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Biomedical subjects

D Zhu

Publications and source records attributed to D Zhu.

At least 127 records · Page 7Linked to original sources
PubMed1999

Automated purification and quantification of oligonucleotides.

We have developed automated methods for the trityl-on purification and quantification of synthetic oligonucleotides. Oligonucleotide purification is by solid-phase extraction cartridges using Amberchrom CG-50 resin on an XYZ-axis robotic system. Quantification is by OD260nm using an online UV-visible spectrophotometer with sipper. The purification of 20 oligonucleotides requires 5 min of user set-up time, plus 20 min per sample of robot time. For a 15-25-mer at the 40 nmol scale of synthesis, the method gives a yield of 2.8 ODs from a load of 10.1 OD, i.e., a 28% average yield. Oligonucleotides purified by this method have proven to be successful for primers for automated DNA sequencing.

Autoanalysis↗
PubMed1999

Nimodipine inhibits calcium-independent nitric oxide synthase activity in transient focal cerebral ischemia rats and cultured mouse astroglial cells.

The effect of nimodipine on nitric oxide synthase (NOS) activities in brains in transient focal cerebral ischemia rats, in cultured mouse neurons and astroglial cells and bovine brain capillary endothelial cells (BCECs) was investigated. The administration of nimodipine (3 mg.kg(-1), p.o., twice a day, for 3 days) before middle cerebral artery (MCA) occlusion significantly reduced infarct size, decreased nitrite/nitrate (NOx) content and inhibited Ca2+-independent NOS activity in the infarct area. Nimodipine inhibited the Ca2+-independent NOS activity induced by lipopolysaccharide (LPS) + tumor necrosis factor alpha (TNF alpha) in mouse astroglial cells with an IC50 value of 0.036+/-0.003 mM and Ca2+-dependent NOS activity in mouse neurons with an IC50 value of 0.047+/-0.003 mM, but did not affect Ca2+-dependent NOS activity in BCECs. The inhibition of Ca2+-independent NOS activity by nimodipine in astroglial cells was competitive with respect to L-arginine. Nimodipine also inhibited the induction of Ca2+-independent NOS activity in vitro. These results suggest that nimodipine in addition to its cerebral vasodilating effect may protect brain from ischemic neuronal damage through modifying NOS activity.

Animals↗
PubMed1999

DNA vaccines against haematological malignancies.

DNA vaccines against cancer have to activate an inadequate or damaged immune system in order to attack residual cancer cells. Although the potential problem of tolerance may be overcome by transplantation, provision of high levels of T-cell help is likely to be an important factor in stimulating effective immune pathways. The fusion gene approach appears to provide the required help, and offers a rational design for raising both antibody and T-cell mediated attack against lymphoma and myeloma, which express idiotypic antigen at the cell surface or as a secreted protein respectively. Intriguingly, preliminary data indicate that the fusion gene approach promotes antibody responses against a different cell surface tumour antigen, CEA. Strategies for using DNA vaccines to induce attack on processed peptides bound to MHC class I molecules are also being developed. We hope and anticipate that all categories of tumour antigen may be susceptible to this powerful new technology. The critical clinical requirement, however, will be to treat the presenting tumour with maintenance or restoration of immune capacity. We await results of the preliminary clinical trials with great interest.

Animals↗
PubMed1999

Enhancing effects of silkworm expressed recombinant human macrophage colony-stimulating factor on hematopoietic recovery of irradiation-injured mice.

OBJECTIVE: To study the hematopoietic enhancing effects of recombinant human macrophage colony-stimulating factor (rhM-CSF) expressed in silkworm. METHOD: Balb/c mice were irradiated with sublethal dose of 60 Co gamma-rays and then administered intraperitoneally with silkworm expressed rhM-CSF (1000 U per individual for 7 days) in treatment group or with normal saline (for 7 days as well) in control group. The hematopoietic recovery of irradiation mice was observed by comparing peripheral white blood cell (WBC) counts, differential counts of WBC and bone marrow hematopoietic progenitor cell colony forming assay in soft agar at different time after irradiation. RESULTS: The total WBC counts (x 10(9)/L) of treatment group at day 15 and 20 after irradiation(3.42 +/- 1.20, 5.56 +/- 2.50, respectively) were significantly higher than those of control group (2.03 +/- 0.90, 3.72 +/- 2.30; both P < 0.05). On days 10, 15, 20, 25 and 30 after irradiation, the monocyte counts (x 10(9)/L) of treatment group (0.08 +/- 0.06, 0.16 +/- 0.10, 0.48 +/- 0.35, 0.47 +/- 0.21 and 0.33 +/- 0.17, respectively) were all significantly higher than those of control group (0.025 +/- 0.016, 0.05 +/- 0.04, 0.23 +/- 0.16, 0.33 +/- 0.19 and 0.17 +/- 0.13; all P < 0.05). On days 15, 20 and 25, the granulocyte count (X 10(9)/L) of treatment group (1.03 +/- 0.61, 2.18 +/- 1.19 and 3.28 +/- 1.09) were also higher than those of control group (0.62 +/- 0.37, 1.40 +/- 0.99 and 2.20 +/- 0.74; all P < 0.05). On day 9 after irradiation, the bone marrow CFU-GM yield of control group (19 +/- 11/10(6) cells) was significantly lower than that of treatment group (78 +/- 30/10(6) cells, P < 0.05). CONCLUSION: rhM-CSF expressed in silkworm could accelerate hematopoietic recovery in irradiated mice.

Animals↗
PubMed1999

Serum leptin levels and adiposity in adult Chinese: a preliminary observation.

OBJECTIVE: To establish normal serum leptin levels in Chinese and investigate the relationship between serum leptin levels and body fat, gender, age and androgen. METHODS: Serum leptin levels were measured in 77 lean (BMI < 25) and 28 overweight or obese (BMI > or = 25) subjects by a radioimmunoassay (RIA) method. RESULTS: The serum leptin levels in lean Chinese were 2.15 +/- 1.46 ng/ml in male and 7.85 +/- 3.60 ng/ml in female, which are similar to those of Caucasians, while in overweight or obese ones, the levels were 4.87 +/- 3.47 ng/ml and 16.59 +/- 6.92 ng/ml respectively, lower than those in Caucasians. A 2-3 times higher leptin concentrations were found in women than in men in both conditions. Even when the number of lean males was expanded to 79 subjects aged from 17-80 years, no significant leptin-age relationship was found. Despite 25% of obese subjects manifested a relative deficiency of leptin, as a whole, leptin levels in both men and women were significantly correlated with BMI (r = 0.69, P < 0.001 in male and r = 0.63, P < 0.001 in female). CONCLUSIONS: Serum leptin levels in Chinese lean people are similar to those in Caucasians and in both lean and obese groups, the leptin levels are correlated with BMI, suggesting that the great majority of the obese patients are resistant to endogenous leptin. Those who are relatively deficient of leptin may become a group of good candidates for leptin treatment in the future.

Adult↗
PubMed1999

[Surveillance of bacterial resistance in Shanghai in 1998].

OBJECTIVE: To investigate the resistance of clinical bacterial isolates as a reference for rational use of antibiotics. METHODS: Using Kirby-Bauer method for bacterial susceptibility testing and NCCLS 1997 as assessment criteria. RESULTS: The number of Gram positive organisms was increasing in clinical isolates. Among the Staphylococci aureus (SA) and coagulase negative Staphylococci (CNS), methicillin resistant SA (MRSA) and methicillin resistant CNS (MRCNS) accounted for 70.7% and 54.9% respectively, but no vancomycin resistant strain was found. Vancomycin-resistant Enterococci was about 5%. Resistant rates of Klebsiella spp, Serratia spp, Enterobacter spp and Acinetobacter spp against third generation cephalosporins and fluoroquinolones increased markedly. The resistant rates of most Enterobacteriaceae species against imipenem were increasing gradually, while the resistant rates of Pseudomonas aeruginosa against ceftazidime, cefoperazone and imipenem were significantly higher as compared with the data in 1994. CONCLUSION: The urgent need of restricting the misuse of antibiotics.

Anti-Bacterial Agents↗
PubMed1999

[Determination of oleanolic acid and ursolic acid in spica Prunellae by derivative GC method].

OBJECTIVE: To develop a GC method to determine the oleanolic acid and ursolic acid in Spica Prunellae. METHOD: Before GC analysis, the sample was derivatized with CH2N2 solution. The GC conditions were as follows: comumn-10% SE-30(2m x 3mm) and column temperature -270 degrees C. RESULT: The two constituents were well separated and had good linearity in the range of 0.0025-0.4000 mg/ml. The average recoveries and RSD of oleanolic acid were 93.53% and 3.5%, 94.18% and 3.0% respectively. CONCLUSION: The method is good for determining oleanolic acid and ursolic acid in Chinese medicines.

Chromatography, Gas↗
PubMed1999

[Beaded molecule imprinted polymer for stereo isomer separation].

Beaded molecule imprinted polymer (MIP) was made by suspension polymerization. Particles with the size of 50-70 microns in diameter were collected and evaluated in HPLC mode to separate stereo isomers. Stereo isomers cinchonine and cinchonidine were successfully discriminated with selectivity factor of 2.89 and resolution factor of 0.76. Stereo selectivity of the MIP was found to come from both the interaction between the analyte and carboxyl group on the MIP and the similarity between the stereo structure of imprinted molecule and the MIP. The thermal analysis results showed that the MIP had high thermal stability with initial thermal decomposition temperature of 320 degrees C. The pore volume of the MIP was 0.1849 mL/g, the specific surface area was 126.84 sqm/g and the average pore diameter was 5.8 nanometer. Scanning electron microscopy showed that MIP had perfect spherical morphology.

Chromatography, High Pressure Liquid↗
PubMed1999

[Separation and characterization of oligodeoxynucleotide by fast protein liquid chromatography].

The antisense drug synthesized by DNA synthesizer is an oligodeoxynucleotide (ODN). The purity of the drug should be determined effectively. Because the ODNs are molecules with negative charges at pH 12, they can be separated according to the quantity of charges with them on the ion-exchange column. On anion exchange column, MONO-Q, the ODNs ranged from 19 bases to 21 bases can be separated successfully by gradient elution of NaCl solution from 0 mol/L to 1.8 mol/L. The experiment proved that the fast protein liquid chromatography is a very useful tool for the determination of the antisense drug.

Chromatography, Liquid↗
PubMed1999

[C677T genetic polymorphism of methylenetetrahydrofolate reductase in premature coronary heart disease].

OBJECTIVE: Methylenetetrahydrofolate reductase (MTHFR) is an important factor responsible for hyperhomocysteinemia. The relation of MTHFR gene C677T polymorphism and premature coronary heart disease was studied. METHODS: MTHFR C677T genetic polymorphisms in 67 patients with premature coronary heart disease were detected by PCR-RFLP technique. RESULTS: In case group, the frequency of T homogenic type was 34.3% (23/67), heterogenic type 43.3% (29/67) and C homogenic type 22.4% (15/67). T allele frequency was 55.9% (75/134) while C allele frequency 44.1% (59/134) in case group. There were significant differences in MTHFR genotype and allele frequencies between cases and controls (chi 2 = 6.82 and 5.41 respectively, P < 0.05). CONCLUSIONS: It was suggested that MTHFR gene C677T mutation was a possible risk factor of Chinese premature coronary heart disease.

Adult↗
PubMed1998

Molecular characterization of Saccharomyces cerevisiae Delta3, Delta2-enoyl-CoA isomerase.

We report here the identification of the Saccharomyces cerevisiae peroxisomal Delta3,Delta2-enoyl-CoA isomerase, an enzyme that is essential for the beta-oxidation of unsaturated fatty acids. The yeast gene YLR284C was identified in an in silico screen for genes that contain an oleate response element, a transcription factor-binding site common to most fatty acid-induced genes. Growth on oleic acid resulted in a significant increase in YLR284C mRNA, demonstrating that it is indeed an oleate-induced gene. The deduced product of YLR284C contains a type 1 peroxisomal targeting signal-like sequence at its C terminus and localizes to the peroxisome in a PEX8-dependent manner. Removal of YLR284C from the S. cerevisiae genome eliminated growth on oleic acid, but had no effect on peroxisome biogenesis, indicating a role for YLR284C in fatty acid metabolism. Cells lacking YLR284C had no detectable Delta3,Delta2-enoyl-CoA isomerase activity, and a bacterially expressed form of this protein catalyzed the isomerization of 3-cis-octenoyl-CoA to 2-trans-octenoyl-CoA with a specific activity of 16 units/mg. We conclude that YLR284C encodes the yeast peroxisomal Delta3,Delta2-enoyl-CoA isomerase and propose a new name, ECI1, to reflect its enoyl-CoA isomerase activity.

Amino Acid Sequence↗
PubMed1998

Kashin-Beck osteoarthropathy in rural Tibet in relation to selenium and iodine status.

BACKGROUND AND METHODS: Kashin-Beck disease is a degenerative osteoarticular disorder that is endemic to certain areas of Tibet, where selenium deficiency is also endemic. Because selenium is involved in thyroid hormone metabolism, we studied the relation among the serum selenium concentration, thyroid function, and Kashin-Beck disease in 575 subjects 5 to 15 years of age in 12 villages around Lhasa, Tibet, including 1 control village in which no subject had Kashin-Beck disease. Clinical, radiologic, and biochemical data were collected. RESULTS: Among the 575 subjects, 280 (49 percent) had Kashin-Beck disease, 267 (46 percent) had goiter, and 7 (1 percent) had cretinism. Of the 557 subjects in whom urinary iodine was measured, 66 percent had a urinary iodine concentration of less than 2 microg per deciliter (157 nmol per liter; normal, 5 to 25 microg per deciliter [394 to 1968 nmol per liter]). The mean urinary iodine concentration was lower in subjects with Kashin-Beck disease than in control subjects (1.2 vs. 1.8 microg per deciliter [94 vs. 142 nmol per liter], P<0.001) and hypothyroidism was more frequent (23 percent vs. 4 percent, P=0.01). Severe selenium deficiency was documented in all villages; 38 percent of subjects had serum concentrations of less than 5 ng per milliliter (64 nmol per liter; normal, 60 to 105 ng per milliliter [762 to 1334 nmol per liter]). When age and sex were controlled for in a multivariate analysis, low urinary iodine, high serum thyrotropin, and low serum thyroxine-binding globulin values were associated with an increased risk of Kashin-Beck disease, but a low serum selenium concentration was not. CONCLUSIONS: In areas where severe selenium deficiency is endemic, iodine deficiency is a risk factor for Kashin-Beck disease.

Adolescent↗
PubMed1998

CD44 isoform-cytoskeleton interaction in oncogenic signaling and tumor progression.

CD44, a major hyaluronan receptor, exists as several isoforms and is widely distributed in different cells and tissues. The isoforms of CD44, such as CD44s (the standard form), CD44E (the epithelial form) and CD44v (variant isoforms) (arise from differential splicing of one to ten (or eleven) variable exons that encode portions of the membrane proximal extracellular domain. The molecular diversity of CD44 isoforms is further compounded by differential biosynthetic processes and post-translational modifications [e.g. N-/O-glycosylation or glycosaminoglycan (GAG) addition]. This structural arrangement, which occurs within either the invariant region or the extracellular domain of the variant region, is important for CD44-mediated communication between extracellular matrix materials [ECM-hyaluronic acid (HA), collagen and fibronectin] and intracellular protein components (e.g cytoskeletal proteins and various regulatory enzymes). The 15 amino acid sequence [e.g. NSGNGAVEDRKPSGL (in human) or NGGNGTVEDRKPSEL (in mouse)] residing in the cytoplasmic domain of CD44 isoforms is the ankyrin-binding domain of this family of transmembrane glycoproteins. Biochemical analyses plus in vitro mutagenesis indicate that the ankyrin-binding domain is required for CD44-mediated "outside-in" and "inside-out" cell activation events. Furthermore, CD44s-cytoskeleton interaction is tightly coupled with signal transducing molecules (e.g. p185HER2 or Src kinases) during oncogenic signaling. Moreover, the transmembrane linkage between CD44v isoforms (CD44v10 and CD44v3) and the cytoskeleton up-regulates invasive and metastatic-specific tumor phenotypes [e.g. matrix degradation (MMPs) activities, tumor cell invasion and migration]. These findings strongly suggest that the interaction between CD44 isoforms and the cytoskeleton plays a pivotal role in the onset of oncogenesis and tumor progression.

Animals↗
PubMed1998

Analysis of VH genes in follicular and diffuse lymphoma shows ongoing somatic mutation and multiple isotype transcripts in early disease with changes during disease progression.

Investigations of VH gene mutational patterns in B-cell tumors are often performed at an arbitrary time point of disease. To assess the effects of disease progression, tumor-derived VH genes have been monitored from presentation through treatment and relapse in one patient with follicle center lymphoma (FCL), and two patients with primary diffuse large B-cell lymphoma (DLCL). The patient with FCL and one patient with DLCL both achieved clinical remission, although this was only partial in the FCL. However, both subsequently relapsed, and the second patient with DLCL was refractory to radiotherapy and chemotherapy. In each case, the tumor-derived VH sequence was identified, and the CDR3 "clonal signature" was used to track tumor cell sequences in subsequent biopsies. All cases showed somatic mutations, with intraclonal heterogeneity evident at presentation, and some sequences were aberrant. The VH sequences of the DLCL which responded to treatment became homogeneous at relapse. The sequences of both the FCL and the refractory DLCL remained heterogeneous. In all cases, transcripts of multiple Ig isotypes could be identified, and there was immunophenotypic evidence for expression of several Ig isotypes. The case of refractory DLCL had identifiable transcripts from IgM, IgD, IgA, IgG, and IgE, but appeared to lose the ability to produce alternative isotype transcripts and protein at the late stage of disease. These cases indicate that VH gene analysis can be used to probe tumor cell behavior in cases of lymphoma and that perturbations caused by therapy and disease progression can occur.

Amino Acid Sequence↗
PubMed1998

Analysis of the forces which stabilize the active conformation of urokinase-type plasminogen activator.

It was recently proposed that hydrophobic interactions control the active conformation of serine proteases in the trypsin family (Hedstrom et al. (1996) Biochemistry 35, 4515-23) rather than a charge interaction with Asp next to the active site Ser, as formerly believed. In the present study, certain site-directed mutants of the serine protease zymogen pro-urokinase (pro-UK) and its two-chain enzymatic derivative urokinase (UK) were characterized. The results provide information on the structure-function of the catalytic domain of pro-UK/UK, which is relevant to this controversy. Mutations at Asp355(c194), which eliminated its charge, induced a 6250-fold reduction in the catalytic activity of UK. By contrast, reducing the hydrophobicity at the neoterminal Ile159(c16) of UK had relatively little effect. However, when both the hydrophobicity and the size of the side chain were reduced by a glycine substitution at this position, a major reduction (9090-fold) in the catalytic efficiency of UK occurred. This effect was related to the smaller side chain increasing the cavity and the flexibility of the N-terminus and thereby interfering with its charge interaction with Asp355(c194). A similar mechanism, rather than a change in hydrophobicity, is believed also to explain the reduction in the stabilization energy of the activation domain observed in a trypsin mutant by Hedstrom et al. (1996). Although hydrophobic interaction facilitated the charge interaction with Asp355(c194), the latter was the primary force which stabilized the active conformation of UK. The charge interaction with Asp355(c194) was also found to be the principal determinant of the intrinsic catalytic activity of single-chain pro-UK. Additionally, the findings confirmed that the KM of pro-UK for its natural substrate was significantly lower than that of UK. Since this same phenomenon was also seen with each of the mutants, the substrate binding pocket of these single-chain zymogens was better formed than that of their two-chain, enzymatic derivatives.

Aspartic Acid↗