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1.  A Fourier Transform Infrared Spectroscopy Analysis of Carious Dentin from Transparent Zone to Normal Zone 
Caries research  2014;48(4):320-329.
It is well known that caries invasion leads to the differentiation of dentin into zones with altered composition, collagen integrity and mineral identity. However, understanding of these changes from the fundamental perspective of molecular structure has been lacking so far. In light of this, the present work aims to utilize Fourier transform infrared spectroscopy (FTIR) to directly extract molecular information regarding collagen's and hydroxyapatite's structural changes as dentin transitions from the transparent zone (TZ) into the normal zone (NZ). Unembedded ultrathin dentin films were sectioned from carious teeth, and an FTIR imaging system was used to obtain spatially resolved FTIR spectra. According to the mineral-to-matrix ratio image generated from large-area low-spectral-resolution scan, the TZ, the NZ and the intermediate subtransparent zone (STZ) were identified. High-spectral-resolution spectra were taken from each zone and subsequently examined with regard to mineral content, carbonate distribution, collagen denaturation and carbonate substitution patterns. The integrity of collagen's triple helical structure was also evaluated based on spectra collected from demineralized dentin films of selected teeth. The results support the argument that STZ is the real sclerotic layer, and they corroborate the established knowledge that collagen in TZ is hardly altered and therefore should be reserved for reparative purposes. Moreover, the close resemblance between the STZ and the NZ in terms of carbonate content, and that between the STZ and the TZ in terms of being A-type carbonate-rich, suggest that the mineral that initially occludes dentin tubules is hydroxyapatite newly generated from odontoblastic activities, which is then transformed into whitlockite in the demineralization/remineralization process as caries progresses.
PMCID: PMC4422165  PMID: 24556607
Carbonate type; Carious dentin; Collagen cross-link; Collagen triple helix; Fourier transform infrared spectroscopy imaging; Mineral content; Subtransparent zone; Transparent zone
2.  Genetic variants in the SOX6 gene are associated with bone mineral density in both Caucasian and Chinese populations 
Given the biological function of SOX6 and recent genome-wide association finding, we performed a fine-mapping association analyses to investigate the relationship between SOX6 and BMD both in Caucasian and Chinese populations. We identified many single-nucleotide polymorphisms (SNPs) within or near the SOX6 gene to be significantly associated with hip bone mineral density (BMD).
SOX6 gene is an essential transcription factor in chondrogenesis and cartilage formation. Recent genome-wide association studies (GWAS) detected a SNP (rs7117858) located at the downstream of SOX6 significantly associated with hip BMD.
Given the biological function of SOX6 and the GWAS finding, we considered SOX6 as a new candidate for BMD and osteoporosis. Therefore, in this study, we performed a fine-mapping association analyses to investigate the relationship between SNPs within and near the SOX6 gene and BMD at both hip and spine. A total of 301 SNPs were tested in two independent US Caucasian populations (2,286 and 1,000 unrelated subjects, respectively) and a Chinese population (1,627 unrelated Han subjects).
We confirmed that the previously reported rs7117858-A was associated with reduced hip BMD, with combined P value of 2.45×10−4. Besides this SNP, we identified another 19 SNPs within or near the SOX6 gene to be significantly associated with hip BMD after false discovery rate adjustment. The most significant SNP was rs1347677 located at the intron 3 (P=3.15×10−7). Seven additional SNPs in high linkage disequilibrium with rs1347677 were also significantly associated with hip BMD. SNPs in SOX6 showed significant skeletal site specificity since no SNP was detected to be associated with spine BMD.
Our study identified many SNPs in the SOX6 gene associated with hip BMD even across different ethnicities, which further highlighted the importance of the SOX6 gene influencing BMD variation and provided more information to the understanding of the genetic architecture of osteoporosis.
PMCID: PMC4171834  PMID: 21625884
Association; BMD; Osteoporosis; SOX6
3.  Histone demethylase Kdm4b functions as a co-factor of C/EBPβ to promote mitotic clonal expansion during differentiation of 3T3-L1 preadipocytes 
Cell Death and Differentiation  2012;19(12):1917-1927.
CCAAT/enhancer-binding protein (C/EBP) β is required for both mitotic clonal expansion (MCE) and terminal adipocyte differentiation of 3T3-L1 preadipocytes. Although the role of C/EBPβ in terminal adipocyte differentiation is well defined, its mechanism of action during MCE is not. In this report, histone demethylase Kdm4b, as well as cell cycle genes Cdc45l (cell division cycle 45 homolog), Mcm3 (mini-chromosome maintenance complex component 3), Gins1 (GINS complex subunit 1) and Cdc25c (cell division cycle 25 homolog c), were identified as potential C/EBPβ target genes during MCE by utilizing promoter-wide chromatin immunoprecipitation (ChIP)-on-chip analysis combined with gene expression microarrays. The expression of Kdm4b is induced during MCE and its induction is dependent on C/EBPβ. ChIP, Electrophoretic Mobility Shift Assay (EMSA) and luciferase assay confirmed that the promoter of Kdm4b is bound and activated by C/EBPβ. Knockdown of Kdm4b impaired MCE. Furthermore, Kdm4b interacted with C/EBPβ and was recruited to the promoters of C/EBPβ-regulated cell cycle genes, including Cdc45l, Mcm3, Gins1, and Cdc25c, demethylated H3K9me3 and activated their transcription. These findings suggest a novel feed forward mechanism involving a DNA binding transcription factor (C/EBPβ) and a chromatin regulator (Kdm4b) in the regulation of MCE by controlling cell cycle gene expression.
PMCID: PMC3504706  PMID: 22722334
3T3-L1 preadipocytes; adipocyte differentiation; mitotic clonal expansion; C/EBPβ; Kdm4b; ChIP-on-chip
4.  Nitrogen-corrected True Metabolizable Energy and Amino Acid Digestibility of Chinese Corn Distillers Dried Grains with Solubles in Adult Cecectomized Roosters 
This study was conducted to evaluate chemical composition, nitrogen-corrected true metabolizable energy (TMEn) and true amino acids digestibility of corn distillers dried grains with solubles (DDGS) produced in China. Twenty five sources of corn DDGS was collected from 8 provinces of China. A precision-fed rooster assay was used to determine TMEn and amino acids digestibility with 35 adult cecectomized roosters, in which each DDGS sample was tube fed (30 g). The average content of ash, crude protein, total amino acid, ether extract, crude fiber and neutral detergent fiber were 4.81, 27.91, 22.51, 15.22, 6.35 and 37.58%, respectively. TMEn of DDGS ranged from 1,779 to 3,071 kcal/kg and averaged 2,517 kcal/kg. Coefficient of variation for non-amino acid crude protein, ether extract, crude fiber and TMEn were 55.0, 15.7, 15.9 and 17.1%, respectively. The average true amino acid digestibility was 77.32%. Stepwise regression analysis obtained the following equation: TMEn, kcal/kg = −2,995.6+0.88×gross energy+49.63×a* (BIC = 248.8; RMSE = 190.8; p<0.01). Removing gross energy from the model obtained the following equation: TMEn, kcal/kg = 57.88×ether extracts+87.62×a* (BIC = 254.3, RMSE = 223.5; p<0.01). No correlation was found between color scores and lysine true digestibility (p>0.05). These results suggest that corn DDGS produced in China has a large variation in chemical composition, and gross energy and a* value can be used to generate TMEn predict equation.
PMCID: PMC4093243  PMID: 25049858
Amino Acid; Corn Distillers Dried Grains with Solubles; Nitrogen-corrected True Metabolizable Energy; Rooster
5.  A genomewide scan for quantitative trait loci underlying areal bone size variation in 451 Caucasian families 
Journal of Medical Genetics  2006;43(11):873-880.
Bone size is an important determinant of bone strength and is under strong genetic control.
To identify quantitative trait loci (QTL) for areal bone size variation, a large‐scale genomewide linkage scan was carried out in 451 Caucasian families.
Participants and methods
Of 4124 people with phenotypes, 3899 were genotyped with 410 microsatellite markers. Multipoint linkage analyses were carried out in the entire sample, as well as in men and women separately. Potential epistatic interactions between identified genomic regions were also assessed.
Several potentially important genomic regions were identified, such as 8q24 for hip bone size (logarithm of the ratio of the odds that two loci are linked (LOD) 3.27) and 2p24 (LOD 2.04) for spine bone size. 8q24 may also interact with 19p13 to affect hip bone size. Several sex‐specific QTL were also detected, such as 14q21 (LOD 2.94) for wrist bone size in women and 16q12 (LOD 2.19) for hip bone size in men.
Together with previous findings, this study has further delineated the genetic basis of bone size and laid a foundation for future studies to eventually elucidate the mechanisms of bone size regulation and associated fracture risks.
PMCID: PMC2563191  PMID: 16825438
7.  P02-08. Enhancement of HIV-1 DNA vaccine immunogenicity by BCG-PSN, a novel adjuvant 
Sun, J | Liu, Y | Li, D | Hou, J | Xu, Z | Fan, W | Fu, J | Liu, Y | Shao, Y
Retrovirology  2009;6(Suppl 3):P13.
PMCID: PMC2767615
Neuroscience  2005;130(2):383-388.
Investigations using Drosophila melanogaster have shown that the circadian clock gene period can influence behavioral responses to cocaine, and the mouse homologues, mPer1 and mPer2, modulate cocaine sensitization and reward. In the present study, we applied DNAzyme targeting mPer1 to interfere the expression of mPer1 in CNS in mice and studied the role of mPer1 on morphine dependence. We found that the DNAzyme could attenuate the expression of mPer1 in CNS in mice. Mice treated with DNAzyme and morphine synchronously did not show preference to the morphine-trained side, whereas the control group did. In contrast, mice treated with DNAzyme after morphine showed preference to the morphine-trained side as well as the control group did. These results indicate that drug dependence seems to be influenced at least partially by mPer1, but mPer1 cannot affect morphine dependence that has been formed.
PMCID: PMC2656444  PMID: 15664694
drug dependence; DNAzyme; learning and memory; circadian; i.c.v.
10.  Cardioprotective effects of peroxisome proliferator activated receptor γ activators on acute myocarditis: anti-inflammatory actions associated with nuclear factor κB blockade 
Heart  2005;91(9):1203-1208.
Objective: To test the hypothesis that activation of peroxisome proliferator activated receptor γ (PPAR-γ) reduces experimental autoimmune myocarditis (EAM) associated with inhibitor κB (IκB) α induction, blockade of nuclear factor κB (NF-κB), and inhibition of inflammatory cytokine expression.
Methods: EAM was induced in Lewis rats by immunisation with porcine cardiac myosin. PPAR-γ activators 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) and pioglitazone (PIO) were administered to rats with EAM.
Results: Enhanced PPAR-γ expression was prominently stained in the nuclear and perinuclear regions of infiltrating inflammatory cells. Administration of 15d-PGJ2 and PIO greatly reduced the severity of myocarditis and suppressed myocardial mRNA and protein expression of inflammatory cytokines in rats with EAM. In addition, treatment with PPAR-γ activators enhanced IκB concentrations in the cytoplasmic fractions and nuclear fractions from inflammatory myocardium. Concurrently, NF-κB was greatly activated in myocarditis; this activation was blocked in the 15d-PGJ2 treated and PIO treated groups.
Conclusions: PPAR-γ may have a role in the pathophysiology of EAM. Because an increase in IκB expression and inhibition of translocation of the NF-κB subunit p65 to the nucleus in inflammatory cells correlated with the protective effects of PPAR-γ activators, these results suggest that PPAR-γ activators act sequentially through PPAR-γ activation, IκB induction, blockade of NF-κB activation, and inhibition of inflammatory cytokine expression. These results suggest that PPAR-γ activators such as 15d-PGJ2 and PIO may have the potential to modulate human inflammatory heart diseases such as myocarditis.
PMCID: PMC1769084  PMID: 15774612
myocarditis; immunity; PPAR-γ; NF-κB; cytokine; inflammation
11.  Torsional ultrasound modality for hard nucleus phacoemulsification cataract extraction 
Zeng, M | Liu, X | Liu, Y | Xia, Y | Luo, L | Yuan, Z | Zeng, Y | Liu, Y
The British Journal of Ophthalmology  2008;92(8):1092-1096.
To evaluate the efficacy and safety of phacoemulsification using torsional modality with different parameter settings for hard nucleus cataract extraction.
A prospective, randomised clinical study.
A clinical practice study conducted at the Cataract Service, Zhongshan Ophthalmic Center, Sun-Yat-Sen University, and Guangzhou. One eye each from 198 consecutive patients with cataract density grade IV according to the Emery–Little system classification system, requiring phacoemulsification and intraocular lens implantation, was included. Eyes were randomly assigned to the Linear Torsional combined with Ultrasound power group (Linear Tor+US group, n = 66), 100% Fixed Torsional group (Fixed Tor group, n = 65) and conventional Ultrasound burst group (US group, n = 67). All surgeries were performed by a single experienced surgeon and outcomes evaluated by another surgeon masked to treatment. Intraoperative parameters were Ultrasound Time (UST), Cumulative Dissipated Energy (CDE) and surgical complications. Patients were examined on post-op days 1, 7 and 30. Postoperative outcomes were final best corrected visual acuity (BCVA), average central and incisional corneal thickness and central endothelial cell counts.
The mean UST was lower in the Fixed Tor group than in the US group and in the Lin US+Tor group (p⩽0.0001). The mean CDE was lower in the Lin Tor+US group and in the Fixed Tor group than in the US group (p⩽0.0001). Comparing with the two Tor group, the US group had a lower average BCVA on post-op 1, 7 (p⩽0.0001) and 30 (p>0.01), greater average central corneal and incisional thickness on days 1, 7 (p⩽0.0001) and 30 (p>0.01), and higher average corneal endothelial cell losses on day 7 and 30 days (p⩽0.0001).
Torsional combined with ultrasound power or high fixed torsional amplitude can yield more effective hard nucleus phacoemulsification than conventional ultrasound modality.
PMCID: PMC2569137  PMID: 18567650
12.  A genome-wide linkage scan for bone mineral density in an extended sample: evidence for linkage on 11q23 and Xq27 
Journal of Medical Genetics  2004;41(10):743-751.
Background: Osteoporosis is a major public health problem, mainly quantified by low bone mineral density (BMD). The majority of BMD variation is determined by genetic effects. A pilot whole genome linkage scan (WGS) was previously reported in 53 white pedigrees with 630 subjects. Several genomic regions were suggested to be linked to BMD variation.
Objective: To substantiate these previous findings and detect new genomic regions.
Methods: A WGS was conducted on an extended sample where the size was almost tripled (1816 subjects from 79 pedigrees). All the subjects were genotyped with 451 microsatellite markers spaced ∼8.1 cM apart across the human genome. Two point and multipoint linkage analyses were carried out using the variance component method.
Results: The strongest linkage signal was obtained on Xq27 with two point LOD scores of 4.30 for wrist BMD, and 2.57 for hip BMD, respectively. Another important region was 11q23, which achieved a maximum LOD score of 3.13 for spine BMD in multipoint analyses, confirming the results on this region in two earlier independent studies. Suggestive linkage evidence was also found on 7p14 and 20p12.
Conclusions: Together with the findings from other studies, the current study has further delineated the genetic basis of bone mass and highlights the importance of increasing sample size to confirm linkage findings and to identify new regions of linkage.
PMCID: PMC1735607  PMID: 15466007
14.  Bone Marrow–derived Cell Therapy for Oral Mucosal Repair after Irradiation 
Journal of Dental Research  2014;93(8):813-820.
Oral mucositis (ulcer) is a serious and painful side effect for patients with head and neck cancer following radiation therapy. However, current clinical strategies cannot efficiently prevent the occurrence of oral mucositis. In this study, we investigated whether bone marrow–derived cells (BMDCs) prevented the occurrence and/or decreased the severity of radiation-induced oral mucositis. Fresh concentrated BMDCs from male C3H mice were transplanted intravenously into female mice after tongue irradiation. For 14 days postirradiation, the changes of body weight and the time courses of ulceration were observed. Until the ulcer reached maximum size (7 days postirradiation), macroscopic and histologic analyses of harvested tongues were performed to detect the behavior of donor BMDCs. Between 2 and 5 days postirradiation, BMDCs-transplanted mice showed more expression of stem cell markers (c-Kit, Sca-1) and EGFR and fewer apoptotic cells when compared with nontransplanted control mice (irradiation group). On day 7, there were fewer and smaller ulcers observed in the BMDCs-transplanted group. Tongues of these mice had preserved their epithelial thickness, and regenerative activities (blood vessels formation, cell proliferation) were higher than they were in the irradiation group. Fluorescently labeled BMDCs were not detected in tongue epithelium but rather in connective tissue (dermis) just below the basal cell layer. These findings suggest that exogenous BMDCs behave to reduce radiogenic oral mucositis in a paracrine manner.
PMCID: PMC4293762  PMID: 24980658
cell transplantaion; stem cells; radiation injury; epithelial tissue; paracrine effect; mucosal regeneration
15.  Addition of Grape Seed Extract Renders Phosphoric Acid a Collagen-stabilizing Etchant 
Journal of Dental Research  2014;93(8):821-827.
Previous studies found that grape seed extract (GSE), which is rich in proanthocyanidins, could protect demineralized dentin collagen from collagenolytic activities following clinically relevant treatment. Because of proanthocyanidin’s adverse interference to resin polymerization, it was believed that GSE should be applied and then rinsed off in a separate step, which in effect increases the complexity of the bonding procedure. The present study aimed to investigate the feasibility of combining GSE treatment with phosphoric acid etching to address the issue. It is also the first attempt to formulate collagen-cross-linking dental etchants. Based on Fourier-transformed infrared spectroscopy and digestion assay, it was established that in the presence of 20% to 5% phosphoric acid, 30 sec of GSE treatment rendered demineralized dentin collagen inert to bacterial collagenase digestion. Based on this positive result, the simultaneous dentin etching and collagen protecting of GSE-containing phosphoric acid was evaluated on the premise of a 30-second etching time. According to micro-Raman spectroscopy, the formulation containing 20% phosphoric acid was found to lead to overetching. Based on scanning and transmission electronic microscopy, this same formulation exhibited unsynchronized phosphoric acid and GSE penetration. Therefore, addition of GSE did render phosphoric acid a collagen-stabilizing etchant, but the preferable phosphoric acid concentration should be <20%.
PMCID: PMC4126223  PMID: 24935065
dental acid etching; demineralized dentin matrix; cross-linking reagents; proanthocyanidins; microbial collagenase; biode gradation
16.  Determination of visual prognosis in children with open globe injuries 
Liu, X | Liu, Z | Liu, Y | Zhao, L | Xu, S | Su, G | Zhao, J
Eye  2014;28(7):852-856.
To determine the predictive factors of visual outcomes in children with open globe injury and to give guidance to reduce the incidence of open globe injury.
One hundred and forty eyes of 137 consecutive open globe injury patients, who were treated at the Eye Center of Second Bethune Hospital affiliated with Jilin University between August 2005 and August 2012, were retrospectively analyzed. Data recorded included demographic characteristics, causes of injury, location and extent of injury, presenting visual acuity, detailed ocular anterior and posterior segment evaluations, details of primary and subsequent surgeries, and postoperative complications and outcomes. The follow-up data included the most recent best-corrected visual acuity, complications, and the duration of follow-up.
Of the 137 patients, there were 116 (84.7%) boys and 21 (15.3%) girls. Their ages ranged between 3 and 17 years old (mean=11.57±4.19 years old). Sixty (43.8%) children had a right eye injury, whereas 74 (54.0%) had a left eye injury. Only three (2.2%) children suffered bilateral eye injury. Living utensils, industrial tools, and fireworks contributed to the most common causes of open globe injury. Eighty-one (59.1%) had sharp force injuries, 23 (16.8%) had blunt injuries, and 33 (24.1%) had missile injuries.
Unfavorable visual outcomes were related to a younger age at presentation, poor presenting visual acuity, injuries caused by blunt or missile objects, posterior wound location, hyphema, vitreous hemorrhage, and surgical intervention of pars plana vitrectomy.
PMCID: PMC4094803  PMID: 24833181
17.  Heat stress induced apoptosis is triggered by transcription-independent p53, Ca2+ dyshomeostasis and the subsequent Bax mitochondrial translocation 
Scientific Reports  2015;5:11497.
In this study, We demonstrated that Bax mitochondrial translocation plays a vital role in the initiation of the mitochondrial signaling pathway upon activation by heat stress. In addition, both p53 mitochondrial translocation and Ca2+ signal mediated MPTP opening activate Bax mitochondrial translocation. Employing pifithrin-α (a p53 mitochondrial translocation inhibitor) and CsA (a permeability transition pore (MPTP) inhibitor), we found that heat stress induced Bax mitochondrial translocation was significantly inhibited in cells pretreated with both PFT and CsA. Furthermore, we demonstrated that generation of reactive oxygen species (ROS) is a critical mediator in heat stress induced apoptosis and that the antioxidant MnTBAP significantly decreased heat stress induced p53 mitochondrial translocation and Ca2+ signal mediated MPTP opening, as well as the subsequent Bax mitochondrial translocation and activation of the caspase cascade. Taken together, our results indicate that heat stress induces apoptosis through the mitochondrial pathway with ROS dependent mitochondrial p53 translocation and Ca2+ dyshomeostasis, and the ensuing intro Bax mitochondrial translocation as the upstream events involved in triggering the apoptotic process observed upon cellular exposure to heat stress.
PMCID: PMC4478470  PMID: 26105784
19.  Prolactin Specifically Regulates Citrate Oxidation and m-Aconitase of Rat Prostate Epithelial Cells 
The prostate gland of many animals, including humans, produces and secretes extremely high levels of citrate. To achieve this function, prostate secretory epithelial cells possess unique metabolic properties that permit accumulation and ultimate secretion (net citrate production) of citrate. Mounting evidence continues to support the concept that prostate epithelial cells possess a limiting mitochondrial (m)-aconitase activity that minimizes citrate oxidation and results in the accumulation of citrate synthesized by the cells, Recent studies have revealed that prolactin (PRL) stimulates net citrate production of rat lateral prostate (RLP), The mechanism of this PRL effect has not been established. The current studies were concerned with the possibility that PRL might be involved in the regulation of citrate oxidation and m-aconitase of prostate cells. Studies were conducted with RLP, RVP (rat ventral prostate), RDP (rat dorsal prostate), and kidney cells, The results showed that PRL in vitro and in vivo decreased citrate utilization and the level of m-aconitase in RLP cells, and conversely increased citrate utilization and m-aconitase in RVP cells, Furthermore, PRL had no effect on either RDP or kidney cells. The effects of PRL on both citrate utilization and m-aconitase of RLP and RVP were abolished by cycloheximide and actinomycin. Mitochondrial studies revealed that PRL decreased citrate oxidation of RLP and increased citrate oxidation of RVP, but had no effect on isocitrate oxidation. In conclusion, these studies establish that PRL has a physiological role in the regulation of citrate oxidation in prostate, and that this action is associated with PRL regulation of the biosynthesis of m-aconitase. Furthermore, the effects of PRL are cell-specific and targeted at m-aconitase.
PMCID: PMC4467526  PMID: 8609829
20.  Mitochondrial Aconitase Gene Expression Is Regulated by Testosterone and Prolactin in Prostate Epithelial Cells 
The Prostate  2000;42(3):196-202.
m-aconitase catalyzes the first step leading to the oxidation of citrate via the Krebs cycle. It is a constituitive enzyme in virtually all mammalian cells, found in excess, and is considered to be a regulatory or regulated enzyme. In contrast to these general relationships, prostate secretory epithelial cells possess a uniquely limiting mitochondrial (m-) aconitase which minimizes the oxidation of citrate. This permits the unique prostate function of accumulating and secreting extraordinarily high levels of citrate. Previous animal studies demonstrated that testosterone and prolactin regulate the level of m-aconitase specifically in citrate-producing prostate cells. The present studies were conducted to determine if testosterone and prolactin regulated the expression of the m-aconitase gene in prostate cells, and to determine the effect of the hormones on human prostate cells.
The studies were conducted with freshly prepared rat ventral, rat lateral, and pig prostate epithelial cells, and with the human malignant cell lines LNCaP and PC-3. The effects of 1 nM testosterone and 3 nM prolactin on the level of m-aconitase mRNA and on the transcription rate of m-aconitase were determined.
The studies revealed that both prolactin and testosterone increase the levels of m-aconitase mRNA and the transcription rates of m-aconitase in rat ventral prostate cells, pig prostate cells, and human malignant prostate cells (LNCaP and PC-3). In contrast, both hormones decreased the level of m-aconitase mRNA and repressed m-aconitase gene transcription in rat lateral prostate cells. The hormonal regulation of m-aconitase corresponded with the levels of m-aconitase enzyme, m-aconitase activity, and citrate oxidation.
In addition to the constitutive expression of m-aconitase, the m-aconitase gene is testosterone- and prolactin-regulated in specifically targeted prostate cells. The hormonal regulation of m-aconitase gene expression and biosynthesis of m-aconitase provide a regulatory mechanism for the oxidation of citrate, and consequently, the level of net citrate production by prostate. The hormonally increased expression and biosynthesis of m-aconitase in human malignant cells might be involved in the increased citrate oxidation associated with the development of true malignant cells in prostate cancer.
PMCID: PMC4464833  PMID: 10639190
mitochondrial aconitase; prostate cells; aconitase gene; testosterone; prolactin; citrate oxidation; prostate cancer
21.  Dietary intake, plasma homocysteine, and repetitive element DNA methylation in the Multi-Ethnic Study of Atherosclerosis (MESA) 
Background and Aims
DNA methylation of repetitive elements may explain the relations among dietary intake, hyperhomocysteinemia, and cardiovascular disease risk. We investigated associations of methyl micronutrient intake and plasma total homocysteine with LINE-1 and Alu methylation in a cross-sectional study of 987 adults aged 45–84 y who participated in the Multi-Ethnic Study of Atherosclerosis (MESA) Stress Study.
Methods and Results
DNA methylation was estimated using pyrosequencing technology. A 120-item food frequency questionnaire was used to ascertain daily intake of folate, vitamin B12, vitamin B6, zinc, and methionine. Plasma total homocysteine was quantified using a fluorescence polarization immunoassay. Associations of micronutrient intake and homocysteine with LINE-1 and Alu methylation were examined using linear regression. Adjusted differences in %5-methylated cytosines (%5mC) were examined by categories of predictors using multivariable linear regression models. Intake of methyl-donor micronutrients was not associated with DNA methylation. After adjustment for covariates, each 3 μmol/L increment of homocysteine corresponded with 0.06 (−0.01, 0.13) %5mC higher LINE-1 methylation. Additionally, BMI was positively associated with LINE-1 methylation (P trend=0.03). Participants with BMI ≥40 kg/m2 had 0.35 (0.03, 0.67) %5mC higher LINE-1 than those with normal BMI. We also observed a 0.10 (0.02, 0.19) %5mC difference in Alu methylation per 10 cm of height. These associations did not differ by sex.
Dietary intake of methyl-donor micronutrients was not associated with measures of DNA methylation in our sample. However, higher BMI was related to higher LINE-1 methylation, and height was positively associated with Alu methylation.
PMCID: PMC4037331  PMID: 24477006
22.  Vascular tone pathway polymorphisms in relation to primary open-angle glaucoma 
Eye  2014;28(6):662-671.
Vascular perfusion may be impaired in primary open-angle glaucoma (POAG); thus, we evaluated a panel of markers in vascular tone-regulating genes in relation to POAG.
We used Illumina 660W-Quad array genotype data and pooled P-values from 3108 POAG cases and 3430 controls from the combined National Eye Institute Glaucoma Human Genetics Collaboration consortium and Glaucoma Genes and Environment studies. Using information from previous literature and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, we compiled single-nucleotide polymorphisms (SNPs) in 186 vascular tone-regulating genes. We used the ‘Pathway Analysis by Randomization Incorporating Structure' analysis software, which performed 1000 permutations to compare the overall pathway and selected genes with comparable randomly generated pathways and genes in their association with POAG.
The vascular tone pathway was not associated with POAG overall or POAG subtypes, defined by the type of visual field loss (early paracentral loss (n=224 cases) or only peripheral loss (n=993 cases)) (permuted P≥0.20). In gene-based analyses, eight were associated with POAG overall at permuted P<0.001: PRKAA1, CAV1, ITPR3, EDNRB, GNB2, DNM2, HFE, and MYL9. Notably, six of these eight (the first six listed) code for factors involved in the endothelial nitric oxide synthase activity, and three of these six (CAV1, ITPR3, and EDNRB) were also associated with early paracentral loss at P<0.001, whereas none of the six genes reached P<0.001 for peripheral loss only.
Although the assembled vascular tone SNP set was not associated with POAG, genes that code for local factors involved in setting vascular tone were associated with POAG.
PMCID: PMC4058608  PMID: 24603425
23.  Rapid genomic DNA changes in allotetraploid fish hybrids 
Wang, J | Ye, L H | Liu, Q Z | Peng, L Y | Liu, W | Yi, X G | Wang, Y D | Xiao, J | Xu, K | Hu, F Z | Ren, L | Tao, M | Zhang, C | Liu, Y | Hong, Y H | Liu, S J
Heredity  2015;114(6):601-609.
Rapid genomic change has been demonstrated in several allopolyploid plant systems; however, few studies focused on animals. We addressed this issue using an allotetraploid lineage (4nAT) of freshwater fish originally derived from the interspecific hybridization of red crucian carp (Carassius auratus red var., ♀, 2n=100) × common carp (Cyprinus carpio L., ♂, 2n=100). We constructed a bacterial artificial chromosome (BAC) library from allotetraploid hybrids in the 20th generation (F20) and sequenced 14 BAC clones representing a total of 592.126 kb, identified 11 functional genes and estimated the guanine–cytosine content (37.10%) and the proportion of repetitive elements (17.46%). The analysis of intron evolution using nine orthologous genes across a number of selected fish species detected a gain of 39 introns and a loss of 30 introns in the 4nAT lineage. A comparative study based on seven functional genes among 4nAT, diploid F1 hybrids (2nF1) (first generation of hybrids) and their original parents revealed that both hybrid types (2nF1 and 4nAT) not only inherited genomic DNA from their parents, but also demonstrated rapid genomic DNA changes (homoeologous recombination, parental DNA fragments loss and formation of novel genes). However, 4nAT presented more genomic variations compared with their parents than 2nF1. Interestingly, novel gene fragments were found for the iqca1 gene in both hybrid types. This study provided a preliminary genomic characterization of allotetraploid F20 hybrids and revealed evolutionary and functional genomic significance of allopolyploid animals.
PMCID: PMC4434252  PMID: 25669608
24.  Computed tomography–based distribution of involved lymph nodes in patients with upper esophageal cancer 
Li, M. | Liu, Y. | Xu, L. | Huang, Y. | Li, W. | Yu, J. | Kong, L.
Current Oncology  2015;22(3):e178-e182.
Delineating the nodal clinical target volume (ctvn) remains a challenging task in patients with cervical or upper thoracic esophageal carcinoma (ec). In particular, the extent of the lymph area that should be included in the irradiation field remains controversial. In the present study, the extent of the ctvn was determined based on the incidence of lymph node involvement mapped by computed tomography (ct) imaging.
Our study included 468 patients who were diagnosed with cervical and upper thoracic ec and who received staging information between June 2005 and April 2011. The anatomic distribution of metastatic regional lymph nodes was mapped using ct images and grouped using the levels established by the Radiation Therapy Oncology Group. The probability of the various groups being involved was examined. If a lymph node group had a probability of 10% or more of being involved, it was considered at high risk for metastasis, and elective treatment as part of the ctvn was recommended.
Lymph node involvement was mapped by ct in 256 patients (54.7%). Not all lymph node groups should be included in the ctvn. For cervical lesions, the involved lymph nodes were located mainly between the hyoid bone and the arcus aortae; the recommended ctvn should consist of the neck lymph nodes at levels iii and iv (supraclavicular group) and thoracic groups 2 and 3P. In upper thoracic ec patients, most of the involved lymph nodes were distributed between the cricoid cartilage and the subcarinal area; the ctvn should cover the supraclavicular group and thoracic nodal groups 2, 3P, 4, 5, and 7.
Our ct-based study indicates a specific distribution and incidence of metastatic lymph node groups in patients with cervical and upper thoracic ec. The results suggest that regional lymph node groups should be electively included in the ctvn for precise radiation administration.
PMCID: PMC4462540  PMID: 26089729
Esophageal squamous cell carcinoma; lymph node metastasis; computed tomography; radiation; clinical target volume
25.  Short-term weight-centric effects of tea or tea extract in patients with metabolic syndrome: a meta-analysis of randomized controlled trials 
Zhong, X | Zhang, T | Liu, Y | Wei, X | Zhang, X | Qin, Y | Jin, Z | Chen, Q | Ma, X | Wang, R | He, J
Nutrition & Diabetes  2015;5(6):e160-.
To evaluate the weight-centric effect of tea or tea extract in participants with metabolic syndrome (MetS), we performed electronic searches in PubMed, EmBase and the Cochrane Library to identify eligible randomized controlled trials (RCTs) comparing tea or tea extract vs a control group. A direct meta-analysis using random-effects model was conducted to pool the standardized mean difference regarding body mass index (BMI), body weight and waist circumference. Study quality was assessed by using the Jadad scale. Pre-specified subgroup and sensitivity analyses were conducted to explore potential heterogeneity. A total of five RCTs involving 338 adult individuals were included. Given the high heterogeneity observed in the overall pooled analysis, we separated the included subjects into two subgroups. Ingestion of tea or tea extract significantly reduced BMI (subgroup 1: −1.60, 95% confidence interval (CI), −2.05 to −1.14; subgroup 2: −0.40, 95% CI, −0.69 to −0.12) and body weight (subgroup 1: −4.14, 95% CI, −4.85 to −3.43; subgroup 2: −0.35, 95% CI, −0.68 to −0.02). This meta-analysis suggests that tea or tea extract has favorable weight-centric effects in MetS patients. Additional large RCTs specifically designed to evaluate the effect on anthropometric measurements are needed to further confirm these findings.
PMCID: PMC4491851  PMID: 26075637

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