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Biomedical subjects

D Lu

Publications and source records attributed to D Lu.

At least 73 records · Page 4Linked to original sources
PubMed2001

A bicistronic retroviral vector to introduce drug resistance genes into human umbilical cord blood CD34+ cells to improve combination chemotherapy tolerance.

OBJECTIVE: To study whether human umbilical cord blood CD34+ cells transduced with human aldehyde dehydrogenase class-1 (ALDH-1) and multidrug resistance gene (MDR1) have increases resistance to 4-Hydroperoxycyclo-phosphamide (4-HC) and P-glycoprotein effluxed drugs. METHODS: A bicistronic retroviral vector G1Na-ALDH1-IRES-MDR1 was constructed and used to transfect the packaging cell lines GP + E86 and PA317 by LipofectAMINE method, using the medium containing VCR and 4-HC agents for cloning selection and ping-ponging supernatant infection between the ecotropic producer clone and the amphotropic producer clone, we obtained high titer amphotropic PA317 producing cells with high titers up to 5.6 x 10(5) CFU/ml. Cord blood CD34+ cells were transfected repeatedly with supernatant of retrovirus containing human ALDH-1 and MDR1cDNA under the stimulation of hemopoietic growth factors. RESULTS: Bicistronic retroviral vector construction was verified by restriction endonuclease analysis. Polymerase chain reaction (PCR), reverse transcription (RT)-PCR, Southern blot, Northern blot, fluorescenceactivated cell sorting (FACS) method and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analyses showed that dual drug resistance genes have been integrated into the genomic DNA of cord blood CD34+ cells and expressed efficiently. The transgenes recipient cells confered 4-fold stronger resistance to 4-HC and 5.5 to 7.2-fold P-glycoprotein effluxed drug than untransduced cells. CONCLUSION: The bicistronic retroviral vector-mediated transfer of two different types of drug resistance genes into human cord blood CD34+ cells and co-expression provided an experimental foundation for improving combination chemotherapy tolerance in tumor clinical trial.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed2001

Low power density microwave radiation induced early changes in rabbit lens epithelial cells.

OBJECTIVE: To determine whether low power density microwave radiation can induce irreversible changes in rabbit lens epithelial cells (LECs) and the mechanisms of the changes. METHODS: One eye of each rabbit was exposed to 5 mW/cm2 or 10 mW/cm2 power density microwaves for 3 hours, while the contralateral eye served as a control. Annexin V-propidium iodide (PI) two-color flow cytometry (FCM) was used to detect the early changes in rabbit lens epithelial cells after radiation. RESULTS: Lots of rabbit LECs were in the initial phase of apoptosis in the 5 mW/cm2 microwave radiation group. A large number of cells became secondary necrotic cells, and severe damage could be found in the group exposed to 10 mW/cm2 microwave radiation. CONCLUSION: Low power densities of microwave radiation (5 mW/cm2 and 10 mW/cm2) can induce irreversible damage to rabbit LECs. This may be the non-thermal effect of microwave radiation.

Animals↗
PubMed2001

[Association of single nucleotide polymorphisms in code region of beta2-adrenoceptor gene with hypertension in Chinese population].

OBJECTIVE: To detect single nucleotide polymorphisms(SNPs) existing in code region of beta(2)-adrenoceptor(2-AR) gene and to investigate association of the identified SNPs with essential hypertension in Chinese Han population. METHODS: Beta(2)-AR gene was sequenced with fluorescent labelling automatic sequencing method in unrelated Chinese Han population from Dabie Mountain in Anhui Province. Genotype of the SNPs were typed with PCR-RFLP method. RESULTS: Two SNPs were identified in length of 774bp, at position + 1053 with G-->C substitution and + 1239 with A-->G substitution respectively. The frequency of genotype of the two SNPs complied well with the Hardy-Weinberg equilibrium in normal group. Distribution of genotype AA, GA, GG of the SNPs at locus + 1239 in hypertension group was significantly different from that in normal group (chi(2) = 6.70, df = 2, P < 0.05). No significant difference was observed in distribution of genotypes of the SNPs at locus + 1053 between the two groups. CONCLUSION: These results indicate that the SNPs at locus + 1239 of beta(2)-AR gene is associated with EH. The SNPs at position + 1053 was not linked to hypertension.

Aged↗
PubMed2001

Dynamic changes of interleukin-1, interleukin-6 and tumor necrosis factor in intermingled skin graft in burned rats.

OBJECTIVE: To compare the dynamic changes of interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor (TNF) in intermingled skin graft with those in other types of skin grafts in rats. METHODS: A 10%-15% third-degree burn was created in 180 Sprague-Dawley (SD) rats. After removing the scar, skin grafts were performed on the open wounds immediately with autoskin (aus, n=54), alloskin (als, n=54) and intermingled skin (n=36). That is to say, in the intermingled skin graft, a big piece of alloskin (mals) was grafted first, and 3 days later, small pieces of autoskin (maus) wer e embedded in the alloskin. The rest 36 rats were taken as the controls. And the biological activities of IL-1, IL-6 and TNF in graft sheets in each group were detected after skin graft. RESULTS: The levels of IL-1, IL-6 and TNF in the aus group decreased steadily after their initial elevations, whereas in the als group they increased significantly and kept on the peak level in the later phases. In the in termingled group, there appeared a lowest IL-1 level in the mals and a highest one in the maus simultaneously at 7 (4) days (The number out of parenthesis is t he days after transplanting with alloskin sheets, and the number in parenthesis is the days after embedding autoskin sheets in the intermingled skin graft. Similarly hereinafter.) after skin graft (P<0.01), and the high level in the maus abruptly decreased at 14 (11) days after skin graft. At exactly the same phase on day 7 (4), a prominent peaked IL-6 in the mals occurred. In the later phases, the levels of TNF remained relatively low both in the m als and in the maus. From day 7 (4) on, each cytokine fluctuation in the mals synchronized with that in the maus. The longer the post transplantation period lasted, the more the positive cytokine correlated between the mals and the maus. CONCLUSIONS: The low levels of IL-1 and TNF may be important f actors to lighten the intensity of local rejection in the intermingled skin graft. The temporarily peaked IL-6 is both an inducer which induces the production of local IL-1 receptor antagonists and soluble TNF receptors and a signal which indicates a local enhancement of Th(2) cells. The mild rejection process and th e synchronized cytokine level during the later phases suggest a possible chimerism between the mals and the maus.

Animals↗
PubMed2001

Treatment of traumatic brain injury in female rats with intravenous administration of bone marrow stromal cells.

OBJECTIVE: To study the effect of bone marrow stromal cells administered intravenously to female rats subjected to traumatic brain injury. METHODS: We injected marrow stromal cells harvested from male rat bone marrow (n = 24) into the tail vein of the female rat (n = 8) 24 hours after traumatic brain injury; the rats were killed at Day 7 or 14 after treatment. The neurological function of the rats was evaluated using the rotarod test and the neurological severity score. The distribution of the male donor cells in brain, heart, lung, kidney, liver, muscle, spleen, and bone marrow of the female recipient rats was measured by identifying Y chromosome-positive cells using fluorescent in situ hybridization. RESULTS: We found that marrow stromal cells injected intravenously significantly reduced motor and neurological deficits compared with control groups by Day 15 after traumatic brain injury (P < 0.05, analysis of covariance for repeated measures). The transplanted cells preferentially engrafted into the parenchyma of the injured brain and expressed the neuronal marker NeuN and the astrocytic marker glial fibrillary acidic protein. Marrow stromal cells were also found in other organs in female rats subjected to traumatic brain injury without any obvious adverse effects. CONCLUSION: These data suggest that the intravenous administration of marrow stromal cells may be a promising therapeutic strategy that warrants further investigation for patients with traumatic brain injury.

Animals↗
PubMed2001

[Inhibition of proliferation and induction of apoptosis by simvastatin in K562 leukemic cell line].

OBJECTIVE: To investigate the anti-apoptotic mechanism and explore approach to inhibiting proliferation and inducing apoptosis of chronic myclogenous leukemia (CML) cells. METHODS: K562 cell line was used to evaluate the effects of simvastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, and the combination of simvastatin with chemotherapeutic agents on the proliferation and apoptosis of CML cells. RESULTS: Simvastatin could significantly inhibit proliferation and induce apoptosis of K562 cells, and could increase the sensitivity of K562 cells to chemotherapeutic agents. Addition of mevalonate, the immediate product of HMG-CoA, could completely reverse this effect. CONCLUSION: Simvastatin inhibited proliferation and induced apoptosis of K562 cells through inhibiting the metabolic pathway of mevalonate. It is promising that HMG-CoA reductase inhibitors may be an effective chemotherapeutic approach to the treatment of CML.

Antineoplastic Agents↗
PubMed2001

[Relationship between soluble Fas ligand levels and complications after allogeneic bone marrow transplantation].

OBJECTIVE: To evaluate the implications of soluble Fas ligand (sFasL) in acute graft-versus-host disease (aGVHD) and discriminating symptoms of aGVHD from those of infection. METHODS: Plasma levels of sFasL were assessed in 84 plasma samples from 13 patients after allogeneic BMT using a sandwich enzyme-linked immunological assay (ELISA). Plasma sFasL levels of the patients before BMT and at different time points in the post-BMT period were measured. The results were analysed for correlation with aGVHD and infections. RESULTS: Plasma sFasL levels were significantly higher in patients with grade II - IV aGVHD than that in those with grade 0 - I aGVHD (P = 0.02). There was no statistic difference in plasma sFasL levels between the infectious and non-infectious patients. In the seven grade II - IV aGVHD patients, the plasma sFasL levels pre-BMT were much lower than that in the six grade 0 - I aGVHD patients. CONCLUSIONS: sFasL may be useful for the diagnosis of aGVHD and for differentiating aGVHD from other BMT related complications. The high level of plasma sFasL pre-BMT may be of importance in decreasing the occurrence of aGVHD after BMT.

Adolescent↗
PubMed2001

[Improvement of combination chemotherapy tolerance of human umbilical cord blood CD(34)(+) cells transducted with double drug resistance genes by a bicistronic retroviral vector].

OBJECTIVE: To explore whether human umbilical cord blood hematopoietic progenitor cells transduced with human aldehyde dehydrogenase class 3 (ALDH3) and multidrug resistance gene (MDR1) could increase resistance to 4-hydroxycyclophosphamide (4-HC) and P-glycoprotein effluxed drugs. METHODS: A bicistronic retroviral vector G1Na-ALDH3-IRES-MDR1 cDNA was constructed and transfected the packaging cell lines GP + E86 and PA317 by LipofectAMINE method, using the medium containing VCR and 4-HC for cloning selection and ping-ponging supernatant infection between ecotropic producer clone and amphotropic producer clone, cord blood CD(34)(+) cells were enriched with a high-gradient magnetic cell sorting system (MACS), and then repeatedly transfected with supernatant of retrovirus containing human ALDH3 and MDR1 cDNA under stimulation of hematopoietic growth factors. PCR, RT-PCR, Southern blot, Northern blot, FACS and MTT assay were used to evaluate the transfection and expression of the double genes. RESULTS: The purity of cord blood CD(34)(+) cells was approximately 91% and the recovery rate was 72%. The highest titer of recombinant amphotropic retrovirus in the supernatant was up to 6.5 x 10(5) CFU/ml. The efficiency of gene transduction was 18%, 20% and 16.7% tested by colony formation, PCR and FACS, respectively. Rhodamine 123 efflux showed 16% transduced cells with P-gp function. No helper virus was found by both nested PCR and rescue assay. The MTT analysis showed a 3.5 to 6.8-fold increase of resistance of transducted cells to cyclophosphamide and P-glycoprotein effluxes drug as compared with the nontransduced cells. CONCLUSION: The efficiency and co-expression of this dual genes transfer system provided a foundation for ameliorating combination chemotherapy toxicity in clinical trial.

ATP Binding Cassette Transporter, Subfamily B↗
PubMed2001

[Cantonese population data of the tetranucleotide repeat locus D7S809 and its application in paternity test].

The tetranucleotide repeat locus D7S809 was investigated in Cantonese population by polymerase chain reaction (PCR) amplification, subsequent polyacryramide gel electrophoresis and silver staining. 14 alleles and 50 genotypes were detected in 190 sample individuals. All alleles differed in size by 4 bp repeat. No interalleles were found. The heterozygosity, power of discrimination and chance of non-paternal exclusion were 0.8613, 0.9645 and 0.7184 respectively. No significant deviations from Hardy-Weinberg equilibrium were found. The D7S809 was a highly polymorphic, stable and accurate type locus and had been successfully applied to 100 paternity test cases.

Alleles↗
PubMed2001

[Cloning and expression of VHB gene in D-arabitol producing yeast].

Recombinant plasmid pVgb-EX2 containing Vitreoscilla hemoglobin gene vgb and formaldehyde resistant gene SFA1 was constructed and transformed into D-arabitol producing yeast strain Saccharomyces sp. X-62. The fact that the amount of VHb in transformant cells was considerably higher than that in control cells indicated that gene vgb was expressed in transformant cells. D-arabitol productivity and yield of fermentation by transformants were improved. The most improvement of D-arabitol productivity in repeat experiments reached 27.3%. It appeared that the fermentation productivity of D-arabitol was relative to the amount of VHb in cells under experimental conditions.

Bacterial Proteins↗
PubMed2001

Identified circadian rhythm genes of ciliary epithelium with differential display.

PURPOSE: To identify differential genes expressed in the rabbit ciliary epithelium during the circadian cycle of aqueous flow. METHODS: Total RNA from ciliary epithelium of rabbits at 8 AM (light on 1 hour) and 8 PM (light off 1 hour) were compared by differential display reverse transcription-polymerase chain reaction (DD RT-PCR), using 6% denaturing polyacrylamide electrophoresis, choose differential display bands, cut and reamplify with the same primer, clone and sequence. Search the database of Genbank, prolong them with 5' RACE and 3' RACE technique then clone, sequence and search database of Genbank. RESULTS: 93 Significant differences gene expression were detected between light on and light off in the rabbit ciliary epithelium. CONCLUSION: Differential display is a powerful tool to screen differentially expressed genes in circadian rhythm of ciliary epithelium.

Animals↗
PubMed2001

[Successful engraftment of HLA-identical sibling cord blood transplantation in an adult with chronic myelogenous leukemia].

OBJECTIVE: To explore the feasibility of cord blood transplantation (CBT) for the treatment of adult hematological malignancies and its long-term hematopoiesis reconstitution and transplantation-related complications. METHODS: An 18 years old patient (body weight 75 kg) with chronic myelogenous leukemia in first chronic phase received HLA-identical sibling CBT after conditioning with modified busulfan/cyclophosphamide (Bu/CTX) regimen. The transplanted number of nucleated cells was 1.73 x 10(7)/kg of body weight, and that of CD34+ cells 2. 7 x 10(5)/kg. Cyclosporin A and methylprednisolone were given as prophylaxis against graft versus-host disease (GVHD). RESULTS: The neutrophil count rose to above 0.5 x 10(9)/L on day 18 and platelet count exceeded 50 x 10(9)/L on day 36. Gene analysis showed that bone marrow cells had completely changed to donor's type on day 80. The patient was diagnosed with grade IV acute hepatic GVHD complicated with CMV infection because of severe jaundice on day 90. After the administration of additional immunosuppressive agents, antiviral agents, plasma exchange and in vitro billirubin adsorption, the complications were well controlled. In the follow-up of 24 months', the patient's general condition is good without obvious hepatic dysfunction and Ph chromosome and bcr/abl fusion gene of bone marrow cells were persistently negative. CONCLUSION: It is the first case reported in China that adult patient with leukemia has been successfully treated by allogeneic CBT, and this indicates that CBT is feasible in the treatment of adult patient with leukemia.

Adolescent↗
PubMed2000

Overexpression of a modified human malonyl-CoA decarboxylase blocks the glucose-induced increase in malonyl-CoA level but has no impact on insulin secretion in INS-1-derived (832/13) beta-cells.

The long-chain acyl-CoA (LC-CoA) model of glucose-stimulated insulin secretion (GSIS) holds that secretion is linked to a glucose-induced increase in malonyl-CoA level and accumulation of LC-CoA in the cytosol. We have previously tested the validity of this proposal by overexpressing goose malonyl-CoA decarboxylase (MCD) in INS-1 cells, but these studies have been criticized due to: 1) the small insulin secretion response (2-4-fold) of the INS-1 cells used; 2) unknown contribution of the ATP-sensitive K(+) (K(ATP)) channel-independent pathway of GSIS in INS-1 cells, which has been implicated as the site at which lipids regulate insulin granule exocytosis; and 3) deletion of the N-terminal mitochondrial targeting sequence, but not the C-terminal peroxisomal targeting sequence in the goose MCD construct, raising the possibility that a significant fraction of the overexpressed enzyme was localized to peroxisomes. To address these outstanding concerns, INS-1-derived 832/13 cells, which exhibit robust K(ATP) channel-dependent and -independent pathways of GSIS, were treated with a new adenovirus encoding human MCD lacking both its mitochondrial and peroxisomal targeting sequences (AdCMV-MCD Delta 5), resulting in large increases in cytosolic MCD activity. Treatment of 832/13 cells with AdCMV-MCD Delta 5 completely blocked the glucose-induced rise in malonyl-CoA and attenuated the inhibitory effect of glucose on fatty acid oxidation. However, MCD overexpression had no effect on K(ATP) channel-dependent or -independent GSIS in 832/13 cells. Furthermore, combined treatment of 832/13 cells with AdCMV-MCD Delta 5 and triacsin C, an inhibitor of long chain acyl-CoA synthetase that reduces LC-CoA levels, did not impair GSIS. These findings extend our previous observations and are not consistent with the LC-CoA hypothesis as originally set forth.

Adenoviridae↗
PubMed2000

Spinal cord injury in rat: treatment with bone marrow stromal cell transplantation.

We tested the hypothesis that transplantation of bone marrow stromal cells (MSCs) into the spinal cord after a contusion injury promotes functional outcome. Rats (n = 31) were subjected to a weight driven implant injury. MSCs or phosphate buffered saline was injected into the spinal cord 1 week after injury. Sections of tissue were analyzed by double-labeled immunohistochemistry for MSC identification. Functional outcome measurements using the Basso-Beattie-Bresnehan score were performed weekly to 5 weeks post-injury. The data indicate significant improvement in functional outcome in animals treated with MSC transplantation compared to control animals. Scattered cells derived from MSCs expressed neural protein markers. These data suggest that transplantation of MSCs may have a therapeutic role after spinal cord injury.

Animals↗
PubMed2000

The role of the central zinc fingers of transcription factor IIIA in binding to 5 S RNA.

In the nine-zinc finger Xenopus transcription factor TFIIIA the central group of fingers, fingers 4 to 7, have been shown to bind to 5 S RNA. In this study, we have attempted to assess the role of this region of the TFIIIA molecule in more detail than hitherto. High-resolution footprinting by RNases A and CV1 has been used to probe the binding to 5 S RNA of three TFIIIA peptides Tf(1-6), Tf(4-6) and Tf(4-7), consisting of fingers 1 to 6, 4 to 6, and 4 to 7, respectively, and of full-length TFIIIA. The results pinpoint the outer margins of binding of the central fingers within helices IV and II of TFIIIA. A comparison of the footprints reveals that the presence of finger 7 affords protection at positions C19 and U55, the distal portion of helix II and the proximal portion of loop B. In addition, our footprints suggest that the central fingers bind in the same manner, whether in an isolated group or in the intact TFIIIA molecule. In a companion study, we have determined the binding affinities of Tf(4-6) and Tf(4-7) for full-length and three truncated 5 S RNA molecules, the latter selected on the basis of the regions of the 5 S RNA molecule known to be important in the binding of TFIIIA. The analysis uses only fully active protein involved in the binding and the results are consistent with the corresponding footprinting results. This is the first time that a detailed study of the binding site of one particular zinc finger to RNA has been reported; the results should be of use in the design of 5 S RNA molecules and TFIIIA peptides for structural studies of the interaction between zinc fingers and RNA.

Amino Acid Sequence↗
PubMed2000

Identification of the residues in the extracellular region of KDR important for interaction with vascular endothelial growth factor and neutralizing anti-KDR antibodies.

The kinase domain receptor (KDR) of vascular endothelial growth factor (VEGF) is the main human receptor responsible for the angiogenic activity of VEGF. The extracellular region of KDR is comprised of seven immunoglobulin-like domains, of which the first three have been shown to be required for ligand binding. We have previously described antibodies directed against the extracellular region of KDR, including MAB383 and MAB664, which were shown to block the binding of VEGF to the receptor and to inhibit both VEGF-induced mitogenesis of human endothelial cells in vitro and tumor growth in vivo. Here we generated a series of KDR deletion mutants consisting of truncated extracellular regions and mapped out the domain(s) responsible for binding to VEGF and the neutralizing anti-KDR antibodies. All neutralizing antibodies were found to require domain 3 for efficient binding. Alanine-scanning mutagenesis of domain 3 identified two different sets of five residues, Ile(256), Asp(257), Glu(261), Leu(313), and Thr(315) and Tyr(262), Pro(263), Ser(264), Ser(265), and Lys(266), that were critical for binding to MAB383 and MAB664, respectively. Combination of alanine mutations affecting both MAB383 and MAB664 binding resulted in a variant that also lost binding to VEGF. These results suggest that the residues within this region of domain 3 are critical for VEGF binding. Our studies provide a basis for the mechanism of action of our anti-KDR antibodies and establish a functional foundation for the development of other classes of antagonists to the receptor.

Alanine↗
PubMed2000

Two cases of AML (M2) with a t(8;19)(q22;q13): a new cytogenetic variant.

"Simple" variants of the t(8;21) translocation involving chromosome 8 and a chromosome other than number 21 are rare. To our knowledge, only t(3;8)(q29;q22), t(8;11)(q22;q13), t(8;16)(q22;q24), t(8;20)(q22;p13), and t(8;22) have been reported in the literature. This paper describes for the first time two patients with acute myelogenous leukemia with a consistent t(8;19)(q22;q13) translocation. Their myelograms were compatible with the FAB-M2 subtype. The blasts from case 2 expressed CD34, CD33, CD13, and CD19. Karyotype analyses were performed on bone marrow cells using R- and G-banding at presentation. A t(8;19)(q22;q13) translocation was found in 28/30 metaphases for case 1 and in 23/25 metaphases for case 2. The latter case also had a deletion of chromosome 9, del(9)(q12q22) as an additional abnormality. Reverse transcriptase-polymerase chain reaction study revealed no AML1/ETO fusion transcript in case 2. Dual-color fluorescence in situ hybridization (FISH) assay using two probes (BAC92 and YAC412A4) convincingly demonstrated that the chromosomal material from 8q was translocated onto 19q rather than 19p in case 2. Thus, we consider t(8;19)(q22;q13) a true "simple" variant of t(8;21), and assume that a fusion gene resulting from the t(8;19) may contain the ETO gene located at 8q22 and an unknown partner gene from 19q13, which probably is a new transcription factor, whose molecular entity warrants further study.

Adolescent↗