Biomedical subjects

D Yu

Publications and source records attributed to D Yu.

At least 55 records · Page 3Linked to original sources

PubMed2001

E1A: tumor suppressor or oncogene? Preclinical and clinical investigations of E1A gene therapy.

In the late 1980s, we have shown that the E1A gene can downregulate HER-2/neu overexpression, thus reversing the tumorigenic and metastatic phenotype. Further, E1A can function as a tumor suppressor gene by inducing apoptosis and inhibiting metastasis. At The University of Texas M. D. Anderson Cancer Center, we have been investigating the adenovirus type 5 E1A gene as a potential therapeutic gene in breast and ovarian cancer since 1995 by using cationic liposome as gene delivery system. In this chapter, we recount our development of E1A as a therapeutic gene.

Adenovirus E1A Proteins↗

PubMed2001

Mechanisms of ErbB2-mediated paclitaxel resistance and trastuzumab-mediated paclitaxel sensitization in ErbB2-overexpressing breast cancers.

The ErbB2 gene encodes a transmembrane growth factor receptor that belongs to the ErbB receptor tyrosine kinase subfamily. ErbB2 protein is overexpressed in approximately 30% of breast cancers. Although controversies exist, data from our laboratory and from clinical trials of trastuzumab indicate that ErbB2 overexpression confers chemoresistance to certain chemotherapeutic agents such as paclitaxel. One of the molecular mechanisms of ErbB2-mediated paclitaxel resistance is that overexpression of the ErbB2 receptor leads to deregulation of the G2/M cell cycle check-point that inhibits paclitaxel-induced apoptosis. Several promising ErbB2-targeting strategies have now been developed to conquer the adverse consequences of ErbB2 overexpression such as paclitaxel resistance. Among these, trastuzumab has brought great promise. We have recently shown that trastuzumab can effectively sensitize ErbB2-overexpressing breast cancer cells to paclitaxel by reversing the antiapoptotic function of ErbB2. Our studies provide additional support for chemotherapy combined with trastuzumab for ErbB2-overexpressing breast cancers, and it may bring insights into designing more effective and specific therapies that could offer great benefits to patients.

Animals↗

PubMed2001

Effect of chemical modifications of cytosine and guanine in a CpG-motif of oligonucleotides: structure-immunostimulatory activity relationships.

Oligodeoxynucleotides containing unmethylated CpG-motifs stimulate the innate immune system, including inducing B-cell proliferation and cytokine production. However, the mechanism of immunostimulation by CpG-oligonucleotides and the precise structural requirements and specific functional groups of cytosine and guanine necessary for recognition of and interaction with protein/receptor factors that are responsible for immune stimulation have not been elucidated. We sought to understand the critical role of each functional group of the cytosine and guanine moieties in a CpG-motif in inducing immunostimulatory activity. To this end, we examined structure-immunostimulatory activity relationships of phosphorothioate oligodeoxynucleotides (PS-oligos) containing YpG- and CpR-motifs (Y and R stand for pyrimidine and purine analogues, respectively). The PS-oligos containing a YpG-motif in which the natural deoxycytidine was replaced with deoxy-5-hydroxycytidine or deoxy-N4-ethylcytidine showed immunostimulatory activity. Substitution of deoxycytidine with a deoxy-5-methylisocytidine, deoxyuridine, or deoxy-P-base-nucleoside in the YpG-motif completely abolished the immunostimulatory activity, similar to the results observed with deoxy-5-methylcytidine. In the case of PS-oligos containing a CpR-motif, 7-deazaguanine substitution for natural guanine showed immunostimulatory activity similar to that of a parent PS-oligo. These studies suggest that the 2-keto, 3-imino and 4-amino groups of cytosine, and the 1-imino, 2-amino and 6-keto groups of guanine in a CpG-motif are important for the immunostimulatory activity of CpG-PS-oligos. The absence of N7 on guanine of the CpG-motif does not affect immunostimulatory activity significantly. These studies suggest that it is possible to develop YpG- and CpR-motifs as an alternative to CpG-motifs in PS-oligos for immunostimulatory studies.

Animals↗

PubMed2001

Immunostimulatory activity of CpG oligonucleotides containing non-ionic methylphosphonate linkages.

Bacterial DNA and synthetic oligodeoxynucleotides containing unmethylated CpG-motifs in a particular sequence context activate vertebrate immune cells. We examined the significance of negatively charged internucleoside linkages in the flanking sequences 5' and 3' to the CpG-motif on immunostimulatory activity. Cell proliferation and secretion of IL-12 and IL-6 in mouse spleen cell cultures, and spleen weights of mice increased significantly when a non-ionic linkage was placed at least four or more internucleoside linkages away from the CpG-motif in the 5'-flanking sequence. When the non-ionic linkage was placed closer than three internucleoside linkages in the 5'-flanking sequence to the CpG-motif, immunostimulatory activity was suppressed compared with that observed with the unmodified parent oligo. In general, the placement of non-ionic linkage in the 3'-flanking sequence to the CpG-motif either did not affect or slightly increased immunostimulatory activity compared with the parent oligo. These results have significance in understanding CpG oligonucleotide-receptor interactions and the development of potent immunomodulatory agents.

Adjuvants, Immunologic↗

PubMed2001

Prostate-specific targeting using PSA promoter-based lentiviral vectors.

The prostate-specific antigen (PSA) promoter is known to be highly tissue specific. Although its tissue specificity has been confirmed, its efficiency of gene transcription is significantly lower compared to known nonspecific viral promoters. These lower levels of promoter activity therefore pose a problem when developing an efficacious gene vector for prostate cancer gene therapy. Thus, selecting an appropriate therapeutic gene and vector system to carry the gene driven by the PSA promoter (PSAP) is important. In the studies described here, a human immunodeficiency virus (HIV)-1-based lentiviral vector carrying either the enhanced green fluorescent protein (EGFP) reporter or the diphtheria toxin A (DTA) gene was constructed. The results demonstrate that the PSA promoter in a lentiviral vector drives genes in prostate cells with satisfactory efficacy and specificity. The tissue-specific expression of the DTA protein efficiently eradicates LNCaP prostate cells in culture. We also infected prostate cancer cells and control cells carried by nude mice with the EGFP lentiviral vector. Significant numbers of EGFP-positive LNCaP cells were detected in all the mice bearing these tumors, but no EGFP-positive control cells were detected in any other mouse tissue. The high levels of expression in prostate cells, compared with the low levels of background expression in other cells, show that the PSAP-lentiviral vector could be a potential useful tool for gene therapy of metastatic prostate cancer.

Animals↗

PubMed2001

Is equity being sacrificed? Willingness and ability to pay for schistosomiasis control in China.

Decentralization of the health care system in China has led to an increasing need for income generation at all operational levels, both for curative services and for public health programmes. In general, people have accepted the costs of curative services, although the impact of charges on health-seeking behaviour has yet to be assessed. Public health programmes present particular problems in terms of revenue generation, however, because of the less direct impact of these activities on individual health and well-being. In this paper, we report the results of a cross-sectional study of willingness and ability to pay for schistosomiasis control. Questionnaires were administered to household heads of six representative villages in the Dongting Lake Region of Hunan Province, China. A total of 628 valid questionnaires were analyzed. The yearly mean household expenditure on schistosomiasis diagnosis and treatment was RMB 59.50 +/- 146.04 Yuan (US$1 = 8 RMB Yuan), accounting for 0.94% of the total yearly household income. Most household heads (514, 82%) thought schistosomiasis was the greatest health threat in their communities, but only 30.9% of them were willing to pay for screening, diagnosis and treatment of the infection. On the other hand, 72.3% of the respondents were willing to undertake volunteer work for control.

Adult↗

PubMed2001

Percutaneous, intramuscular neuromuscular electrical stimulation for the treatment of shoulder subluxation and pain in chronic hemiplegia: a case report.

This case report describes the first survivor with chronic stroke who was treated with percutaneous, intramuscular neuromuscular electrical stimulation (NMES) for shoulder subluxation and pain. The patient developed shoulder subluxation and pain within 2 mo of his stroke. After discharge from acute inpatient rehabilitation, he developed shoulder and hand pain, which was treated with subacromial bursa steroid injection and ibuprofen with eventual resolution. The patient remained clinically stable until approximately 15 mo after his stroke-when he developed severe shoulder pain associated with shoulder abduction, external rotation, and downward traction. The patient could not tolerate transcutaneous NMES because of the pain of stimulation. At approximately 17 mo post-stroke, the patient's posterior deltoid, middle deltoid, and supraspinatus muscles were percutaneously implanted with intramuscular electrodes. After 6 wk of percutaneous, intramuscular NMES treatment, marked improvements in shoulder subluxation and pain, and modest improvements in activities of daily living and motor function were noted. One year after the onset of treatment, the patient remained pain free, but subluxation had recurred. However, the patient was able to volitionally reduce the subluxation by abducting his shoulder. The patient remained pain free for up to 40 mo after the initiation of percutaneous, intramuscular NMES treatment. This case report demonstrates the feasibility of using percutaneous, intramuscular NMES for treating shoulder subluxation and pain in hemiplegia.

Electric Stimulation Therapy↗

PubMed2001

Evidence for an important role of WRKY DNA binding proteins in the regulation of NPR1 gene expression.

The Arabidopsis NPR1 gene is a positive regulator of inducible plant disease resistance. Expression of NPR1 is induced by pathogen infection or treatment with defense-inducing compounds such as salicylic acid (SA). Transgenic plants overexpressing NPR1 exhibit enhanced resistance to a broad spectrum of microbial pathogens, whereas plants underexpressing the gene are more susceptible to pathogen infection. These results suggest that regulation of NPR1 gene expression is important for the activation of plant defense responses. In the present study, we report the identification of W-box sequences in the promoter region of the NPR1 gene that are recognized specifically by SA-induced WRKY DNA binding proteins from Arabidopsis. Mutations in these W-box sequences abolished their recognition by WRKY DNA binding proteins, rendered the promoter unable to activate a downstream reporter gene, and compromised the ability of NPR1 to complement npr1 mutants for SA-induced defense gene expression and disease resistance. These results provide strong evidence that certain WRKY genes act upstream of NPR1 and positively regulate its expression during the activation of plant defense responses. Consistent with this model, we found that SA-induced expression of a number of WRKY genes was independent of NPR1.

Arabidopsis↗

PubMed2001

Computer-aided dynamic simulation of microwave-induced thermal distribution in coagulation of liver cancer.

To develop a method of dynamic three-dimensional (3-D) simulation of thermal distribution in ultrasound-guided microwave coagulation therapy of liver cancer and to verify its accuracy. The specific absorption rate (SAR) values were established by measuring the temperature in equivalent phantom tests. Those values were different under different power output condition. Dynamic 3-D temperature distributions were reconstructed with a finite-element model. Testing and rectification were performed through animal experiments and clinical trials, respectively. The temperature curves in the experiments corresponded well with simulated ones in vitro--91.4% and 88.9% using single and double electrodes, respectively. The measured coagulated boundary and simulated temperature boundary had a good correspondence in 85.7% of the specimens. In both in vivo experiments and clinical trials, blood perfusion influenced the rise in temperature significantly. Temperature curves between the simulations and actual measured results showed good correspondence--67.8% (19/28) in the patients with hepatocellular carcinoma. Distance between electrodes and combined thermal distributions were both optimized with computer-aided simulation during simultaneous two-electrode coagulation. The results demonstrated that computer-aided simulation of microwave thermal distribution is an accurate and reliable method which provides a theoretical and technical basis for controlling coagulated tissue volume and placement of the electrodes during microwave coagulation therapy of liver cancer.

Animals↗

PubMed2001

Variation in the dominant period during ventricular fibrillation.

Time-varying periodicities are commonly observed in biological time series. In this paper, we discuss three different algorithms to detect and quantify change in periodicity. Each technique uses a sliding window to estimate periodic components in short subseries of a longer recording. The three techniques we utilize are based on: 1) standard Fourier spectral estimation; 2) an information theoretic adaption of linear (autoregressive) modeling; and 3) geometric properties of the embedded time series. We compare the results obtained from each of these methods using artificial data and experimental data from swine ventricular fibrillation (VF). Spectral estimates have previously been applied to VF time series to show a time-dependent trend in the dominant frequency. We confirm this result by showing that the dominant period of VF, following onset, first decreases to a minimum and then rises to a plateau. Furthermore, our algorithms detect longer period correlations which may indicate the presence of additional periodic oscillations or more complex nonlinear structure. We show that in general this possibly nonlinear structure is most apparent immediately after the onset of VF.

Algorithms↗

PubMed2001

Herpes simplex virus DNA cleavage and packaging proteins associate with the procapsid prior to its maturation.

Packaging of DNA into preformed capsids is a fundamental early event in the assembly of herpes simplex virus type 1 (HSV-1) virions. Replicated viral DNA genomes, in the form of complex branched concatemers, and unstable spherical precursor capsids termed procapsids are thought to be the substrates for the DNA-packaging reaction. In addition, seven viral proteins are required for packaging, although their individual functions are undefined. By analogy to well-characterized bacteriophage systems, the association of these proteins with various forms of capsids, including procapsids, might be expected to clarify their roles in the packaging process. While the HSV-1 UL6, UL15, UL25, and UL28 packaging proteins are known to associate with different forms of stable capsids, their association with procapsids has not been tested. Therefore, we isolated HSV-1 procapsids from infected cells and used Western blotting to identify the packaging proteins present. Procapsids contained UL15 and UL28 proteins; the levels of both proteins are diminished in more mature DNA-containing C-capsids. In contrast, UL6 protein levels were approximately the same in procapsids, B-capsids, and C-capsids. The amount of UL25 protein was reduced in procapsids relative to that in more mature B-capsids. Moreover, C-capsids contained the highest level of UL25 protein, 15-fold higher than that in procapsids. Our results support current hypotheses on HSV DNA packaging: (i) transient association of UL15 and UL28 proteins with maturing capsids is consistent with their proposed involvement in site-specific cleavage of the viral DNA (terminase activity); (ii) the UL6 protein may be an integral component of the capsid shell; and (iii) the UL25 protein may associate with capsids after scaffold loss and DNA packaging, sealing the DNA within capsids.

Animals↗

PubMed2001

Potentiation of antitumor activity of irinotecan by chemically modified oligonucleotides.

Co-administration of synthetic chemically modified oligonucleotides with irinotecan, a selective topoisomerase I inhibitor, provided a significant enhancement in the antitumor activity of irinotecan. The enhancement of antitumor activity of irinotecan with co-administration of chemically modified oligonucleotides was observed in several tumor models--pancreatic cancer (Panc-1), colon cancer (HCT-116) and melanoma (A375). Inhibition of tumor growth in all three models required the co-administration of irinotecan and chemically modified oligonucleotides, but was independent of the nucleotide sequence of the oligonucleotides. The potentiation of antitumor activity was dependent on the dose of irinotecan and chemically modified oligonucleotides administered. The enhancement of antitumor activity of irinotecan was also observed by co-administration of a phosphorothioate oligonucleotide, however, to a lesser extent than did chemically modified oligonucleotides, suggesting that metabolic stability of the oligonucleotide contributes to the enhancement of antitumor activity seen with irinotecan. The co-administration of dextran sulfate sodium with irinotecan showed insignificant potentiation of antitumor activity of irinotecan, suggesting that the enhancement of antitumor activity of irinotecan observed was not a result of polyanionic characteristic of oligonucleotides. Co-administration of irinotecan and chemically modified oligonucleotides did not result in increased toxicity in the tumor models studied. Potentiation of antitumor activity of irinotecan observed with co-administration of oligonucleotides suggests that the oligonucleotides affect the pharmacokinetics and/or metabolism of irinotecan. The use of chemically modified oligonucleotides together with irinotecan may increase the therapeutic index of irinotecan in cancer patients and continued development of such agents should be considered.

Animals↗

PubMed2001

The Cockayne syndrome group B DNA repair protein as an anti-cancer target.

Cells from individuals with Cockayne syndrome (CS) have a defect in transcription-coupled DNA repair (TCR), which rapidly corrects certain DNA lesions located on the transcribed strand of active genes. Despite this DNA repair defect, individuals with CS (of which there are two complementation groups, CSA and CSB) do not demonstrate an elevated incidence of cancer. Recently, we demonstrated that disruption of the CSB gene reduces the spontaneous tumor rate in cancer predisposed Ink4a/ARF-/- mice as well as causing their embryo fibroblasts to proliferate more slowly and be more sensitive to UV-induced apoptosis. In the present study we characterized phosphorothioate backbone antisense oligodeoxynucleotides (AOs) that reduced the levels of CSB mRNA in A2780/CP70 ovarian carcinoma cells. The AOs caused the cells to proliferate more slowly and made them more sensitive to either cisplatin or oxaliplatin. The AOs also enhanced the cytotoxicity of hydrogen peroxide and gamma-radiation, both of which can induce oxidative DNA lesions, which are subject to TCR. The AOs did not potentiate the cytotoxicity of topotecan, which induces DNA strand breaks. Chemically modified ( ) AOs (MBOs) targeting CSB were able to potentiate the anti-tumor effect of cisplatin against A2780/CP70 tumor xenografts formed in nude mice. The MBOs enabled a non-toxic (3 mg/kg) dose of cisplatin to have the same degree of anti-tumor efficacy as a more toxic (5 mg/kg) cisplatin dose. Collectively, these results suggest that the CSB gene product may be viewed as an anti-cancer target.

Animals↗

PubMed2001

Antiapoptotic activity is dispensable for insulin-like growth factor I receptor-mediated clonogenic radioresistance after gamma-irradiation.

PURPOSE: The purpose of this study was to evaluate the relationship between apoptotic activity and clonogenic radiosensitivity in vitro using an insulin-like growth factor I receptor (IGF-IR) signaling model, which is known to exert tumorigenic and antiapoptotic effects. EXPERIMENTAL DESIGN: We used mouse embryo fibroblast cell lines expressing either human IGF-IR [R+(Wt) and R+] or the marker gene alone [R-(puro)]; these cell lines were derived from R- cells, which are deficient in IGF-IR. After gamma-irradiation, apoptotic activity was determined by the presence of DNA fragmentation and caspase-3-, -8-, and -9-like activities. Clonogenic radiosensitivity was determined by a colony-forming assay. RESULTS: R+(Wt) and R+ cells expressed similar levels of IGF-IR, transducing phosphorylation signals to major downstream substrates on insulin-like growth factor I stimulation. R+ cells were resistant to the induction of apoptosis after gamma-irradiation; however, both R+(Wt) and R-(puro) cells demonstrated significant DNA fragmentation and increase in caspase-3-, -8-, and -9-like activities. Both R+(Wt) and R+ cells were radioresistant (to a similar extent) compared with R-(puro) cells as measured by a colony-forming assay. Clonogenic radioresistance was not influenced by the inhibition of Akt/protein kinase B through treatment with wortmannin at low concentrations specifically inhibiting phosphatidylinositol 3'-kinase. CONCLUSIONS: Our findings indicate that apoptotic activity does not necessarily predict clonogenic survival after exposure to ionizing radiation. This study provides clinical implications in the evaluation of apoptotic activities observed during the course of radiotherapy to predict accurate tumor response or local control.

Androstadienes↗

PubMed2001

Isolation and purification of guinea pig inner ear antigens by preparative polyacrylamide gel electrophoresis.

OBJECTIVE: To isolate and purify the subcomponents of guinea pig inner ear antigens for further study on the autoimmunity of the inner ear. METHODS: Inner ear tissues were homogenized in phosphate buffered saline containing 0.1% SDS, and then frozen and defrosted repeatedly to extract inner ear antigens. Preparative polyacrylamide gel electrophoresis was used to separate the subcomponents of inner ear antigens. Following electrophoresis, the protein bands were localized by rapid staining and destaining. RESULTS: The major protein bands were clearly distinct when 3 mg of crude inner ear antigens was loaded, and the three major subcomponents (31, 42-45 and 60 kD proteins) accounted for about 25.99%, 21.91% and 21.10%, respectively. CONCLUSION: Preparative polyacrylamide gel electrophoresis can be used to purify the major subcomponents of inner ear antigens.

Animals↗

PubMed2001

Anticancer effect of a lentiviral vector capable of expressing HIV-1 Vpr.

A lentiviral vector capable of expressing the HIV-1 vpr gene (Vpr lentiviral vector) was constructed, and its in vivo anticancer effect was determined against cutaneous tumors derived from the AT-84 oral cancer cells in immunocompetent mice. A single intratumoral injection of the Vpr lentiviral vector not only significantly reduced the primary tumor volume but also completely regressed tumors in >40% of animals. More interestingly, the mice of which the primary tumors were completely regressed by the Vpr lentiviral vector were additionally protected from a secondary challenge of AT-84 cells. These data suggest that the Vpr lentiviral vector elicits its anticancer activity in part by the activation of the immune system. The above suggestion is additionally supported by the failure of the lentiviral vector to demonstrate anticancer activity in immunocompromised nude or SCID mice. The Vpr lentiviral vector offers a powerful new strategy for cancer gene therapy and may be useful for the control of solid tumors, such as human oral squamous cell carcinomas.

Animals↗

PubMed2001

Antisense anti-MDM2 oligonucleotides as a novel therapeutic approach to human breast cancer: in vitro and in vivo activities and mechanisms.

The mouse double minute 2 (MDM2) oncogene has been suggested as a target for cancer therapy. It is amplified or overexpressed in many human cancers, including breast cancer, and MDM2 levels are associated with poor prognosis of several human cancers, including breast cancer, ovarian cancer, osteosarcoma, and lymphoma. In the present study, we investigated the functions of MDM2 oncogene in the growth of breast cancer and the potential value of MDM2 as a drug target for cancer therapy by inhibiting MDM2 expression with a specific antisense antihuman-MDM2 oligonucleotide (oligo). The selected antisense mixed-backbone oligo was evaluated for its in vitro and in vivo antitumor activity in human breast cancer models: MCF-7 cell line containing wild-type p53 and MDA-MB-468 cell line containing mutant p53. In MCF-7 cells, p53 and p21 levels were elevated, resulting from specific inhibition of MDM2 expression by the antisense oligo (AS). In MDA-MB-468 cells, after inhibition of MDM2 expression, p21 levels were elevated, although p53 levels remained unchanged. After i.p. administration of the antisense anti-MDM2 oligo, in vivo antitumor activity occurred in a dose-dependent manner in nude mice bearing MCF-7 or MDA-MB-468 xenografts. In both models, in vivo synergistically or additive therapeutic effects of MDM2 inhibition and the clinically used cancer chemotherapeutic agents irinotecan, 5-fluorouracil, and paclitaxel (Taxol) were observed. These results suggest that MDM2 have a role in tumor growth through both p53-dependent and p53-independent mechanisms. We speculate that MDM2 inhibitors, such as ASs, have a broad spectrum of antitumor activities in human breast cancers, regardless of p53 status. This study should provide a basis for future development of anti-MDM2 ASs as cancer therapeutic agents used alone or in combination with conventional chemotherapeutics.

Animals↗

PubMed2001

[Clinical research of pasinizid on retreated sputum positive pulmonary tuberculosis in senilities].

OBJECTIVE: To observe and evaluate the effect and safety of pasinizid on treatment of the senile pulmonary tuberculosis with bacteriological positive. METHODS: 115 cases of the senile pulmonary tuberculosis with bacteriological positive were divided into therapy group(group A) and control group(group B1 and B2) randomly. Chemotherapy regimen: group A were treated by 3DZL2E/5DL2E mainly composed of pasinizid; group B1 were treated by 3HPZL2/5HL2E(abbreviated HP regimen); group B2 were treated by 3DZL2E/5DL2E(abbreviated SH regimen). RESULTS: When the chemotherapy was over, the sputum negative conversion rates were 92%, 72%, 71% in group A, group B1 and group B2 respectively. The conversion rate in group A was significantly higher than that in group B1 and group B2 (P < 0.05); focal absorption rates were 75%, 50%, 50% respectively, while the vanishing rates of cavity were 85%, 46%, 50% respectively. The rate in group A was significantly higher than that in group B1 and B2(P < 0.05); Rates of side effects were 17%, 58%, 18% respectively. The rates in group A and group B2 were apparently lower than that in group B1(P < 0.01). Bacteriological relapse rate of 2 years were 5%, 19%, 18%, respectively. The rate in group A was lower than that in group B1 and B2 (P < 0.01). CONCLUSIONS: The effect of pasinizid on senile retreated sputum positive pulmonary tuberculosis was better than HP and SH regimen. Due to its lower rates of side effects, it is recommended for clinical usage.

Aminosalicylic Acid↗