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Biomedical subjects

F Fang

Publications and source records attributed to F Fang.

At least 91 records · Page 5Linked to original sources
PubMed2000

[The research development of single nucleotide polymorphism].

Single nucleotide polymorphism(SNP) is a novel set of polymorphism based on the single nucleotide variations at the genomic level, which distributes widely across the human genome and has high polymorphism information content (PIC). Now, tens of thousands SNPs have been discovered among the human genome and are introduced by many biological medicine networks. With the development of identification and analysis technique for SNP, especially the combination with DNA chips, they, as the third generation of genetic markers, are gradually fitable for fine-scale mapping of diseases, especially the complex diseases susceptibility genes and will eventually substitute for the most commonly used microsatellite markers presently.

Genetic Markers↗
PubMed2000

[Changes of AC/cAMP system and phosphorylation regulation of adenylate cyclase activity in brain regions from morphine-dependent mice].

OBJECTIVE: To further understand the changes of AC/cAMP system in the brain regions from morphine-dependent mice. METHODS: By inducing morphine dependence in mice, we observed changes in AC/cAMP signal system, the phosphorylation regulation of adenylate cyclase (AC) activity in brain regions and effect of protein kinase A (PKA) inhibitor on the development of morphine dependence. RESULTS: (1) In morphine-dependent mice, AC activity, cAMP contents, and cytosolic PKA activity in striatum, hippocampus, and cerebral cortex were significantly higher than those of control. But there were no similar changes in cerebellum, and PKA inhibitor injected intracerebroventricular 15 min prior to morphine injection could inhibit the changes of AC activity; (2) These changes described above were not observed in mice treated with naloxone 30 min prior to daily morphine injection; (3) In striatum and cerebral cortex of morphine-dependent mice, level of AC phosphorylation in vitro was apparently higher as compared to control group. It indicated that the level of AC phosphorylation in vivo was decreased in morphine-dependent mice; (4) PKA inhibitor was given to mice intracerebroventricular (i.c.v.) 15 min. prior to daily morphine injection could prevent the development of morphine dependence in mice. CONCLUSIONS: The up-regulation of AC/cAMP-PKA signal system in some brain regions occurred in the state of morphine dependence and the effect was mediated via specific activation of opiate receptors. Furthermore, that PKA affecting AC phosphorylated state and leading to the increase of AC activity suggest that the increase of PKA activities during chronic morphine treatment could lead to the positive feedback regulation in AC/cAMP system and further potentiate the AC/cAMP system, and these may be one of important mechanisms by which chronic opiate induce dependence in target neurons.

Adenylyl Cyclases↗
PubMed1999

Treatment of hepatitis caused by cytomegalovirus with allitridin injection--an experimental study.

To demonstrate the anti-cytomegalovirus (CMV) activity of allitridin injection (AI), an active anti-infection component of garlic, the in vitro effects of AI on human CMV (HCMV) AD169 and 7 newly isolated strains from patients and its in vivo effect on mice of murine CMV (MCMV) hepatitis were assessed. It was found that plaque reduction rate reached 63.5% after infected cells were treated with cell maximal tolerable concentration (7.5 micrograms/ml) of AI. Meanwhile, flow cytometric analysis for effect of AI on expression of HCMV immediate-early antigen (IEA) showed that IEA inhibition rate of 7 isolated strains was in the range from 43.3% to 66.7%, with a mean of 58.4%, similar to that of AD169 strain (60.5%). On the other hand, in vivo anti-CMV activity of AI was evaluated in terms of liver pathological changes, liver function and viral replication. Six model mice with MCMV hepatitis received the treatment of AI for 2 weeks. The severity of liver damage, levels of sALT and MCMV IE genes expression in liver tissues in the treated mice were significantly lower than those of the corresponding untreated controls. Our results showed that AI had an obvious anti-CMV activity both in vitro and in vivo.

Allyl Compounds↗
PubMed1999

A model system of primary murine hepatocytes infected by murine cytomegalovirus.

In order to establish a model system of the murine hepatocyte infection by murine cytomegalovirus (MCMV), the primary cultured murine hepatocytes were obtained in a modified low-serum medium system by a non-perfusion method, and then infected by Smith strain MCMV. Infected hepatocytes showed characteristic cytopathic effect (CPE) at 30 h after infection, in which a large number of viral particles was found and ultrastructures were destroyed (as revealed by disappearance of bile canalicula and organelles) under the electron microscope and MCMV immediate-early genes were detected by in situ hybridization. Meanwhile, infected cells produced albumin significantly less than corresponding uninfected controls. On the contrary, uninfected controls simultaneously cultured under the same conditions showed normal function and ultrastructure (glycogen rosettes, bile canalicula, wheel-like mitochondria and well-developed rough and smooth endoplasmic reticula). These results demonstrated that a model system of primary cultured murine hepatocytes infected by MCMV was successfully set up.

Animals↗
PubMed1999

Phenotype of mice and macrophages deficient in both phagocyte oxidase and inducible nitric oxide synthase.

The two genetically established antimicrobial mechanisms of macrophages are production of reactive oxygen intermediates by phagocyte oxidase (phox) and reactive nitrogen intermediates by inducible nitric oxide synthase (NOS2). Mice doubly deficient in both enzymes (gp91(phox-/-)/NOS2(-/-)) formed massive abscesses containing commensal organisms, mostly enteric bacteria, even when reared under specific pathogen-free conditions with antibiotics. Neither parental strain showed such infections. Thus, phox and NOS2 appear to compensate for each other's deficiency in providing resistance to indigenous bacteria, and no other pathway does so fully. Macrophages from gp91(phox-/-)/NOS2(-/-) mice could not kill virulent Listeria. Their killing of S. typhimurium, E. coli, and attenuated Listeria was markedly diminished but demonstrable, establishing the existence of a mechanism of macrophage antibacterial activity independent of phox and NOS2.

Abscess↗
PubMed1999

[Evidence for involvement of NO/NOS-cGMP signal system in morphine dependence].

The present study was undertaken to observe changes in cGMP contents, calcium-dependent and non-calcium-dependent NOS activities in brain regions isolated from morphine-dependent mice as well as the effect of NOS inhibitor (L-NMMA) on the development of this dependence. It was found that (1) cGMP contents in cerebellum, striatum, hippocampus and cerebral cortex were significantly decreased. (2) Calcium-dependent NOS activity was noticeably increased in striatum and cerebral cortex, which was inhibited by PKA inhibitor. No similar changes were found in cerebellum and hippocampus. Changes of non-calcium-dependent NOS activity did not occur in morphine-dependent mice brain. (3) In the striatum and cerebral cortex of morphine-dependent mice, the level of 150 kD protein phosphorylation in vitro was noticeably decreased, which was inhibited by IP20 (PKA inhibitor). (4) NOS inhibitor injected (icv) 15 min prior to daily morphine injection could prevent the development of morphine dependence. (5) All the changes above were not observed in mice treated with naloxone 30 min prior to daily morphine injection. Our data suggest that the reduction of cGMP contents and the increase of calcium-dependent NOS activity in striatum and cerebral cortex isolated from morphine-dependent mice may be mediated by opioid receptors and involved in the development of morphine-dependence. Why the increase of NOS activity was in association with the reduction of cGMP contents remains to be answered and it implies that the effect of NO/NOS involved in morphine-dependence may be produced through other mechanisms other than those producing cGMP signal. NOS phosphorylation in some other brain regions, which may be regulated by PKA, probably contributes to the increase of NOS activity in morphine-dependent mice.

Animals↗
PubMed1999

Studies on renin-2 gene in transgenic rats.

OBJECTIVE: To determine the function of, in vivo, renin and its role in the pathogenesis of hypertension. METHODS: A renin-2 gene restriction map was constructed by endonuclease digestion and Southern blotting hybridization. Transgenic rats were produced via microinjection method. RESULTS: The 24 kb fragments containing mouse full-length ren-2 and its flanking sequence were cleaved by single enzymes (EcoR I, Kpn I and BamH I) and combined enzymes (EcoR I/Kpn I, Kpn I/BamH I and BamH I/EcoR I), respectively. The digests were electrophoresed in 0.8% agarose plates and transferred onto NC membranes. Radioactive 735 bp and 1400 bp probes obtained from half and full-length renin-1 cDNA were used in southern blotting hybridization. According to the electrophoresis and hybridization patterns, a ren-2 restriction map was constructed. 1603 fertilized rat ova after injection with purified 24 kb renin-2 gene were implanted into the oviducts of 81 pseudopregnant recipients in about 20 ova per female rat. 306 progenies were obtained from 50 foster mothers (average of pregnancies was 56.6%). 248 survived pups were identified by PCR analysis and Southern hybridization, and eight positive rats were found to be the transgenic rats (founder, F). All of them carried long fragments (24 kb) of renin-2 gene with normal blood pressure. Preliminary breeding and screening were carried out in the founder. Total survival pups (17.8%) and overall efficiencies (1%) were harvested as the same as those reported in the literatures. A systemic observation and the problems occurred during production of transgenic rats were also described besides the technique procedure used in this study. CONCLUSIONS: Mapping of full-length murine ren-2 can be used in investigation of the structure and function of the gene. The results denoted that the ren-2 transgenic rats were successfully established in this study and the technique used in the production of transgenic rats was proved to be valid in leading to wide-spread application of transgenic technique to many other related researches.

Animals↗
PubMed1999

[Nucleotide sequence analysis for high variance region of hepatitis C virus in patients with hemodialysis].

OBJECTIVE: To investigate the route of transmission for hepatitis C virus (HCV) infection in patients on hemodialysis (HD). METHODS: Nucleotide sequence of high variance region (HVR) of HCV was analyzed with nucleotide sequencing technique in 15 HD patients, and its homology was compared. RESULTS: Comparison of the nucleotide sequences of HCV HVR in 15 HD patients showed that their homologies in isolates 8, 12 and 14 reached 97.50%, 95.00% in isolates 4, 5 and 10, and more than 80.00% in isolates 1, 3 - 8 and 10 - 14. Clinical data showed that all these patients were hemodialyzed in the same room with the same HD machine. Isolates 2 and all others were 61.25% - 66.25% in homology, and 57.50% - 67.50% in isolates 9 and all others, and both the patients had histories of large-quantity blood transfusion. CONCLUSION: Blood transfusion was the primary risk factor for HCV infection in HD patients, but iatrogenic transmission of HCV through HD environment could also exist.

Adult↗
PubMed1999

[Simulation of gas chromatographic peak motion process].

Solute motion process under different temperature, pressure and/or flow program conditions is simulated with the aid of numerical calculation of the differential equation. It is demonstrated that there exist some differences between the normalized average linear velocity and the normalized local linear velocity. As time develops, they intercept, and may not reach the column end at the same time, thus may result in errors in retention time simulation. In constant pressure process, where the compressibility factor (j) does not change, simulation results in a model index of 1, and no model difference can be found when the integrate equation is used. In pressure varying process, where j changes by time, simulation results in an model index which deviates to 1, and positive (for a j decrease process) or negative (for a j increase process) error can be found when the integrate equation is used. The numerical solution of the differential equation is singular under given programs, which corresponds to a specific space line in z, t, u coordinates.

Chromatography, Gas↗
PubMed1999

[Role of glutathione S-transferase pi in tumor drug resistance].

OBJECTIVE: To establish the cytologic expressing system for the glutathione S-transferase pi (GST-pi) cDNA. METHODS: The pSV-GT, a recombinant plasmid containing rat GST-pi cDNA and the pSV-neo, a vector plasmid, were transfected the HeLa cells respectively and then G418 resistant colonies were isolated. The degree of the expression of GST-pi mRNA in the resultant cell lines was estimated by in situ hybridization using Digoxin-labelled probe cDNA. Cytotoxicity of variant anticancer drugs on the transfected cells were estimated by MTT assays. RESULTS: Two subclones, HeLa/pSV-GT, and HeLa/pSV-neo, which transfected respectively by pSV-GT and pSV-neo, were established. HeLa/pSV-GT expressed significantly high degree of the GST-pi mRNA, where as both the HeLa/pSV-neo and the HeLa cell lines showed the similar but negligible degree of the GST-pi mRNA expression. Moreover, HeLa/pSV-GT cells were insensitive to adriamycin, mitomycin C, and cisplatinum as compared to HeLa/pSV-neo. Concentrations of adriamycin producing 50% inhibition (IC50) to HeLa/pSV-GT was 70.13 micrograms/ml, highly elevated 6.78-fold of that of HeLa/pSV-neo, to which IC50 was 10.34 micrograms/ml.IC50 of mitomycin and cisplatinum to HeLa/pSV-GT were 10.95 and 16.52 micrograms/ml respectively, slightly but significantly higher than those to HeLa/pSV-neo, which were 7.48 and 13.70 micrograms/ml. But the cytotoxicity of vincristine on the two cell lines were not significantly different. CONCLUSIONS: These findings suggeste that HeLa/pSV-GT cell line, after being transfected with GST-pi cDNA, acquired drug-resistance property and the overexpression of GST-pi mRNA may contribute to the multidrug resistance of HeLa/pSV-GT cells. This cell line, as a stable cytogenetic system, will contribute more for the observation of GST-pi and drug resistance.

Antineoplastic Agents↗
PubMed1999

[Effects of long-term morphine exposure on the cAMP system and c-Fos phosphorylation in differentiated SH-SY5Y cells].

OBJECTIVE: To further understand the effects of long-term morphine exposure on the cAMP system and c-Fos phosphorylation in differentiated SH-SY5Y human neuroblastoma cells. METHODS: Cellular changes of cAMP, PKA and c-Fos were detected by protein competitive conjunction, enzyme activity and isotope incorporation methods respectively. RESULTS: (1) Long-term exposure (2 min-36 h) to morphine (100 mumol/L) could induce the biphasic changes in cAMP contents. Treament for 2 min to 1 h, morphine caused rapid and siginificant decrease of the cAMP level and then gradully recovered and apparently increased at 36 h. At that time, naloxone added to the incubation media caused an overshoot of cellular cAMP; (2) Long-term exposure to morphine could also induce the biphasic changes in cytosolic PKA activity. This is consistent with the changes of cAMP during the chronic treatment of cells with morphine. But no changes were observed in membrane PKA activity; (3) In morphine dependent-like cells decreased c-Fos phosphorylation level was observed. PKA inhibitor could significantly inhibit this change; (4) Concomitant administration of naloxone could block the changes in PKA activity and c-Fos phosphorylation described above. CONCLUSIONS: The up-regulation of cAMP system in differentiated SH-SY5Y cells may be involved in the development of morphine dependent and in morphine dependent-like SH-SY5Y cells and PKA was suggested to regulate c-Fos dephosphorylation through activating phosphatase and then activate some genes transcription, which might be one of the important mechanism regardingas cellular adaptive responses underlying dependence to opioid drugs.

Cyclic AMP↗
PubMed1999

[Experimental studies in vitro on the alpha-fetoprotein-specific promoter mediated target gene therapy of hepatocellular carcinoma].

OBJECTIVE: To investigate the alpha-fetoprotein (AFP)-specific promoter mediated target gene therapy of hepatocellular carcinoma in vitro. METHODS: Retroviral vector (LX2.2CD) in which the cytosine deaminase (CD) gene was driven by the 2.2 kb recombinant human AFP TRS. After transfecting three human hepatoma cell lines and one non-hepatoma cell line with LX2.2 CD, anti-G418 clones integrated CD gene were selected, and inhibitory experiment of cell growth was performed. RESULTS: 5-fluorocytosine (5FC) could confer the chemosensitivity to transduced AFP-producing hepatoma cells (HuH-7 and huH-1/c1-2), but not to AFP-nonproducing hepatoma cells (HLE) or nonhepatoma cells (GLC). On the other hand, when transfecting the above four tumor cell lines with another retroviral vector pCD2 (CD gene was driven by 5'LTR internal promoter), no cell selection was found in 5FC-induced cell grow inhibition. CONCLUSIONS: Recombinant retrovival transfer of the CD gene under the control of the AFP TRS followed by 5FC may well be a promising trageted gene therapy for hepatoma.

Antineoplastic Agents↗
PubMed1998

Antinociception produced by microinjection of morphine in the rat periaqueductal gray is enhanced in the foot, but not the tail, by intrathecal injection of alpha1-adrenoceptor antagonists.

Antinociception produced by microinjection of morphine in the ventrolateral periaqueductal gray is mediated in part by alpha2-adrenoceptors in the spinal cord dorsal horn. However, several recent reports demonstrate that microinjection of morphine in the ventrolateral periaqueductal gray inhibits nociceptive responses to noxious heating of the tail by activating descending neuronal systems that are different from those that inhibit the nociceptive responses to noxious heating of the feet. More specifically, alpha2-adrenoceptors appear to mediate the antinociception produced by morphine using the tail-flick test, but not that using the foot-withdrawal or hot-plate tests. The present study extended these findings and determined the role of alpha1-adrenoceptors in mediating the antinociceptive effects of morphine microinjected into the ventrolateral periaqueductal gray using both the foot-withdrawal and the tail-flick responses to noxious radiant heating in lightly anesthetized rats. Intrathecal injection of selective antagonists was used to determine whether the antinociceptive effects of morphine were modulated by alpha1-adrenoceptors. Injection of the selective alpha1-adrenoceptor antagonists prazosin or WB4101 potentiated the increase in the foot-withdrawal response latency produced by microinjection of morphine in the ventrolateral periaqueductal gray. In contrast, either prazosin or WB4101 partially reversed the increase in the tail-flick response latency produced by morphine. These results indicate that microinjection of morphine in the ventrolateral periaqueductal gray modulates nociceptive responses to noxious heating of the feet by activating descending neuronal systems that are different from those that inhibit the nociceptive responses to noxious heating of the tail. More specifically, alpha1-adrenoceptors mediate a pro-nociceptive action of morphine using the foot-withdrawal response, but in contrast, alpha1-adrenoceptors appear to mediate part of the antinociceptive effect of morphine determined using the tail-flick test.

Adrenergic alpha-Antagonists↗
PubMed1998

Tumor necrosis factor alpha, interleukin-6, interleukin-8, and interferon alpha in children with viral hepatitis.

To investigate the cellular immunological changes in children with viral hepatitis, interleukin 6 (IL-6), interleukin 8 (IL-8), interferon alpha (IFN alpha), and tumor necrosis factor alpha (TNF alpha) in supernatant of cultured peripheral blood mononuclear cells (PBMCs) of 49 children with hepatitis A, B or C were measured. The levels of IL-6, IL-8, TNF alpha in PBMCs of the 3 viral hepatitis groups were increased and the level of IFN alpha decreased as compared with those of normal control group. But there were no significant differences among the 3 viral hepatitis groups. It was concluded that cellular immunological disorders were related to the onset and the induced damage of the viral hepatitis in children.

Adolescent↗
PubMed1998

A 16.6-kilodalton protein in the Cyanobacterium synechocystis sp. PCC 6803 plays a role in the heat shock response.

The low molecular weight (LMW) heat shock protein (HSP) gene hsp16.6 was identified and cloned from the unicellular cyanobacterium Synechocystis sp. PCC 6803 through comparisons of genomic sequences and conserved gene sequences of the LMW HSPs. Hsp16.6 was isolated using PCR and cloned into the pGEMT plasmid. Hsp16.6 showed a significant increase in transcription after heat shock at 42 degreesC that indicated hsp16.6 was a heat shock gene. To determine the role that hsp16.6 plays in the heat shock response, a mutant Synechocystis cell line was generated. Cell growth and oxygen evolution rates of wild type and mutant cells were compared after heat shock. Results showed significantly decreased cell growth rates and a 40% reduction in oxygen evolution rates in mutants after heat shock treatments. These data indicate a protective role for hsp16.6 in the heat shock response.

Amino Acid Sequence↗
PubMed1998

Antimalarial activity of radicicol, heptelidic acid and other fungal metabolites.

In the course of our screening program for artemisinin-like antimalarial compounds from microorganisms, seven fungal metabolites such as radicicol and heptelidic acid were identified as active compounds. Some of them exhibited antimalarial activity in vitro against the human malaria parasite Plasmodium falciparum to the extent of approximately 1/10 as potent as artemisinin. Radicicol was moderately active in vivo against Plasmodium berghei in mice.

Animals↗