IL-12 and TNF-beta mRNA expression in brain tissue sections appeared early and peaked at the height of clinical signs in both acute and PR-EAE, consistent with a disease-promoting role for these cytokines. High levels of IL-6 mRNA-expressing cells were present in CNS and lymph node cells in PR-EAE, while almost absent in acute EAE. In contrast, IL-10 was very low in PR-EAE but strongly expressed in acute EAE, in particular during clinical recovery. Regulatory changes of IL-6 and IL-10 both systemically and within the CNS, but with temporal differences between compartments, seem pivotal for development of PR-EAE in DA rats. These findings could have relevance for pathogenesis and treatment of MS.

Animals↗
PubMed1997

Activation of the cloned human kappa opioid receptor by agonists enhances [35S]GTPgammaS binding to membranes: determination of potencies and efficacies of ligands.

Activation of kappa receptors inhibits adenylate cyclase, enhances K+ conductance and reduces Ca++ conductance via pertussis toxin-sensitive G proteins. We recently cloned a human kappa opioid receptor and stably expressed it in Chinese hamster ovary (CHO) cells. In this study, the effects of activation of the human kappa receptor by agonists on [35S]GTPgammaS binding to CHO cell membranes were examined. The presence of GDP and Mg++ was essential for the kappa agonist (-)-U50,488H-induced increase in [35S]GTPgammaS binding to be observed and the optimal concentration was 3 microM and 5 mM, respectively. The presence of 100 mM Na+ was necessary to produce the maximal signal-to-background ratio. (-)U50,488H-induced increase in [35S]GTPgammaS binding was time- and tissue concentration-dependent. (-)U50,488H increased [35S]GTPgammaS binding in a dose-dependent manner with an EC50 of 3.1 nM. (+)-U50,488H had no effect, which indicates that this effect is stereospecific. Naloxone (1 microM) or norbinaltorphimine (10 nM) shifted the dose-response curve of (-)-U50,488H to the right by 100-fold. These results indicate that enhancement of [35S]GTPgammaS binding by (-)-U50,488H is a kappa receptor-mediated event. Pretreatment of the cells with pertussis toxin, but not cholera toxin, abolished the (-)-U50,488H-induced increase in [35S]GTPgammaS binding, which indicates the involvement of Gi and/or Go proteins. [35S]GTPgammaS binding induced by (-)-U50,488H had a Kd value of 0.34 +/- 0.08 nM and a Bmax value of 431 +/- 29 fmol/mg protein. The rank order of potencies of opioid ligands tested in stimulating [35S]GTPgammaS binding was dynorphin A 1-17 > (+/-)-ethylketocyclazocine > beta-funaltrexamine, (-)-U50,488H, tifluadom > nalorphine > pentazocine, nalbuphine > buprenorphine. Dynorphin A 1-17, (+/-)-ethylketocyclazocine, (-)-U50,488H, tifluadom and beta-funaltrexamine were full agonists, but nalorphine and pentazocine were partial agonists producing maximal responses of 68% and 23% of those of full agonists, respectively. Nalbuphine and buprenorphine had low levels of agonist activities. Norbinaltorphimine and naloxone were antagonists devoid of activities. Enhancement of [35S]GTPgammaS binding by kappa agonists provides a simple functional measure for receptor activation and can be used for determination of potencies and efficacies of opioid ligands at the kappa receptor.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed1997

QTL analysis to study the association between leaf size and abscisic acid accumulation in droughted rice leaves and comparisons across cereals.

Plants accumulate abscisic acid (ABA) under droughted conditions. Genetic variation in the accumulation of ABA in deteched and partially dehydrated leaves of rice has previously been reported, and this was found to be associated with variation in leaf size (smaller leaves made more ABA). Correlation analysis failed to distinguish clearly between a causal relationship between the two traits and close genetic between loci controlling the traits. Here we present a detailed genetic analysis of ABA accumulation in detached and partially dehydrated rice leaves. using a population of F2 plants generated from the lowland x upland cross IR20 (high-ABA) x 63-83 (low-ABA) which was mapped with RFLP and AFLP markers. Several highly significant quantitative trait loci (QTLs) for ABA accumulation and leaf weight were identified. Only one of the minor QTLs for ABA accumulation (accounting for only 4% of the phenotypic variance) was coincident with any QTLs for leaf size such that the high-ABA allele was associated with smaller leaves. This analysis, therefore, showed that the association previously found between ABA accumulation and leaf size was probably largely due to genetic linkage and not to a direct effect of leaf size on ABA accumulation or vice versa. Because of the importance of ABA accumulation in regulating responses of plants to drought stress and the effects of plant size on the rate of development of stress, QTLs for drought-induced ABA accumulation, leaf size and tiller number were compared between rice and wheat. In particular, a possible location in rice was sought for a homoeologue of the major wheat vernalization responsive gene, Vrn1, as this gene is also associated with major effects on leaf size, tiller number and ABA accumulation in wheat. The likelihood of homoeologous loci regulating ABA accumulation, leaf size and tiller number in the two crops is discussed.

Abscisic Acid↗
PubMed1997

Bayesian adaptive alignment and inference.

Sequence alignment without the specification of gap penalties or a scoring matrix is attained by using Bayesian inference and a recursive algorithm. This procedure's recursive algorithm sums over all possible alignments on the forward step to obtain normalizing constants essential to Bayesian inferences, and samples from the exact posterior distribution on the backward step. Since both terminal and intervening unrelated subsequences will often be excluded from an alignment, the resulting alignments may be seen as extensions of local alignments. An alignment's significance is assessed using the Bayesian evidence. A shuffling simulation shows that Bayesian evidence against the null hypothesis tends to be a conservative measure of significance compared to classical p-values. An application to proteins from the GTPase superfamily shows that the posterior distribution of the number of gaps is often flat and that the posterior distribution of the evolutionary distance is often flat and sometimes bimodal. An alignment of 1GIA with 1ETU shows good correspondence with a structural alignment.

Algorithms↗
PubMed1997

[Cloning of BRCA1 cDNA and detection of BRCA1 mRNA expression in breast cancer cells].

The fragment of BRCA1 cDNA obtained by reverse transcription and polymerase chain reaction(RT-PCR) was inserted into plasmid pUC118 and demonstrated by DNA sequencing. Nucleotide sequence analysis demonstrated that the cloned cDNA for BRCA1 includes zinc finger domain. Two differences in nucleotides were found as compared with the sequence published. One occurs at nucleotide number 409 where Creplaced by A (Asp-->Glu). Another difference occurs at nucleotide number 879 where A replaced by T (samesense mutation). In order to further study the relationship-between BRCA1 function and breast cancer, the probe was prepared from the recombinant plasmid and then hybridized to total RNAs from 6 cases of breast cancer. Compared with normal cells, the expression level of BRCA1 mRNA was normal in 4, decreased markedly in 1, and in one patient there was no any expression of BRCA1 mRNA at all. The results suggested that the expression of BRCA1 mRNA was relatively low in some breast cancer cells.

Amino Acid Sequence↗
PubMed1997

Studies on multiple forms of maltotetraose-forming amylase from Alcaligenes sp.

Zymograms of the cultural supernatant of Alcaligenes sp. showed three bands, the major one being G4A-1 and the minor two, G4A-2 and G4A-3. Based on the electrophoretic homogeneity of the purified three bands and the enzymatic activities identified by a thin layer chromatography of the soluble starch hydrolysates, all the three bands were confirmed to be maltotetraose-forming amylase but in multiple forms. Neither glycosidase nor protease activities could be detected in the culture (only very weak protease activities were observed at 48 hours after cultivation), which indicate that the two enzymes were not involved in the amylase multiple-form formation. Only the relative amount of the two minor bands (but not the multiple-form pattern) was changed when the initial pH of the medium varied from 6.5-8.5. An addition of 0.3% glucose raised the yield of G4A-2 and G4A-3.

Alcaligenes↗
PubMed1997

Comparison of acid anhydrides with carboxylic acids in enantioselective enzymatic esterification of racemic menthol.

Optical resolution of racemic menthol has been efficiently achieved by lipase-catalyzed enantioselective esterification in an organic solvent. The performance of the reaction using an acid anhydride as an acyl donor was compared with that using its corresponding free acid. The reactivities of acid anhydrides were found to be higher than their corresponding free acids, but acid anhydrides were also found to be easily hydrolyzed into free acids under the catalysis of the same enzyme. The existence of a too-high concentration of an acid anhydride in a micro-aqueous reaction system will cause dehydration and thus deactivation of the enzyme, and will enhance non-selective esterification of a chiral alcohol, which will reduce the optical purity of the product. All these drawbacks, however, could be effectively overcome in a semi-batch reaction system into which propionic anhydride was continuously fed. This system showed some advantages over a batch reaction system using free propionic acid: the reaction time of dl-menthol was shortened by half, the stability of the enzyme was much enhanced, and the optical purity of the product (l-menthyl ester) was kept at a similarly high level (> 98% ee).

Acids↗
PubMed1997

[Usage of double immuno-enzyme labeled staining method in forensic pathology].

In this paper we report a kind of double immunoenzyme labelled staining method (single enzyme method). The principle of the method is that HPR will show blue-black color when it meet DAB-CoCL2/H2O2, while it will show brown color as it meet DAB/H2O2, two colors constract very remarkably. This method was used to detect the antigens of hearts and brains which obtained from forensic autopsy. The results showed that the method was sensitive, simple and economical. One of the advantages is that the stained sections can keep for a long time. Thus, is very practical for forensic pathology.

Brain↗
PubMed1997

[Epidemiological studies on 321 children with congenital diaphragmatic hernia in China].

Perinatal babies delivered in hospitals with gestation of 28 weeks to seven days after birth were monitored by National Birth Defects Monitoring Network from September 1986 to December 1993 to study the prevalence of congenital diaphragmatic hernia in China and the risk factors contributing to their prognosis. Results showed that 321 cases of congenital diaphragmatic hernia in 4,777,220 perinatal babies were detected with an incidence of 0.7 per ten thousand, a case-fatality ratio of 84.1 percent and, fetal death and stillbirth accounting for 28.1 percent of the total perinatal deaths. There was significant difference in birth weight between perinatal deaths and survival perinatal. And, 54.4 percent of the cases complicated with other congenital deformities and severe deformities in heart, lung and central nervous system accounted for the most proportion of them, which were the most important factors contributing to death. A case-fatality ratio of cases with single malformation was 74.7 percent and that of those with multiple malformation 92 percent, with a statistically significant difference. The cases diagnosed antenatally accounted for 11.3 percent of the total, and multiple malformation accounted for a large proportion of them. It indicated that proportion of antenatal diagnosis for congenital diaphragmatic hernia deformity was lower, and their perinatal case-fatality ratio was higher and their prognosis worse.

Abnormalities, Multiple↗
PubMed1996

The PML and PML/RARalpha domains: from autoimmunity to molecular oncology and from retinoic acid to arsenic.

Acute promyelocytic leukemia (APL) is specifically associated to a t(15; 17) translocation which fuses a gene encoding a nuclear receptor for retinoic acid, RARalpha, to a previously unknown gene PML. The PML protein is localized in the nucleus on a specific domain of unknown function (PML nuclear bodies, NB) previously detected with autoimmune sera from patients with primary biliary cirrhosis (PBC). These bodies are nuclear matrix-associated and all of their identified components (PML, Sp100, and NDP52) are sharply upregulated by interferons. We show that autoantibodies against both PML and Sp100 are usually associated in sera with multiple nuclear dot anti-nuclear antibodies and demonstrate that PML is an autoantigen, not only in PBC, but also in other autoimmune diseases. In APL, the PML/RARalpha fusion interferes with both the retinoic acid (RA) response and PML localization on nuclear bodies, but the respective contribution of each defect to leukemogenesis is unclear. RA induces the terminal differentiation of APL blasts, yielding to complete remissions, and corrects the localization of NB antigens. Arsenic trioxide (As2O3) also induces remissions in APL, seemingly through induction of apoptosis. We show that in APL, As2O3 leads to the rapid reformation of PML bodies. Thus, both agents correct the defect in NB antigen localization, stressing the role of nuclear bodies in the pathogenesis of APL.

Animals↗