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Biomedical subjects

Jun Li

Publications and source records attributed to Jun Li.

At least 361 records · Page 20Linked to original sources

A genetic variation map for chicken with 2.8 million single-nucleotide polymorphisms.

We describe a genetic variation map for the chicken genome containing 2.8 million single-nucleotide polymorphisms (SNPs). This map is based on a comparison of the sequences of three domestic chicken breeds (a broiler, a layer and a Chinese silkie) with that of their wild ancestor, red jungle fowl. Subsequent experiments indicate that at least 90% of the variant sites are true SNPs, and at least 70% are common SNPs that segregate in many domestic breeds. Mean nucleotide diversity is about five SNPs per kilobase for almost every possible comparison between red jungle fowl and domestic lines, between two different domestic lines, and within domestic lines--in contrast to the notion that domestic animals are highly inbred relative to their wild ancestors. In fact, most of the SNPs originated before domestication, and there is little evidence of selective sweeps for adaptive alleles on length scales greater than 100 kilobases.

Alleles↗

Conduction switching of photochromic molecules.

We report a theoretical study of single molecule conduction switching of photochromic dithienylethene molecules. The light-induced intramolecular transformation drives a swapping of the highest occupied molecular orbital and lowest unoccupied molecular orbital between two distinct conjugated paths. The shuffling of single and double bonds produces a significant conductance change when the molecule is sandwiched between metal electrodes. We model the switching event using quantum molecular dynamics and the conductance changes using Green's function electronic transport theory. We find large on-off conductance ratios (between 10 and over 100) depending on the side group outside the switching core.

Journal Article↗

ISSLS prize winner: LMP-1 upregulates intervertebral disc cell production of proteoglycans and BMPs in vitro and in vivo.

STUDY DESIGN: Experiments using both in vitro tissue culture and in vivo rabbit methods were used to study the effect of Lim Mineralization Protein-1 (LMP-1) on intervertebral disc (IVD) cell production of proteoglycans and bone morphogenetic proteins (BMPs). OBJECTIVES: To determine the effect of LMP-1 overexpression in IVD cells on the production of proteoglycans and BMPs both in vitro and in vivo and to show that LMP-1 mediates the control of proteoglycan production through its action on BMPs. SUMMARY OF BACKGROUND DATA: Because BMPs are known to increase proteoglycan synthesis and LMP-1 is known to upregulate BMPs in certain cells, it was hypothesized that LMP-1 may increase proteoglycan production in IVD cells. METHODS: DMMB, real-time polymerase chain reaction, and ELISA methods were used to quantitate proteoglycan, mRNA, and protein levels, respectively. Noggin was used to block the effect of the adenovirus carrying LMP-1 (AdLMP-1) on proteoglycan synthesis. In vivo experiments using intradiscal AdLMP-1 injection were performed with New Zealand White rabbits. Three weeks later, the mRNA levels of LMP-1, aggrecan, BMP-2, and BMP-7 were measured. RESULTS: In vitro experiments revealed that the sulfated glycosaminoglycan (sGAG) and aggrecan mRNA levels were significantly increased with AdLMP-1 treatment. Similarly, BMP-2 and BMP-7 mRNA and protein levels increased significantly, but BMP-4 and BMP-6 levels were unchanged. Noggin blocked the upregulation of proteoglycan by AdLMP-1. In vivo discs injected with AdLMP-1 had significantly elevated levels of LMP-1, BMP-2, and BMP-7 mRNA. CONCLUSIONS: LMP-1 overexpression increases disc cell production of proteoglycan, BMP-2, and BMP-7. LMP-1 mediates the control of proteoglycan production through its action on BMP.

Adaptor Proteins, Signal Transducing↗

Catecholamines are involved in the mechanism of the urinary alcohol level cycle in rats fed ethanol intragastrically at a constant rate.

Studies have indicated that blood alcohol levels cycle exists when ethanol is fed continuously using the intragastric feeding rat model of early alcoholic liver disease. The aim of the present study was to determine the role played by catecholamines in the pathogenesis of the blood alcohol cycling observed when ethanol is fed at a constant rate. The rats were tested at the peaks and troughs of the urinary alcohol level (UAL) cycle and the results were compared with controls. Blood catecholamine levels were markedly increased at the peaks, but not at the troughs. Propranolol, a beta adrenergic blocker, attenuated the amplitude of the cycle. Phenoxybenzamine, an alpha blocker disrupted the cycle and elevated ethanol to fatal levels. The results indicate that both alpha and beta adrenergic mechanisms are required for the cycle to occur.

Animals↗

Structural basis of constitutive activity and a unique nucleotide binding mode of human Pim-1 kinase.

Pim-1 kinase is a member of a distinct class of serine/threonine kinases consisting of Pim-1, Pim-2, and Pim-3. Pim kinases are highly homologous to one another and share a unique consensus hinge region sequence, ER-PXPX, with its two proline residues separated by a non-conserved residue, but they (Pim kinases) have <30% sequence identity with other kinases. Pim-1 has been implicated in both cytokine-induced signal transduction and the development of lymphoid malignancies. We have determined the crystal structures of apo Pim-1 kinase and its AMP-PNP (5'-adenylyl-beta,gamma-imidodiphosphate) complex to 2.1-angstroms resolutions. The structures reveal the following. 1) The kinase adopts a constitutively active conformation, and extensive hydrophobic and hydrogen bond interactions between the activation loop and the catalytic loop might be the structural basis for maintaining such a conformation. 2) The hinge region has a novel architecture and hydrogen-bonding pattern, which not only expand the ATP pocket but also serve to establish unambiguously the alignment of the Pim-1 hinge region with that of other kinases. 3) The binding mode of AMP-PNP to Pim-1 kinase is unique and does not involve a critical hinge region hydrogen bond interaction. Analysis of the reported Pim-1 kinase-domain structures leads to a hypothesis as to how Pim kinase activity might be regulated in vivo.

Adenylyl Imidodiphosphate↗

Icosahedral gold cage clusters: M@Au12- (M=V, Nb, and Ta).

We report the observation and characterization of a series of stable bimetallic 18-valence-electron clusters containing a highly symmetric 12-atom icosahedral Au cage with an encapsulated central heteroatom of Group VB transition metals, M@Au(12) (-) (M=V,Nb,Ta). Electronic and structural properties of these clusters were probed by anion photoelectron spectroscopy and theoretical calculations. Characteristics of the M@Au(12) (-) species include their remarkably high binding energies and relatively simple spectral features, which reflect their high symmetry and stability. The adiabatic electronic binding energies of M@Au(12) (-) were measured to be 3.70+/-0.03, 3.77+/-0.03, and 3.76+/-0.03 eV for M=V, Nb, and Ta, respectively. Comparison of density-functional calculations with experimental data established the highly symmetric icosahedral structures for the 18-electron cluster anions, which may be promising building blocks for cluster-assembled nanomaterials in the form of stoichiometric [M@Au(12) (-)]X(+) salts.

Journal Article↗

4,5-dimethylthio-4'-[2-(9-anthryloxy)ethylthio]tetrathiafulvalene, a highly selective and sensitive chemiluminescence probe for singlet oxygen.

4,5-Dimethylthio-4'-[2-(9-anthryloxy)ethylthio]tetrathiafulvalene has been designed and synthesized as a highly selective and sensitive chemiluminescence (CL) probe for singlet oxygen ((1)O(2)). The design strategy for the probe is directed by the idea of photoinduced electron-transfer process and carried out through the incorporation of electron-rich tetrathiafulvalene unit into a reactive luminophore of anthracene specific for (1)O(2). Upon reaction with reactive oxygen species (ROS), such as hydrogen peroxide, hypochlorite, superoxide, hydroxyl radical, or (1)O(2), the probe exhibits both strong CL response to and high selectivity for (1)O(2) only, rather than the other ROS. This remarkable CL property permits (1)O(2) to be distinguished easily from the other ROS and makes the probe possible to be used widely for (1)O(2) detection in many chemical and biological systems and even in light water (H(2)O) environments. This applicability has been demonstrated by monitoring the (1)O(2) generation in a metal-catalyzed decomposition system of tert-butyl hydroperoxide. Moreover, the CL reaction mechanism of the present system is also discussed, clearly confirming that the introduction of electron-rich tetrathiafulvalene into the 9-position of anthracene can greatly activate its reactivity toward (1)O(2).

Journal Article↗

Evidence and characteristics of putative human alpha recombination hotspots.

Understanding recombination rate variation is very important for studying genome diversity and evolution, and for investigation of phenotypic association and genetic diseases. Recombination hotspots have been observed in many species and are well studied in yeast. Recent study demonstrated that recombination hotspots are also a ubiquitous feature of the human genome. But the nature of human hotspots remains largely unknown. We have developed and validated a novel computational method for testing the existence of hotspots as well as for localizing them with either unphased or phased genotyping data. To study the characteristics of hotspots within or close to genes, we scanned for unusually high levels of recombination using the European population samples in the SeattleSNPs database, and found evidence for the existence of human alpha hotspots similar to those of yeast. This type of hotspots, found at promoter regions, accounts for about half of the total detected and appears to depend on some specific transcription factor binding sites (such as CGCCCCCGC). These characteristics can explain the observed weak correlation between hotspots and GC-content, and their variation may contribute to the diversity of hotspot distribution among different individuals and species. These long-sought putative human alpha recombination hotspots should deserve further experimental investigations.

Binding Sites↗

Miniaturized multiplex label-free electronic chip for rapid nucleic acid analysis based on carbon nanotube nanoelectrode arrays.

BACKGROUND: Reducing cost and time is the major concern in clinical diagnostics, particularly in molecular diagnostics. Miniaturization technologies have been recognized as promising solutions to provide low-cost microchips for diagnostics. With the recent advancement in nanotechnologies, it is possible to further improve detection sensitivity and simplify sample preparation by incorporating nanoscale elements in diagnostics devices. A fusion of micro- and nanotechnologies with biology has great potential for the development of low-cost disposable chips for rapid molecular analysis that can be carried out with simple handheld devices. APPROACH: Vertically aligned multiwalled carbon nanotubes (MWNTs) are fabricated on predeposited microelectrode pads and encapsulated in SiO2 dielectrics with only the very end exposed at the surface to form an inlaid nanoelectrode array (NEA). The NEA is used to collect the electrochemical signal associated with the target molecules binding to the probe molecules, which are covalently attached to the end of the MWNTs. CONTENT: A 3 x 3 microelectrode array is presented to demonstrate the miniaturization and multiplexing capability. A randomly distributed MWNT NEA is fabricated on each microelectrode pad. Selective functionalization of the MWNT end with a specific oligonucleotide probe and passivation of the SiO2 surface with ethylene glycol moieties are discussed. Ru(bpy)2+ -mediator-amplified guanine oxidation is used to directly measure the electrochemical signal associated with target molecules. SUMMARY: The discussed MWNT NEAs have ultrahigh sensitivity in direct electrochemical detection of guanine bases in the nucleic acid target. Fewer than approximately 1000 target nucleic acid molecules can be measured with a single microelectrode pad of approximately 20 x 20 microm2, which approaches the detection limit of laser scanners in fluorescence-based DNA microarray techniques. MWNT NEAs can be easily integrated with microelectronic circuitry and microfluidics for development of a fully automated system for rapid molecular analysis with minimum cost.

Algorithms↗

[Association between serotonin 2C gene polymorphisms and attention deficit hyperactivity disorder comorbid or not comorbid with learning disorder].

OBJECTIVE: To investigate the relationship between two HTR2C gene polymorphisms, that is C-759T and G-697C polymorphisms, and attention deficit hyperactivity disorder (ADHD) comorbid or not comorbid learning disorder (LD). METHODS: Blood samples were taken from 189 trios with probands of ADHD comorbid LD (ADHD+LD) and 299 trios with probands of ADHD not comorbid LD (ADHD-LD). DNA was extracted and PCR was performed to amplify the fragments containing both C-759T and G-697C polymorphisms. Aci I was used to detect different alleles of the two polymorphisms. Allele- based and haplotype- based TDT analysis were used to test the association of the two polymorphisms of HTR2C gene and ADHD-LD and ADHD+LD. RESULTS: -759C(chi(2)=6.961, P=0.008), -697G(chi(2)=8.346, P=0.004), as well as -759C/-697G haplotype were over- transmitted(chi(2)=9.000, P=0.002 7), while haplotype -759T/-697C was under- transmitted(chi(2)=7.784, P=0.005 3) to probands with ADHD-LD. No biased transmission of any allele and haplotype were found in families with probands of ADHD+LD. CONCLUSION: ADHD-LD and ADHD+LD are different at the level of HTR2C gene polymrohisms of C-759T and G-697C. HTR2C is related to ADHD-LD, while not related to ADHD+LD.

Attention Deficit Disorder with Hyperactivity↗

FR167653 attenuates murine immunological liver injury.

AIM: To study the effect of FR167653 on immunological liver injury (ILI) in mice. METHODS: ILI was established by tail vein injection of 2.5 mg Bacillus Calmette-Guerin (BCG), and 10 d later with 10 mg lipopolysaccharide (LPS) in 0.2 mL saline (BCG plus LPS). Alanine aminotransferase (ALT), aspartate aminotransferase (AST) in sera and malondialdehyde (MDA), glutathione peroxidase (GSHpx) contents in liver homogenates were assayed by spectrophotometry. The levels of tumor necrosis factor-alpha (TNF-alpha) and nitric oxide (NO) levels in sera were determined using ELISA. Interleukin-1 (IL-1) produced by peritoneal macrophages was determined by the method of (3)H-infiltrated cell proliferation. The nuclear factor-kappa B (NF-kappaB) p65 in liver tissue was analyzed with reverse transcription polymerase chain reaction (RT-PCR). Liver samples collected were stained with hematoxylin and eosin. RESULTS: FR167653 (50, 100, 150 mg/kg) could significantly decrease the serum transaminase (ALT, AST) activity and MDA content in liver homogenate, and improve reduced GSHpx level of liver homogenate. Liver histopathological examination showed FR167653 (100, 150 mg/kg) significantly reduced inflammatory cells infiltration and liver cells necrosis. FR167653 (50, 100, 150 mg/kg) significantly lowered TNF-alpha and NO levels in serum, and IL-1 produced by peritoneal macrophages. Moreover, expression of NF-kappaB mRNA in liver tissue of ILI induced by BCG plus LPS was significantly reduced by FR167653. CONCLUSION: All results showed that FR167653 had significant inhibitory action on ILI in mice.

Animals↗

A new approach to random mutagenesis in vitro.

Random mutagenesis is a powerful tool for studying the effects of a large number of permutations of a particular DNA sequence and its encoded products. Here we describe a new strategy of conducting in vitro random mutagenesis using ethyl methane sulfonate (EMS). The Bacillus aprN18 gene, coding for a serine protease with fibrinolytic activity, was used as a target gene. To study the mutations of the coding region, rather than the whole plasmid, the 1.4 kb gene fragment was cut out from an expression plasmid and treated with 10 mM EMS at 37 degrees C for 1 h. The treated fragment was then ligated back into the original expression vector and a library of random mutants was constructed in a protease-deficient Bacillus subtilis strain. A plate assay-based screening method was used to select for mutant clones with altered enzyme activity, and the change of activity was then confirmed by a semi-quantitative enzyme assay using liquid culture supernatant. The inserts of five clones with altered enzyme activity were randomly chosen for sequencing analysis. Among the point mutations detected, GC --> AT transition accounts for 42.1%, AT --> GC transition 34.2% and GC/CG transversion 23.7%, respectively. To our knowledge this is the first application of EMS for in vitro mutagenesis of a defined DNA sequence.

Amino Acid Sequence↗

Hyperphosphorylation of rat liver proteasome subunits: the effects of ethanol and okadaic acid are compared.

In experimental alcoholic liver disease, protein degradation by the ATP-ubiquitin-proteasome pathway is inhibited. Failure of the proteasome to eliminate cytoplasmic proteins leads to the accumulation of oxidized and otherwise modified proteins. One possible explanation for the inhibition of the proteasome is hyperphosphorylation of proteasome subunits. To examine this possibility, the 26S proteasomes from the liver of rats fed ethanol and a pair-fed control were studied by isolating the proteasomes in a purified fraction. The effect of ethanol on the phosphorylation of proteasomal subunits was compared with the hyperphosphorylation of the proteasomes caused by okadaic acid given to rats in vivo. Ethanol ingestion caused an inhibition of the chymotrypsin-like activity of the purified proteasome. The 2D electrophoresis and Western blot analysis of the purified 20S and 26S proteasomes from the ethanol-fed rats indicated that hyperphosphorylation of proteasomal subunits had occured. The proteasomal alpha type subunits C9/alpha3 and C8/alpha7 were hyperphosphorylated compared to the controls. Chymotrypsin-like activity was also inhibited by okadaic acid treatment similar to ethanol feeding. The 26S proteasome fraction examined by isoelectric focusing gel revealed many hyperphosphorylated bands in the proteasomes from the okadaic acid treated and the ethanol fed rat livers compared with the controls. In conclusion hyperphosphorylation of the proteasome subunits occurs in the ethanol treated proteasomal subunits which could be one mechanism of the inhibition of the 26S proteasome caused by ethanol feeding.

Adenosine Triphosphatases↗