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1.  Specificity Protein 1 Regulates Gene Expression Related to Fatty Acid Metabolism in Goat Mammary Epithelial Cells 
Specificity protein 1 (SP1) is a ubiquitous transcription factor that plays an important role in controlling gene expression. Although important in mediating the function of various hormones, the role of SP1 in regulating milk fat formation remains unknown. To investigate the sequence and expression information, as well as its role in modulating lipid metabolism, we cloned SP1 gene from mammary gland of Xinong Saanen dairy goat. The full-length cDNA of the SP1 gene is 4376 bp including 103 bp of 5'UTR, 2358 bp of ORF (HM_236311) and 1915 bp of 3'UTR, which is predicted to encode a 786 amino acids polypeptide. Phylogenetic tree analysis showed that goat SP1 has the closest relationship with sheep, followed by bovines (bos taurus, odobenus and ceratotherium), pig, primates (pongo, gorilla, macaca and papio) and murine (rattus and mus), while the furthest relationship was with canis and otolemur. Expression was predominant in the lungs, small intestine, muscle, spleen, mammary gland and subcutaneous fat. There were no significant expression level differences between the mammary gland tissues collected at lactation and dry-off period. Overexpression of SP1 in goat mammary epithelial cells (GMECs) led to higher mRNA expression level of peroxisome proliferator-activated receptor-γ (PPARγ) and lower liver X receptor α (LXRα) mRNA level, both of which were crucial in regulating fatty acid metabolism, and correspondingly altered the expression of their downstream genes in GMECs. These results were further enhanced by the silencing of SP1. These findings suggest that SP1 may play an important role in fatty acid metabolism.
PMCID: PMC4307335  PMID: 25594872
dairy goat; mammary epithelial cells; specificity protein 1; fatty acid metabolism
2.  Cobalt Protoporphyrin Pretreatment Protects Human Embryonic Stem Cell-Derived Cardiomyocytes From Hypoxia/Reoxygenation Injury In Vitro and Increases Graft Size and Vascularization In Vivo 
These experiments demonstrate that ex vivo pretreatment of human embryonic stem cell-derived cardiomyocytes with a single dose of cobalt protoporphyrin before intramyocardial implantation more than doubled resulting graft size and improved early graft vascularization in acutely infarcted hearts. These findings open the door for delivery of these, or other, stem cells during acute interventional therapy following myocardial infarction or ischemia.
Human embryonic stem cell-derived cardiomyocytes (hESC-CMs) can regenerate infarcted myocardium. However, when implanted into acutely infarcted hearts, few cells survive the first week postimplant. To improve early graft survival, hESC-CMs were pretreated with cobalt protoporphyrin (CoPP), a transcriptional activator of cytoprotective heme oxygenase-1 (HO-1). When hESC-CMs were challenged with an in vitro hypoxia/reoxygenation injury, mimicking cell transplantation into an ischemic site, survival was significantly greater among cells pretreated with CoPP versus phosphate-buffered saline (PBS)-pretreated controls. Compared with PBS-pretreated cells, CoPP-pretreated hESC-CM preparations exhibited higher levels of HO-1 expression, Akt phosphorylation, and vascular endothelial growth factor production, with reduced apoptosis, and a 30% decrease in intracellular reactive oxygen species. For in vivo translation, 1 × 107 hESC-CMs were pretreated ex vivo with CoPP or PBS and then injected intramyocardially into rat hearts immediately following acute infarction (permanent coronary ligation). At 1 week, hESC-CM content, assessed by quantitative polymerase chain reaction for human Alu sequences, was 17-fold higher in hearts receiving CoPP- than PBS-pretreated cells. On histomorphometry, cardiomyocyte graft size was 2.6-fold larger in hearts receiving CoPP- than PBS-pretreated cells, occupying up to 12% of the ventricular area. Vascular density of host-perfused human-derived capillaries was significantly greater in grafts composed of CoPP- than PBS-pretreated cells. Taken together, these experiments demonstrate that ex vivo pretreatment of hESC-CMs with a single dose of CoPP before intramyocardial implantation more than doubled resulting graft size and improved early graft vascularization in acutely infarcted hearts. These findings open the door for delivery of these, or other, stem cells during acute interventional therapy following myocardial infarction or ischemia.
PMCID: PMC4039456  PMID: 24736402
Cell therapy; Infarct repair; Human embryonic stem cell; Heme oxygenase-1; Preconditioning; Acute myocardial infarction
3.  Lipoprotein Lipase, Tissue Expression and Effects on Genes Related to Fatty Acid Synthesis in Goat Mammary Epithelial Cells 
Lipoprotein lipase (LPL) serves as a central factor in hydrolysis of triacylglycerol and uptake of free fatty acids from the plasma. However, there are limited data concerning the action of LPL on the regulation of milk fat synthesis in goat mammary gland. In this investigation, we describe the cloning and sequencing of the LPL gene from Xinong Saanen dairy goat mammary gland, along with a study of its phylogenetic relationships. Sequence analysis showed that goat LPL shares similarities with other species including sheep, bovine, human and mouse. LPL mRNA expression in various tissues determined by RT-qPCR revealed the highest expression in white adipose tissue, with lower expression in heart, lung, spleen, rumen, small intestine, mammary gland, and kidney. Expression was almost undetectable in liver and muscle. The expression profiles of LPL gene in mammary gland at early, peak, mid, late lactation, and the dry period were also measured. Compared with the dry period, LPL mRNA expression was markedly greater at early lactation. However, compared with early lactation, the expression was lower at peak lactation and mid lactation. Despite those differences, LPL mRNA expression was still greater at peak, mid, and late lactation compared with the dry period. Using goat mammary epithelial cells (GMEC), the in vitro knockdown of LPL via shRNA or with Orlistat resulted in a similar degree of down-regulation of LPL (respectively). Furthermore, knockdown of LPL was associated with reduced mRNA expression of SREBF1, FASN, LIPE and PPARG but greater expression of FFAR3. There was no effect on ACACA expression. Orlistat decreased expression of LIPE, FASN, ACACA, and PPARG, and increased FFAR3 and SREBF1 expression. The pattern of LPL expression was similar to the changes in milk fat percentage in lactating goats. Taken together, results suggest that LPL may play a crucial role in fatty acid synthesis.
PMCID: PMC4284735  PMID: 25501331
LPL gene; lactation; goat mammary epithelial cells; Orlistat
4.  Decoding the androgen receptor splice variants 
In the past five years, multiple structurally and functionally distinct androgen receptor (AR) splice variants have been decoded and characterized. The mature transcripts for the majority of the fully decoded AR splice variants contain a transcribed “intronic” sequence, capable of encoding a short variant-specific peptide to replace the AR ligand-binding domain (LBD). Functionally, AR splice variants represent a diverse group of molecules often demonstrating cell context-specific genomic functions that may or may not be coupled with the functions of the canonical full-length AR (AR-FL). However, the full spectrum of their functional diversity and the underlying mechanistic basis remains very poorly characterized. In clinical specimens derived from men treated with a variety of hormone therapy regimens, AR splice variants are almost always expressed at detectable, yet lower levels when compared to that of AR-FL. In spite of the collective in vitro data supporting the putative role of AR splice variants in therapeutic resistance to hormone therapies, the extent to which AR splice variants mediate resistance to each individual regimen is not known and awaits thorough investigations in a clinically relevant setting using specimens from men undergoing treatments. Among the AR splice variants, AR-V7 is more abundantly and frequently expressed in castration-resistant prostate cancer (CRPC) and remains the most important variant identified so far. The relative importance of different AR molecules, including AR-FL, should be functionally dissected in the setting of castration-resistant prostate cancer, particularly in tumors resistant to more potent inhibitors of AR-FL recently approved by the FDA. In this review, we will focus on the discovery and characterization of AR splice variants, their putative functions and roles in mediating constitutively active AR signaling, and key areas of investigation that are necessary to establish their clinical relevance.
PMCID: PMC4209743  PMID: 25356377
Androgen receptor (AR); AR Splice Variants; AR signaling; full-length AR (AR-FL); prostate cancer; castration-resistant prostate cancer (CRPC)
5.  A direct real-time polymerase chain reaction assay for rapid high-throughput detection of highly pathogenic North American porcine reproductive and respiratory syndrome virus in China without RNA purification 
Porcine reproductive and respiratory syndrome virus (PRRSV), and particularly its highly pathogenic genotype (HP-PRRSV), have caused massive economic losses to the global swine industry.
To rapidly identify HP-PRRSV, we developed a direct real-time reverse transcription polymerase chain reaction method (dRT-PCR) that could detect the virus from serum specimen without the need of RNA purification. Our dRT-PCR assay can be completed in 1.5 h from when a sample is received to obtaining a result. Additionally, the sensitivity of dRT-PCR matched that of conventional reverse transcription PCR (cRT-PCR) that used purified RNA. The lowest detection limit of HP-PRRSV was 6.3 TCID50 using dRT-PCR. We applied dRT-PCR assay to 144 field samples and the results showed strong consistency with those obtained by cRT-PCR. Moreover, the dRT-PCR method was able to tolerate 5-20% (v/v) serum.
Our dRT-PCR assay allows for easier, faster, more cost-effective and higher throughput detection of HP-PRRSV compared with cRT-PCR methods. To the best of our knowledge, this is the first report to describe a real-time RT-PCR assay capable of detecting PRRSV in crude serum samples without the requirement for purifying RNA. We believe our approach has a great potential for application to other RNA viruses.
Electronic supplementary material
The online version of this article (doi:10.1186/2049-1891-5-45) contains supplementary material, which is available to authorized users.
PMCID: PMC4198619  PMID: 25324970
Highly pathogenic; Porcine reproductive and respiratory syndrome virus; Real-time RT-PCR
6.  GOLPH2 and MYO6:Putative Prostate Cancer Markers Localizedtothe Golgi Apparatus 
The Prostate  2008;68(13):1387-1395.
Malignant transformation is often accompanied by morphological and functional alterations in subcellular organelles. The Golgi apparatus is a subcellular structure primarily involved in modification and sorting of macromolecules for secretion and transport to other cellular destinations. Molecular alterations associated with the Golgi apparatus may take place during prostate carcinogenesis but such alterations have not been documented.
To demonstrate that the Golgi apparatus undergoes alterations during prostate carcinogenesis, we examined the expression and localization of two candidate molecules, Golgi phosphoprotein 2 (GOLPH2) and myosin VI (MYO6), both overexpressed in prostate cancer as initially identified by expression microarray analysis.
Elevated GOLPH2 expression in prostate cancers was validated through real-time RT-PCR, Western blot, and tissue microarray analysis, and its Golgi localization in surgical prostate cancer tissues confirmed using two-color immunofluorescence. In addition, distinctive juxtanuclear MYO6 staining pattern consistent with Golgi localization was observed in surgical prostate cancer tissues. Two-color immunofluorescence revealed intensive Golgi-specific staining for both GOLPH2 and myosin VI in prostate cancer cells but not in the adjacent normal prostate epithelium.
We show that the Golgi apparatus in prostate cancer cells differs from the normal Golgi by elevated levels of two molecules, GOLPH2 and MYO6. These results for the first time demonstrated consistent cancer cell-specific alterations in the molecular composition of the Golgi apparatus. Such alterations can be explored for discovery of novel prostate cancer biomarkers through targeted organellar approaches.
PMCID: PMC4124602  PMID: 18543251
prostate cancer; GOLPH2; myosin VI; Golgi
7.  Relaxor-PbTiO3 Single Crystals for Various Applications 
Piezoelectric materials lie at the heart of electromechanical devices. Applications include actuators, ultrasonic imaging, high intensity focused ultrasound, underwater ultrasound, nondestructive evaluation transducer, pressure sensors, and accelerometers, to name a few. In this work, the advantages and disadvantages of relaxor-PbTiO3-based single crystals are discussed, based on the requirements (figure of merit) of various applications, with emphasis on recent developments of the shear properties of single crystals as a function of temperature and applied fields.
PMCID: PMC4105699  PMID: 25004527
8.  Definition of a FoxA1 Cistrome that is Crucial for G1-S Phase Cell-Cycle Transit in Castration-Resistant Prostate Cancer 
Cancer research  2011;71(21):6738-6748.
The enhancer pioneer transcription factor FoxA1 is a global mediator of steroid receptor (SR) action in hormone-dependent cancers. In castration-resistant prostate cancer (CRPC), FoxA1 acts as an androgen receptor co-factor to drive G2-M phase cell-cycle transit. Here we describe a mechanistically distinct SR-independent role for FoxA1 in driving G1-S phase cell-cycle transit in CRPC. By comparing FoxA1 binding sites in prostate cancer cell genomes, we defined a co-dependent set of FoxA1-MYBL2 and FoxA1-CREB1 binding sites within the regulatory regions of the Cyclin E2 and E2F1 genes that are critical for CRPC growth. Binding at these sites upregulate the Cyclin E2 and Cyclin A2 genes in CRPC but not in earlier stage androgen-dependent prostate cancer (ADPC), establishing a stage-specific role for this pathway in CRPC growth. Mechanistic investigations indicated that FoxA1, MYBL2 or CREB1 induction of histone H3 acetylation facilitated nucleosome disruption as the basis for co-dependent transcriptional activation and G1-S phase cell-cycle transit. Our findings establish FoxA1 as a pivotal driver of the cell-cycle in CRPC which promotes G1-S phase transit as well as G2-M phase transit through two distinct mechanisms.
PMCID: PMC4081454  PMID: 21900400
FoxA1; cistrome; cell-cycle G1-S progression; castration-resistant prostate cancer
9.  Photosynthetic and Canopy Characteristics of Different Varieties at the Early Elongation Stage and Their Relationships with the Cane Yield in Sugarcane 
The Scientific World Journal  2014;2014:707095.
During sugarcane growth, the Early Elongation stage is critical to cane yield formation. In this study, parameters of 17 sugarcane varieties were determined at the Early Elongation stage using CI-301 photosynthesis measuring system and CI-100 digital plant canopy imager. The data analysis showed highly significant differences in leaf area index (LAI), mean foliage inclination angle (MFIA), transmission coefficient for diffused light penetration (TD), transmission coefficient for solar beam radiation penetration (TR), leaf distribution (LD), net photosynthetic rate (PN), transpiration rate (E), and stomatal conductance (GS) among sugarcane varieties. Based on the photosynthetic or canopy parameters, the 17 sugarcane varieties were classified into four categories. Through the factor analysis, nine parameters were represented by three principal factors, of which the cumulative rate of variance contributions reached 85.77%. A regression for sugarcane yield, with relative error of yield fitting less than 0.05, was successfully established: sugarcane yield = −27.19 − 1.69 × PN + 0.17 ×  E + 90.43 × LAI − 408.81 × LD + 0.0015 × NSH + 101.38 ×  D (R2 = 0.928**). This study helps provide a theoretical basis and technical guidance for the screening of new sugarcane varieties with high net photosynthetic rate and ideal canopy structure.
PMCID: PMC4090427  PMID: 25045742
10.  Cultivar Evaluation and Essential Test Locations Identification for Sugarcane Breeding in China 
The Scientific World Journal  2014;2014:302753.
The discrepancies across test sites and years, along with the interaction between cultivar and environment, make it difficult to accurately evaluate the differences of the sugarcane cultivars. Using a genotype main effect plus genotype-environment interaction (GGE) Biplot software, the yield performance data of seven sugarcane cultivars in the 8th Chinese National Sugarcane Regional Tests were analyzed to identify cultivars recommended for commercial release. Fn38 produced a high and stable sugar yield. Gn02-70 had the lowest cane yield with high stability. Yz06-407 was a high cane yield cultivar with poor stability in sugar yield. Yz05-51 and Lc03-1137 had an unstable cane yield but relatively high sugar yield. Fn39 produced stable high sugar yield with low and unstable cane production. Significantly different sugar and cane yields were observed across seasons due to strong cultivar-environment interactions. Three areas, Guangxi Chongzuo, Guangxi Baise, and Guangxi Hechi, showed better representativeness of cane yield and sugar content than the other four areas. On the other hand, the areas Guangxi Chongzuo, Yunnan Lincang, and Yunnan Baoshan showed strong discrimination ability, while the areas Guangxi Hechi and Guangxi Liuzhou showed poor discrimination ability. This study provides a reference for cultivar evaluation and essential test locations identification for sugarcane breeding in China.
PMCID: PMC4055468  PMID: 24982939
11.  Modulation of the Cellular Distribution of Human Cytomegalovirus Helicase by Cellular Factor Snapin 
Journal of Virology  2013;87(19):10628-10640.
Controlled regulation of genomic DNA synthesis is a universally conserved process for all herpesviruses, including human cytomegalovirus (HCMV), and plays a key role in viral pathogenesis, such as persistent infections. HCMV UL105 is believed to encode the helicase of the DNA replication machinery that needs to localize in the nuclei, the site of viral DNA synthesis. No host factors that interact with UL105 have been identified. In this study, we show that UL105 specifically interacts with Snapin, a human protein that is predominantly localized in the cytoplasm and associated with cellular vesicles. UL105 was found to interact with Snapin in both the yeast two-hybrid screen and coimmunoprecipitation experiments in HCMV-infected cells. The nuclear and cytoplasmic levels of UL105 were decreased and increased in cells overexpressing Snapin, respectively, while the levels of UL105 in the nuclei and cytoplasm were increased and decreased in cells in which the expression of Snapin was downregulated with anti-Snapin small interfering RNA (siRNA) molecules, respectively. Furthermore, viral DNA synthesis and progeny production were decreased in cells overexpressing Snapin and increased in the anti-Snapin siRNA-treated cells, respectively. Our results provide the first direct evidence to suggest that Snapin interacts with UL105 and alters its cellular distribution, leading to modulation of viral DNA synthesis and progeny production. Our study further suggests that modulation of the cellular distribution of viral helicase by Snapin may represent a possible mechanism for regulating HCMV genomic DNA synthesis, a key step during herpesvirus lytic and persistent infections.
PMCID: PMC3807405  PMID: 23885069
12.  Rapamycin promoted thrombosis and platelet adhesion to endothelial cells by inducing membrane remodeling 
BMC Cell Biology  2014;15:7.
Recently, evidence indicated that the rapamycin-eluting stent which was used worldwide may contribute to an increased risk for thrombosis. On the contrary, other researchers found it was safe. Thus, it is necessary to clarify the effect of rapamycin on thrombosis and the corresponding mechanisms.
The effects of rapamycin in vivo were evaluated by modified deep vein thrombosis animal model. The platelets were from healthy volunteers and the platelet-endothelium (purchased from ATCC) adhesion in cultured endothelial cells was assessed. Membrane rufflings in endothelial cells were examined by confocal and electron microscope. Thrombus formation increased in rats that were injected with rapamycin. Electron microscope analysis exhibited microvilli on the rapamycin-treated endothelium in rats. Rapamycin enhanced membrane ruffling in human umbilical vein endothelial cells (HUVECs) and adhesion of platelets to HUVECs. The platelet-HUVECs adhesion was attenuated when cells were treated with cytochalacin B. Inhibition of autophagy by 3-methyladenine led to suppression of membrane ruffles in HUVECs and augmentation of platelet-endothelial adhesion.
In conclusion, we found that endothelial membrane remodeling induced by rapamycin is crucial for the adhesion of platelets to endothelial cells and thereby for thrombosis in vivo, and that the endothelial membrane remodeling is autophagy dependent.
PMCID: PMC3936831  PMID: 24564184
Thrombosis; Membrane remodeling; Endothelial cell; Platelet
13.  Tasquinimod is an Allosteric Modulator of HDAC4 Survival Signaling within the Compromised Cancer Microenvironment 
Cancer research  2012;73(4):1386-1399.
Tasquinimod is an orally active anti-angiogenic drug that is currently in Phase III clinical trials for the treatment of castration resistant prostate cancer. However, the target of this drug has remained unclear. In this study we applied diverse strategies to identify the histone deacetylase HDAC4 as a target for the anti-angiogenic activity of tasquinimod. Our comprehensive analysis revealed allosteric binding (Kd 10–30 nM) to the regulatory Zn2+ binding domain of HDAC4 which locks the protein in a conformation preventing HDAC4/N-CoR/HDAC3 complex formation. This binding inhibited co-localization of N-CoR/HDAC3, thereby inhibiting deacetylation of histones and HDAC4 client transcription factors, such as HIF-1α, which are bound at promoter/enhancers where epigenetic reprogramming is required for cancer cell survival and angiogenic response. Through this mechanism, tasquinimod is effective as a monotherapeutic agent against human prostate, breast, bladder, and colon tumor xenografts, where its efficacy could be further enhanced in combination with a targeted thapsigargin prodrug (G202) that selectively kills tumor endothelial cells. Together, our findings define a mechanism of action of tasquinimod and offer a perspective on how its clinical activity might be leveraged in combination with other drugs that target the tumor microenvironment.
PMCID: PMC3578133  PMID: 23149916
14.  Prevalence, awareness, treatment and control of dyslipidemia among adults in Northwestern China: the cardiovascular risk survey 
The aim of this study was to estimate the prevalence, awareness, treatment, and control of dyslipidemia in Xinjiang, China.
Stratified sampling method was used to select a representative sample of the general population including Chinese Han, Uygur, and Kazak in this geographic area. Seven cities were chosen. Based on the government records of registered residences, one participant was randomly selected from each household. The eligibility criterion for the study was ≥ 35 years of age.
A total of 14,618 participants (5,757 Han, 4,767 Uygur, and 4,094 Kazak), were randomly selected from 26 villages in 7 cities. The prevalence of dyslipidemia was 52.72% in the all participants. The prevalence of dyslipidemia was higher in Han than that in the other two ethnic (58.58% in Han, 48.27% in Uygur, and 49.60% in Kazak, P < 0.000). The prevalence of dyslipidemia was higher in men than that in women (56.4% vs. 49.3%, P < 0.000). Among the participants with dyslipidemia, the proportion of those who aware, treat, control of dyslipidemia were 53.67%, 22.51%, 17.09% in Han, 42.19%, 27.78%, 16.20% in Uygur, 37.02%, 21.11%, 17.77% in Kazak.
Dyslipidemia is highly prevalent in Xinjiang. The proportion of participants with dyslipidemia who were aware, treated, and controlled is unacceptably low. These results underscore the urgent need to develop national strategies to improve the prevention, detection, and treatment of dyslipidemia in Xinjiang.
PMCID: PMC3895843  PMID: 24393232
Dyslipidemia; Prevalence; Awareness; Treatment; Control
15.  The substantial hospitalization burden of influenza in central China: surveillance for severe, acute respiratory infection, and influenza viruses, 2010–2012 
Published data on influenza in severe acute respiratory infection (SARI) patients are limited. We conducted SARI surveillance in central China and estimated hospitalization rates of SARI attributable to influenza by viral type/subtype.
Surveillance was conducted at four hospitals in Jingzhou, China from 2010 to 2012. We enrolled hospitalized patients who had temperature ≥37·3°C and at least one of: cough, sore throat, tachypnea, difficulty breathing, abnormal breath sounds on auscultation, sputum production, hemoptysis, chest pain, or chest radiograph consistent with pneumonia. A nasopharyngeal swab was collected from each case-patient within 24 hours of admission for influenza testing by real-time reverse transcription PCR.
Of 17 172 SARI patients enrolled, 90% were aged <15 years. The median duration of hospitalization was 5 days. Of 16 208 (94%) SARI cases tested, 2057 (13%) had confirmed influenza, including 1427 (69%) aged <5 years. Multiple peaks of influenza occurred during summer, winter, and spring months. Influenza was associated with an estimated 115 and 142 SARI hospitalizations per 100 000 during 2010–2011 and 2011–2012 [including A(H3N2): 55 and 44 SARI hospitalizations per 100 000; pandemic A(H1N1): 33 SARI hospitalizations per 100 000 during 2010–2011; influenza B: 26 and 98 hospitalizations per 100 000], with the highest rate among children aged 6–11 months (3603 and 3805 hospitalizations per 100 000 during 2010–2011 and 2011–2012, respectively).
In central China, influenza A and B caused a substantial number of hospitalizations during multiple periods each year. Our findings strongly suggest that young children should be the highest priority group for annual influenza vaccination in China.
PMCID: PMC4177798  PMID: 24209711
China; disease burden; influenza; seasonality
16.  Complete Genome Sequence of a Human Enterovirus 71 Strain Isolated in Wuhan, China, in 2010 
Genome Announcements  2013;1(6):e01112-13.
The complete genome sequence of a human enterovirus 71 strain (EV71/wuhan/3018/2010), which was isolated in Wuhan in 2010, was amplified by a reverse transcription-PCR method and sequenced. Phylogenetic analysis based on the complete genome sequence classified this strain into subgenogroup A.
PMCID: PMC3873616  PMID: 24371206
17.  Seasonal Variation of the Canopy Structure Parameters and Its Correlation with Yield-Related Traits in Sugarcane 
The Scientific World Journal  2013;2013:801486.
Population structure determines sugarcane yield, of which canopy structure is a key component. To fully understand the relations between sugarcane yield and parameters of the canopy structure, 17 sugarcane varieties were investigated at five growth stages. The results indicated that there were significant differences between characterized parameters among sugarcane populations at different growth stages. During sugarcane growth after planting, leaf area index (LAI) and leaf distribution (LD) increased, while transmission coefficient for diffuse radiation (TD), mean foliage inclination angle (MFIA), transmission coefficient for solar beam radiation penetration (TR), and extinction coefficient (K) decreased. Significant negative correlations were found between sugarcane yield and MFIA, TD, TR, and K at the early elongation stage, while a significant positive correlation between sugarcane yield and LD was found at the same stage. A regression for sugarcane yield, with relative error of yield fitting less than 10%, was successfully established: sugarcane yield = 2380.12 + 46.25 × LD − 491.82 × LAI + 1.36 × MFIA + 614.91 × TD − 1908.05 × TR − 182.53 ×  K + 1281.75 × LD − 1.35 × MFIA + 831.2 × TR − 407.8 ×  K + 8.21 × MFIA − 834.50 × TD − 1695.49 ×  K  (R2 = 0.94**).
PMCID: PMC3888768  PMID: 24453909
18.  Ecological Adaption Analysis of the Cotton Aphid (Aphis gossypii) in Different Phenotypes by Transcriptome Comparison 
PLoS ONE  2013;8(12):e83180.
The cotton aphid, Aphis gossypii Glover, is a destructive insect pest worldwide; it directly or indirectly damages (virus transmission) 300 species of host plants. Knowledge of their ecologically adaptive mechanisms at the molecular level may provide an essential and urgent method to effectively control this pest. However, no transcriptome information is available for the cotton aphid and sequence data are scarce. Therefore, we obtained transcriptome data.
To facilitate such a study, two cotton aphid transcriptomes at different growth stages of cotton, seedling and summer, were sequenced. A total of 161,396 and 66,668 contigs were obtained and assembled into 83,671 and 42,438 transcripts, respectively. After combining the raw date for both transcriptomes, the sequences were reassembled into 66,695 transcripts, and 52,160 were annotated based on BLASTX analyses. Comparison of the transcriptomes revealed that summer presented less challenges for the cotton aphids than the seedling stage of cotton. In total, 58 putative heat shock protein genes and 66 candidate cytochrome p450 genes were identified with BLASTX.
Our results form a basis for exploring the molecular mechanisms of ecological adaption in the cotton aphid. Our study also provides a baseline for the exploration of abiotic stress responses. In addition, it provides large-scale sequence information for further studies on this species.
PMCID: PMC3871566  PMID: 24376660
19.  Astragalus Polysaccharide Suppresses Skeletal Muscle Myostatin Expression in Diabetes: Involvement of ROS-ERK and NF-κB Pathways 
Objective. The antidiabetes drug astragalus polysaccharide (APS) is capable of increasing insulin sensitivity in skeletal muscle and improving whole-body glucose homeostasis. Recent studies suggest that skeletal muscle secreted growth factor myostatin plays an important role in regulating insulin signaling and insulin resistance. We hypothesized that regulation of skeletal muscle myostatin expression may be involved in the improvement of insulin sensitivity by APS. Methods. APS was administered to 13-week-old diabetic KKAy and nondiabetic C57BL/6J mice for 8 weeks. Complementary studies examined APS effects on the saturated acid palmitate-induced insulin resistance and myostatin expression in C2C12 cells. Results. APS treatment ameliorated hyperglycemia, hyperlipidemia, and insulin resistance and decreased the elevation of myostatin expression and malondialdehyde production in skeletal muscle of noninsulin-dependent diabetic KKAy mice. In C2C12 cells in vitro, saturated acid palmitate-induced impaired glucose uptake, overproduction of ROS, activation of extracellular regulated protein kinases (ERK), and NF-κB were partially restored by APS treatment. The protective effects of APS were mimicked by ERK and NF-κB inhibitors, respectively. Conclusion. Our study demonstrates elevated myostatin expression in skeletal muscle of type 2 diabetic KKAy mice and in cultured C2C12 cells exposed to palmitate. APS is capable of improving insulin sensitivity and decreasing myostatin expression in skeletal muscle through downregulating ROS-ERK-NF-κB pathway.
PMCID: PMC3880770  PMID: 24454989
21.  Clinical significance of Girdin expression detected by immunohistochemistry in non-small cell lung cancer 
Oncology Letters  2013;7(2):337-341.
Girdin protein has been implicated in cell migration and proliferation control. Previous evidence has confirmed that Girdin is a pivotal protein during cancer progression. To date, no evidence has been identified for the clinical significance of Girdin expression in non-small cell lung cancer (NSCLC). The current study aimed to investigate the expression and clinical significance of Girdin protein in NSCLC. In total, 36 tumor samples were obtained from patients undergoing surgery for NSCLC at The 309th Hospital of Chinese People’s Liberation Army (Beijing, China). The protein expression of Girdin was determined by immunohistochemistry analysis and the levels of Girdin protein were significantly higher in tumor samples than in distal normal lung tissue. A significant correlation was identified between Girdin overexpression and blood vessel infiltration of the tumor (P=0.013). Furthermore, analysis found that the Girdin-high phenotype was not associated with higher Ki-67 score. Girdin protein was frequently overexpressed in NSCLC and expression of Girdin was associated with blood vessel infiltration. The results of the present study suggest that Girdin should be considered as a potential marker for the prognosis of NSCLC; however, future studies are required to confirm theses results.
PMCID: PMC3881701  PMID: 24396443
Girdin; non-small cell lung cancer; Ki-67; metastasis
22.  Main Factors Influencing the Efficient Propagation of Virulent or Attenuated Strains of Japanese encephalitis virus in BHK-21 Cells 
Japanese encephalitis virus (JEV) causes severe viral encephalitis in humans and some other mammalian animals. Highly efficient culture of the virus is critical for antigen preparation, vaccine production and other basic researches. We have investigated the influence of a number of variables such as the strain virulence, the state of the host cells, medium composition, infection method and others on the proliferation of distinct JEV strains in BHK-21 cells. The results showed that two distinct strategies are needed for the propagation of virulent or attenuated JEV strains. The most critical variables were the method of infection, and especially the density of the host cell. Our studies indicate the general approaches to the in vitro culture of other JEV strains using BHK-21 cell line.
PMCID: PMC3550778  PMID: 24293822
JEV; In vitro culture; TCID50; BHK-21 cell line
23.  The Effect of Apigenin on Pharmacokinetics of Imatinib and Its Metabolite N-Desmethyl Imatinib in Rats 
BioMed Research International  2013;2013:789184.
The purpose of this study was to determine the effect of apigenin on the pharmacokinetics of imatinib and N-desmethyl imatinib in rats. Healthy male SD rats were randomly divided into four groups: A group (the control group), B group (the long-term administration of 165 mg/kg apigenin for 15 days), C group (a single dose of 165 mg/kg apigenin), and D group (a single dose of 252 mg/kg apigenin). The serum concentrations of imatinib and N-desmethyl imatinib were measured by HPLC, and pharmacokinetic parameters were calculated using DAS 3.0 software. The parameters of AUC(0−t), AUC(0−∞), Tmax, Vz/F, and CLz/F for imatinib in group B were different from those in group A (P < 0.05). Besides, MRT(0−t) and MRT(0−∞) in groups C and D differed distinctly from those in group A as well. The parameters of AUC(0−t) and Cmax for N-desmethyl imatinib in group C were significantly lower than those in group A (P < 0.05); however, compared with groups B and D, the magnitude of effect was modest. Those results indicated that apigenin in the short-term study inhibited the metabolism of imatinib and its metabolite N-desmethyl imatinib, while in the long-term study the metabolism could be accelerated.
PMCID: PMC3863468  PMID: 24369535
24.  Effects of recombinant human interleukin-10 on Treg cells, IL-10 and TGF-β in transplantation of rabbit skin 
Molecular Medicine Reports  2013;9(2):639-644.
The current study aimed to investigate the rejection and survival time of grafted skin, and the changes of Treg cells, interleukin 10 (IL-10) and transforming growth factor-β (TGF-β) in peripheral blood following skin transplantation with recombinant human interleukin-10 (rhIL-10) or cyclosporin A (CsA), as well as the role of IL-10 in immunological rejection mechanisms. A total of 36 rabbits were divided into two groups. The skin of a donor rabbit was transplanted onto the back of one receptor rabbit. Receptors were randomly divided into six groups, including rhIL-10 low-dose (5 μg/kg/d), rhIL-10 high-dose (10 μg/kg/d), CsA low-dose (5 mg/kg/d), CsA high-dose (10 mg/kg/d), rhIL-10 (5 μg/kg/d) and CsA (5 mg/kg/d) and negative control normal saline (NS; 1 ml/d). All groups received intramuscular drug injection for ten days, beginning one day prior to skin transplantation surgery. Following transplantation, each rabbit’s peripheral blood was collected at different times. The changes of CD4+CD25+ regulatory T cells, IL-10 and TGF-β were determined by flow cytometry and enzyme-linked immunosorbent assay. When compared with the control group, the rejection and survival times of the experimental groups were longer following skin graft. Compared with the two CsA groups and the control group, the proportion of CD4+CD25+ regulatory T cells of rhIL-10 groups was significantly upregulated on the 4th and 7th days following surgery. However, TGF-β levels were not significantly different. Data suggested that the concentration of IL-10 was positively correlated with the proportion of CD4+CD25+ regulatory T cells. In addition, IL-10 may delay the rejection time of rabbit skin transplantation and prolong the survival time. Thus, the role of IL-10 in inhibited allograft rejection may be associated with CD4+CD25+ regulatory T cells and IL-10, and may be independent of TGF-β.
PMCID: PMC3896515  PMID: 24270972
cytokines; graft rejection; interleukin-10; Treg cells
25.  MiR-103 Controls Milk Fat Accumulation in Goat (Capra hircus) Mammary Gland during Lactation 
PLoS ONE  2013;8(11):e79258.
Milk is the primary source of nutrition for young mammals including humans. The nutritional value of milk is mainly attributable to fats and proteins fractions. In comparison to cow milk, goat milk contains greater amounts of total fat, including much higher levels of the beneficial unsaturated fatty acids. MicroRNAs (miRNAs), a well-defined group of small RNAs containing about 22 nucleotides (nt), participate in various metabolic processes across species. However, little is known regarding the role of miRNAs in regulating goat milk composition. In the present study, we performed high-throughput sequencing to identify mammary gland-enriched miRNAs in lactating goats. We identified 30 highly expressed miRNAs in the mammary gland, including miR-103. Further studies revealed that miR-103 expression correlates with the lactation. Further functional analysis showed that over-expression of miR-103 in mammary gland epithelial cells increases transcription of genes associated with milk fat synthesis, resulting in an up-regulation of fat droplet formation, triglyceride accumulation, and the proportion of unsaturated fatty acids. This study provides new insight into the functions of miR-103, as well as the molecular mechanisms that regulate milk fat synthesis.
PMCID: PMC3823599  PMID: 24244462

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