Biomedical subjects

F Huang

Publications and source records attributed to F Huang.

At least 19 recordsLinked to original sources

PubMed2002

A 588-gene microarray analysis of the peripheral blood mononuclear cells of spondyloarthropathy patients.

OBJECTIVES: To identify genes which are more highly expressed in the peripheral blood mononuclear cells (PBMC) of patients with spondyloarthropathy (SpA), rheumatoid arthritis (RA) and psoriatic arthritis (PsA), in comparison to normal subjects. METHODS: A 588-gene microarray was used as a screening tool to select a panel of such genes from PBMC of these subjects and of normal subjects. Results were then validated by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The following genes were more highly expressed in arthritis patients than in normal subjects: macrophage differentiation marker MNDA (myeloid nuclear differentiation antigen), MRP8 and MRP14 (migratory inhibitory factor-related proteins); signalling molecules JAK3 (janus kinase 3) and MAP kinase p38 (mitogen-activated protein kinase); receptors TNFR2/p75, C-C-chemokine receptor type 1 (CCR1), C-X-C-chemokine receptor type 4 (CXCR4) and integrin beta1; and the cytokines/chemokines interleukin (IL) 1beta and IL-8. Expression of CXCR4 was unexpectedly high among all arthritis subjects. Using RT-PCR, ELISA and immunohistology, expression of stromal cell-derived factor 1 (SDF-1) was demonstrated in arthritis joints. CONCLUSIONS: The CXCR4/SDF-1 is a potential pro-inflammatory axis for RA, PsA and SpA.

Adolescent↗

PubMed2002

Design, pharmacokinetic, and pharmacodynamic evaluation of soft anticholinergics based on tropyl alpha-phenylcyclopentylacetate.

Four new soft anticholinergic agents based on tropyl alpha-phenylcyclopentylacetate, 15a, 15b, 18a, and 18b, were designed and synthesized. Receptor binding studies on the cloned human muscarinic receptors indicated that the new soft anticholinergic agents possessed moderate potency as pKi ranged from 6.7 to 7.6. Mydriatic studies in rabbit eyes revealed that the duration of the action of the new soft anticholinergics (8.5-11.0 h) were shorter than that of atropine (about 24 h) under pharmacodynamic equivalent dose, and one of them, 18a, showed even shorter than that of tropicamide. In addition, after unilateral administration, significant dilation of pupil in the control eyes was observed with tropicamide and atropine but not with soft drugs, suggesting the systemic activity of soft drugs was minimal. With their soft nature, the new soft anticholinergics displayed much shorter protective effect against carbachol-induced bradycardia (about 30 min) than atropine (at least 60 min) in rats. In vitro and in vivo pharmacokinetic studies demonstrated that the soft anticholinergics were rapidly hydrolyzed into the corresponding inactive metabolites once they were introduced into the systemic circulation.

Algorithms↗

PubMed2001

The initial steps of biogenesis of cyanobacterial photosystems occur in plasma membranes.

During oxygenic photosynthesis in cyanobacteria and chloroplasts of plants and eukaryotic algae, conversion of light energy to biologically useful chemical energy occurs in the specialized thylakoid membranes. Light-induced charge separation at the reaction centers of photosystems I and II, two multisubunit pigment-protein complexes in the thylakoid membranes, energetically drive sequential photosynthetic electron transfer reactions in this membrane system. In general, in the prokaryotic cyanobacterial cells, the thylakoid membrane is distinctly different from the plasma membrane. We have recently developed a two-dimensional separation procedure to purify thylakoid and plasma membranes from the genetically widely studied cyanobacterium Synechocystis sp. PCC 6803. Immunoblotting analysis demonstrated that the purified plasma membrane contained a number of protein components closely associated with the reaction centers of both photosystems. Moreover, these proteins were assembled in the plasma membrane as chlorophyll-containing multiprotein complexes, as evidenced from nondenaturing green gel and low-temperature fluorescence spectroscopy data. Furthermore, electron paramagnetic resonance spectroscopic analysis showed that in the partially assembled photosystem I core complex in the plasma membrane, the P700 reaction center was capable of undergoing light-induced charge separation. Based on these data, we propose that the plasma membrane, and not the thylakoid membrane, is the site for a number of the early steps of biogenesis of the photosynthetic reaction center complexes in these cyanobacterial cells.

Cell Membrane↗

PubMed2001

[Analysis of inflammation related gene expression spectrum in ankylosing spondylitis patients using cDNA microarray].

OBJECTIVE: To test the difference of gene expression patterns in peripheral blood mononuclear cells (PBMCs) among patients with ankylosing spondylitis (AS) or rheumatoid arthritis (RA) and healthy volunteers and to find potential discriminating genes. METHODS: The PBMC gene expression spectra of 7 patients with active AS, 6 patients with active RA, and 7 healthy volunteers were tested by microarray assay with 588 target gene filters. Semi-quantitative RT-PCR was performed to test the expression rates of thirteen inflammation related genes which had been with high expression rates in at least 4 arthritis patients by microarray assay, among 22 AS patients, 8 RA patients, and 7 healthy volunteers. RESULTS: The expression rates of inflammation related genes by cDNA microarray assay in healthy volunteer group, AS group, and RA group were 8.4%, 32.5% and 44.8% respectively (P < 0.001). The positive rate in RA patients was much higher than that in AS patients (P < 0.012). A significant increase of positive rate of these genes was also noticed in 2 active pulmonary tuberculosis patients. The expression rates of the 13 genes which were positive in at least 4 arthritis patients tested by semiquantitative RT-PCR in 22 AS patients, 8 RA patients and 7 healthy volunteers were very similar to those expression rates tested by cDNA microarray. CONCLUSION: The gene expression profiles of PBMC among AS patients are significantly abnormal and are significantly different from those among RA patients. The high expression of the 13 cytokines is not specific to AS.

Adolescent↗

PubMed2001

Characterization of the rat GRIK5 kainate receptor subunit gene promoter and its intragenic regions involved in neural cell specificity.

The GRIK5 (glutamate receptor ionotropic kainate-5) gene encodes the kainate-preferring glutamate receptor subunit KA2. The GRIK5 promoter is TATA-less and GC-rich, with multiple consensus initiator sequences. Transgenic mouse lines carrying 4 kilobases of the GRIK5 5'-flanking sequence showed lacZ reporter expression predominantly in the nervous system. Reporter assays in central glial (CG-4) and non-neural cells indicated that a 1200-base pair (bp) 5'-flanking region could sustain neural cell-specific promoter activity. Transcriptional activity was associated with the formation of a transcription factor IID-containing complex on an initiator sequence located 1100 bp upstream of the first intron. In transfection studies, deletion of exonic sequences downstream of the promoter resulted in reporter gene activity that was no longer neural cell-specific. When placed downstream of the GRIK5 promoter, a 77-bp sequence from the deleted fragment completely silenced reporter expression in NIH3T3 fibroblasts while attenuating activity in CG-4 cells. Analysis of the 77-bp sequence revealed a functional SP1-binding site and a sequence resembling a neuron-restrictive silencer element. The latter sequence, however, did not display cell-specific binding of REST-like proteins. Our studies thus provide evidence for intragenic control of GRIK5 promoter activity and suggest that elements contributing to tissue-specific expression are contained within the first exon.

5' Flanking Region↗

PubMed2001

FKBP12 binding modulates ryanodine receptor channel gating.

The ryanodine receptor (RyR1)/calcium release channel on the sarcoplasmic reticulum of skeletal muscle is comprised of four 565,000-dalton RyR1s, each of which binds one FK506 binding protein (FKBP12). RyR1 is required for excitation-contraction coupling in skeletal muscle. FKBP12, a cis-trans peptidyl-prolyl isomerase, is required for the normal gating of the RyR1 channel. In the absence of FKBP12, RyR1 channels exhibit increased gating frequency, suggesting that FKBP12 "stabilizes" the channel in the open and closed states. We now show that substitution of a Gly, Glu, or Ile for Val2461 in RyR1 prevents FKBP12 binding to RyR1, resulting in channels with increased gating frequency. In the case of the V2461I mutant RyR1, normal channel function can be restored by adding FKBP12.6, an isoform of FKBP12. These data identify Val2461 as a critical residue required for FKBP12 binding to RyR1 and demonstrate the functional role for FKBP12 in the RyR1 channel complex.

Amino Acid Sequence↗

PubMed2001

Urinary levels of interleukin-8 (IL-8) and disease activity in patients with IgA nephropathy.

Using quantitative sandwich ELISA, we studied 27 patients with IgA nephropathy to determine whether the levels of urinary IL-8 might reflect the disease activity. The levels of urinary IL-8 in patients with advanced stage IgA nephropathy were significantly higher than those in the patients with the mild stage of this disease, or in the healthy controls. The results showed a positive significant correlation between the levels of IL-8 and disease activity, i.e., between levels of urinary protein and urinary casts. A significant correlation between levels of urinary IL-8 and tubular function damage was also found. It was thus suggested that measurement of urinary IL-8 might be useful in evaluating the degree of renal injuries and/or prognosis in patients with IgA nephropathy.

Adult↗

PubMed2001

Expression of host defense scavenger receptors in spondylarthropathy.

OBJECTIVE: Reactive arthritis (ReA) is postulated to be caused by a defective host defense against gram-negative bacteria. HLA-B27 could play a role in this process, but does not account for the many HLA-B27 negative patients. The objective of this study was to test the expression of 3 macrophage scavenger receptors (SRs) that are responsible for innate immunity against gram-negative bacteria: SR class A type I (SR-AI), SR-AII, and the macrophage receptor with collagenous structure (MARCO). We postulate that defects in such receptors might also contribute to the host risk factors that increase the predisposition to ReA and perhaps other subtypes of spondylarthropathy (SpA). METHODS: Peripheral blood, synovial fluid, and synovial tissue samples were obtained from patients with recent Salmonella infection, ReA, other SpA, and rheumatoid arthritis (RA). The expression of SRs receptors was assessed by semiquantitative reverse transcriptase-polymerase chain reaction. RESULTS: Evaluation of the peripheral blood mononuclear cells (PBMC) from 4 patients who were recently infected with Salmonella, showed that PBMC from 2 patients who developed ReA expressed positive levels of MARCO, while PBMC from 2 patients who recovered from infection without sequelae did not. The synovial fluid mononuclear cells (SFMC) from some ReA patients expressed MARCO, but the levels were only moderate. The level of MARCO in the SFMC from the SpA patient group was low. In marked contrast, MARCO expression was high in almost all samples of RA SFMC. These findings also extended to synovial tissues. CONCLUSION: Expression of the host defense gene MARCO was susceptible to modulation, not only during infections, but also in the inflammatory arthritis conditions RA and SpA. MARCO is a variable to be considered as a candidate factor that might contribute to ReA.

Adolescent↗

PubMed2001

The phase-shift mutation in the glucocorticoid receptor gene: potential etiologic significance of neuroendocrine mechanisms in lupus nephritis.

BACKGROUND: Glucocorticoid (GC), a mediator of the hypothalamo-pituitary-adrenal axis, has been found to play an important role in maintaining the stability of immune endo-environment of the body. The pathogenesis of lupus nephritis, an autoimmune disease, is thought to be related to the intrinsic hyposensitivity to GC secreted by adrenal gland, and impairs the regulation of the immuno-neuro-endocrine axis. METHODS: To test this hypothesis, we examined the response of 39 clinic patients with lupus nephritis to GC and analyzed the molecular structure and function of the GC receptor (GR) on peripheral blood mononuclear cells. RESULTS: There was no difference in the level of ACTH, GC and ligand affinity of GR between the patients and the controls. The GR number on mononuclear cells of lupus patients was lower than that of the controls. There was no difference in GR number between the patients with heterogeneous response, i.e. sensitive, dependent and resistance, to GC. The analysis of exon 9 of the GC receptor with PCR-amplified single strand conformation polymorphism (PCR-SSCP) method showed the polymorphism in exon 9 of GC receptor in 8 of the 39 lupus nephritis patients. DNA sequence analysis revealed an adenine insertion at the 2439 base pair of the GC receptor gene. This phase-shift mutation caused an additional 20 amino acids being translated into protein of GC receptor. CONCLUSION: The decreased number of GC receptor and the molecular variation of GR on mononuclear cells could explain the phenomenon of GC resistance, potentially to endogenous GC, which suggested an etiological significance of neuro-endocrine-immune mechanism in lupus nephritis. This may be useful in the design of lupus nephritis therapy.

Adolescent↗

PubMed2001

The study of spectrosgraphic properties and molecular recognition of cali.

The absorption spectrum and fluorescence spectrum of calix[n]arenes are studied systematically. The self-association of calix[n]arenes is proposed and the self-association constant is also obtained. The recognition properties of calix[n]arenes are studied and the inclusion constant is calculated.

Calixarenes↗

PubMed2001

Trafficking of yellow-fluorescent-protein-tagged mu1 subunit of clathrin adaptor AP-1 complex in living cells.

Clathrin adaptor protein AP-1 complex is thought to function in forming clathrin-coated vesicles at the trans-Golgi network (TGN) and mediating transport of cargo between the TGN and endosomes. To study trafficking of AP-1 in living cells, yellow fluorescent protein (YFP) was inserted in the middle of mu1 A subunit of AP-1. When expressed in a tetracycline-dependent manner in HeLa cells, YFP-mu1 was efficiently incorporated into the AP-1 complex, replacing endogenous mu1 in most of cellular AP-1. Time-lapse imaging revealed that YFP-mu1/AP-1 departs from TGN as isolated vesicles and spherical structures, or varicosities, associated with fine tubular processes. Typically, several vesicles or varicosities were seen moving sequentially along the same 'tracks' from TGN to cell periphery. These data suggest that AP-1 may function after formation of Golgi transport intermediates in facilitating their intracellular movement. Mutagenesis of YFP-mu1 determined that the structural requirements for its binding to tyrosine-containing sequence motifs are similar to those previously defined in mu2 subunit of AP-2. Moreover, the carboxyl-terminal half of mu2 could replace the corresponding fragment of mu1 without loss of the ability of the resulting mu1-YFP-mu2 chimeric protein to incorporate into AP-1 and bind tyrosine-containing motifs. Mutations that abolish binding capacity for tyrosine motifs did not mistarget AP-1 in the cell, suggesting that AP-1 interactions with this type of sorting signals are not essential for membrane docking of AP-1 at the TGN. Altogether, this study demonstrates that YFP-tagged mu1 protein can serve as a useful tool for visualizing the dynamics of AP-1 in living cells and for the structure-function analysis of mu1-cargo interactions.

Bacterial Proteins↗

PubMed2001

Adenovirus-mediated expression of pig alpha(1, 3) galactosyltransferase reconstructs Gal alpha(1, 3) gal epitope on the surface of human tumor cells.

Gal alpha(1, 3) Gal (gal epitope) is a carbohydrate epitope and synthesized in large amount by alpha(1, 3) galactosyltransferase [alpha(1, 3) GT] enzyme on the cells of lower mammalian animals such as pigs and mice. Human has no gal epitope due to the inactivation of alpha(1, 3) GT gene but produces a large amount of antibodies (anti-Gal) which recognize Gal alpha(1, 3) Gal structures specifically. In this study, a replication-deficient recombinant adenoviral vector Ad5sGT containing pig alpha(1, 3) GT cDNA was constructed and characterized. Adenoviral vector-mediated transfer of pig alpha(1, 3) GT gene into human tumor cells such as malignant melanoma A375, stomach cancer SGC-7901, and lung cancer SPC-A-1 was reported for the first time. Results showed that Gal epitope did not increase the sensitivity of human tumor cells to human complement-mediated lysis, although human complement activation and the binding of human IgG and IgM natural antibodies to human tumor cells were enhanced significantly after Ad5sGT transduction. Appearance of gal epitope on the human tumor cells changed the expression of cell surface carbohydrates reacting with Ulex europaeus I (UEA I) lectins, Vicia villosa agglutinin (VVA), Arachis hypogaea agglutinin (PNA), and Glycine max agglutinin (SBA) to different degrees. In addition, no effect of gal epitope on the growth in vitro of human tumor cells was observed in MTT assay.

Adenoviridae↗

PubMed2001

GABA transporter 1 transcriptional starting site exhibiting tissue specific difference.

GABA transporter 1(GAT1) takes important roles in multiple physiological processes through the uptake and release of GABA, but the regulation of GAT1 gene expression in different tissues is rarely known. To address the question, first, 5' Rapid amplification of cDNA end (RACE) was used to determine GAT1 transcriptional starting sites in neonatal mouse cerebral cortex and intestine, adult mouse brain and adult rat testis. The products of 5'RACE were confirmed by DNA sequencing. We found that the transcript of GAT1 in neonatal mouse cerebral cortex and adult mouse brain starts at the same site (inside of exon 1), while in mouse intestine, GAT1 starts transcription in intron 1, and in rat testis, the transcript of GAT1 has an additional untranslation exon to the 5' direction.

Aging↗

PubMed2001

Spondyloarthropathies in eastern Asia.

The association between HLA-B27 and the spondyloarthropathies (SpAs) is so strong that it is supposed that the HLA-B27 molecule plays a pathogenetic role. In whites and Indonesians, the frequency of HLA-B27 is about 10%; in Chinese it is about 8%; and in Japanese it is only about 1%. The prevalence of SpA in the Chinese is at least 0.2%, but in native Indonesians, Philippinos, and Malaysians, SpA is rarely seen. Twenty-three subtypes (B*2701-B*2723) have been distinguished. These subtypes are not equally distributed over the world. In most countries the distribution of the subtypes among HLA-B27 SpA patients is the same as that among the normal HLA-B27-positive population. In China, the subtype B*2704 is frequent and the prevalence of SpA is high. Native Indonesians, however, mostly have subtype B*2706, and SpA is rarely seen in this population. It was shown that B*2706, probably like B*2709 in Sardinia, is not associated with SpA. The difference between the SpA-associated and non-SpA-associated subtypes is limited to only two amino acid residues (114 and 116) at the bottom of the peptide-binding groove of HLA-B27. This small difference between health and disease rewards studies for different peptide-binding capacities and may help us characterize the peptides that are involved in the pathogenesis of SpA. The differences in disease associations in these countries also have clinical implications. In Southeast Asia, HLA-B27 typing without subtyping has less clinical usefulness than in parts of the world where B*2706 is rarely seen. When native Indonesians, Malaysians, or Philippinos are suspected of having ankylosing spondylitis or a related SpA, it is worth asking if they had white or Chinese ancestors. If native HLA-B27-positive Indonesians (with subtypes other than B*2706) develop SpA, the clinical features are not different from those in other parts of the world. In the Chinese population on the mainland and in Taiwan, juvenile SpA is frequently seen. The onset is often a peripheral arthritis or enthesitis.

Asian People↗

PubMed2001

Protein phosphatase 2B inhibitor potentiates endothelial PKC activity and barrier dysfunction.

Serine/threonine (Ser/Thr) protein phosphatases (PPs) are implicated in the recovery from endothelial barrier dysfunction caused by inflammatory mediators. We hypothesized that Ser/Thr PPs may regulate protein kinase C (PKC), a critical signaling molecule in barrier dysfunction, in the promotion of barrier recovery. Western analysis indicated that bovine pulmonary microvascular endothelial cells (BPMECs) expressed the three major Ser/Thr PPs, PP1, PP2A, and PP2B. Pretreatment with 100 ng/ml of FK506 (a PP2B inhibitor) but not with the PP1 and PP2A inhibitors calyculin A or okadaic acid potentiated the thrombin-induced increase in PKC phosphotransferase activity. FK506 also potentiated thrombin-induced PKC-alpha but not PKC-beta phosphorylation. FK506 but not calyculin A or okadaic acid inhibited recovery from the thrombin-induced decrease in transendothelial resistance. Neither FK506 nor okadaic acid altered the thrombin-induced resistance decrease, whereas calyculin A potentiated the decrease. Downregulation of PKC with phorbol 12-myristate 13-acetate rescued the FK506-mediated inhibition of recovery, which was consistent with the finding that the thrombin-induced phosphorylation of PKC-alpha was reduced during the recovery phase. These results indicated that PP2B may play a physiologically important role in returning endothelial barrier dysfunction to normal through the regulation of PKC.

Animals↗

PubMed2001

Receptor binding studies of soft anticholinergic agents.

Receptor binding studies were performed on 24 soft anticholinergic agents and 5 conventional anticholinergic agents using 4 cloned human muscarinic receptor subtypes. The measured pK(i) values of the soft anticholinergic agents ranged from 6.5 to 9.5, with the majority being in the range of 7.5 to 8.5. Strong correlation was observed between the pK(i) s determined here and the pA 2 values measured earlier in guinea pig ileum contraction assays. The corresponding correlation coefficients (r2) were 0.80, 0.73, 0.81, and 0.78 for pK(i) (m1), pK(i) (m2), pK(i) (m3), and pK(i) (m4), respectively. Quantitative structure-activity relationship (QSAR) studies were also performed, and good characterization could be obtained for the soft anticholinergics containing at least 1 tropine moiety in their structure. For these compounds, the potency as measured by the pK i values was found to be related to geometric, electronic, and lipophilicity descriptors. A linear regression equation using ovality (O(e)), dipole moment (D), and a calculated log octanol-water partition coefficient (QLogP) gave reasonably good descriptions (r = 0.88) for the pK(i) (m3) values.

Animals↗

PubMed2001

[Using fluorescence PCR analysis for early diagnosis and carriers detection of Chinese Wilson's disease].

OBJECTIVE: To diagnose the pre-symptomatic cases of Chinese Wilson's disease(WD) and detect the potential carriers by using fluorescence PCR technique. METHODS: Screening the high frequency spot Arg778Leu of WD gene mutation in 66 Chinese WD patients, 55 healthy family members and 30 controls, and selecting 3 random samples (2 from WD patients, 1 from controls) for DNA sequencing to testify the accuracy of fluorescence PCR. RESULTS: Among 66 Chinese WD cases, 5 were found homozygous for mutation of Arg778Leu and 21 were compound heterozygous; the gene mutation rate was totally 39.4%. Out of 55 healthy family members, 12 were found heterozygous, and it was confirmed that 11 of these 12 individuals were WD gene carriers but not pre-symptomatic patients. The results of direct DNA sequencing consisted with those results detected by fluorescence PCR. CONCLUSION: The Arg778Leu mutation in exon 8 is the high frequency spot of Chinese Wilson's disease gene. Fluorescence PCR analysis is a rapid, accurate gene diagnostic method and demonstrates a high detecting rate.

Adolescent↗

PubMed2001

[Genetic polymorphism of eight STR loci in the Han population in Henan province].

OBJECTIVE: To investigate the allele frequencies of eight short tandem repeats(STR) loci: TH01, FES, D19S400, D7S820, D16S539, D20S161, D3S1545 and D5S818 in Han population in Henan province. METHODS: DNA was extracted with phenol-chloroform from EDTA-blood samples of the unrelated individuals in Henan province and amplified with PCR technique. The PCR product was analyzed with the undenatured PAGE vertical electrophoresis and silver-stain. RESULTS: The authors got the frequencies of the eight loci. The heterozygosities of the eight loci are 0.66, 0.67, 0.80, 0.76, 0.79, 0.79, 0.78 and 0.78; the discrimination powers are 0.83, 0.83, 0.94, 0.91, 0.93, 0.93, 0.92 and 0.92. CONCLUSION: The heterozygosities of the eight loci are high and the frequencies are in good agreement with Hardy-Weinberg equilibrium, so the eight loci can be used in individual identification testing.

Alleles↗