Biomedical subjects
Publications and source records attributed to S Liang.
The toxic peptides in Amanita virosa and Amanita verna from various region were analyzed by a reversed-phase high-performance liquid chromatography. It is showed that there were great differences in the concentrations of toxic peptides between species and strains of A. virosa and A. verna.
Immuonosuppressive drugs FK506 and rapamycin block a number of signal transduction pathways in eukaryotic systems. The 12 kDa FK506 binding protein (FKBP12) mediates the action of both FK506 and rapamycin against their functional targets. In this report, we cloned, sequenced and characterized a gene encoding FKBP12 in Vicia faba (VfFKBP12). While VfFKBP12 is highly homologous to animal and yeast FKBP12, it does not mediate the action of FK506 and rapamycin. There are unique features in plant FKBP12 sequences that cause the variation in their function. One lies in the domain that is critical for interaction with calcineurin (CaN), the mammalian and yeast target of FKBP12-FK506 complex. Protein-protein interaction assays revealed a low-affinity and unstable VfFKBP12-FK506-CaN ternary complex. In the genetic assay, VfFKBP12 did not restore the sensitivity of yeast FKBP12 mutant to rapamycin or FK506, supporting that plant FKBP12-ligand complexes are unable to block the function of the drug target. Also unique to plant FKBP12 proteins, a pair of cysteines is spatially adjacent to potentially form disulfide linkage. Treatment of VfFKBP12 with reductant dithiothreitol (DTT) abolished the formation of VfFKBP12-FK506-CaN ternary complex. Site-directed mutagenesis to substitute one of the cysteines, Cys26, with Ser produced a similar effect as DTT treatment. These results indicate that an intramolecular disulfide bond is a novel structural feature required for the low-affinity interaction between plant FKBP12 and CaN. In conclusion, plant FKBP12 proteins have evolved structural changes that modify their protein-protein interacting domains and cause loss of function against the drug targets.
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K-ras oncogene mutations were detected with PCR-SSCP-DNA direct sequencing technique in 40 cases of lung cancer. The result of PCR-SSCP silver staining indicated that the mutational rate was 30% (12/40), all mutations were observed in lung adenocarcinoma and its mutational rate was 44% (12/27). The result of DNA direct sequencing showed that 11 of the 12 positive samples screened by PCR-SSCP had mutation and 90% of K-ras mutations were in codon 12. The mutation was mainly G-->T transversion and G-->A transition. The study suggested that SSCP silver staining analysis is very useful in screening large amount of samples simultaneously and PCR-SSCP-DNA direct sequencing method is quite efficient for the detection of oncogene mutation.
OBJECTIVE: To investigate the difference between cerebellar medulloblastoma (MB) and primary cerebral small cell tumor in histogenesis, morphologic features and biological behavior. METHODS: 210 cases of MB and 9 cases of small cell tumor of cerebrum were observed with histologic and immunohistochemical techniques. RESULTS: Both tumors were composed of primitive cells with focal evidence of glial and/or neuronal differentiation. In 63.2% of MB and five-eighths of small cell tumor of cerebrum coexpressed GFAP and Syn. Both types of tumors were highly malignant. The overall 1 year survival rates were 34.63% and 25.65% respectively. The survival rate was lower for patients with high proliferative index, with necrosis or without receiving radiation therapy. CONCLUSION: MB is similar to other small cell tumors of CNS in morphological features, specific marker expression and biological behavior. These tumors can be classified as primitive neuro-ectodermal tumors (PNET).
This paper reports the results of identification of powdered Bungarus muliteinctus and its adulterants (Natrix annularis; Dinodon rufozonatum and Bungarus fascitus). A comparative table of and a key to the characteristics are given.
DNA was extracted from human bones remained for 3 to 15 years by using the method developed in our laboratory. A pair of X-Y homologous primer were used to amplify the specific DNA fragments of Amelogenin gene. All the 35 samples yielded correct results. Our method is rapid, sensitive and reliable for the sex determination of skeletal remains.
This paper presents a convenient long-distance ECG communication system. It is made up of portable ECG recorder, hospital work station and public telephone network. This paper also introduces a kind of modulation and demodulation method in sound coupling communication-pulse edge modulation and demodulation. This system has rational structure, convenient operation and strong management power. A Philips 80C552 used as CPU in the recoder which is portable, cheap and reliable.
Trough pharmacologic test on seral cholesterol and teral triglycerin in normal mouse fed by high fat food and by hperlipidemia mouse, it is found that "Sen Yang Formulae" has a very good effect on lowering cholesterol and triglycerin, and it also has good effect on promoting quantity of HDL-C, ratio of HDL-C/TC, lowering quantity of LDL-C and improving condition of blood viscosity and fluidity. The results indicate that it is effective on prevention and treatment of hyperlipidemia and benefits at prevention and treatment of arteriosclerosis and coronary heart disease.
Given the immanent gene expression mapping covering whole genomes during development, health and disease, we seek computational methods to maximize functional inference from such large data sets. Is it possible, in principle, to completely infer a complex regulatory network architecture from input/output patterns of its variables? We investigated this possibility using binary models of genetic networks. Trajectories, or state transition tables of Boolean nets, resemble time series of gene expression. By systematically analyzing the mutual information between input states and output states, one is able to infer the sets of input elements controlling each element or gene in the network. This process is unequivocal and exact for complete state transition tables. We implemented this REVerse Engineering ALgorithm (REVEAL) in a C program, and found the problem to be tractable within the conditions tested so far. For n = 50 (elements) and k = 3 (inputs per element), the analysis of incomplete state transition tables (100 state transition pairs out of a possible 10(15)) reliably produced the original rule and wiring sets. While this study is limited to synchronous Boolean networks, the algorithm is generalizable to include multi-state models, essentially allowing direct application to realistic biological data sets. The ability to adequately solve the inverse problem may enable in-depth analysis of complex dynamic systems in biology and other fields.
In this paper we show how Boolean genetic networks could be used to address complex problems in cancer biology. First, we describe a general strategy to generate Boolean genetic networks that incorporate all relevant biochemical and physiological parameters and cover all of their regulatory interactions in a deterministic manner. Second, we introduce "realistic Boolean genetic networks" that produce time series measurements very similar to those detected in actual biological systems. Third, we outline a series of essential questions related to cancer biology and cancer therapy that could be addressed by the use of "realistic Boolean genetic network" modeling.
The biological activity of the transcription factor NF-kappaB is mainly controlled by the IkappaB proteins IkappaBalpha and IkappaBbeta, which restrict NF-kappaB in the cytoplasm and enter the nucleus where they terminate NF-kappaB-dependent transcription. In this paper we describe the cloning and functional characterization of mouse IkappaBepsilon. Mouse IkappaBepsilon contains 6 ankyrin repeats required for its interaction with the Rel proteins and is expressed in different cell types where we found that it is up-regulated by NF-kappaB inducers, as is the case for IkappaBalpha and human IkappaBepsilon. IkappaBepsilon functions as a bona fide IkappaB protein by restricting Rel proteins in the cytoplasm and inhibiting their in vitro DNA binding activity. Surprisingly, IkappaBepsilon did not inhibit transcription of genes regulated by the p50/p65 heterodimer efficiently, such as the human interferon-beta gene. However, IkappaBepsilon was a strong inhibitor of interleukin-8 expression, a gene known to be regulated by p65 homodimers. In addition, IkappaBepsilon appears to function predominantly in the cytoplasm to sequester p65 homodimers, in contrast with the other two members of the family, IkappaBalpha and IkappaBbeta, which also function in the nucleus to terminate NF-kappaB-dependent transcriptional activation.
A new derivatizing reagent, acetyl isothiocyanate (AITC), is applied for C-terminal peptide sequencing. It has been successfully used to sequence six C-terminal residues of a synthetic peptide at low nanomole levels. According to the mechanism study of the derivatization of C-terminal amino acid with various reagents, the derivatizing reagents (R-N=C=S or SCN-) were classified into three types: type I, the ionic compound (e.g., HSCN, NH4SCN, KSCN); type II, R is a good leaving group (e.g., TMS-ITC, TBSn-ITC); type III, R is reactive (e.g., AITC, BITC, DPP-ITC). Type III reagents are superior to other reagents because their double-function of activation and derivatization. Unlike type I and type II reagents, type III reagent chemistry does not require oxazolinone formation which can cause racemization. Compared with benzoyl isothiocyanate, diphenyl phosphoroisothiocyanatidate, trimethylsilyl isothiocyanate, and ammonium thiocyanate, AITC is the most effective derivatizing reagent. As a type III reagent, AITC possesses some features such as no need for oxazolinone formation, no requirement for separate activation step, high reactivity, easy preparation, and low absorption at 260-270 nm. Different reaction conditions were investigated for optimization and the chemical mechanism of AITC chemistry is illustrated. A convenient and efficient approach for synthesis of amino acid thiohydantoins as reference standards has also been developed.
A novel and efficient method to prepare amino acid thiohydantoins, which are required as reference standards for development of C-terminal protein sequencing, is reported. Amino acid thiohydantoins were prepared using a straightforward method involving reaction of 20 free amino acids with acetyl chloride as activating reagent and trimethylsilyl isothiocyanate (TMS-ITC) as derivatizing reagent. The products were characterized by HPLC, uv spectra, amino acid analysis, MS, and NMR. Different reaction conditions were investigated and the chemical mechanism of the formation of amino acid thiohydantoins was illustrated.
Although immunosuppressant immunophilin ligands promote neurite outgrowth in vitro, their neurotrophic activities are clearly independent of their immunosuppressive activity. In the present report, a novel nonimmunosuppressive immunophilin ligand, GPI-1046 (3-(3-pyridyl)-1-propyl (2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate+ ++) is described. In vitro, GPI-1046 bound to FK506 binding protein-12 and elicited neurite outgrowth from sensory neuronal cultures with picomolar potency with maximal effects comparable to nerve growth factor. In vivo, GPI-1046 stimulated the regeneration of lesioned sciatic nerve axons and myelin levels. In the central nervous system, GPI-1046 promoted protection and/or sprouting of serotonin-containing nerve fibers in somatosensory cortex following parachloroamphetamine treatment. GPI-1046 also induced regenerative sprouting from spared nigrostriatal dopaminergic neurons following 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxicity in mice or 6-hydroxydopamine (6-OHDA) toxicity in rats. The rotational abnormality in 6-OHDA treated rats was alleviated by GPI-1046. These neurotrophic actions in multiple models suggest therapeutic utility for GPI-1046 in neurodegenerative diseases.
The three-dimensional structure in aqueous solution of native huwentoxin-I, a neurotoxin from the venom of the spider Selenocosmia huwena, has been determined from two-dimensional H NMR data recorded at 500 and 600 MHz. Structural constraints consisting of interproton distances inferred from NOEs and dihedral angles from spin-spin coupling constants were used as input for distance geometry calculation with the program XPLOR 3.1. The best 10 structures have NOE violations < 0.3 A, dihedral violations < 2 degrees, and pairwise root-mean-square differences of 1.08 (+/- 0.20) A over backbone atoms (N, C alpha, C). The molecule adopts a compact structure consisting of a small triple-stranded antiparallel beta-sheet and five beta-turns. A small hydrophobic patch consisting of Phe 6, Trp 28, and Trp 31 is located on one side of the molecule. All six lysine residues are distributed on the molecular surface. The three disulfide bridges are buried within the molecule. The structure contains an "inhibitor cystine knot motif" which is adopted by several other small proteins, such as omega-conotoxin, agatoxin IVA, and gurmarin.
Current literature suggests that a massive influx of Ca2+ into the cells of the CNS induces cell damage associated with traumatic brain injury (TBI). Using an in vitro model for stretch-induced cell injury developed by our laboratory, we have investigated the role of extracellular Ca2+ in astrocyte injury. The degree of injury was assessed by measurement of propidium iodide uptake and release of lactate dehydrogenase. Based on results of in vivo models of TBI developed by others, our initial hypothesis was that decreasing extracellular Ca2+ would result in a reduction in astrocyte injury. Quite unexpectedly, our results indicate that decreasing extracellular Ca2+ to levels observed after in vivo TBI increased astrocyte injury. Elevating the extracellular Ca2+ content to twofold above physiological levels (2 mM) produced a reduction in cell injury. The reduction in injury afforded by Ca2+ could not be mimicked with Ba2+, Mn2+, Zn2+, or Mg2+, suggesting that a Ca(2+)-specific mechanism is involved. Using 45Ca2+, we demonstrate that injury induces a rapid influx of extracellular Ca2+ into the astrocyte, achieving an elevation in total cell-associated Ca2+ content two- to threefold above basal levels. Pharmacological elevation of intracellular Ca2+ levels with the Ca2+ ionophore A23187 or thapsigargin before injury dramatically reduced astrocyte injury. Our data suggest that, contrary to popular assumptions, an elevation of total cell-associated Ca2+ reduces astrocyte injury produced by a traumatic insult.
The mammalian high-mobility-group protein I(Y) [HMG I(Y)], while not a typical transcriptional activator, is required for the expression of many eukaryotic genes. HMG I(Y) appears to recruit and stabilize complexes of transcriptional activators through protein-DNA and protein-protein interactions. The protein binds to the minor groove of DNA via three short basic repeats, preferring tracts of adenines and thymines arranged on the same face of the DNA helix. However, the mode by which these three basic repeats function together to recognize HMG I(Y) binding sites has remained unclear. Here, using deletion mutants of HMG I(Y), DNase I footprinting, methylation interference, and in vivo transcriptional assays, we have characterized the binding of HMG I(Y) to the model beta-interferon enhancer. We show that two molecules of HMG I(Y) bind to the enhancer in a highly cooperative fashion, each molecule using a distinct pair of basic repeats to recognize the tandem AT-rich regions of the binding sites. We have also characterized the function of each basic repeat, showing that only the central repeat accounts for specific DNA binding and that the presence of a second repeat bound to an adjacent AT-rich region results in intramolecular cooperativity in binding. Surprisingly, the carboxyl-terminal acidic tail of HMG I(Y) is also important for specific binding in the context of the full-length protein. Our results present a detailed examination of HMG I(Y) binding in an important biological context, which can be extended not only to HMG I(Y) binding in other systems but also to the binding mode of many other proteins containing homologous basic repeats, which have been conserved from bacteria to humans.