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1.  The loss of taste genes in cetaceans 
BMC Evolutionary Biology  2014;14(1):218.
Five basic taste modalities, sour, sweet, bitter, salt and umami, can be distinguished by humans and are fundamental for physical and ecological adaptations in mammals. Molecular genetic studies of the receptor genes for these tastes have been conducted in terrestrial mammals; however, little is known about the evolution and adaptation of these genes in marine mammals.
Here, all five basic taste modalities, sour, sweet, bitter, salt and umami, were investigated in cetaceans. The sequence characteristics and evolutionary analyses of taste receptor genes suggested that nearly all cetaceans may have lost all taste modalities except for that of salt.
This is the first study to comprehensively examine the five basic taste modalities in cetaceans with extensive taxa sampling. Our results suggest that cetaceans have lost four of the basic taste modalities including sour, sweet, umami, and most of the ability to sense bitter tastes. The integrity of the candidate salt taste receptor genes in all the cetaceans examined may be because of their function in Na+ reabsorption, which is key to osmoregulation and aquatic adaptation.
Electronic supplementary material
The online version of this article (doi:10.1186/s12862-014-0218-8) contains supplementary material, which is available to authorized users.
PMCID: PMC4232718  PMID: 25305673
Cetacean; Taste genes; Pseudogenization; Molecular evolution
2.  Blood Lead Levels and Associated Factors among Children in Guiyu of China: A Population-Based Study 
PLoS ONE  2014;9(8):e105470.
Children's health problems caused by the electronic waste (e-waste) lead exposure in China remains. To assess children's blood lead levels (BLLs) in Guiyu of China and investigate risk factors of children's elevated BLLs in Guiyu.
Material and Methods
842 children under 11 years of age from Guiyu and Haojiang were enrolled in this population-based study during 2011–2013. Participants completed a lifestyle and residential environment questionnaire and their physical growth indices were measured, and blood samples taken. Blood samples were tested to assess BLLs. Children's BLLs between the two groups were compared and factors associated with elevated BLLs among Guiyu children were analyzed by group Lasso logistic regression model.
Children living in Guiyu had significant higher BLLs (7.06 µg/dL) than the quantity (5.89 µg/dL) of Haojiang children (P<0.05). Subgroup analyses of BLLs exceeding 10 µg/dL showed the proportion (24.80%) of high-level BLLs for Guiyu children was greater than that (12.84%) in Haojiang (P<0.05). Boys had greater BLLs than girls, irrespectively of areas (P<0.05). The number of e-waste piles or recycling workshops around the house (odds ratio, 2.28; 95% confidence interval [CI], 1.37 to 3.87) significantly contributed to the elevated BLLs of children in Guiyu, and girls had less risk (odds ratio, 0.51; 95% CI, 0.31 to 0.83) of e-waste lead exposure than boys.
This analysis reinforces the importance of shifting e-waste recycling piles or workshops to non-populated areas as part of a comprehensive response to e-waste lead exposure control in Guiyu. To correct the problem of lead poisoning in children in Guiyu should be a long-term mission.
PMCID: PMC4138148  PMID: 25136795
3.  Dynamic Transcription of Long Non-Coding RNA Genes during CD4+ T Cell Development and Activation 
PLoS ONE  2014;9(7):e101588.
Recent evidence shows that long non-coding RNA (LncRNA) play important regulatory roles in many biology process, including cell development, activation and oncogenesis. However, the roles of these LncRNAs in the development and activation of CD4+ T cells, which is an important component of immune response, remain unknown.
To predict the function of LncRNA in the development and activation of CD4+ T cells, first, we examined the expression profiles of LncRNAs and mRNAs in CD4−CD8− (DN), CD4+CD8+ (DP), CD4+CD8−, and activated CD4+CD8− T cells in a microarray analysis and verified these results by real time PCRs (qPCR). We found that the expression of hundreds of LncRNAs significantly changed in each process of developmental transition, including DN into DP, DP into CD4+CD8−, and CD4+CD8− into activated CD4+ T cells. A Kendall distance analysis suggested that the expression of LncRNAs in DN, DP, CD4+CD8− T cells and activated CD4+ T cells were correlated with the expression of mRNAs in these T cells. The Blat algorithm and GO analysis suggested that LncRNAs may exert important roles in the development and activation of CD4+ T cells. These roles included proliferation, homeostasis, maturation, activation, migration, apoptosis and calcium ion transportation.
The present study found that the expression profiles of LncRNAs in different stages of CD4+ T cells are distinguishable. LncRNAs are involved in the key biological process in CD4+ T cell development and activation.
PMCID: PMC4086894  PMID: 25003630
4.  Inhibitory effects of subcutaneous tumors in nude mice mediated by low-frequency ultrasound and microbubbles 
Oncology Letters  2014;7(5):1385-1390.
The aim of the present study was to investigate the sonication effects of 21-kHz ultrasound (US) with microbubbles (MBs) on the subcutaneous prostate tumors of nude mice. In total, 15 tumor-bearing nude mice were divided into three groups: The control group, the low-frequency US group and the US+MB group. The MBs used were from US contrast agent SonoVue. The parameters of the US were as follows: 21 kHz, 26 mW/cm2 and a 40% duty cycle (2 sec on, 3 sec off) for 3 min, once every other day for 2 weeks. Color Doppler flow imaging, hematoxylin and eosin (HE) staining, immunoblotting and transmission electron microscopy (TEM) were used to evaluate the results. Following 2 weeks of treatment, the blood flow signal disappeared in the US+MB group only, and the tumor size was smaller when compared with the control and US groups. For the immunoblotting, the intensity of cyclooxygenase-2 and vascular endothelial growth factor in the US+MB group was lower compared with the other two groups. Tumor necrosis was present and the nucleus disappeared upon HE staining in the US+MB group. Upon TEM analysis, increased cytoplasmic vacuolation and dilatation of the perinuclear cisternae of the tumor cells were found in the US+MB group. In the control and US groups, the tumors had intact vascular endothelia and vessel lumens. However, lumen occlusion of the vessels was observed in the US+MB group. In conclusion, 21-kHz low-intensity US with MBs may result in vessel occlusion and growth inhibitory effects in the subcutaneous tumors of nude mice.
PMCID: PMC3997662  PMID: 24765142
ultrasound; low frequency; microbubbles; cavitation
5.  Cost-Utility of Lateral-Flow Urine Lipoarabinomannan for Tuberculosis Diagnosis in HIV-infected African Adults 
Inpatient hospitals in South Africa and Uganda
To evaluate the cost-effectiveness of a lateral flow urine lipoarabinomannan (LAM) test when added to existing strategies for tuberculosis (TB) diagnosis in HIV-infected adults (CD4+ T-cell counts<100 cells/μL) with symptoms of active TB.
Decision-analytic cost-utility model with the primary outcome being the incremental cost-effectiveness ratio (ICER), expressed in 2010 US dollars per disability-adjusted life year (DALY) averted, from the perspective of a public-sector TB control program.
Results and Conclusion
For every 1000 patients tested, adding lateral-flow urine LAM generated 80 incremental appropriate TB treatments and averted 224 DALYs. Estimated cost-utility was $353 per DALY averted (95% uncertainty range: $192–$1161) in South Africa and $86 per DALY averted (95% uncertainty range: $49–$239) in Uganda, reflecting the lower treatment costs in Uganda. Cost-utility was most sensitive to assay specificity, cost of TB treatment, life expectancy after TB cure, and cohort TB prevalence but did not rise above $1500 per DALY averted in South Africa under any one-way sensitivity analysis. The probability of acceptability was >99.8% at a per-DALY willingness-to-pay threshold equal to the per-capita gross domestic product in South Africa ($7275) and Uganda ($509).
PMCID: PMC3918209  PMID: 23485389
6.  miR394 and LCR are involved in Arabidopsis salt and drought stress responses in an abscisic acid-dependent manner 
BMC Plant Biology  2013;13:210.
MicroRNAs (miRNAs) are a class of short, endogenous non-coding small RNAs that have ability to base pair with their target mRNAs to induce their degradation in plants. miR394a/b are conserved small RNAs and its target gene LCR (LEAF CURLING RESPONSIVENESS) encodes an F-box protein (SKP1-Cullin/CDC53-F-box) but whether miR394a/b and its target gene LCR are involved in regulation of plant response to abscisic acid (ABA) and abiotic stresses is unknown.
Mature miR394 and precursor miR394a/b are shown to be slightly induced by ABA. By contrast, LCR expression is depressed by ABA. Analysis of LCR and its promoter (pLCR::GUS) revealed that LCR is expressed at all development stages. MIR394a/b over-expression (35S::MIR394a/b) and lcr (LCR loss of function) mutant plants are hypersensitive to salt stress, but LCR over-expressing (35S::m5LCR) plants display the salt-tolerant phenotype. Both 35S::MIR394a/b and lcr plants are highly tolerant to severe drought stress compared with wild-type, but 35S::m5LCR plants are susceptible to water deficiency. Over-expression of MIR394a/b led to ABA hypersensitivity and ABA-associated phenotypes, whereas 35S::m5LCR plants show ABA resistance phenotypes. Moreover, 35S::MIR394a/b plants accumulated higher levels of ABA-induced hydrogen peroxide and superoxide anion radicals than wild-type and 35S::m5LCR plants. Expressions of ABA- and stress-responsive genes, ABI3, ABI4, ABI5, ABF3, and ABF4 are up-regulated in MIR394a/b over-expressing plants but down-regulated in 35S::m5LCR plants. Over-expression of MIR394a in abi4-1 or abi5-1 background resulted in loss of ABA-sensitivity in 35S::MIR394a plants.
The silencing of LCR mRNA by miR394 is essential to maintain a certain phenotype favorable for the adaptive response to abiotic stresses. The contrasting phenotypes of salt and drought responses may be mediated by a functional balance between miR394 and LCR. If the balance is perturbed in case of the abiotic stress, an identical phenotype related to the stress response occurs, resulting in either ABA sensitive or insensitive response. Thus, miR394-regulated LCR abundance may allow plants to fine-tune their responses to ABA and abiotic stress.
PMCID: PMC3870963  PMID: 24330668
miR394; LCR; Abscisic acid; Salinity; Drought; Arabidopsis
7.  Intervention Effects of Ganoderma Lucidum Spores on Epileptiform Discharge Hippocampal Neurons and Expression of Neurotrophin-4 and N-Cadherin 
PLoS ONE  2013;8(4):e61687.
Epilepsy can cause cerebral transient dysfunctions. Ganoderma lucidum spores (GLS), a traditional Chinese medicinal herb, has shown some antiepileptic effects in our previous studies. This was the first study of the effects of GLS on cultured primary hippocampal neurons, treated with Mg2+ free medium. This in vitro model of epileptiform discharge hippocampal neurons allowed us to investigate the anti-epileptic effects and mechanism of GLS activity. Primary hippocampal neurons from <1 day old rats were cultured and their morphologies observed under fluorescence microscope. Neurons were confirmed by immunofluorescent staining of neuron specific enolase (NSE). Sterile method for GLS generation was investigated and serial dilutions of GLS were used to test the maximum non-toxic concentration of GLS on hippocampal neurons. The optimized concentration of GLS of 0.122 mg/ml was identified and used for subsequent analysis. Using the in vitro model, hippocampal neurons were divided into 4 groups for subsequent treatment i) control, ii) model (incubated with Mg2+ free medium for 3 hours), iii) GLS group I (incubated with Mg2+ free medium containing GLS for 3 hours and replaced with normal medium and incubated for 6 hours) and iv) GLS group II (neurons incubated with Mg2+ free medium for 3 hours then replaced with a normal medium containing GLS for 6 hours). Neurotrophin-4 and N-Cadherin protein expression were detected using Western blot. The results showed that the number of normal hippocampal neurons increased and the morphologies of hippocampal neurons were well preserved after GLS treatment. Furthermore, the expression of neurotrophin-4 was significantly increased while the expression of N-Cadherin was decreased in the GLS treated group compared with the model group. This data indicates that GLS may protect hippocampal neurons by promoting neurotrophin-4 expression and inhibiting N-Cadherin expression.
PMCID: PMC3634853  PMID: 23637882
8.  Development of a Non-Invasive Method, Multiplex Methylation Specific PCR (MMSP), for Early Diagnosis of Nasopharyngeal Carcinoma 
PLoS ONE  2012;7(11):e45908.
Increasing evidence demonstrated that inactivation of tumor suppressor genes (TSGs) by aberrant promoter methylation is an early event during carcinogenesis. Aiming at developing early diagnostic or prognostic tools for various tumors, we took an EBV-associated tumor, nasopharyngeal carcinoma (NPC), as a model and developed a powerful assay based on “multiplex methylation specific-PCR (MMSP)”. The MMSP assay was designed to detect tumor-specific methylation status of several NPC-related genes and was capable of acquiring multiplex information simultaneously through a single PCR reaction with the tiny tumor DNA derived from the direct body fluid close to the primary tumor. In this study, we collected paired nasopharyngeal (NP) swabs and NPC biopsies from 49 NPC patients and twenty noncancerous controls. A panel of markers including two EBV, and two cellular TSG markers were applied in this NPC-specific-MMSP assay. We optimized the working condition of MMSP so that it provides information equal to that from the corresponding separate PCRs. The results showed that MMSP patterns of NPC swab were largely consistent with those of corresponding biopsies and significantly distinguished themselves from those of 20 noncancerous volunteers. Among the 69 samples (49 NPCs and 20 normal controls), the sensitivity of detecting NPC from NP swabs is 98%. The specificity is as high as 100%. In conclusion, being characterized by its noninvasiveness, high reproducibility and informativeness, MMSP assay is a reliable and potential diagnostic tool for NPC. It paves the way for the development of population screening and early diagnosis approaches for various tumor types.
PMCID: PMC3489875  PMID: 23144779
9.  Chronic Activation of the G Protein-Coupled Receptor 30 with Agonist G-1 Attenuates Heart Failure 
PLoS ONE  2012;7(10):e48185.
G protein-coupled receptor (GPR) 30 is a novel estrogen receptor. Recent studies suggest that activation of the GPR30 confers rapid cardioprotection in isolated rat heart. It is unknown whether chronic activation of GPR30 is beneficial or not for heart failure. In this study we investigated the cardiac effect of sustained activation or inhibition of GPR30. Female Sprague–Dawley rats were divided into 7 groups #2Q1: sham surgery (Sham), bilateral ovariectomy (OVX), OVX+estrogen (E2), OVX+isoproterenol (ISO), OVX+ISO+G-1, OVX+ISO+E2+G15, OVX+ISO+E2. ISO (85 mg/kg×17 day, sc) was given to make the heart failure models. G-1(120 µg/kg·d×14 day) was used to activate GPR30 and G15 (190 µg/kg·d×14 day) was used to inhibit GPR30. Concentration of brain natriuretic peptide in serum, masson staining in isolated heart, contractile function and the expression of β1 and β2- adrenergic receptor (AR) of ventricular myocytes were also determined. Our data showed that ISO treatment led to heart failure in OVX rats. G-1 or E2 treatment decreased concentration of brain natriuretic peptide, reduced cardiac fibrosis, and enhanced contraction of the heart. Combined treatment with β1 (CGP20712A) and β2-AR (ICI118551) antagonist abolished the improvement of myocardial function induced by G-1. We also found that chronic treatment with G-1 normalized the expression of β1-AR and increased the expression of β2-AR. Our results indicate that chronic activation of the GPR30 with its agonist G-1 attenuates heart failure by normalizing the expression of β1-AR and increasing the expression of β2-AR.
PMCID: PMC3482180  PMID: 23110207
10.  Ammine(2,2′-bipyridine-κ2 N,N′)silver(I) nitrate: a dimer formed by π–π stacking and ligand-unsupported Ag⋯Ag inter­actions 
Reaction of AgNO3 and 2,2′-bipyridine (bipy) under ultrasonic treatment gave the title compound, [Ag(C10H8N2)(NH3)]NO3. The crystal structure consists of dimers formed by two symmetry-related AgI–bipy monomers connected through intra-dimer π–π stacking and ligand-unsupported Ag⋯Ag inter­actions. A crystallographic C2 axis passes through the mid-point of and is perpendicular to the Ag⋯Agi(−x + 1, y, −z + ) axis. In addition, each AgI cation is coordinated by one chelating bipy ligand and one ammine ligand, giving a trigonal coordination environment capped by the symmetry-equivalent Ag atom. Mol­ecules are assembled by Ag⋯Ag, π–π, hydrogen-bond (N—H⋯O and C—H⋯O) and weak Ag⋯π inter­actions into a three-dimensional framework. Comparing the products synthesized under different mechanical treatments, we found that reaction conditions have a significant influence on the resulting structures. The luminescence properties of the title compound are also discussed.
PMCID: PMC2855567  PMID: 20203399
11.  Bis[diamminesilver(I)] 5-nitro­iso­phthalate monohydrate 
In the title compound, [Ag(NH3)2]2(C8H3NO6)·H2O, the cations have an almost linear coordination geometry with two ammine ligands and inter­act with the water mol­ecules [Ag⋯Owater = 2.725 (4) and 2.985 (4) Å]. In the crystal, N—H⋯O and O—H⋯O hydrogen bonds, combined with weak (lone pair)⋯π [O⋯centroid distance = 3.401 (4) Å] and π–π stacking [centroid–centroid distance = 3.975 (3) Å] inter­actions, stabilize the three-dimensional supra­molecular network.
PMCID: PMC2983872  PMID: 21580507
12.  Tetra­aqua­tetra­kis{μ3-3,3′-[(E,E)-ethane-1,2-diylbis(nitrilo­methyl­idyne)]benzene-1,2-diolato}octa­zinc(II) N,N-dimethyl­formamide hexa­solvate 
The asymmetric unit of the title compound [Zn8(C16H12N2O4)4(H2O)4]·6C3H7NO, consists of eight ZnII cations, four tetra­valent anionic ligands, L 4− (L 4− = 3,3′-(1E,1′E)-(ethane-1,2-diylbis(azan-1-yl-1-yl­idene))bis­(methan-1-yl-1-yl­idene)dibenzene-1,2-bis­(olate), four coordinated water mol­ecules and six N,N-dimethyl­formamide solvate mol­ecules. The coordination complex comprises an octa­nuclear ZnII unit with its ZnII centers coordinated in two discrete distorted square-pyramidal geometries. Four ZnII atoms each coordinate to two nitro­gen atoms and two phenolate oxygen atoms from an individual L 4− ligand and one coordinated water mol­ecule. The other four ZnII atoms each bind to five phenolate oxygen atoms from three different L 4− ligands. In the crystal structure, the ZnII complex unit, coordinated water mol­ecules and dimethyl­formamide solvate mol­ecules are linked via O—H⋯O and C—H⋯O hydrogen bonds. Mol­ecules are connected by additional inter­molecular O—H⋯O and C—H⋯O hydrogen bonds, forming an extensive three dimensional framework.
PMCID: PMC2972079  PMID: 21578596
13.  5-Bromo-2,3-dihydro-1H-cyclo­penta­[a]naphthalen-1-one 
The title compound, C13H9BrO, has been synthesized by the intra­molecular Friedel–Crafts reaction of 1-(1-bromo-4-naphth­yl)-3-chloro­propan-1-one. There are two approximately planar [maximum deviations of 0.8 (2) and 0.4 (2) Å in the two mol­ecules] molecules in the asymmetric unit. The dihedral angle between their mean planes is 19.72 (8)°. Weak inter­molecular C—H⋯O hydrogen bonding is present in the crystal structure.
PMCID: PMC2970092  PMID: 21577580
14.  Poly[(μ2-4,4′-bipyridine)bis­(μ4-5-tert-butyl­isophthalato)bis­(μ3-5-tert-butyl­isophthalato)di-μ3-hydroxido-penta­zinc(II)] 
The asymmetric unit of the title compound, [Zn5(C12H12O4)4(OH)2(C10H8N2)]n, consists of three ZnII ions (one of which is located on a twofold rotation axis), two 5-tert-butyl­isophthalate ligands, one 4,4′-bipyridine ligand and one hydroxide group. The five ZnII ions form a penta­nuclear zinc cluster, which is further bridged by ten organic ligands, forming two-dimensional sheets. The central zinc ion of the cluster has site symmetry 2 and is octahedrally coordinated in a N2O4 donor set, whereas the other four zinc atoms are tetrahedrally coordinated by four O atoms. The coordination modes for the 5-tert-butyl­isophthalates are bis­(bidentate) or bidentate-monodentate. Hydrogen bonds are formed between adjacent sheets through the hydroxide groups and the O atoms of the monodentate carboxyl­ate groups. The two tert-butyl groups are disordered over two positions with ratios of 0.64 (2):0.36 (2) and 0.85 (3):0.15 (3).
PMCID: PMC2969937  PMID: 21577432
15.  4H-Cyclo­penta­[def]phenanthren-4-one 
In the title compound, C15H8O, the asymmetric unit contains four independent mol­ecules and crystallizes with aromatic π–π stacking inter­actions[centroid–centroid distances = 3.5326 (18) Å].
PMCID: PMC2970027  PMID: 21577520
16.  3′,6′-Bis(diethyl­amino)-2-phenyl­spiro[isoindoline-1,9′-xanthen]-3-one 
The title compound, C34H35O2N3, was synthesized by the reaction of 2-[3,6-bis­(diethyl­amino)-9H-xanthen-9-yl]benzoyl chloride with aniline. In the mol­ecular structure, the dihedral angles between the isoindoline and xanthene planes and between the isoindoline and benzene planes are 86.9 (3) and 47.0 (2)°, respectively. The mol­ecular packing in the crystal structure is stabilized by weak C—H⋯O hydrogen bonding.
PMCID: PMC2969317  PMID: 21582767
17.  Benzo[a]fluoren-11-one 
The mol­ecule of the title compound, C17H10O, is nearly planar, the largest deviation from the mean plane being 0.06 Å. The crystal structure is governed by π–π inter­actions, with centroid–centroid distances ranging from .559 to 3.730 Å.
PMCID: PMC2962100  PMID: 21203182

Results 1-17 (17)