PubMed Health⌕ Search

Biomedical subjects

H Fan

Publications and source records attributed to H Fan.

At least 91 records · Page 5Linked to original sources
PubMed1999

Tandemization of a subregion of the enhancer sequences from SRS 19-6 murine leukemia virus associated with T-lymphoid but not other leukemias.

Most simple retroviruses induce tumors of a single cell type when infected into susceptible hosts. The SRS 19-6 murine leukemia virus (MuLV), which originated in mainland China, induces leukemias of multiple cellular origins. Indeed, infected mice often harbor more than one tumor type. Since the enhancers of many MuLVs are major determinants of tumor specificity, we tested the role of the SRS 19-6 MuLV enhancers in its broad disease specificity. The enhancer elements of the Moloney MuLV (M-MuLV) were replaced by the 170-bp enhancers of SRS 19-6 MuLV, yielding the recombinants DeltaMo+SRS(+) and DeltaMo+SRS(-) M-MuLV. M-MuLV normally induces T-lymphoid tumors in all infected mice. Surprisingly, when neonatal mice were inoculated with DeltaMo+SRS(+) or DeltaMo+SRS(-) M-MuLV, all tumors were of T-lymphoid origin, typical of M-MuLV rather than SRS 19-6 MuLV. Thus, the SRS 19-6 MuLV enhancers did not confer the broad disease specificity of SRS 19-6 MuLV to M-MuLV. However, all tumors contained DeltaMo+SRS M-MuLV proviruses with common enhancer alterations. These alterations consisted of tandem multimerization of a subregion of the SRS 19-6 enhancers, encompassing the conserved LVb and core sites and adjacent sequences. Moreover, when tumors induced by the parental SRS 19-6 MuLV were analyzed, most of the T-lymphoid tumors had similar enhancer alterations in the same region whereas tumors of other lineages retained the parental SRS 19-6 MuLV enhancers. These results emphasize the importance of a subregion of the SRS 19-6 MuLV enhancer in induction of T-cell lymphoma. The relevant sequences were consistent with crucial sequences for T-cell lymphomagenesis identified for other MuLVs such as M-MuLV and SL3-3 MuLV. These results also suggest that other regions of the SRS 19-6 MuLV genome contribute to its broad leukemogenic spectrum.

3T3 Cells↗
PubMed1999

[Determination of 10-HDA in honeybee body by HPLC].

In the present work we found that in the honeybee body there exists an unsaturated fatty acid, trans-10-hydroxy-2-decenoic acid (10-HDA), which was known only to be present in royal jelly. We established the analytical method of 10-HDA in honeybee body by HPLC and simplified the extraction method of 10-HDA. In the optimum conditions the linear range of detection was 10-1,000 ng, the correlation coefficient was 0.9998, the recovery was 96.5%-99.2% and the detectable limit was 0.53 microgram/g.

Animals↗
PubMed1999

[Determination of nervonic acid in oral tonic by GC].

A method for determination of nervonic acid in oral tonic has been developed by gas chromatography. The samples were derivatized by BF3-CH3OH. Dinonyl phthalate was used as the internal standard. The calibration curve was linear over a range from 0.3 g/L to 3 g/L with a correlation coefficient of 0.9994. The average recovery of the method was 97.04% and the RSD was 0.90%. The method has been successfully applied to quality control of nervonic acid in oral tonic.

Administration, Oral↗
PubMed1999

[Genetic diversity among Chinese peanut rhizobia by rep-PCR analysis].

Repetitive sequences(repetitive extragenic palindromic[REP], and enterobacterial repetitive intergenic consensus [ERIC]) with the polymerase chain reaction(PCR) were used to fingerprint pure DNA extracted from 79 bradyrhizobial strains. These strains included 59 peanut rhizobia [Bradyrhizobium sp. (Arachis)] isolated from root nodules of 24 peanut (Arachis hypogaea L) cultivars from 23 sites in 11 provinces in China, 6 peanut rhizobial strains from other nations, and 14 reference strains of other bradyrhizobia. All the strains were clustered at the level of 11%, 2%, and 11, 24 clusters were obtained at the similarity of 50% with REP-PCR and ERIC-PCR fingerprints, respectively. The results showed that significant genomic diversity exists within the peanut rhizobia from China. Also they suggested that there are different distributions of REP and ERIC in genomic DNA of the peanut rhizobia. The combined results, REP plus ERIC data, were found between the both above. The genomic diversity seems not to correlate with their host and geographic origins. Our results supported this technique is a useful tool for genotypic characterization and identification of rhizobia as well as ecological studies.

Arachis↗
PubMed1998

The mammalian ribonucleotide reductase R2 component cooperates with a variety of oncogenes in mechanisms of cellular transformation.

Ribonucleotide reductase, which is composed of the two protein components R1 and R2, is a highly regulated enzyme activity that is essential for DNA synthesis and repair. Recent studies have shown that elevated expression of the rate-limiting R2 component increases Raf-1 protein activation and mitogen-activated protein kinase activity and acts as a novel malignancy determinant in cooperation with H-ras and rac-1. We show that R2 cooperation in cellular transformation extends to a variety of oncogenes with different functions and cellular locations. Anchorage-independent growth of cells transformed with v-fms, v-src, A-raf, v-fes, c-myc, and ornithine decarboxylase was markedly enhanced when the R2 component of ribonucleotide reductase was overexpressed. In addition, we observed that elevated R2 expression conferred on c-myc-transformed NIH 3T3 cells an increased tumorigenic potential in immunoincompetent mice. Taken together, these observations demonstrate that the R2 protein is not only a rate-limiting component for ribonucleotide reduction but that it is also capable of acting in cooperation with a variety of oncogenes to determine transformation and tumorigenic potential.

3T3 Cells↗
PubMed1998

[Association between angiotensin II type I receptor gene and human essential hypertension].

OBJECTIVE: To determine whether the angiotensin II type I receptor gene might be implicated in human essential hypertension. METHODS: This was a case-control study on hypertension and normal blood pressure in Chinese DNA abstracted samples from 51 cases of essential hypertension and 74 normal controls were analysed by polymerase chain reaction,digestion of restriction enzyme and electrophesis. RESULTS: The genotype frequency of 1166A/1166C was 0. 196 (10/51) in the essential hypertension group, 0.081 (6/74) in the control group. The genotype frequency of 1166C/1166C was 0 in both groups. The gene frequency of 1166C was 0.098 in the essential hypertension group, 0.041 in the controls. There was a significant increase in allelic frequency of 1166C in hypertension. CONCLUSION: The results indicate that the increase in allelic frequency of 1166C is a risk factor and hence suggest that the change in the angiotensin II type I receptor gene is associated with essential hypertension.

Adolescent↗
PubMed1998

Differential behavior of the Mo + PyF101 enhancer variant of Moloney murine leukemia virus in rats and mice.

The Mo + PyF101 enhancer variant of Moloney murine leukemia virus (M-MuLV) has been very useful in investigating M-MuLV leukemogenesis. When inoculated subcutaneously (s.c.) into neonatal mice, Mo + PyF101 M-MuLV is attenuated for development of disease. Previous studies in mice infected with wild-type M-MuLV have revealed several important preleukemic events, including development of splenic hyperplasia, defects in bone marrow hematopoiesis, and in vivo generation of MCF viruses that arise by recombination in the uninfected mouse. Mo + PyF101 M-MuLV is defective in inducing these effects after s.c. inoculation. In the experiments reported here, a study of Mo + PyF101 M-MuLV infection in rats was carried out. Wild-type M-MuLV is leukemogenic in rats, but infected rats do not form MCF recombinants since they lack the necessary endogenous polytropic envelope sequences. Since Mo + PyF101 M-MuLV's leukemogenic defect is correlated with a failure to generate MCF recombinants, it seemed possible that wild-type M-MuLV might not have a leukemogenic advantage over Mo + PyF101 M-MuLV in rats, where MCF recombinants cannot form. Neonatal Fisher F344 rats were inoculated s.c. or intraperitoneally by wild-type and Mo + PyF101 M-MuLVs. Surprisingly, Mo + PyF101 M-MuLV was completely deficient in leukemogenesis in rats when inoculated by either route while wild-type M-MuLV induced lymphoma with the predicted time course. The leukemogenic defect for Mo + PyF101 M-MuLV resulted from a pronounced defect for establishing infection in rats. Further studies of wild-type M-MuLV in rats indicated that infection was confined almost exclusively to the thymus at early times. In mice wild-type M-MuLV establishes substantial infection in other hematopoietic organs such as spleen and bone marrow as well. Thymic infection was also correlated with a decrease in thymic cellularity at early times.

Animals↗
PubMed1998

Purification and structural characterization of insulin and glucagon from the bichir Polypterus senegalis (Actinopterygii: Polypteriformes).

The Polypteriformes (bichirs and reedfish) are a family of ray-finned fishes of ancient lineage. Insulin has been isolated from an extract of the pancreas and upper gastrointestinal tract of the bichir Polypterus senegalis and its primary structure established as A-chain: Gly-Ile-Val-Glu-Gln-Cys-Cys-Asp-Thr-Pro10-Cys-Ser- Leu-Tyr-Asp-Leu-Glu-Asn-Tyr-Cys20-Asn: B-chain: Ala-Ala-Asn-Arg-His-Leu-Cys-Gly-Ser-His10-Leu-Val- Glu-Ala-Leu-Tyr-Leu-Val-Cys-Gly20-Asn-Arg-Gly-Phe- Phe-Tyr-Ile-Pro-Ser-Lys30-Met. Despite the fact that Polypterus insulin contains several unusual structural features that are not found in insulins from other jawed fish (Asp at A-8, Thr at A-9, Arg at B-4, Asn at B-21, Ile at B-27, Met at B-31), all the residues in human insulin that are involved in receptor binding, dimerization, and hexamerization have been conserved. A comparison of the structures of insulins from a range of species indicates that Polypterus insulin most closely resembles paddlefish insulin II (seven amino acid substitutions). In contrast, Polypterus glucagon (His-Ser- Gln-Gly-Thr-Phe-Thr-Asn-Asp-Tyr10-Thr-Lys-Tyr- Gln-Asp-Ser-Arg-Arg-Ala-Gln20-Asp-Phe-Val-Gln- Trp-Leu-Met-Ser-Asn) most closely resembles the glucagons from the gar Lepisosteus spatula and the bowfin Amia calva (four amino acid substitutions). The data are consistent with the conclusion based on comparison of morphological characteristics that the Polypterids are the most basal living group of the Actinopterygians with evolutionary connections to both the Acipenserids and the Neopterygians.

Amino Acid Sequence↗
PubMed1998

Total energy expenditure and physical activity as assessed by the doubly labeled water method in Swedish adolescents in whom energy intake was underestimated by 7-d diet records.

Swedish children and adolescents may be adopting a sedentary lifestyle with low energy expenditures and intakes, but no quantitative data are available. The purpose of the present study in 50 adolescents aged 15 y was to investigate whether assessment of total energy expenditure (TEE) and physical activity level (PAL) by the doubly labeled water method and indirect calorimetry and estimation of energy intake by a 7-d diet record would indicate physical inactivity. The boys' (n = 25) mean weight was 112% and the girls' (n = 25) was 109% of Swedish reference values from 1976; the mean height of both boys and girls was 102% of those reference values. Mean TEE in the boys and girls, 13.82 +/- 1.90 and 10.70 +/- 1.59 MJ/d, and mean PAL (TEE/basal metabolic rate), 1.89 +/- 0.16 and 1.79 +/- 0.22, respectively, were nonsignificantly higher than corresponding figures from other published studies. Mean energy intake as a percentage of TEE was 81.9 +/- 17.9% in the boys and 78.3 +/- 16.4% in the girls. Significant negative correlations were found both between energy intake as a percentage of TEE and percentage body fat and between energy intake as a percentage of TEE and body mass index. These results add to the evidence that 7-d diet records underestimate energy intake in adolescents, particularly those with a tendency for overweight and increased body fat. The results support indications of a trend of increasing body weight and height in Swedish adolescents, but conflict with the presumptions of low physical activity, low energy expenditure, and low energy intake. These results support the view that current recommendations for energy intake during adolescence are too low.

Adipose Tissue↗
PubMed1998

Molecular methods for detecting t(11;14) translocations in mantle-cell lymphomas.

The t(11;14)(q13;q32) and its molecular counterpart, bcl1/JH, are characteristic of mantle-cell lymphomas (MCL). Molecular detection of the translocation is useful in diagnosis and classification, and also shows promise in detecting minimal residual disease. The purpose of this study was to determine the frequency of detecting bcl1/JH by polymerase chain reaction (PCR) compared with Southern blot analysis in cases proven by cytogenetic analysis to harbor t(11;14). Southern blot analysis using two probes targeting the major translocation cluster (MTC) and a third probe targeting the p94 region was performed, along with PCR using two different bcl1 MTC primers, on 18 cases of MCL known to have t(11;14). Southern blot analysis revealed bcl1 rearrangement in 13 of 18 cases (72%), 12 with MTC breakpoints and 1 with a p94 breakpoint. The 2.1-kb MTC probe "b" was superior to the smaller 700-bp probe "a" in detecting these rearrangements. The MTC translocation was identified by PCR in 10 of 12 cases, and both primer sets that were tested performed equally well. This study illustrates the frequency with which molecular methods detect known t(11;14) translocations in MCLs. These results may help clinical laboratory scientists optimize their procedure for detecting bcl1 translocations by molecular methods at initial diagnosis and for purposes of detecting minimal residual disease.

Blotting, Southern↗
PubMed1998

In vivo footprinting of the enhancer sequences in the upstream long terminal repeat of Moloney murine leukemia virus: differential binding of nuclear factors in different cell types.

The enhancer sequences in the Moloney murine leukemia virus (M-MuLV) long terminal repeat (LTR) are of considerable interest since they are crucial for virus replication and the ability of the virus to induce T lymphomas. While extensive studies have identified numerous nuclear factors that can potentially bind to M-MuLV enhancer DNA in vitro, it has not been made clear which of these factors are bound in vivo. To address this problem, we carried out in vivo footprinting of the M-MuLV enhancer in infected cells by in vivo treatment with dimethyl sulfate (DMS) followed by visualization through ligation-mediated PCR (LMPCR) and gel electrophoresis. In vivo DMS-LMPCR footprinting of the upstream LTR revealed evidence for factor binding at several previously characterized motifs. In particular, protection of guanines in the central LVb/Ets and Core sites within the 75-bp repeats was detected in infected NIH 3T3 fibroblasts, Ti-6 lymphoid cells, and thymic tumor cells. In contrast, factor binding at the NF-1 sites was found in infected fibroblasts but not in T-lymphoid cells. These results are consistent with the results of previous experiments indicating the importance of the LVb/Ets and Core sequences for many retroviruses and the biological importance especially of the NF-1 sites in fibroblasts and T-lymphoid cells. No evidence for factor binding to the glucocorticoid responsive element and LVa sites was found. Additional sites of protein binding included a region in the GC-rich sequences downstream of the 75-bp repeats (only in fibroblasts), a hypersensitive guanine on the minus strand in the LVc site (only in T-lymphoid cells), and a region upstream of the 75-bp repeats. These experiments provide concrete evidence for the differential in vivo binding of nuclear factors to the M-MuLV enhancers in different cell types.

3T3 Cells↗
PubMed1998

Tumorigenic potential of a recombinant retrovirus containing sequences from Moloney murine leukemia virus and feline leukemia virus.

A recombinant retrovirus, termed MoFe2-MuLV, was constructed in which the U3 region of T-lymphomagenic Moloney murine leukemia virus (Mo-MuLV) was replaced by that of FeLV-945, a provirus of unique long terminal repeat (LTR) structure identified only in non-T-cell, non-B-cell lymphomas of the domestic cat. The LTR of FeLV-945 is unusual in that it contains only a single copy of the transcriptional enhancer followed 25 bp downstream by a 21-bp sequence in triplicate in tandem. Infectivity of MoFe2-MuLV was demonstrated in vitro in SC-1 cells and in vivo in neonatal NIH-Swiss mice. Tumors occurred in MoFe2-MuLV-infected animals following a latency period of 4 to 10 months (average, 6 months). The results of Southern blot analysis of the T-cell receptor beta locus demonstrated that all tumors were lymphomas of T-cell origin. MoFe2-MuLV LTRs were amplified by PCR from tumor DNA and were characterized by nucleotide sequence analysis. LTRs from the tumors that occurred with relatively shorter latency predominantly retained the original MoFe2-MuLV sequence intact and unaltered. Tumors that occurred with relatively longer latency contained LTRs that also retained the 21-bp sequence triplication characteristic of the original virus but had acquired various duplications of enhancer sequences. The repeated identification of enhancer duplications in late-appearing tumors suggests that the duplication affords a selective advantage, although apparently not in the efficient induction of T-cell lymphoma. Proto-oncogenes known to be targets of insertional mutagenesis in the majority of Mo-MuLV-induced tumors or in feline non-T-cell, non-B-cell lymphomas were shown not to be rearranged in any tumor examined. Mink cell focus-inducing (MCF) proviral DNA was readily detectable in some, but not all, tumors. The presence or absence of MCF did not correlate with the kinetics of tumor induction. These studies indicate that the single-enhancer, triplication-containing FeLV LTR, typical of non-T-cell, non-B-cell lymphomas in cats, is competent in the induction of T-cell lymphoma in mice. The findings suggest that the mechanism of MoFe2-MuLV-mediated lymphomagenesis may differ from that of Mo-MuLV-mediated disease, considering the possible involvement of novel oncogenes and the variable presence of MCF recombinants.

Animals↗
PubMed1998

5' processing of tRNA precursors can Be modulated by the human La antigen phosphoprotein.

Eukaryotic precursor (pre)-tRNAs are processed at both ends prior to maturation. Pre-tRNAs and other nascent transcripts synthesized by RNA polymerase III are bound at their 3' ends at the sequence motif UUUOH [3' oligo(U)] by the La antigen, a conserved phosphoprotein whose role in RNA processing has been associated previously with 3'-end maturation only. We show that in addition to its role in tRNA 3'-end maturation, human La protein can also modulate 5' processing of pre-tRNAs. Both the La antigen's N-terminal RNA-binding domain and its C-terminal basic region are required for attenuation of pre-tRNA 5' processing. RNA binding and nuclease protection assays with a variety of pre-tRNA substrates and mutant La proteins indicate that 5' protection is a highly selective activity of La. This activity is dependent on 3' oligo(U) in the pre-tRNA for interaction with the N-terminal RNA binding domain of La and interaction of the C-terminal basic region of La with the 5' triphosphate end of nascent pre-tRNA. Phosphorylation of La is known to occur on serine 366, adjacent to the C-terminal basic region. We show that this modification interferes with the La antigen's ability to protect pre-tRNAiMet from 5' processing either by HeLa extract or purified RNase P but that it does not affect interaction with the 3' end of pre-tRNA. These findings provide the first evidence to indicate that tRNA 5'-end maturation may be regulated in eukaryotes. Implications of triphosphate recognition is discussed as is a role for La phosphoprotein in controlling transcriptional and posttranscriptional events in the biogenesis of polymerase III transcripts.

Adenosine Triphosphatases↗
PubMed1998

[Detection of type 6 human herpesvirus (HHV-6) specific DNA sequence in lymphoma tissues by PCR].

OBJECTIVE: To study the relationship between human herpesvirus-6 (HHV-6) and lymphoma in China. METHODS: The specimens of paraffin-embedded lymphoma tissue, leukemia cell lines and normal blood donor peripheral blood lymphocytes(PBL) were examined for the presence of HHV-6 DNA sequence by polymerase chain reaction (PCR). RESULTS: Of the 52 cases lymphoma studied, HHV-6 DNA sequence was detected in 21 cases; of 43 normal PBL, HHV-6 DNA sequence was detected in 8 cases. The difference was statistically significant (P < 0.05). HHV-6 DNA sequence was also present in two leukemia cell lines. CONCLUSION: The results suggest that HHV-6 may be related to lymphoma in China.

DNA, Viral↗
PubMed1998

[Effects of ligustrazine on myocardium and coronary artery].

OBJECTIVE: To study the effects of ligustrazine on mechanical electricity in guinea pig ventricular papillary muscles and on contraction of pig coronary artery. METHOD: Standard glass microelectrode technology; electrophysiological method; and blood vessel perfusion were used. RESULT: 1. ligustrazine(3-100 mumol/L) suppressed the action potential duration (APD) and the force of contraction (FC) of the fast action potentials (FAP), as well as the action potential amplitude (APA), APD, FC and the maximal upstroke velocity(Vmax) of the slow action potentials (SAP). 2. ligustrazine (3.0 mmol/L) induced SAP and contraction, but failed in beta-adrenoreceptor blocked muscles. Elevation of[Ca2+]o increased the action potential amplitude (APA). 3. ligustrazine(10(-5)-5 x 10(-3) mol/L) decreased the contractile response to high KCl in pig coronary artery. CONCLUSION: Ligustrazine might affect the inward current of myocardial cells through beta-adrenoreceptor, and the diastolizing effect of ligustrazine on coronary artery might have nothing to do with the exciting beta-adrenoreceptor.

Action Potentials↗
PubMed1998

[Indirect determination of sulphur in plant tissues by flame atomic absorption spectrometry].

The sulphur in plant tissues was transformed into SO4(2-) by digestion and it reacted on BaCrO4 to produce CrO4(2-) by exchange reaction, then the sulphur quantity in the plant was indirectly determined by flame atomic absorption spectrometry (FAAS). Various conditions affecting the quantative determination of SO4(2-) with this method were studied and the interference of PO4(3-), which existed in the digestion solution, could be overcome by addition of sufficient calcium. The results obtained by this method are consistent with those obtained by ICP-AES.

Ammonium Hydroxide↗