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Biomedical subjects

F He

Publications and source records attributed to F He.

At least 37 records · Page 2Linked to original sources

Differences in composition and mucosal adhesion of bifidobacteria isolated from healthy adults and healthy seniors.

Fifty-one Bifidobacterium strains were isolated from the feces of healthy adults (30-40 years old) and seniors (older than 70 years of age). B. adolescentis, B. breve, B. infantis, and B. longum were isolated from the healthy adults and B. adolescentis and B. longum from elderly subjects. The tested bacteria bound, in vitro, to intestinal mucus in a strain dependent manner. The strains isolated from healthy adults, and especially B. adolescentis, bound better to intestinal mucus than those isolated from seniors. These results indicate that the mucosal adhesive properties of the human Bifidobacterium flora were reduced with the aging of the host. This shift to a Bifidobacterium flora with reduced adhesive abilities may explain the decrease in bifidobacteria levels in the intestinal microflora of aging people.

Adult↗

Gas-phase cleavage of PTC-derivatized electrosprayed tryptic peptides in an FT-ICR trapped-ion cell: mass-based protein identification without liquid chromatographic separation.

Condensed phase protein sequencing typically relies on N-terminal labeling with phenylisothiocyanate ("Edman" reagent), followed by cleavage of the N-terminal amino acid. Similar Edman degradation has been observed in the gas phase by collision-activated dissociation of the N-terminal phenyl thiocarbamoyl protonated peptide [1] to yield complementary b1 and y(n-1) fragments, identifying the N-terminal amino acid. By use of infrared multiphoton (rather than collisional) activation, and Fourier transform ion cyclotron resonance (rather than quadrupole) mass analysis, we extend the method to direct analysis of a mixture of tryptic peptides. We validate the approach with bradykinin as a test peptide, and go on to analyze a mixture of 25 peptides produced by tryptic digestion of apomyoglobin. A b1+ ion is observed for three of the Edman-derivatized peptides, thereby identifying their N-terminal amino-acids. Search of the SWISS-PROT database gave a single hit (myoglobin, from the correct biological species), based on accurate-mass FT-ICR MS for as few as one Edman-derivatized tryptic peptide. The method is robust-it succeeds even with partial tryptic digestion, partial Edman derivatization, and partial MS/MS IRMPD cleavage. Improved efficiency and automation should be straightforward.

Amino Acid Sequence↗

Highly sensitive spectrofluorimetric determination of trace amounts of lead with a new fluorescent reagent, 2-hydroxy-1-naphthaldehydene-8-aminoquinoline.

A new fluorescent reagent, 2-hydroxy-1-naphthaldehydene-8-aminoquinoline (HNAAQ), was synthesized. The fluorescent reaction of this reagent with lead was also studied. Based on this chelation, a highly sensitive spectrofluorimetric method was developed for the determination of trace amounts of lead in a water-ethanol (5 + 1, v/v) medium at pH = 9.4. Under these conditions, the Pb-HNAAQ complex has excitation and emission maxima at 398 and 450 nm, respectively. The linear range of the method is from 0 to 100 microg l(-1) and detection limit is 0.28 microg l(-1) of lead. Interference of other ions was studied. In strongly basic media most interfering metal ions form precipitates of hydroxides or oxides and can be removed efficiently through filtration while the residual cations can be removed with a cation-exchange resin. Hence, the selectivity of the method can be increased considerably. The procedure can easily be performed and affords good precision and accuracy. The method was successfully applied to the determination of lead in wheat and rice flour.

Aminoquinolines↗

cDNA transfection of amino-terminal fragment of urokinase efficiently inhibits cancer cell invasion and metastasis.

Focusing of urokinase-type plasminogen activator (uPA) to the cell surface via binding to its specific receptor (uPAR, CD87) is critical for tumor invasion and metastasis. Consequently, the inhibition of uPA-uPAR interaction on the cell surface might be a promising anti-invasion and anti-metastasis strategy. We examined the effects of cDNA transfection of the human uPA amino-terminal fragment (ATF) on invasion and metastasis of cancer cells. First, a highly metastatic human lung giant-cell carcinoma cell line (PG), used as the target cell for evaluation of this effect, was demonstrated to express both uPA and uPAR. Then, ATF, which contains an intact uPAR binding site but is catalytically inactive, was designed as an antagonist of uPA-uPAR interaction and was transfected into PG cells. [(3)H]-Thymidine incorporation and cell growth curves indicated that expressed ATF did not affect the proliferation of transfected cells. However, analysis by scanning electron microscopy revealed that ATF changed the host cells from the typical invasive phenotype to a noninvasive one. Correspondingly, the modified Boyden chamber test in vitro showed that ATF expression significantly decreased the invasive capacity of transfected cells. Furthermore, in the spontaneous metastasis model, it was confirmed in vivo that expressed ATF remarkably inhibited lung metastasis of implanted ATF-transfected PG cells. In summary, autocrine ATF could act as an antagonist of uPA-uPAR interaction, and ATF cDNA transfection could efficiently inhibit the invasion and metastasis of the cancer cells. Inhibition of uPA-uPAR interaction on the cell surface might be a promising anti-invasion and anti-metastasis strategy.

Animals↗

Hepatopoietin acts as an autocrine growth factor in hepatoma cells.

Hepatopoietin (HPO) is a novel human hepatotrophic factor. Its known function is mainly limited to supporting liver regeneration. Recently, it was shown by our laboratory that HPO acts as a mitogen for hepatoma cell lines and that there are HPO-specific receptors on the surface of these cells (Wang, G., et al., J Biol Chem 1999;274:11469-11472), indicating that HPO might be involved in oncogenesis in the liver. To study this hypothesis, we first conducted experiments in vitro to identify the existence of an autocrine loop of HPO/HPO receptor in hepatoma cell lines. It was demonstrated that HPO was actually expressed by hepatoma cells, such as HepG2, Bel 7402, and SMMC-7721, and secreted into the culture medium. Furthermore, it was shown that HPO-neutralizing antibody has an inhibitory effect on the uptake of tritiated thymidine by hepatoma cells. The results strongly suggest that HPO acts as an autocrine factor for hepatoma cells in vitro.

Aged↗

Effect of dimethoate on the function and expression of nicotinic acetylcholine receptor in primary skeletal muscle cell culture.

To investigate the molecular mechanism of intermediate myasthenia syndrome (IMS), we analyzed the toxic effects of the representative organophosphate dimethoate on the function and expression of the nicotinic acetylcholine receptor (nAChR) in primary skeletal muscle cell culture. The results showed that the expression of nAChR on the muscle cell membrane was significantly increased after cells were exposed to dimethoate (130 microM). AChR function measured by carbachol-induced (22)Na+ influx demonstrated that dimethoate may inhibit the nAChR function either by binding to a noncompetitive site and changing the conformational state of nAChR or by blocking the nAChR channel directly. This study also demonstrated that dimethoate could rapidly induce the expression of c-fos, with a maximal effect at about 40 min, and c-fos might act as a transcriptional factor in regulating the expression of nAChR in the primary skeletal muscle cell culture following organophosphate exposure.

Animals↗

Gene expression profiling in human fetal liver and identification of tissue- and developmental-stage-specific genes through compiled expression profiles and efficient cloning of full-length cDNAs.

Fetal liver intriguingly consists of hepatic parenchymal cells and hematopoietic stem/progenitor cells. Human fetal liver aged 22 wk of gestation (HFL22w) corresponds to the turning point between immigration and emigration of the hematopoietic system. To gain further molecular insight into its developmental and functional characteristics, HFL22w was studied by generating expressed sequence tags (ESTs) and by analyzing the compiled expression profiles of liver at different developmental stages. A total of 13,077 ESTs were sequenced from a 3'-directed cDNA library of HFL22w, and classified as follows: 5819 (44.5%) matched to known genes; 5460 (41.8%) exhibited no significant homology to known genes; and the remaining 1798 (13.7%) were genomic sequences of unknown function, mitochondrial genomic sequences, or repetitive sequences. Integration of ESTs of known human genes generated a profile including 1660 genes that could be divided into 15 gene categories according to their functions. Genes related to general housekeeping, ESTs associated with hematopoiesis, and liver-specific genes were highly expressed. Genes for signal transduction and those associated with diseases, abnormalities, or transcription regulation were also noticeably active. By comparing the expression profiles, we identified six gene groups that were associated with different developmental stages of human fetal liver, tumorigenesis, different physiological functions of Itoh cells against the other types of hepatic cells, and fetal hematopoiesis. The gene expression profile therefore reflected the unique functional characteristics of HFL22w remarkably. Meanwhile, 110 full-length cDNAs of novel genes were cloned and sequenced. These novel genes might contribute to our understanding of the unique functional characteristics of the human fetal liver at 22 wk.

Cloning, Molecular↗

Adhesion of Bifidobacterium spp. to human intestinal mucus.

Twenty-four Bifidobacterium strains were examined for their ability to bind to immobilized human and bovine intestinal mucus glycoproteins. Each of the tested bacteria exhibited its characteristic adhesion to human and bovine fecal mucus. No significant differences were found among the taxonomic species. Among the tested bacteria, B. adolescentis, B. angulatum, B. bifidum, B. breve, B. catenulatum, B. infantis, B. longum and B. pseudocatenulatum adhered to human fecal mucus better than bovine fecal mucus, while the binding of B. animalis and B. lactis was not preferential. These results suggest that the mucosal adhesive properties of bifidobacteria may be a strain dependent feature, and the mucosal binding of the human bifidobacteria may be more host specific.

Animals↗

Modulation of human humoral immune response through orally administered bovine colostrum.

Eighteen healthy volunteers were randomized into two treatment groups and consumed liquid prepackaged bovine colostrum whey and placebo for 7 days. On days 1, 3 and 5, an attenuated Salmonella typhi Ty21a oral vaccine was given to all subjects to mimic an enteropathogenic infection. The circulating antibody secreting cells and the expression of phagocytosis receptors of the subjects before and after oral immunization were measured with the ELISPOT assay and flow cytometry. All subjects responded well to the vaccine. No significant differences were observed in ELISPOT values for IgA, IgG, IgM, Fcgamma and CR receptor expression on neutrophils and monocytes between the two groups. There was a trend towards greater increase in specific IgA among the subjects receiving their vaccine with bovine colostrum. These results suggest that bovine colostrum may possess some potential to enhance human special immune responses.

Administration, Oral↗

Comparison of mucosal adhesion and species identification of bifidobacteria isolated from healthy and allergic infants.

Fifty bifidobacteria strains were isolated from fecal samples of allergic and age matched healthy infants. Allergic infants were found to have an adult type Bifidobacterium flora with high levels of Bifidobacterium adolescentis. Healthy infants had a typical infant Bifidobacterium flora with high levels of Bifidobacterium bifidum. These isolates were tested for their adhesive properties to human intestinal mucus. The adhesion of the fecal bifidobacteria from healthy infants was significantly higher (P<0.0001) than for allergic infants. This suggests a correlation between allergic disease and the composition of the intestinal bifidobacteria flora which has reduced adhesive abilities to the intestinal mucus. Therefore, dietary supplementation of bifidobacteria typical for healthy infants, may be beneficial in the treatment of allergic disorders.

Bacterial Adhesion↗

Absence of Dbp2p alters both nonsense-mediated mRNA decay and rRNA processing.

Dbp2p, a member of the large family of DEAD-box proteins and a yeast homolog of human p68, was shown to interact with Upf1p, an essential component of the nonsense-mediated mRNA decay pathway. Dbp2p:Upf1p interaction occurs within a large conserved region in the middle of Upf1p that is largely distinct from its Nmd2p and Sup35/45p interaction domains. Deletion of DBP2, or point mutations within its highly conserved DEAD-box motifs, increased the abundance of nonsense-containing transcripts, leading us to conclude that Dbp2p also functions in the nonsense-mediated mRNA decay pathway. Dbp2p, like Upf1p, acts before or at decapping, is predominantly cytoplasmic, and associates with polyribosomes. Interestingly, Dbp2p also plays an important role in rRNA processing. In dbp2Delta cells, polyribosome profiles are deficient in free 60S subunits and the mature 25S rRNA is greatly reduced. The ribosome biogenesis phenotype, but not the mRNA decay function, of dbp2Delta cells can be complemented by the human p68 gene. We propose a unifying model in which Dbp2p affects both nonsense-mediated mRNA decay and rRNA processing by altering rRNA structure, allowing specific processing events in one instance and facilitating dissociation of the translation termination complex in the other.

Blotting, Northern↗

Upf1p, Nmd2p, and Upf3p regulate the decapping and exonucleolytic degradation of both nonsense-containing mRNAs and wild-type mRNAs.

In Saccharomyces cerevisiae, rapid degradation of nonsense-containing mRNAs requires the decapping enzyme Dcp1p, the 5'-to-3' exoribonuclease Xrn1p, and the three nonsense-mediated mRNA decay (NMD) factors, Upf1p, Nmd2p, and Upf3p. To identify specific functions for the NMD factors, we analyzed the mRNA decay phenotypes of yeast strains containing deletions of DCP1 or XRN1 and UPF1, NMD2, or UPF3. Our results indicate that Upf1p, Nmd2p, and Upf3p regulate decapping and exonucleolytic degradation of nonsense-containing mRNAs. In addition, we show that these factors also regulate the same processes in the degradation of wild-type mRNAs. The participation of the NMD factors in general mRNA degradation suggests that they may regulate an aspect of translation termination common to all transcripts.

Adaptor Proteins, Signal Transducing↗

Evidence of evolving carbaryl resistance in western corn rootworm (Coleoptera: Chrysomelidae) in areawide-managed cornfields in north central Kansas.

Susceptibility of adult populations of the western corn rootworm, Diabrotica virgifera virgifera LeConte, to carbaryl was determined by a survey in 1996 before the implementation of an areawide management program near Scandia in north central Kansas. Subsequently, the susceptibility of western corn rootworm adults to carbaryl has been monitored throughout the program from 1997 to 2000 in both control and managed areas. In 1996, adults were highly susceptible to carbaryl with a mean LC50 value of 0.64 microg/vial. This value was comparable to those for adults collected from other regions within Kansas. However, adult susceptibility to carbaryl decreased rapidly within the managed area, where the cucurbitacin- carbaryl-based bait SLAM has been used as the primary tool to control adults in this project since 1997. In 1999, adults collected from the managed area were 9- and 20-fold less susceptible to carbaryl at the LC50 and LC90 levels, respectively, than those evaluated in 1996. In contrast, adults collected from the control area were only 2- and 3-fold less susceptible to carbaryl at the LC50 and LC90 levels, respectively, than adults evaluated in 1996. Although field adult populations of western corn rootworm were relatively low in 2000, evaluations showed trends similar to those in 1999 regarding their carbaryl susceptibility in the managed and control areas. These results provide evidence that western corn rootworm has been evolving carbaryl resistance rapidly in response to the use of SLAM in areawide-managed cornfields near Scandia.

Animals↗

[The influence of hydraulic characteristics on wastewater purifying efficiency in vertical flow constructed wetlands].

The hydraulic characteristics of vertical flow constructed wetlands was investigated, and the influence of these characteristics on wastewater purifying efficiency was studied as an emphasis. The results of the experiments in small scale plots and medium scale plots showed the plant roots, with the physical and biological effects, play a significant role in hydraulics and the wastewater purifying efficiency. The other characteristics such as retention time, effluent velocity and influent load also directly affect the wastewater purifying efficiency of constructed wetland. To get a better understanding of the hydraulics will be crucial to the improvement of purifying efficiency and facilitate the practical use of constructed wetland.

Sewage↗

The role of sexual related Y gene detection in the diagnosis of patients with gonadal dysgenesis.

OBJECTIVE: To clarify the role of sexual related Y (SRY) gene detection in the diagnosis of gonadal dysgenesis. METHODS: Sixteen cases of gonadal dysgenesis were included in this study: 5 with androgen insensitivity syndrome, 1 with 17-alpha-hydroxylase deficiency, 4 with true hermaphrodite, 2 with 45, X/46, XY gonadal dysgenesis, 1 with 45, X gonadal dysgenesis, 1 with XY pure gonadal dysgenesis, 1 with testicular regression, and 1 XY female who gave birth to a normal baby. SRY gene was detected by using polymerase chain reaction (PCR) in blood and gonad samples and by direct sequencing of the SRY motif. RESULTS: Among the 16 cases, 15 were blood SRY positive, among which 13 (86.7%) showed the presence of testicular tissue, and 2 showed ovaries without testicular tissue. One SRY negative case showed the presence of testicular tissue. In 3 cases, SRY detection in gonadal tissue correlated with pathological findings but not with blood karyotype. The correlation between peripheral blood SRY and the pathology of the gonads was 81.25% and the correlation between the presence of peripheral blood Y chromosome and pathology of the gonads was 68.75%. Sequencing of the SRY motif in an XY female who gave birth to a normal baby showed no mutation. CONCLUSIONS: SRY detection is more sensitive and specific than blood karyotype in the prediction of the presence of testicular tissue. Peripheral blood karyotype does not necessarily reflect gonadal type. There may be testicular related factors other than the SRY gene.

DNA↗

[Experimental study of anti-metastasis effect of urokinase amino-terminal fragment gene on human breast cancer cells].

OBJECTIVE: To explore the suppressive effects of urokinase amino-terminal fragment (ATF) gene on metastatic potential of human breast cancer cell line MCF-7. METHODS: A pcDNA3-ATF plasmid containing ATF cDNA under CMV promotor/enhancer control was constructed and transfected into MCF-7 cells by lipofectin. The expression of of uPA/uPAR and ATF in MCF-7 cells were analyzed by RT-PCR and Western blot. The effect of ATF expression on invasiveness in vitro, tumorigenesis and metastasis in vivo of MCF-7 cell was investigated. RESULTS: MCF-7 cells displayed an overexpression of uPA/uPAR. Expression of ATF was detected after ATF gene-transfection. The invasive capacity of ATF gene-transfected MCF-7 cells was decreased significantly. Although the tumorigenesis was not affected, the in vivo metastasis of ATF gene-transfected MCF-7 cells was remarkably inhibited. CONCLUSION: Suppression of invasiveness and metastasis of ATF-transfected MCF-7 cells is perhaps due to a competitive inhibition of interaction with endogenous uPA/uPAR.

3T3 Cells↗

[Detection of DNA crosslinks with comet assay].

A simple and rapid method for the detection of DNA cross-links was explored. DNA cross-links was induced by formaldehyde (FA) in TK6 cells. Hydrogen peroxide was used as a standard DNA breakage agent. Two protocols on detecting DNA cross-links with comet assay were tested and compared. The results showed that with increasing FA, the length of DNA migration caused by H2O2 was decreased. When the level of FA was 1000 mumol/L, no any tailed cell could be observed, the migration of DNA was inhibited completely even H2O2 was used. The detection of DNA cross-link could be tested by both protocols. However, when the DNA cross-link agent (FA) was added to the cells before the treatment with DNA breakage agent (H2O2), the result of the scheme was much better than that in the opposite order. The former scheme was much easy to be manipulated, with no influence from DNA repair and had better dose-response relationship in the detection of DNA cross-links. The comet assay is indeed a sensitive, convenient and rapid method for the detection of DNA cross-link.

Comet Assay↗