Biomedical subjects

H Zhou

Publications and source records attributed to H Zhou.

At least 217 records · Page 12Linked to original sources

PubMed2001

[The expression of growth factors in filtering area following sclerectomy and the interaction between growth factors and interferon].

OBJECTIVE: To study the expression of beta-Fibroblastic Growth Factor (bFGF) and Transforming Growth Factor beta 1 (TGF beta 1) in filtering area following sclerectomy and the interaction between growth factors and interferon alpha-2b (IFN alpha-2b). METHODS: Immunohistochemical technique were applied to test the expression bFGF and TGF beta 1 in frozen sections of filtration area following sclerectomy on postoperative day three, five, seven and 14 in white rabbits, respectively. 16 eyes of eight rabbits in all were randomly and equally divided into two groups. The experimental eyes had subconjunctivally been administered IFN alpha-2b 5 x 10(5) IU 0.2 ml each time at filtering bleb following operation whereas the control eyes hadn't taken. RESULTS: In control group, positive cell and substance expressing bFGF and TGF beta 1 on postoperative day three were revealed in the filtration area, on postoperative day five and seven, they were gradually increased. On the postoperative day 14, all significant cells and substance markly decreased. In experimental group only small amount of positive cell and bFGF or TGF beta 1-expressing substance were sparely distributed in the filtration area. CONCLUSION: The filtration area after glaucoma filtering surgery was "bathed" in growth factors such as bFGF and TGF beta 1 which well-known play an important role in stimulating wound healing response, IFN alpha-2b had an inhibitory effect on bFGF or TGF beta 1 expression in wound environment.

Animals↗

PubMed2001

[Ethyl acetate synthesis on-line measurement by Raman spectroscopy].

We report an on-line Raman spectroscopy measurement with fiber probe. In the experiment, the real-time spectrum measurement of ethyl acetate synthesis reaction with ethanol and acetic acid was carried out by using a homemade Raman spectrometer. The time variation of integrated spectrum peak intensities of reaction product and raw materials are obtained. From the integrated peak intensities, the concentrations of the reactants can be deduced and the ending point of reaction under this experimental condition can be determined.

Acetates↗

PubMed2001

[Color reaction of chromogenic reagent O-hydroxybenzenediazoaminoazobenzene with thallium (III) and its application].

The chromogenic reaction of the O-hydroxybenzenediazoaminoazobenzene (O-HDAA) with thallium was studied in this paper. In the presence of Triton X-100 and SDBS and in ammonia medium of 0.54-1.1 mol.L-1, O-HDAA reacts with thallium (III) to form a stable red complex with molar radio of 1:5. The apparent molar absorptivity is 1.4 x 10(5) L.mol-1.cm-1 at the maximum absorption wavelength of 520 nm. Beer's law is obeyed in the concentration range of 0-600 micrograms.L-1 Tl(III). The method has been applied to the direct determination of trace amounts of thallium in waste water and in geological samples by using polyurethane foam to absorb and separate Tl(III) with satisfactory results.

Color↗

PubMed2001

[Expression of D-animo acid oxidase gene in K562 cells and the cytotoxicity of D-alanine to the cells].

OBJECTIVE: To explore the feasibility of expression of R. gracilis D-amino acid oxidase(DAAO) gene in leukemia cell line K562 and the cytotoxicity of D-Alanine to the cells. METHODS: DAAO cDNA was cloned into retroviral vector pLSN and pLDAAOSN was generated. The vector was then packaged with phiNXA and the virus titer was measured with NIH3T3 cells. Leukemia cell line K562 was infected with the viral supernatant. The positive clones were obtained by G418 selection and named KDAAO. PCR and in situ hybridization were used to identify the integration and expression of DAAO gene in KDAAO. KDAAO was treated with different concentrations of D-Alanine. RESULTS: pLDAAOSN was confirmed containing the full-length of DAAO cDNA. Infectious titer generated by the packaging cells was 5.2 x 10(6) cfu/ml. PCR and in situ hybridization analysis showed integration of DAAO gene in KDAAO and expression of DAAO mRNA. Preliminary observation suggested that D-Ala could effectively kill KDAAO. CONCLUSION: DAAO/D-Ala suicide gene system might be useful in cancer gene therapy.

Alanine↗

PubMed2001

[The impact of arsenic trioxide or all-trans retinoic acid treatment on coagulopathy in acute promyelocytic leukemia].

OBJECTIVE: To study the effect of arsenic trioxide (As2O3) or all-trans retinoic acid (ATRA) on coagulopathy in patients with acute promyelocytic leukemia (APL), and the mechanism of hemorrhage in these patients. METHODS: Thrombomodulin (TM) or tissue factor (TF) transcription of mRNA of freshly isolated bone marrow blast from APL patients was detected by semi-quantitative RT-PCR. The parameters of coagulation and cell procoagulation activity (PCA) were assessed in plasmic levels. Bleeding symptom was observed during As2O3 or ATRA treatment. RESULTS: TM expression in the APL cell surface was significantly upregulated from (14.31 +/- 1.60) ng/10(7) to (21.61 +/- 6.82) ng/10(7) cells. The levels of P-selectin, soluble fibrin monomer complex (SFMC) and D-dimer (D-D) decreased after ATRA or As2O3 treatment. Abnormal high expression of TF in APL cell was downregulated in patients treated with ATRA or As2O3. The expression level was (14.81 +/- 6.23) ng/L before treatment, but undetected after 20 days of treatment. In addition, the membrane PCA of fresh APL cells was predominantly FVII-dependent after ATRA or As2O3 treatment. Bleeding symptom was ameliorated during As2O3 or ATRA treatment. CONCLUSION: Bleeding symptom was controlled in patients with APL after As2O3 or ATRA treatment.

Adult↗

PubMed2000

Immunoassay of the MRSA-related toxic protein, leukocidin, with scanning electrochemical microscopy.

Scanning electrochemical microscopy (SECM) was applied to the immunoassay of leukocidin, which is a toxic protein produced by methicillin-resistant Staphylococcus aureus (MRSA), with the intention of developing and early diagnostic for MRSA infection. An antibody-chip for leukocidin was prepared by self-assembling of anti-leukocidin on a protein A-coated glass substrate. A sample solution containing leukocidin was spotted onto the antibody-chip, followed by labeling with horseradish peroxidase (HRP) via a sandwich method. The reduction current of the oxidized form of ferrocenylmethanol generated by the HRP reaction was monitored to view SECM images of the spot of captured leukocidin. The amplitude of reduction current depended on the concentrations of sample solutions used for making spots. This SECM-based immunoassay detects as low as 5.25 pg mL(-1) leukocidin.

Electrochemistry↗

PubMed2000

Akt regulates cell survival and apoptosis at a postmitochondrial level.

Phosphoinositide 3 kinase/Akt pathway plays an essential role in neuronal survival. However, the cellular mechanisms by which Akt suppresses cell death and protects neurons from apoptosis remain unclear. We previously showed that transient expression of constitutively active Akt inhibits ceramide-induced death of hybrid motor neuron 1 cells. Here we show that stable expression of either constitutively active Akt or Bcl-2 inhibits apoptosis, but only Bcl-2 prevents the release of cytochrome c from mitochondria, suggesting that Akt regulates apoptosis at a postmitochondrial level. Consistent with this, overexpressing active Akt rescues cells from apoptosis without altering expression levels of endogenous Bcl-2, Bcl-x, or Bax. Akt inhibits apoptosis induced by microinjection of cytochrome c and lysates from cells expressing active Akt inhibit cytochrome c induced caspase activation in a cell-free assay while lysates from Bcl-2-expressing cells have no effect. Addition of cytochrome c and dATP to lysates from cells expressing active Akt do not activate caspase-9 or -3 and immunoprecipitated Akt added to control lysates blocks cytochrome c-induced activation of the caspase cascade. Taken together, these data suggest that Akt inhibits activation of caspase-9 and -3 by posttranslational modification of a cytosolic factor downstream of cytochrome c and before activation of caspase-9.

Apoptosis↗

PubMed2000

Effect of antibiotics on development in vitro of hamster pronucleate ova.

Antibiotics are commonly added to embryo culture media, but effects on embryo development have not been examined thoroughly. Hamster ova were used to investigate whether penicillin, streptomycin or gentamicin affect embryo development in vitro. Ova were collected 10 h post activation by spermatozoa in vivo and cultured in five treatments: 1) CONTROL: chemically-defined medium HECM-9 with no antibiotics; 2) HECM-9 with 100 IU/mL penicillin; 3) HECM-9 with 50 microg/mL streptomycin; 4) HECM-9 with 10 microg/mL gentamicin and 5) HECM-9 with both 100 IU/mL penicillin and 50 microg/mL streptomycin. Individually, penicillin, streptomycin and gentamicin did not affect embryo development to the 8-cell stage at 58 h post oocyte activation, or morula/blastocyst stages, or blastocysts alone at 82 h post activation. However, when penicillin and streptomycin were both present in the culture medium the percentages of 8-cell embryos at 58 h and blastocysts at 82 h were significantly lower than the control. No antibiotic treatment improved hamster embryo development in vitro. We caution against the use of penicillin and streptomycin together for hamster embryo culture, and show that it is not necessary to include any antibiotics in embryo culture media for up to 72 h if proper sterile technique is used with an oil overlay.

Animals↗

PubMed2000

A new vector system with inducible E2a cell line for production of higher titer and safer adenoviral vectors.

Adenoviral vectors have been used in gene therapy and for vaccination. The major concerns with using adenoviral vectors are the pathogenic potential of the virus backbone and the generation of replication-competent adenovirus that may replicate in an uncontrolled manner, especially in immunocompromised patients. It is important to develop new vectors that are safer for clinical trials while maintaining high titer and efficient transduction. A new adenovirus vector production system was developed, which includes several vector backbone plasmids deleted for E2a and a new cell line expressing both E1 and E2a. The new cell line with the tTA-inducible E2a expression cassette can significantly increase the titer of E1/E2a-deleted vectors by four to five orders of magnitude upon withdrawal of tetracycline. Furthermore, there is no sequence overlap between the vector and the cellular DNA within the E2a open reading frame and downstream, making the generation of virus with wild-type E2a through homologous recombination substantially less likely. The new vector system may improve the safety of vectors for vaccination and cancer therapy and may also provide safer backbones for further vector development, such as helper-dependent and hybrid vectors.

Adenoviridae↗

PubMed2000

Novel fimbrial subunit genes of Dichelobacter nodosus: recombination in vivo or in vitro?

Polymerase chain reaction (PCR) was used to amplify the variable region of the fimbrial subunit encoding gene (fimA) of Dichelobacter nodosus from sheep and goats infected with footrot. Two amplimers (designated X and Y) generated single-strand conformation polymorphism (SSCP) patterns different to those of previously identified serogroups and serotypes. DNA sequencing revealed that these two fragments were novel. The upstream of X (nt 1-183) was identical to serotype M1 while its downstream (nt 223-414) was identical to serotype F1; the upstream of Y (nt 1-116) was identical to serotype E1, whereas its downstream (nt 148-423) was identical to serotype F1. A 14-mer sequence consisting of two partially overlapping Chi-like sequences, 5'-GCTGGTGCTGGTGA-3', was also found in these fragments. Two primer sets with the downstream primer specific for serotype F1 and the upstream primer specific for serotype M or E1, generated PCR products of the expected sizes from the footrot samples from which fragments X and Y were isolated, respectively. These primer sets did not appear to amplify artificially mixed genomic DNA from serotypes M and F1 or E1 and F1. However, when the reactions were re-amplified, PCR recombination artifacts were observed, suggesting that PCR recombination does occur, but at a low frequency. It, therefore, seems more likely that fragments X and Y reflect genuine fimA genes of D. nodosus which have resulted from in vivo DNA recombination rather than from a PCR recombination artifact.

Animals↗

PubMed2000

Coding of border ownership in monkey visual cortex.

Areas V1 and V2 of the visual cortex have traditionally been conceived as stages of local feature representations. We investigated whether neural responses carry information about how local features belong to objects. Single-cell activity was recorded in areas V1, V2, and V4 of awake behaving monkeys. Displays were used in which the same local feature (contrast edge or line) could be presented as part of different figures. For example, the same light-dark edge could be the left side of a dark square or the right side of a light square. Each display was also presented with reversed contrast. We found significant modulation of responses as a function of the side of the figure in >50% of neurons of V2 and V4 and in 18% of neurons of the top layers of V1. Thus, besides the local contrast border information, neurons were found to encode the side to which the border belongs ("border ownership coding"). A majority of these neurons coded border ownership and the local polarity of luminance-chromaticity contrast. The others were insensitive to contrast polarity. Another 20% of the neurons of V2 and V4, and 48% of top layer V1, coded local contrast polarity, but not border ownership. The border ownership-related response differences emerged soon (<25 msec) after the response onset. In V2 and V4, the differences were found to be nearly independent of figure size up to the limit set by the size of our display (21 degrees ). Displays that differed only far outside the conventional receptive field could produce markedly different responses. When tested with more complex displays in which figure-ground cues were varied, some neurons produced invariant border ownership signals, others failed to signal border ownership for some of the displays, but neurons that reversed signals were rare. The influence of visual stimulation far from the receptive field center indicates mechanisms of global context integration. The short latencies and incomplete cue invariance suggest that the border-ownership effect is generated within the visual cortex rather than projected down from higher levels.

Animals↗

PubMed2000

12-O-tetradecanoylphorbol-13-acetate (TPA)-induced c-Jun N-terminal kinase (JNK) phosphatase renders immortalized or transformed epithelial cells refractory to TPA-inducible JNK activity.

c-Jun N-terminal kinase (JNK) regulates gene expression in response to various extracellular stimuli. JNK can be activated by the tumor promoting agent, 12-O-tetradecanoylphorbol-13-acetate (TPA) in normal human oral keratinocytes but not in human keratinocytes that have been immortalized (HOK-16B and HaCaT) or transformed (HOK-16B-Bap-T) nor in a cervical carcinoma cell line (HeLa). The refractory JNK activation response to TPA is not due a defect in the JNK pathway, because JNK can be activated by other stimuli, e.g. UV irradiation and an alkylating agent N-methyl-N'-nitrosoguanidine in these immortalized or transformed cells. More importantly, the refractory JNK and JNKK activation response to TPA can be restored by treatment of the cells with a combination of TPA and a protein-tyrosine phosphatase inhibitor, sodium orthovanadate. Furthermore, pretreatment of cells with TPA partially inhibited UV- or N-methyl-N'-nitrosoguanidine-induced JNK activity. These results suggest that a TPA-inducible, orthovanadate-sensitive protein-tyrosine phosphatase may specifically down-regulate JNK signaling pathway in these immortalized/transformed epithelial cells. In contrast, ERK and p38/Mpk2 are not regulated by this TPA-induced phosphatase. This putative protein-tyrosine phosphatase appears to be JNK pathway-specific.

Amino Acid Sequence↗

PubMed2000

Recombinant respiratory syncytial viruses with deletions in the NS1, NS2, SH, and M2-2 genes are attenuated in vitro and in vivo.

Respiratory syncytial virus (RSV) encodes several proteins that lack well-defined functions; these include NS1, NS2, SH, and M2-2. Previous work has demonstrated that NS2, SH, and M2-2 can each be deleted from RSV genome and thus are considered as accessory proteins. To determine whether RSV can replicate efficiently when two or more transcriptional units are deleted, we removed NS1, NS2, SH, and M2-2 genes individually and in different combinations from an infectious cDNA clone derived from human RSV A2 strain. The following six mutants with two or more genes deleted were obtained: DeltaNS1NS2, DeltaM2-2SH, DeltaM2-2NS2, DeltaSHNS1, DeltaSHNS2, and DeltaSHNS1NS2. Deletion of M2-2 together with NS1 was detrimental to RSV replication. It was not possible to obtain a recombinant RSV when all four genes were deleted. All of the double and triple deletion mutants exhibited reduced replication and small plaque morphology in vitro. Replication of these deletion mutants was more reduced in HEp-2 cells than in Vero cells. Among the 10 single and multiple gene deletion mutants obtained, DeltaM2-2NS2 was most attenuated. DeltaM2-2NS2 formed barely visible plaques in HEp-2 cells and had a reduction of titer of 3 log(10) compared with the wild-type recombinant RSV in infected HEp-2 cells. When inoculated intranasally into cotton rats, all of the deletion mutants were attenuated in the respiratory tract. Our data indicated that the NS1, NS2, SH, and M2-2 proteins, although dispensable for virus replication in vitro, provide auxiliary functions for efficient RSV replication.

Animals↗

PubMed2000

A population-based study of the effectiveness of inferior vena cava filter use among patients with venous thromboembolism.

BACKGROUND: There are few population-based data regarding the effectiveness of inferior vena cava filter use in the prevention of symptomatic pulmonary embolism. OBJECTIVE: To determine the 1-year cumulative incidence of rehospitalization for venous thrombosis or pulmonary embolism among patients with thromboembolism treated with a vena cava filter compared with the incidence in a control population with thromboembolism. PATIENTS AND METHODS: Population-based retrospective analysis of linked hospital discharge abstracts in California. From January 1, 1991, through December 30, 1995, 3632 patients were treated with a filter and 64,333 controls were admitted with a principal diagnosis of venous thromboembolism. RESULTS: Filter-treated patients had significantly greater comorbidity, with a higher frequency of previous pulmonary embolism, recent major bleeding, malignant neoplasm, and stroke. Patients who initially manifested pulmonary embolism were significantly more likely to be rehospitalized for pulmonary embolism than patients with an initial diagnosis of venous thrombosis alone, among filter-treated patients (relative risk, 6.72; 95% confidence interval, 3.61-12.49) and controls (relative risk, 5.30; 95% confidence interval, 4.61-6.10). Risk-adjusted proportional hazards modeling showed no significant difference between filter-treated patients and controls in the relative hazard of rehospitalization for pulmonary embolism. However, filter placement was associated with a significantly higher relative hazard of rehospitalization for venous thrombosis among patients who initially manifested pulmonary embolism (relative hazard, 2.62; 95% confidence interval, 2.09-3.29), but not among those who presented with venous thrombosis (relative hazard, 1.14; 95% confidence interval, 0.92-1.43). CONCLUSIONS: Insertion of a vena cava filter was not associated with a significant reduction in the 1-year incidence of rehospitalization for pulmonary embolism. Use of a filter was associated with a higher incidence of rehospitalization for venous thrombosis, but only among patients who initially manifested pulmonary embolism. A prospective clinical study is needed to determine the efficacy of filter use among patients with pulmonary embolism who do not meet strict guidelines for insertion of a vena cava filter.

Adult↗

PubMed2000

Is heterozygous alpha-1-antitrypsin deficiency type PIZ a risk factor for primary liver carcinoma?

BACKGROUND: It is well known that homozygotes with alpha-1-antitrypsin deficiency type PiZ are associated with an increased risk of chronic liver disease and liver carcinoma. The aim of this study was to determine whether heterozygous PiZ status is a risk factor for liver carcinoma development. METHODS: Three hundred seventeen consecutive primary liver carcinomas and the tumor-bearing liver tissue (tumor series) from adult patients were screened immunohistochemically for hepatocellular PiZ deposits. Liver specimens from 1663 consecutive adult patients (biopsy series) and liver tissue from 1030 consecutive adult autopsies (autopsy series) served as controls. The zygosity status of alpha-1-antitrypsin was verified by analysis of single strand conformational polymorphism and by sequencing DNA extracted from paraffin embedded tissue. RESULTS: The PiZ frequency in the tumor series (5.99%) was significantly higher than in the biopsy series (3.43%) or the autopsy series (1.84%). Cholangiocarcinomas and/or combined hepatocholangiocarcinomas were seen significantly more frequently in PiZ-associated liver carcinomas (57.9%) than in non-PiZ-associated carcinomas (27.2%). Cirrhosis was found in only 3 of the 19 PiZ-associated carcinomas. The remaining 16 livers showed varying stages of fibrosis or normal tissue. All nine cases with PiZ-associated liver carcinoma suitable for genetic analysis showed heterozygous PiZ mutations. CONCLUSIONS: Heterozygotes of type PiZ are associated with an increased risk of primary liver carcinoma. PiZ-associated carcinoma may develop in noncirrhotic liver tissue and without concurrent liver disease, and is frequently characterized by cholangiocellular differentiation. The site specific antibody ATZ11 is a reliable morphologic tool for detecting PiZ individuals.

Aged↗

PubMed2000

Identification and gene organization of three novel members of the IL-1 family on human chromosome 2.

Members of the IL-1 family of cytokines are important in mediating inflammatory responses. The genes encoding IL-1alpha, IL-beta, and the IL-1 receptor antagonist (IL-1Ra) are clustered within 450 kb on human chromosome 2q. By searching the EST databases and sequencing this region of chromosome 2, we have identified three novel genes that show homology to the IL-1 family, which we have named IL-1-related protein 1, 2, and 3 (IL-1RP1, IL-1RP2, and IL-1RP3). All three genes contain a signature motif common to the IL-1 family and appear to be more closely related to IL-1Ra. Similar to the intracellular form of IL-1Ra, these genes lack conventional hydrophobic signal sequences. The expression of these genes appears to be highly restricted to various epithelial cell populations. Our results demonstrate the existence of additional IL-1 gene family members within the previously defined IL-1 cluster and point to this region of chromosome 2 as an evolutionary hotspot for IL-1 gene duplication. These genes may prove to have an important role in inflammatory responses.

Amino Acid Sequence↗

PubMed2000

A low-voltage droplet charging circuit with simulative cell-sorting function for flow cytometer-cell sorter.

BACKGROUND: Flow cytometer cell sorters have become important tools in many biological laboratories. Commercial electrically-deflected cell sorters that deflect wanted cells in electrically charged droplets need high-voltage amplifiers which are expensive and difficult to obtain. Effort was made to build an alternative droplet charging circuit with low-voltage amplifiers that are much easier to get and have more reasonable price. METHODS: A low-voltage charging circuit was designed. Every time a cell was to be separated, a pair of complementary charging pulses were produced: one was positive and the other was negative with equal amplitude. These were enlarged by two low-voltage charging amplifiers to drive two charging electrodes respectively. RESULTS: Due to the effect of addition, the voltage between the two electrodes was double as high as the output of either amplifier. The result of test experiment proved that the cell sorter with low-voltage amplifiers, which was cheaper and easier to obtain, could separate cells as efficiently as the instrument with high-voltage ones that were more expensive and more difficult to make. In addition, a simulative cell-sorting function was provided. CONCLUSIONS: This low-voltage, easily-built and low-price charging circuit for flow cytometer cell sorter is a good alternative to the commonly used high-voltage one, especially to researcher who hopes to build his own personal instrument.

Animals↗

PubMed2000

The Akt proto-oncogene links Ras to Pak and cell survival signals.

The Ras oncogene regulates cellular proliferation, differentiation, transformation, and survival through multiple downstream signals. Ras signals through its effector phosphoinositide 3 (PI3) kinase to the Pak protein kinase (p65(pak)), but the steps from Ras to Pak remain to be elucidated. PI3 kinase can stimulate the small G protein, Rac, a direct activator of Pak, as well as the Akt proto-oncogene, a serine-threonine protein kinase. We found that activated Akt stimulated Pak, whereas a dominant negative Akt inhibited Ras activation of Pak in transfection assays. Akt stimulation of Pak was not inhibited by dominant negative mutants of either Rac or Cdc42 suggesting that Akt activated Pak through a GTPase-independent mechanism. We also developed a novel cell-free system to study Ras activation of Pak. In this system Ras activated Pak only in the presence of a crude cell extract but failed to activate Pak when Akt was immunodepleted from the extract. Akt protects cells from apoptosis through phosphorylation of downstream targets such as the Bcl-2 family member, Bad. We found that activated Pak decreased apoptosis and increased phosphorylation of Bad, whereas dominant negative Pak increased apoptosis and decreased phosphorylation of Bad. These studies define a new oncogene-mediated cell survival signal.

Animals↗