Biomedical subjects

Yi Zhang

Publications and source records attributed to Yi Zhang.

At least 181 records · Page 10Linked to original sources

PubMed2005

Anti-SARS-CoV immunity induced by a novel CpG oligodeoxynucleotide.

To develop CpG oligodeoxynucleotides (CpG ODNs) based therapy for prevention and treatment of severe acute respiratory syndrome (SARS), we selected a novel CpG ODN (BW001), which displays B-type CpG ODN structure feature at the 5' and A-type CpG ODN structure feature at the 3', and tested for its anti-SARS-CoV activity. We found that the supernatants of human PBMCs stimulated by BW001 significantly protected Vero cells from SARS-CoV infection. BW001 could stimulate human PBMCs and pDCs to secrete high level of IFN-alpha and promote human PBMCs and B cells to proliferate. Furthermore, we demonstrated that BW001 could activate CD19+ B cells and CD56+ NK cells in human PBMCs. In addition, BW001 could enhance NK cytotoxicity and IFN-gamma secretion in human PBMCs. Together, BW001 represents a novel type of CpG ODN and may have potential for the development of treatment and prevention for SARS as well as other viral associated diseases.

Adjuvants, Immunologic↗

PubMed2005

Abnormal expression and processing of the proprotein convertases PC1 and PC2 in human colorectal liver metastases.

BACKGROUND: The family of proprotein convertases has been recently implicated in tumorigenesis and metastasis in animal models. However, these studies have not yet been completely corroborated in human tumors. METHODS: Using RT PCR, immunoblot and immunohistochemistry we assessed the presence and the processing patterns of the convertases PC1 and PC2 as well as the PC2 specific chaperone 7B2 in human liver metastases originating from colorectal cancer and compared them to unaffected and normal liver. Furthermore, we assessed the presence and processing profiles of PC1, PC2 and 7B2 in primary colon cancers. RESULTS: mRNA, protein expression, and protein cleavage profiles of proprotein convertases 1 and 2 are altered in liver colorectal metastasis, compared to unaffected and normal liver. Active PC1 protein is overexpressed in tumor, correlating with its mRNA profile. Moreover, the enhanced PC2 processing pattern in tumor correlates with the overexpression of its specific binding protein 7B2. These results were corroborated by immunohistochemistry. The specific and uniform convertase pattern observed in the metastases was present only in a fraction of primary colon cancers. CONCLUSION: The uniformly altered proprotein convertase profile in liver metastases is observed only in a fraction of primary colon cancers, suggesting possible selection processes involving PCs during metastasis as well as an active role of PCs in liver metastasis. In addition, the exclusive presence of 7B2 in metastatic tumors may represent a new target for early diagnosis, prognosis and/or treatment.

Colonic Neoplasms↗

PubMed2005

Patterns of nestin expression in human skin.

Nestin, a member of the sixth class of intermediate filament proteins, has long been known not only as a specific marker for central nervous system stem cells, but also as an indicator of the degree of neural stem cell differentiation. Immunohistochemical analysis showed nestin expression in the epidermis and the upper two-third of the hair follicle in non-balding human scalp skin, but not in the lower third of the follicle. Expression was very weak in the sebaceous gland. The level of nestin mRNA transcription was much higher in the bulge region of the follicle than in the sebaceous gland. These results show that the pattern of nestin expression is related to the differentiation of epidermal stem cells.

Epidermal Cells↗

PubMed2005

Isolation and characterization of 18 genes encoding alpha- and beta-expansins in wheat (Triticum aestivum L.).

Expansins are thought to be key regulators of cell wall extension during plant growth. In this study, we isolated 18 expansin genes from wheat, nine of which encode alpha-expansins while the other nine code for beta-expansins. The cysteine-rich and tryptophan-rich regions of the deduced amino acid sequences of all 18 expansins were highly conserved. Genomic sequences were obtained for 17 of the genes, and their intron patterns were determined. Four (A, C, D, E) of the six intron positions known in expansin genes from other species were found to be occupied in these wheat expansin genes. Five wheat expansin genes were mapped to chromosomes 1L, 2L, 5L and 6L respectively, by in silico and comparative mapping. The 18 wheat expansin genes were expressed in leaf, root and the developing seed. Moreover, it was demonstrated that four beta-expansin genes were up-regulated in the internode tissue in F1 hybrids, suggesting that changes in the regulation of these genes in hybrid might contribute to the heterosis observed in internode length and plant height. We therefore conclude that expansins are encoded by a multigene family in wheat, and could play important roles in growth and development.

Amino Acid Sequence↗

PubMed2005

Circumventing central leptin resistance: lessons from central leptin and POMC gene delivery.

We identified that leptin resistance in aged-obese rats has both peripheral and central components. The central resistance is characterized by diminished hypothalamic leptin receptors and impaired leptin signal transduction. We developed a new model of leptin-induced leptin resistance in which application of the central leptin gene delivery produces unabated hypothalamic leptin over-expression. The chronic central elevation of leptin precipitates leptin resistance in young animals devoid of obesity and exacerbates it in mature or aged animals with obesity. Despite leptin resistance, our aged obese, DIO, and leptin-induced leptin resistant rats were fully responsive to central pharmacological melanocortin activation. We propose that the central leptin resistance resides between leptin receptor and melanocortin receptor activation. Our central POMC gene therapy overcame leptin resistance, producing weight and fat loss and improved insulin sensitivity in obese Zucker and aged rats. This success highlights the central melanocortin system as a useful drug target for combating obesity.

Adiposity↗

PubMed2005

Effects of fluid shear stress on apoptosis of cultured human umbilical vein endothelial cells induced by LPS.

The objective of our research was to reveal the effects of shear stress on the apoptosis of cultured human umbilical vein endothelial cells (HUVECs) induced by lipopolysaccharide (LPS). A parallel-plate flow chamber was used to control the strength and duration of shear stress (SS), and apoptosis was measured by immunocytochemistry and radio-immunoassay. Some important conclusions were drawn. In the stationary state, apoptosis of HUVECs could be induced by LPS (50 microg/ml). An SS of 15 dyn/cm(2) could inhibit the apoptosis induced by LPS. However, an SS of 4 dyn/cm(2) had less effect on the same process. At the same time, the experiment demonstrated that the increase in IL-6 secretion by LPS can be inhibited by two different levels of shear stress. Moreover, the inhibition effect was more obvious under high level stress than under low level. We also found that the effect of shear stress on IL-8 was less effective than on IL-6. This research provides data for understanding the mechanism of the contribution of hemodynamic forces to atherosclerosis.

Apoptosis↗

PubMed2005

BCRP transports dipyridamole and is inhibited by calcium channel blockers.

PURPOSE: We investigated whether dipyridamole and various calcium channel blockers are inhibitors and/or substrates of breast cancer resistance protein (BCRP). METHODS: The effect of dipyridamole and the calcium channel blockers on mitoxantrone efflux by BCRP-overexpressing human embryonic kidney (HEK) cells was determined by flow cytometry. The ability of some of these compounds to reverse BCRP-mediated mitoxantrone resistance was measured by cytotoxicity assays. Transport studies were performed using radiolabeled compounds. RESULTS: Dipyridamole, nicardipine, nitrendipine, and nimodipine effectively inhibited BCRP-mediated mitoxantrone efflux; however, bepridil, diltiazem, and verapamil had no significant effect. Nifedipine is a much weaker BCRP inhibitor compared with other dihydropyridines tested. Nicardipine and dipyridamole were the most potent BCRP inhibitors among the compounds tested with IC50 values of 4.8 +/- 1.3 and 6.4 +/- 0.9 microM, respectively. Nicardipine and dipyridamole also effectively reversed BCRP-mediated mitoxantrone resistance in HEK cells. [3H]Nitrendipine was found not to be transported by BCRP. However, the transport of [3H]dipyridamole by BCRP was observed in both HEK and Madin-Darby canine kidney cells stably expressing the transporter, and this transport was completely abolished by fumitremorgin C, a known BCRP inhibitor. CONCLUSIONS: Dipyridamole and several dihydropyridines are effective BCRP inhibitors, but bepridil, diltiazem, and verapamil are not. We also identified a new BCRP substrate, dipyridamole.

ATP Binding Cassette Transporter, Subfamily G, Mem↗

PubMed2005

[Studies on susceptibility of Pomacea canaliculata of different developmental stages to infection with Angiostrongylus cantonensis].

UNLABELLED: OBJECTIVE To study the susceptibility of Pomacea canaliculata of different developmental stages to Angiostrongylus cantonensis infection. METHODS: P. canaliculata snails breeding at laboratory were divided into four rank (I-IV) according to the weight, and infected with the first stage larvae of A. cantonensis from Fujian Province. Their mortality, infection rate, worm burden, and the size, development and distribution of larvae in snails were examined. RESULTS: Snails at different developmental stages were readily infected with A. cantonensis. The infection rate was between 76% and 100%, with no significant difference among the groups (P>0.05). Snails at earlier developmental stage showed higher mortality. The heaviest worm burden and the largest number of snails harboring more than 100 larvae were found in snails of rank III. In general the larvae in snails showed a synchronous development in the groups. Sizes of the third stage larvae in snails of various ranks were homogeneous. The period before the third stage larva emergence and the time for a peak percentage of the larvae exhibited no significant difference among the four ranks. The larvae widely distributed in various parts of snails, with more in the lung and foot, and larvae from snails of different ranks could all infect rats successfully. CONCLUSION: P. canaliculata of the four ranks can all be infected by the first stage larvae of A. cantonensis. Rank III snails may be better for studying the relationship between P. canaliculata and A. cantonensis. The potential role of young snails in angiostrongyliasis transmission should be recognized.

Angiostrongylus cantonensis↗

PubMed2005

The inhibitory effects of anticoagulation on in vivo gene transfer by adeno-associated viral or adenoviral vectors.

Identifying factors that influence gene transfer efficacy is critical for a successful gene-based clinical study. Here we demonstrate that in vivo AAV-2-mediated gene transfer is efficiently inhibited by unfractionated heparin, but not by a heparin preparation containing mainly low-molecular-weight forms (LMWH). Surprisingly, inhibitors of thrombin or factor Xa (F.Xa) significantly reduced AAV-2 transduction in a dose-dependent manner. These effects were independent of the vector promoter, transgene, or strain of mice. Expression by alternate AAV serotypes 5 and 8 was not affected by anticoagulant drugs, which suggests an AAV-2-specific effect. Moreover, AAV-2-mediated gene expression was diminished in mice with deficiency in thrombin generation (factor IX deficiency) and enhanced in mice with procoagulant phenotype due to factor V Leiden. In addition, inhibitors of F.Xa diminished adenovirus-mediated gene expression. These results demonstrated that coagulation activity itself is critical to ensure optimal viral vector transduction. Since intravascular delivery of vectors often requires the use of anticoagulants, the use of LMWH appears to be safe. These observations are of relevance for approaches using AAV-2 or adenoviral vectors, especially in early phase studies designed to identify the minimum therapeutic doses.

Adenoviridae↗

PubMed2005

Growth-associated protein GAP-43 and L1 act synergistically to promote regenerative growth of Purkinje cell axons in vivo.

Neuronal expression of growth-associated protein 43 (GAP-43) and the cell adhesion molecule L1 has been correlated with CNS axonal growth and regeneration, but it is not known whether expression of these molecules is necessary for axonal regeneration to occur. We have taken advantage of the fact that Purkinje cells do not express GAP-43 or L1 in adult mammals or regenerate axons into peripheral nerve grafts to test the importance of these molecules for axonal regeneration in vivo. Transgenic mice were generated in which Purkinje cells constitutively express L1 or both L1 and GAP-43 under the Purkinje cell-specific L7 promoter, and regeneration of Purkinje cell axons into peripheral nerve grafts implanted into the cerebellum was examined. Purkinje cells expressing GAP-43 or L1 showed minor enhancement of axonal sprouting. Purkinje cells expressing both GAP-43 and L1 showed more extensive axonal sprouting and axonal growth into the proximal portion of the graft. When a predegenerated nerve graft was implanted into double-transgenic mice, penetration of the graft by Purkinje cell axonal sprouts was strongly enhanced, and some axons grew along the entire intracerebral length of the graft (2.5-3.0 mm) and persisted for several months. The results demonstrate that GAP-43 and L1 coexpressed in Purkinje cells can act synergistically to switch these regeneration-incompetent CNS neurons into a regeneration-competent phenotype and show that coexpression of these molecules is a key regulator of the regenerative ability of intrinsic CNS neurons in vivo.

Animals↗

PubMed2005

Differential roles of nuclear and cytoplasmic cyclin-dependent kinase 5 in apoptotic and excitotoxic neuronal death.

Cyclin-dependent kinase 5 (cdk5) is a member of the cyclin-dependent kinase family whose activity is localized mainly to postmitotic neurons attributable to the selective expression of its activating partners p35 and p39. Deregulation of cdk5, as a result of calpain cleavage of p35 to a smaller p25 form, has been suggested to be a central component of neuronal death underlying numerous neurodegenerative diseases. However, the relevance of cdk5 in apoptotic death that relies on the mitochondrial pathway is unknown. Furthermore, evidence that cdk5 can also promote neuronal survival has necessitated a more complex understanding of cdk5 in the control of neuronal fate. Here we explore each of these issues using apoptotic and excitotoxic death models. We find that apoptotic death induced by the DNA-damaging agent camptothecin is associated with early transcription-mediated loss of p35 and with late production of p25 that is dependent on Bax, Apaf1, and caspases. In contrast, during excitotoxic death induced by glutamate, neurons rapidly produce p25 independent of the mitochondrial pathway. Analysis of the localization of p35 and p25 revealed that p35 is mainly cytoplasmic, whereas p25 accumulates selectively in the nucleus. By targeting a dominant-negative cdk5 to either the cytoplasm or nucleus, we show that cdk5 has a death-promoting activity within the nucleus and that this activity is required in excitotoxic death but not apoptotic death. Moreover, we also find that cdk5 contributes to pro-survival signaling selectively within the cytoplasm, and manipulation of this signal can modify death induced by both excitotoxicity and DNA damage.

Animals↗

PubMed2005

A new LAGE-1 peptide recognized by cytolytic T lymphocytes on HLA-A68 tumors.

Antigens encoded by genes of the LAGE family, including LAGE-1 and NY-ESO-1, are of interest for cancer immunotherapy because they are tumor-specific and shared by tumors of different histological types. Several clinical trials are in progress with NY-ESO-1 peptides, protein, recombinant poxviruses, and dendritic cells pulsed with peptides. In this study, CD8 T lymphocytes from an individual without cancer were stimulated with dendritic cells infected with a recombinant avian poxvirus encoding a complete LAGE-1 protein. A CTL clone was isolated that recognized a new LAGE-1 peptide, ELVRRILSR, which corresponds to position 103-111 of the protein sequence. It is presented by HLA-A6801 molecules. When tumor cells expressing LAGE-1 were transfected with HLA-A68, they were lysed by the CTL clone, indicating that the peptide is processed in tumor cells. These results indicate that the LAGE-1.A68 peptide can be used for antitumoral vaccination. We observed also that specific T cells could be detected in a blood sample with a high sensitivity by using an A68/LAGE-1 fluorescent multimer.

Amino Acid Sequence↗

PubMed2005

Electrochemical dechlorination of chloroacetic acids (CAAs) using hemoglobin-loaded carbon nanotube electrode.

Hemoglobin (Hb) was immobilized on carbon nanotube (CNT) electrode to catalyze the dechlorination of chloroacetic acids (CAAs), and the electrocatalytic behaviors of the Hb-loaded electrode for the dechlorination of trichloroacetic acid (TCAA) were studied by cyclic voltammetry and constant-potential electrolysis technique. An Hb-loaded packed-bed flow reactor was also constructed for bioelectrocatalytic dechloriantion of CAAs from drinking water. The results showed that the reduced heme of Hb immobilized on CNT electrode was easily regenerated, and Hb exhibited a stable and high activity for reductive dechlorination of CAAs with significant lowering of overpotential. TCAA could be reduced at -0.450 V (vs. saturated calomel electrode (SCE)) with catalysis of Hb-loaded electrode and its dechlorination was stepwise, following the pathway of TCAA-->dichloroacetic acid (DCAA)-->monochloroacetic acid (MCAA)-->acetic acid. It was also found that all CAAs, e.g., TCAA, DCAA and MCAA, could be dechlorinated completely at -0.450 V. The removal of 30.0 mM TCAA and DCAA is ca. 40% and 31%, respectively, with electrolysis for 100 min at -0.600 V (vs. SCE) using the Hb-loaded packed-bed flow reactor. The dechlorination activities of CAAs follow the decreasing order: TCAA>DCAA>MCAA, and the average current efficiency is over 90%.

Acetates↗

PubMed2005

Galaxy: a platform for interactive large-scale genome analysis.

Accessing and analyzing the exponentially expanding genomic sequence and functional data pose a challenge for biomedical researchers. Here we describe an interactive system, Galaxy, that combines the power of existing genome annotation databases with a simple Web portal to enable users to search remote resources, combine data from independent queries, and visualize the results. The heart of Galaxy is a flexible history system that stores the queries from each user; performs operations such as intersections, unions, and subtractions; and links to other computational tools. Galaxy can be accessed at http://g2.bx.psu.edu.

Biological Evolution↗

PubMed2005

Altered expression of MAP-2, GAP-43, and synaptophysin in the hippocampus of rats with chronic cerebral hypoperfusion correlates with cognitive impairment.

Chronic cerebral hypoperfusion causes cognitive impairment, but the underlying molecular mechanism is not well understood. We used permanent occlusion of bilateral common carotid arteries (2-VO) to induce chronic cerebral hypoperfusion in male Wistar rats. Cognitive impairment and the expression patterns of MAP-2, GAP-43, and synaptophysin were examined. We found that both learning capacity and memory were gradually impaired in the rats with chronic cerebral hypoperfusion concomitant with increased duration of 2-VO treatment. Four weeks of 2-VO treatment resulted in down-regulation of synaptophysin expression at the protein levels, and a further decrease was observed at 10-20 weeks, although mRNA levels remained the same. Ten weeks of 2-VO treatment lead to down-regulation of MAP-2 expression at both the mRNA and protein levels with a further decrease at 20 weeks. Interestingly, GAP-43 mRNA was significantly up-regulated by 2-VO treatment, although the protein levels were not altered. Therefore, the cognitive impairment caused by chronic cerebral hypoperfusion may be partially explained by reduced expression of synaptophysin and MAP-2 at the protein level. The reduction in MAP-2 expression may be attributed to the inhibition of transcription, while the reduction in synaptophysin expression might be due to the inhibition of translation. Up-regulation of GAP-43 mRNA in the rat hippocampus with 2-VO treatment suggests that a compensatory mechanism may antagonize ischemic challenges.

Animals↗

PubMed2005

A peripheral element assembles the compact core structure essential for group I intron self-splicing.

The presence of non-conserved peripheral elements in all naturally occurring group I introns underline their importance in ensuring the natural intron function. Recently, we reported that some peripheral elements are conserved in group I introns of IE subgroup. Using self-splicing activity as a readout, our initial screening revealed that one such conserved peripheral elements, P2.1, is mainly required to fold the catalytically active structure of the Candida ribozyme, an IE intron. Unexpectedly, the essential function of P2.1 resides in a sequence-conserved short stem of P2.1 but not in a long-range interaction associated with the loop of P2.1 that stabilizes the ribozyme structure. The P2.1 stem is indispensable in folding the compact ribozyme core, most probably by forming a triple helical interaction with two core helices, P3 and P6. Surprisingly, although the ribozyme lacking the P2.1 stem renders a loosely folded core and the loss of self-splicing activity requires two consecutive transesterifications, the mutant ribozyme efficiently catalyzes the first transesterification reaction. These results suggest that the intron self-splicing demands much more ordered structure than does one independent transesterification, highlighting that the universally present peripheral elements achieve their functional importance by enabling the highly ordered structure through diverse tertiary interactions.

Base Pairing↗

PubMed2005

[Inhibitory effect of dendritic cells pulsed with MAGE-3 peptide on transplanted murine gastric cancer in mice].

OBJECTIVE: To observe the inhibitory effect of MAGE-3 peptide pulsed DC on transplanted murine gastric cancer in 615 mice. METHODS: The CTL clones directed against MAGE-3 peptide were tested for the ability to lyse the gastric cancer cell line MFC which can express MAGE-3 antigen. In immunoprotection experiment, mice of the study group were immunized with MAGE-3 peptide pulsed DC (DC/MAGE-3) at the dosage of 1 x 10(6) on d0 and d7 by sc inoculation, mice of control groups were immunized with influenza virus peptide pulsed DC (DC/Nup) or unpulsed DC at the same dosage on days as the DC/MAGE-3 group. On d14, all the mice were challenged with sc injections of 5 x 10(5) MFC gastric cancer cells. In immunotherapy experiment, all the mice were sc injected 5 x 10(5) MFC gastric cancer cells on d0, and on d3, d10 mice of each groups were sc inoculated with DC/MAGE-3, DC/Nup or unpulsed DC at the dosage of 1 x 10(6) respectively. All mice were monitored closely with respect to tumor growth and survival times. RESULTS: The CTL clone induce by MAGE-3 peptide could lyse the MFC cells efficiently. Immunization of mice with DC pulsed with MAGE-3 generated partial protective immunity against MFC tumor, as well as significant inhibition of tumor growth in a 3-day tumor model. CONCLUSION: The tumor vaccine with DCs pulsed with MAGE-3 peptide possesses the ability to stimulate tumor specific CTL activity and to establish antitumor immunity when administered in vivo.

Animals↗

PubMed2005

Dimethyl sulfoxide targets phage RNA polymerases to promote transcription.

Dimethyl sulfoxide (DMSO) is a "universal" solvent in pharmaceutical sciences and cell biology. DMSO was previously reported to facilitate in vitro transcription of chromatin and supercoiled plasmid, with the underlying mechanism being attributed to the alteration of the template structure. Here, we demonstrated that low concentrations of DMSO significantly increase the phage polymerase-catalyzed RNA synthesis when the naked and short PCR products were used as templates, suggesting that DMSO promotes transcription through additional mechanism(s). Interestingly, SP6 RNA polymerase was more sensitive to the DMSO stimulation than T7 RNA polymerase, suggesting that the polymerase is an important target for DMSO stimulation of RNA synthesis. Consistent with this finding, we also showed that DMSO dramatically elevated the RNA polymerase activity. This elevated activity is explained by altered polymerase structure as reflected by a change in intrinsic fluorescence. Furthermore, DMSO was shown to simultaneously accumulate both the abortive and full-length transcripts, leading us to conclude that the DMSO-altered polymerase structure primarily encodes an enhanced activity at the stage of transcription initiation. DMSO-induced alteration of the polymerase structure and function highlights a potentially generalized mechanism in interpreting the molecular action of this popular solvent.

DNA-Directed RNA Polymerases↗