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Biomedical subjects

H Chen

Publications and source records attributed to H Chen.

At least 361 records · Page 20Linked to original sources
PubMed2001

Construction of the enhanced yellow fluorescent protein expression vector carrying IFN-gamma gene.

PURPOSE: To construct the enhanced yellow fluorescent protein (EYFP) vector carrying interferon-gamma gene (ifn-gamma) in order to provide an ideal reporter in the expression of ifn-gamma and location of protein in vitro and in vivo. METHOD: According to the nucleotide sequence of ifn-gamma gene, a pair of oligonucleotides was designed as primer whose two end contained nucleotide sequence of EcoR V and Not I restriction endonuclease respectively. The gene encoding for inf-gamma was amplified using PCR technqiue. After the PCR product was retrieved and purified, it was digested with EcoR V and Not I restriction endonuclease, and then cloned into the plasmid pIRES-EYFP. The recombinant plasmid pIRES-EYFPIFN-gamma was identified by restriction endonuclease enzyme analysis and DNA sequence analysis. RESULTS: The ifn-gamma was successfully amplified and verified by partial DNA sequence analysis. The recombinant plasmid was correctly screened. CONCLUSION: The EYFP expression vector carrying ifn-gamma gene was successfully established. This research work has formed a base for monitoring the ifn-gamma gene expression and protein position in living cells.

Bacterial Proteins↗
PubMed2001

[Growth and osteogenesis characteristics of cultured rat marrow stromal cells under bone induction condition].

OBJECTIVE: To investigate the growth and osteogenesis characteristics of cultured rat marrow stromal cells(rMSCs) under bone induction condition. METHODS: rMSCs were isolated from adult rat using density gradient separation method. The rMSCs attachment formed soon after seeding and grew into colonies with the appearance of fibroblastic cells. The osteogenic induction compound of Dex(10(-8) mol/L), beta-GP(10 mmol/L) and AA(50 micrograms/ml) was added to different passaged rMSCs and the proliferation and osteogenic differentiation of them was observed. RESULTS: The induction compound had strong effect on promoting proliferation, especially on that of further passaged rMSCs, and its stimulation effect on osteogenesis was also well proved with the expression of alkaline phosphatase (ALP) rising after one-week induction. Further subculturing from P1 to P6 led to the increase of proliferation activity of rMSCs. After three-week induction, mineral deposits appeared in the culture. CONCLUSION: The lower expression of ALP in P1 rMSCs and the increase of it in further passages or under induction condition suggest the rMSCs in our culture system are mainly undifferentiated osteoprogenitors.

Alkaline Phosphatase↗
PubMed2001

[Study on the construction of DYS385 allelic ladder and the genotype distribution in two populations].

OBJECTIVE: We have designed a new method to produce standard DYS385 allelic ladder in order to solve the problem of the accuracy and standardization of STR-PCR typing in forensic practice. METHODS: Nine different PCR amplified DYS385 allelic fragments were isolated from the gel, eluted into the distilled water and reamplified by PCR. The purified allelic fragments were then blunt-end subcloned individually into the pUC plasmid vectors and transfected into competent E. coli DH5 alpha cells. The sequencing results confirmed that the size and the constructure of the inserts were correct. The recombinant plasmids DNA with 9 inserts were then used as templates for reamplification to generate DYS385 standard ladder. RESULTS: A large quantity of standard DYS385 allelic ladder was obtained, with which the genetic polymorphisms of DYS385 locus both in Chinese Han and German populations were studied. CONCLUSION: The STR standard ladder constructed by this method has high value in the forensic application, and the DYS385 locus is robust for forensic analysis.

Alleles↗
PubMed2001

[Effects of estradiol and isoflavoid on the expression of adhesion molecules on neutrophils].

OBJECTIVE: To elucidate the effects of estradiol and isoflavoid on the expression of adhesion molecules on neutrophils. METHODS: Neutrophils of healthy subjects and ischemic stroke patients were treated with tumor necrosis factor alpha (TNF alpha), in the presence or absence of different concentrations of isoflavoid (WZ1, WZ2) and estradiol (WZ3, WZ4). Flow cytometry was used to detect the expression of CD18 and CD62L on neutrophil surface. RESULTS: 1. 10 ng/ml TNF alpha could activate neutrophils of healthy subjects; it increased the expression of CD18 by 10% on neutrophil surface and shed CD62L from the surface as shown by a 15% decrease of the fluorescence intensity and a 30% decrease of the percentage of positive cell. 2. Isoflavoid (WZ1, WZ2) had no significant effect on the expression of CD18 and CD62L on neutrophils. 3. Pretreatment of neutrophils with estradiol (WZ3, WZ4) could inhibit the activation of neutrophils by TNF alpha, which decreased the fluorescence intensity of CD18 by 8%, increased the fluorescence intensity of CD62L by 15% and increased the percentage of CD62L positive cell by 20%. 4. TNF alpha could activate the neutrophils of ischemic stroke patients strikingly; it increased the fluorescence intensity of CD18 by 20% and decreased the fluorescence intensity and percentage of positive cell of CD62L by 30%, and there was a significant difference when the patients were compared with the healthy subjects. Estradiol had the same effect on the expression of CD18 and CD62L as on those of healthy subjects. CONCLUSION: 1. TNF alpha is a strong activator of neutrophils; it plays an important role in the occurrence and development of ischemic stroke. 2. Isoflavoid has no obvious effect on the expression of adhesion molecules on neutrophils, so its role in protecting the cardiovascular system may come into play not by the way of affecting adhesion molecules expression. 3. Pretreatment of neutrophils with estradiol could protect them from activation by TNF alpha, thus decreasing the expression of adhesion molecules, the adhesion of neutrophils -endothelial cells, and hence the risk of ischemic stroke. However, in case that the neutrophils have been activated by TNF alpha, estradiol has no effect on the expression of adhesion molecules; this implies that estradiol possibly has no valid anti-adhesion therapeutic effect on ischemic stroke.

Adult↗
PubMed2001

Effects of the HOE 694 on transient inward current and Na(+) - Ca(2+) exchange in guinea pig cardiomyocytes.

OBJECTIVE: To determine the effects of HOE 694, a new and potent Na(+)-H+ exchanger blocker, on transient inward current (It(i)) and Na(+) - Ca(2+) exchange during hypoxia-reoxygenation in guinea pig cardiomyocytes. METHODS. Cardiomyocytes were isolated from adult guinea pig ventricle. Experiment was performed in an experimental chamber that allowed the cells to be exposed to a sufficiently low O2 pressure. The cells were subjected to hypoxia and reoxygenation. The ionic currents were studied with patch clamp technique. RESULTS: In the absence of HOE 694, hypoxia-reoxygenation induced It(i) in 12 of 15 experiments; but in cardiomyocytes pretreated with HOE 694 (10 approximately 50 micro mol/L), the incidence of It(i) observed during reoxygenation was reduced to 5 of 11 experiments and 3 of 10 experiments, P < 0.05 vs control respectively. The Na(+) - Ca(2+) exchange current was unaffected by HOE 694 under normoxic condition. However, when cells were pretreated with 10 micromol/L HOE 694 for 10 min, then subjected to hypoxia condition, the Na(+) - Ca(2+) exchange current significantly inhibited. CONCLUSIONS: Blockade of the Na(+)-H+ exchange by HOE 694 could reduce Ca(2+) overload upon hypoxia-reoxygenation, and inhibition of Na(+) - H+ exchange may also indirectly decrease Na(+) - Ca(2+) exchange activity during hypoxia.

Animals↗
PubMed2001

[Luminescence dependence upon concentration and temperature in YPO4:Pr3+].

Under selective excitation 3P0 state of Pr3+ ion in YPO4, the emission spectra and fluorescence decay curves are measured at different concentrations and temperatures. The origin of the spectral line located at 613.2 nm has been discussed and assigned to be the 1D2 (gamma 3)-->3H4 (gamma 5) transition. The different concentration dependence of emission intensities of levels 3P0 and 1D2 has been studied. Using static model, the fluorescence decay curve of 1D2 level is fitted, and the result shows that the cross-relaxation caused by the interaction of dipole-quadrupole between adjacent Pr3+ ions results in the concentration quenching.

Attention↗
PubMed2001

[Characteristics in frequency domain by inductively coupled plasma atomic emission spectrometry with charge injection detection].

The characteristics of two-dimension spectra obtained by inductively coupled plasma atomic emission spectrometry (ICP-AES) with charge injection detection (CID) in frequency domain were studied in the present paper. The measurement spectra were Fourier transformed and the frequency distribution of the spectra was obtained. Results showed that the spectra in frequency domain could be divided into two parts: high frequency and low frequency signals. The later stood for measurement spectra and the former for background and noises. However, the high frequency signals could not be smoothed simply to reduce noises because the background was deteriorated even though the spectral signal did not change significantly.

Mass Spectrometry↗
PubMed2001

[Synthesis of nanocrystalline Y2O3:Eu3+ and study on spectral characteristics].

Nano-Y2O3:Eu3+ was synthesized with oxalic acid as precipitate agent while adding little surfactant controlling the condition of the reaction. We obtained nanoscale powder. The particle diameter was 15-19 nm. The agglomerate size was less 1.0 micron. Compared with the ordinary powder of Y2O3:Eu3+, the peak of emission spectrum of nano-Y2O3:Eu3+ is at lambda em = 612 nm under 254 nm excitation, blue-shift 6 nm. Excitation spectrum has no difference. The phosphor was measured by PMS-3 chromatic detector and its luminescent chromatic coordinate parameters were x = 0.6479, y = 0.3442. The quenching concentration increases from micro-Y2O3:Eu3+'s 6% to 9% and the intensity of luminescence increases with the grain size growing.

Crystallization↗
PubMed2001

[Diagnosis and treatment of congenital biliary duct cyst: twenty-year experience].

OBJECTIVE: To summarize the experience in diagnosis and treatment of congenital biliary duct cyst. METHODS: Clinical data from 108 patients treated from 1980 to 2000 were analyzed retrospectively. RESULTS: Abdominal pain, jaundice and abdominal mass were presented in most pediatric patients. Clinical symptoms in adult patients were non-specific, resulting in delayed diagnosis frequently. Fifty-seven patients (52.7%) had coexistent pancreatic biliary disease. Carcinoma of the biliary duct occurred in 18 patients (16.6%). Ultrasonic examination was performed in 94 patients, ERCP in 46, and CT in 71. All of the patients were correctly diagnosed before operation. Abnormal pancreatobiliary duct junction was found in 39 patients. Before 1985, the diagnosis and classification of congenital biliary duct cyst were established by ultrasonography preoperatively and confirmed during operation. The main procedure was internal drainage by cyst-enterostomy. After 1985, the diagnosis was decided with ERCP and CT, the procedure was cyst excision with Roux-en-Y hepaticojejunostomy. In 1994, we used a new and simplified operative procedure to reduce the risk of malignancy of choledochal cyst. Retrograde infection of the biliary tract the major postoperative complication, could be controlled by the administration of antibiotics. CONCLUSIONS: The concept in diagnosis and treatment of congenital choledochal cyst has been changed greatly. CT and ERCP are of great help in the classification of the disease. Currently, cyst excision with Roux-en-Y hepaticojejunostomy is strongly recommended as the treatment of choice for patients with type I and IV cysts. Piggyback orthotopic liver transplantation is indicated for type V cysts (Caroli's disease) with frequently recurrent cholangitis, resulting in biliary cirrhosis.

Adolescent↗
PubMed2001

[Diagnosis and treatment of paragangliomas in the central nerve system].

OBJECTIVE: To explore the diagnosis and treatment of paragangliomas in the central nerve system. METHODS: 17 patients with paragangliomas in the central nerve system over a 20-year period (1976-1999) were analyzed retrospectively. They included 11 patients with glomus jugular tumors, 1 with carotid body tumor, 1 with supersellar paraganglioma, 2 with spinal involvement, and 2 with superficial hemisphere involvement. RESULTS: Neoplasm was completely resected in 11 patients, subtotally in 3, and partial in 3. The postoperative mortality was 1/17. One patient remained severe disability, yet the others had excellent long-time outcome. CONCLUSIONS: Both the characteristic clinical presentation and neuroradiological manifestation were useful for preoperative diagnosis. Total resection is ideal for paragangliomas. Preoperative embolization should be used routinely to control intraoperative bleeding, and postoperative radiation-therapy would be valuable to control the recurrence of the remaining tumor.

Adult↗
PubMed2001

[Percutaneous radiofrequency ablation of liver malignancies].

OBJECTIVE: To describe the safety and efficacy of percutaneous radiofrequency ablation (PRFA) to treat liver cancer. METHODS: One hundred patients with histologically or clinically confirmed primary or metastatic malignancies were treated percutaneously under ultrasound guidance using LeVeen multipolar array needle electrode and RF 2000 generator between October, 1999 and October, 2000. All patients were followed up to assess complication, treatment response, recurrence and curative effect. RESULTS: PRFA was performed in 76 patients with primary liver cancer and 24 patients with metastatic liver cancer. AFP positive of small hepatic cancer (inoperative) became AFP negative in 75.0% patient (21/28). The rate of completely concrete necrosis of small hepatic malignancies shown by MRI was 85.9%(61/71). CONCLUSIONS: PRFA as a local thermal therapy is a kind of new palliative treatment for hepatic malignancies. It is minimally invasive, safe and effective especially for small hepatic tumors and can also be used in conjunction with TACE for larger ones.

Adult↗
PubMed2001

[Administration of leukemic granulocyte colony-stimulating factor to donors: the changes in donor bone marrow cell composition and the transplantation outcome].

OBJECTIVE: To investigate the change of bone marrow cell composition the effect of graft on hemopoietic reconstitution and the incidence of acute graft-versus-host disease (aGVHD) with granulocyte colony-stimulating factor G-CSF administration to donors before marrow harvesting. METHODS: Twenty-four patients with myeloid leukemia underwent allogeneic bone marrow transplantation. The donors of twelve patients (study group) were given G-CSF 250 microg/d for seven days prior to harvesting. The marrow cell composition, the outcome of subsequent engraftment and the occurrence of aGVHD were compared with those in twelve patients without G-CSF (control group). RESULTS: The bone marrow stimulated by G-CSF yielded higher number of nucleated cells, CD34+ cells, CFU-GM and CFU-MK (P < 0.01). The number of T lymphocyte subsets in the graft stimulated with G-CSF was different from that in the control group. The number of CD4+ cells was decreased and that of CD8+ increased. The CD4+/CD8+ ratio was reduced markedly (P < 0.01). The change of CD34+, CD4+, CD8+ percentage and CFU-GM, CFU-MK proliferation in donor bone marrow with and without G-CSF was compared. The same characteristic change as above graft was observed. Hemopoietic reconstitution in the study group after allo-BMT was accelerated. In the study group the days of granulocyte count exceeding 0.5 x 10(9)/L and platelet count exceeding 20 x 10(9)/L were 16 days (range 11 - 23 days) and 17 days (range 14 - 25 days) (in the control group 20.5 day, range 14 - 29 days and 23 days, range 17 - 32 days P < 0.05). Grade II - IV aGVHD did not occur in patients of the study group but in 3 patients of the control group. CONCLUSION: The bone marrow harvested from donor with G-CSF stimulation can accelerate hemopoietic recovery and decrease the incidence of severe aGVHD. The accelerated hematopoietic recovery is associated with increased number of CD34+, CFU-GM and CFU-MK in the graft.

Adolescent↗
PubMed2000

Roles of Se and NO in apoptosis of hepatoma cells.

Mice inoculated with hepatoma cell (H22) suspension subcutaneously at their right axilla were administered orally with kappa-selenocarrageenan (Se) solution, the inoculated hepatoma's growth was suppressed. Different concentrations of Se solution added in human hepatoma cell line culture could inhibit proliferation and induce apoptosis in hepatoma cells. Meanwhile Se solution could increase the activity of glutathione peroxidase (GSH-Px) in the mice's plasma and the content of NO in the mice's sera and the hepatoma cell culture supernatant as well. Therefore, apoptosis in hepatoma cell induced by Se solution may be associated with the increase in antioxidative activity, the suppression free radical's intervention, and the excessive release of NO by stress.

Animals↗
PubMed2000

Acyl-CoA hydrolysis by the high molecular weight protein 1 subunit of yersiniabactin synthetase: mutational evidence for a cascade of four acyl-enzyme intermediates during hydrolytic editing.

Yersiniabactin (Ybt) synthetase is a three-subunit, 17-domain [7 domains in high molecular weight protein (HMWP)2, 9 in HMWP1, and 1 in YbtE] enzyme producing the virulence-conferring siderophore yersiniabactin in Yersinia pestis. The 350-kDa HMWP1 subunit contains a polyketide synthase module (KS-AT-MT(2)-KR-ACP) and a nonribosomal peptide synthetase module (Cy(3)-MT(3)-PCP(3)-TE). The full-length HMWP1 was heterologously overexpressed in Escherichia coli and purified to near homogeneity. The purified HMWP1 showed thioesterase activity toward acyl-CoAs, such as acetyl-CoA, benzoyl-CoA, and malonyl-CoA, with saturation kinetics and relative catalytic efficiencies of 172:50:1. A chain-releasing thioesterase (TE) activity is ascribed to the C-terminal TE domain, and this was substantiated by the fact that acyl-N-acetylcysteamines were hydrolyzed by the didomain PCP(3)-TE fragment of HMWP1. However, PCP(3)-TE failed to hydrolyze any of the acyl-CoAs, suggesting the TE domain does not recognize CoA moiety, thus the acyl-CoA hydrolysis by HMWP1 must involve other domains. Ser-to-Ala mutants in each of the AT, ACP, PCP(3), and TE domains reduced hydrolysis rates of the two fastest substrates, acetyl-CoA and benzoyl-CoA, by more than two orders of magnitude. Thus, the acyl-CoA hydrolysis activity requires 4 of the 9 domains of HMWP1, and it is consistent with autoacylation of the AT domain active site serine and subsequent passage of the itinerant acyl chain from AT to ACP to PCP(3) to the TE domain, a cascade of four sequential acyl-enzyme intermediates, for hydrolytic turnover. This could represent an editing pathway for this polyketide synthase/nonribosomal peptide synthetase assembly line.

Acyl Coenzyme A↗
PubMed2000

Expression and fast-flow purification of a polyhistidine-tagged myoglobin-like aerotaxis transducer.

A Co(2+)-affinity, fast-flow perfusion chromatography method to purify a polyhistidine-tagged myoglobin-like aerotaxis transducer HemAT-Hs has been developed. The method relies upon a six-histidine affinity tag fused to the C-terminus and N-terminus of HemAT-Hs for expression in the native host, an extremely halophilic Archaeon Halobacterium salinarum, and in the heterologous host Escherichia coli, respectively. The His-tagged HemAT-Hs can be purified rapidly using either low or high ionic strength buffers. Purified His-tagged HemAT-Hs in high or low salt buffers demonstrated no difference in spectral characteristics and retained reversible oxygen binding capacity. This fast-flow Co(2+)-affinity perfusion chromatography provides a simple method for preparation of halophilic heme containing soluble proteins for biophysical and structural studies.

Archaeal Proteins↗
PubMed2000

Sequence and analysis of chromosome 1 of the plant Arabidopsis thaliana.

The genome of the flowering plant Arabidopsis thaliana has five chromosomes. Here we report the sequence of the largest, chromosome 1, in two contigs of around 14.2 and 14.6 megabases. The contigs extend from the telomeres to the centromeric borders, regions rich in transposons, retrotransposons and repetitive elements such as the 180-base-pair repeat. The chromosome represents 25% of the genome and contains about 6,850 open reading frames, 236 transfer RNAs (tRNAs) and 12 small nuclear RNAs. There are two clusters of tRNA genes at different places on the chromosome. One consists of 27 tRNA(Pro) genes and the other contains 27 tandem repeats of tRNA(Tyr)-tRNA(Tyr)-tRNA(Ser) genes. Chromosome 1 contains about 300 gene families with clustered duplications. There are also many repeat elements, representing 8% of the sequence.

Arabidopsis↗
PubMed2000

X-Ray emission following low-energy charge exchange collisions of highly charged ions

K-shell x-ray emission following low-energy charge exchange collisions ( </=11 eV/amu) has been measured for ten bare and hydrogenic ions up to U91+. The data resolve capture into angular momentum states with l = 1 and provide a stringent test of theory. The assumption of Stark mixing of angular momentum states is inappropriate at such energies. Results from detailed calculations give a better description of the data, but significant discrepancies are noted. The measurements show that the hardness ratio of charge-exchange induced K-shell x-ray emission represents a diagnostic of the dynamics of ion-atom interactions in situations where the energy is below 1 keV/amu, e.g., in solar wind collisions.

Journal Article↗