A cross-sectional study was carried out involving a cohort of HIV-infected patients undergoing regular assessment in a tertiary hospital. Eighty-nine patients [mean (± standard deviation) age 42 ± 8 years] were included in the study: 14 patients were antiretroviral therapy (ART)-naïve, while

75 were on ART. Vitamin KU-60019 purchase D insufficiency (VDI) was defined as 25(OH)D < 75 nmol/L; insulin sensitivity was determined using a 2-h continuous infusion of glucose model assessment with homeostasis (CIGMA-HOMA), using the trapezoidal model to calculate the incremental insulin and glucose areas under the curve (AUCins and AUGglu, respectively). Beta cell function was assessed using the disposition index (DI). Abdominal visceral adipose tissue (VAT) and hepatic triglyceride content (HTGC) were measured by magnetic resonance imaging (MRI) and 1-H magnetic resonance spectroscopy. Multivariate linear regression analysis was performed. VDI was associated with insulin resistance (IR), as indicated by a higher

CIGMA-HOMA index (odds ratio 1.1) [1.01–1.2]. This association was independent of the main confounders, such as age, Centers for Disease Control and Prevention (CDC) stage, ART, lipodystrophy, body mass index, VAT:subcutaneous adipose tissue ratio and HTGC, as confirmed by multivariate analysis (B = 12.3; P = 0.01; r2 = 0.7). IR in patients with VDI was compensated by an increase in insulin response. However, beta cell function was lower in see more the VDI subpopulation (33% decrease in DI). VDI in nondiabetic HIV-positive male patients is associated with impaired insulin sensitivity and a decrease in pancreatic beta cell function. “
“We compared the use of computational models developed with and without HIV genotype vs. genotyping itself to predict effective

regimens for patients experiencing first-line virological failure. Two sets of models predicted virological response for 99 three-drug regimens for patients on a failing regimen of two nucleoside/nucleotide reverse transcriptase inhibitors and one nonnucleoside reverse transcriptase inhibitor in the Second-Line study. One set used viral load, CD4 count, genotype, plus treatment history Florfenicol and time to follow-up to make its predictions; the second set did not include genotype. Genotypic sensitivity scores were derived and the ranking of the alternative regimens compared with those of the models. The accuracy of the models and that of genotyping as predictors of the virological responses to second-line regimens were compared. The rankings of alternative regimens by the two sets of models were significantly correlated in 60−69% of cases, and the rankings by the models that use a genotype and genotyping itself were significantly correlated in 60% of cases. The two sets of models identified alternative regimens that were predicted to be effective in 97% and 100% of cases, respectively.

The aetiology and pathogenesis of MIH are PI3K inhibitor still unclear. The ameloblasts’ high sensitivity to relatively insignificant changes such as fever, hypocalcemia, etc., that alter normal cell function during amelogenesis can give rise to permanent morphological consequences[14, 38]. Although a number of risk factors have been related to MIH, the present study has not found any significant association. A cross-sectional design has evident limitations for studying aetiological factors and prospective studies are therefore needed to clarify the aetiology of MIH. This study shows that MIH is a relatively frequent syndrome among schoolchildren (21.8%).

MIH prevalence is high in the child population of this region, although the influence on

treatment needing is mild. A significant association with dental caries was observed, caries indices were significantly higher in the children with MIH than in the healthy children. Prospective studies are therefore needed to clarify the aetiology of MIH. Why this paper is important to paediatric dentists This study found a positive association between MIH and decay and therefore warns paediatric dentists on the increased need buy GW-572016 for treatment of affected children. This study was funded by the University of Valencia 2008 special action grants programme as project number UV-AE-08-2327. The authors would like to thank Dr Ivar Spelid and Dr Karin Weerheijm for their assistance when preparing the array of photographs for calibration purposes. The manuscript was translated into English by Mary Georgina those Hardinge. The authors have no conflict of interest to declare. “
“International Journal of Paediatric Dentistry 2011; 21: 119–125 Background.  In schoolchildren the most commonly decayed primary teeth are molars affecting proximal adjacent surfaces especially. Aim. 

To determine whether a more acidic plaque in response to sucrose challenge is detected in children with more carious lesions. Design.  Plaque pH measurements, using the microtouch technique, were carried out in interproximal spaces between primary molars, in 157 high caries risk children (314 sites and caries status of the 628 proximal surfaces recorded). The area under the curve (AUC5.7 and AUC6.2) was analyzed. Results.  The AUC5.7 and the AUC6.2 showed a statistically significant difference between plaque adjacent to proximal surfaces with or without caries. Differences for AUC5.7 and AUC6.2 were recorded between one decayed surface compared to two decayed surfaces (P < 0.01) whereas a statistical significant difference was only observed for AUC5.7, when the areas under the curve were obtained near one decayed surface compared to two sound surfaces (P = 0.04). Conclusions.

Polyketides are also signal compounds that control the differentiation of Dictyostelium. The polyketide synthases (PKSs) inhibitor, cerulenin, inhibits Dictyostelium differentiation and therefore its development (Serafimidis & Kay, 2005). The diversity this website of the biological activity of polyketides has rendered these secondary metabolites and the PKS genes that regulate their production the focus of biomedical and biopharmaceutical research. The completion of the Dictyostelium genome project revealed that the Dictyostelium genome contains more than 40 PKS genes, indicating that it has huge potential for polyketide production. In addition, the Dictyostelium genome contained two novel hybrid-type PKS

genes (Eichinger et al., 2005; Zucko et al., 2007). This novel structure was known as ‘Steely’. In Steely PKS proteins, the type III PKS domain was fused to

the C-terminus of a multidomain type I PKS (Eichinger et al., 2005; Austin et al., 2006). The two Steely-type PKSs were called SteelyA and SteelyB. SteelyB was reported to be responsible for the production of the stalk-inducing factor DIF-1 and the knockout mutant of stlB lacked DIF-1 (Austin et al., 2006). This stlB mutant aided the elucidation of the functions of DIF-1 in vivo (Saito et al., 2008). GSK-3 beta phosphorylation However, two different reports associated with SteelyA expression pattern and its products have been identified. According to one report in 2006, an in vitro product was identified as pyrone and the stlA gene was expressed maximally in early development before cell aggregation. Another report in 2008 identified 4-methyl-5-pentylbenzene-1,3-diol (MPBD) as the main in vitro product and the stlA gene was found to be expressed only in late development (Austin et al., 2006; Ghosh et al., 2008). In this study, we re-examined the expression pattern of stlA using two different primer sets and observed that it was similar to that in the dictyExpress database and our previous report (Austin et al., 2006; Rot et al., 2009). Furthermore, we used an stlA mutant and showed that one of the in vivo products of SteelyA was MPBD, a differentiation-inducing factor that was

Fossariinae identified in the conditioned medium for a dmtA mutant (Saito et al., 2006). Finally, we observed that MPBD induced the formation of mature spore cells in the fruiting body. The Dictyostelium discoideum Ax2 strain was grown in an axenic medium at 22 °C and was harvested at a density of approximately 5 × 106 cells mL−1. A stlA null strain that we reported previously (Austin et al., 2006) was grown in an axenic medium in the presence of 10 μg mL−1 balsticidin S. The axenically grown cells were washed and were developed at 22 °C on the phosphate buffer (2.7 mM Na2HPO4/10.7 mM K2HPO4 pH 6.2) agar plates at a density of 1–2 × 106 cells cm−2. For reverse transcription (RT)-PCR analysis, developing cells were harvested every 3 h until t21 (late culmination stage) and used for RNA purification.

Polyketides are also signal compounds that control the differentiation of Dictyostelium. The polyketide synthases (PKSs) inhibitor, cerulenin, inhibits Dictyostelium differentiation and therefore its development (Serafimidis & Kay, 2005). The diversity Doramapimod ic50 of the biological activity of polyketides has rendered these secondary metabolites and the PKS genes that regulate their production the focus of biomedical and biopharmaceutical research. The completion of the Dictyostelium genome project revealed that the Dictyostelium genome contains more than 40 PKS genes, indicating that it has huge potential for polyketide production. In addition, the Dictyostelium genome contained two novel hybrid-type PKS

genes (Eichinger et al., 2005; Zucko et al., 2007). This novel structure was known as ‘Steely’. In Steely PKS proteins, the type III PKS domain was fused to

the C-terminus of a multidomain type I PKS (Eichinger et al., 2005; Austin et al., 2006). The two Steely-type PKSs were called SteelyA and SteelyB. SteelyB was reported to be responsible for the production of the stalk-inducing factor DIF-1 and the knockout mutant of stlB lacked DIF-1 (Austin et al., 2006). This stlB mutant aided the elucidation of the functions of DIF-1 in vivo (Saito et al., 2008). PARP activity However, two different reports associated with SteelyA expression pattern and its products have been identified. According to one report in 2006, an in vitro product was identified as pyrone and the stlA gene was expressed maximally in early development before cell aggregation. Another report in 2008 identified 4-methyl-5-pentylbenzene-1,3-diol (MPBD) as the main in vitro product and the stlA gene was found to be expressed only in late development (Austin et al., 2006; Ghosh et al., 2008). In this study, we re-examined the expression pattern of stlA using two different primer sets and observed that it was similar to that in the dictyExpress database and our previous report (Austin et al., 2006; Rot et al., 2009). Furthermore, we used an stlA mutant and showed that one of the in vivo products of SteelyA was MPBD, a differentiation-inducing factor that was

Sclareol identified in the conditioned medium for a dmtA mutant (Saito et al., 2006). Finally, we observed that MPBD induced the formation of mature spore cells in the fruiting body. The Dictyostelium discoideum Ax2 strain was grown in an axenic medium at 22 °C and was harvested at a density of approximately 5 × 106 cells mL−1. A stlA null strain that we reported previously (Austin et al., 2006) was grown in an axenic medium in the presence of 10 μg mL−1 balsticidin S. The axenically grown cells were washed and were developed at 22 °C on the phosphate buffer (2.7 mM Na2HPO4/10.7 mM K2HPO4 pH 6.2) agar plates at a density of 1–2 × 106 cells cm−2. For reverse transcription (RT)-PCR analysis, developing cells were harvested every 3 h until t21 (late culmination stage) and used for RNA purification.

Nevertheless, it is worth noting that IncL/M is the most Selleckchem Cobimetinib frequently found incompatibility group among the Enterobacteriaceae carrying blaDHA-1 genes studied in our setting (96.6%; 28 from 29 isolates) (data not published). Curiously, qnrB4 genes have frequently been linked to the broad-host-range IncL/M plasmids (Carattoli, 2009). The presence of both resistance genes on the same plasmid and the reported increase in PMQR could perhaps account for the increasing

number of isolates harbouring blaDHA-1 genes (Park et al., 2007; Tamang et al., 2008; Strahilevitz et al., 2009). The possibility that blaDHA-1 genes may be mobilized by a vector with a greater capacity to spread could perhaps explain the recently widespread distribution of blaDHA-1 genes. Southern hybridization analysis revealed the colocalization of blaDHA-1 and qnrB resistance genes on the same conjugative plasmid (Fig. 1). In S. marcescens and E. coli donor strains, blaDHA-1 and qnrB genes hybridized to an approximately 70 kb-sized plasmid. Plasmids

coharbouring these resistances in their transconjugants Sunitinib manufacturer were larger than in wild strains; that in the S. marcescens transconjugant was around 190 kb, while that in the E. coli transconjugant was around 250 kb. All plasmids belonged to the IncL/M group (Fig. 1). These discrepancies in the size between donors and their respective transconjugants could be explained by cointegrates formed during the conjugation process (García et al., 2005; Tamang et al., 2008). Care should therefore be taken in molecular epidemiology studies when plasmid size is only estimated in transconjugants

because it could be overestimated. To sum up, this is the first report of an isolate of S. marcescens harbouring a pACBL. The observation of scattered colonies near the edge of the inhibition zones was the only phenotypic method that led us to suspect the presence of a pACBL in a chromosomal AmpC producer. Our results suggest an in vivo horizontal transfer of a plasmid coharbouring blaDHA-1 and qnrB resistance genes between S. marcescens and E. coli isolates. We would like to express our sincere gratitude to Dr Gimeno (Servei de Microbiologia, Fludarabine chemical structure Fundació Puigvert, Barcelona) for providing data patient, Dr Llagostera (Dep. Microbiologia Molecular, UAB, Barcelona) for providing us with the E. coli HB101 (UA6190) strain, A. Alvarado for carefully reading this manuscript and to C. Newey for revising the English. This study was partially supported by the Ministry of Health and Consumer Affairs, Instituto de Salud Carlos III-Feder, Spanish Network for the Research in Infectious Diseases (REIPI/RD06/0008/0013 and RD06/0008/1012) and BFU2008-00995/BMC (Spanish Ministry of Education). “
“Programa de Ingeniería Genómica, Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México. Av.

3%, p < 0.001) compared with those born in North Africa. Overall, 135 (21.3%) pilgrims had traveled and planned to travel outside France, both before and after the pilgrimage. Our results show a complex pattern of international travel in French pilgrims participating in the Hajj of 2010. Two-thirds of them underwent a trip to their country of origin in North Africa, 1 to 4 months before traveling from France to Saudi Arabia and a quarter planned to go back to North Africa after a short stop-over in France, following the Hajj. Vorinostat supplier This reflects

France’s past colonial history in Algeria, Morocco, and Tunisia and the post-colonial migrations. Therefore, French pilgrims arriving to Saudi Arabia may both present with long incubation communicable diseases, acquired in North Africa and short incubation infections acquired in France. In case of acquisition of communicable diseases during their stay in Saudi Arabia, French pilgrims will have the potential to spread infectious disease agents not only in France, but also in North Africa. This was particularly worrying during the Hajj of

2009 regarding the risk of spread of influenza A H1N1 09.6 Collaborative sentinel surveillance networks monitoring disease trends among travelers offer valuable tools for evaluating travel health issues. However, the major multinational sentinel networks addressing travel health issues globally (GeoSentinel and EuroTravNet) are mainly based in industrialized countries and do not include sites in North Africa.7,8 The EuroTravNet center in Marseille captures only few cases of Hajj-associated PI3K inhibitor infectious diseases in returned French

pilgrims, although cohort studies have demonstrated that most pilgrims Ceramide glucosyltransferase departing from Marseille get ill during their stay in Saudi Arabia.9 This is due in part to the mild nature of Hajj-associated diseases that are not likely to be seen at a specialized clinic, but also to the fact that a significant proportion of travelers may exhibit symptoms in North Africa rather than in Marseille. Therefore, surveillance of Hajj-associated infectious diseases in French pilgrims should be coordinated between France and North African countries. In this perspective, collaboration with EpiSouth network, a recently born network aiming to improve communicable disease surveillance in the Mediterranean area could be useful.10 Our study is limited to a small cohort of pilgrims from one large city in France and although it is a tradition in Muslim communities that many pilgrims travel after Hajj in the Middle East and Indian subcontinent,6 our results cannot be extrapolated to all pilgrims. Better linked surveillance for travelers, including pilgrims to the Hajj, is needed by health information system development such as real time electronic reporting, rapid data collection and post-event reporting using mobile phone technology and social networking, and rapid laboratory testing where possible to improve outbreak detection and control.

400, P = 0.033; post hoc t-test with Bonferroni correction, valproic acid vs. control, t9 = 2.852, P = 0.019; sodium butyrate vs. control, t8 = 2.946, selleckchem P = 0.019). These

data indicate that two different drugs sharing an inhibitory activity on HDACs promote VEP acuity recovery. Thus, increasing histone acetylation promoted functional recovery in adult long-term MD rats. To investigate whether the recovery of visual acuity assessed electrophysiologically in long-term MD rats treated with HDAC inhibitors was relevant for rat behavior we devised a longitudinal behavioral assessment of the effect of the treatment on visual acuity (Fig. 2A). We chose to asses the effects of valproic acid because it is an FDA-approved molecule well tolerated by animals even for chronic treatments. In addition, valproic acid is soluble in aqueous buffers and easily crosses the blood–brain barrier. Behavioral visual acuity of the nondeprived eye in long-term MD rats

was assessed using the Prusky visual water task before RS. After RS at P120, visual acuity of the deprived eye was measured to obtain the pretreatment visual acuity value of the amblyopic eye. This procedure lasted ∼10 days. Subsequently, Etoposide price rats were randomly assigned to the groups of treatment with valproic acid or control saline. Daily treatment was performed for 15 days. Then, visual acuity of the long-term deprived eye was reassessed in the same animals. The treatment was continued during the behavioral experiments, resulting on average in a total Amrubicin treatment

duration of 25 days. Examples of the results obtained in a saline-treated and in a valproic acid-treated rat are shown in Fig. 2B-D, respectively. Fig. 3 reports the average visual acuity of the two groups (valproic acid, n = 4; saline, n = 3). Before the treatment the deprived eye of both groups was clearly amblyopic; indeed, its visual acuity was lower than that of the fellow eye (two-way anova, effect of factor ‘MD’, F1,10 = 59.389, P < 0.001; effect of factor ‘group of treatment’, F1,10 = 1.085 P = 0.322; interaction, F1,10 = 2.861 P = 0.122). After the treatment, the amblyopic eye acuity was significantly improved in the group receiving valproic acid, while it remained unchanged in the group receiving saline: two-way anova for the factors ‘type of treatment’ and ‘before or after treatment’ showed an interaction of the factors (F1,5 = 8.323, P = 0.03); post hoc Holm–Sidak indicated that saline and valproic treated groups did not differ before the treatment (t5 = 0.326, P = 0.75) but they significantly differed after the treatment (t5 = 3.6, P = 0.006). Within treatments, acuity of valproic acid-treated rats was significantly different before and after the treatment (t5 = 3.951, P = 0.011) whereas acuity of saline treated rats was not (t5 = 0.394, P = 0.71).

The diagnosis of enamel defects was performed using the Developmental Defects of Enamel (DDE) Index. Through interviews, information was collected on socio-demographic aspects, pregnancy, birthweight, prematurity, and breastfeeding. Statistical analysis was performed using the SPSS program for Windows and involved descriptive analysis, Fisher’s exact test, the chi-square test, and Poisson regression. Results:  The prevalence of developmental defects of enamel was 29.9%. PD-332991 Demarcated opacity was the most frequent type of defect. Children with a history

of very low birthweight had a greater prevalence of enamels defects (PR, 2.7; 95% CI, 1.66–4.61). Prematurity and socio-demographic variables BIBF 1120 cell line were not associated with enamel defects. Conclusion:  Children with a history of very low birthweight had a greater frequency of enamel defects in primary teeth. “
“Objective.  The aim of this study was to assess the influence of sucking habits and facial pattern measurements on the development of anterior open bite (AOB). Methods. 

A case–control study was carried out on 60 children aged 7 and 8 years attending municipal public schools in the city of Recife, Brazil. Data collection included interviews with guardians, oral examinations, and facial growth pattern analysis using cephalometric radiographs. The following cephalometric measurements were assessed: SN.Gn, SN.GoGn, FMA, and Facial Axis. Statistical analyses were performed using the Student’s t-test and Pearson’s chi-square test at a 5% level of significance. Results.  The percentage of children with sucking habits in the case group was much higher than in the control group (53.3%vs 16.7%) (P = 0.003). Children with sucking habits were six times more likely to develop AOB. Regarding the measurements assessed, no statistically significant differences

were observed between groups. Conclusion.  This study found no evidence that variations tuclazepam in cephalometric angles (SN.Gn, FMA, SN.GoGn, and facial axis) are risk factors for AOB. Only sucking habits demonstrated a positive correlation with an increased AOB. “
“Introduction.  It is well established that severe periodontitis clusters in families, but there are no data about the relationship between mothers with chronic periodontitis and their children’s periodontal status. Objective.  To evaluate a risk for periodontal diseases in children of periodontally diseased and healthy mothers. Methods.  Four study groups were included: (I) 20 female patients with untreated generalized severe chronic periodontitis, (II) their children (34), (III) 13 periodontally healthy mothers and (IV) their children (13). Material was collected from years 2004–2006. The clinical examination included registration of visible plaque index, modified gingival index and, bleeding sites on probing.

g. to seeing $5 as opposed to 10 cents), but also reflects something about action. We started out defining an ‘urge’ as ‘how much someone wants something’. Based on the current paradigms

at least, the concept of urge could be refined to ‘how much someone wants something when action is required to get it’. In respect of the need for action, our findings are at selleckchem odds with those of (Kapogiannis et al., 2008), who identified an effect of reward on the motor cortex using paired-pulse TMS in a paradigm in which the participants did not make any response. However, with the task used in their study, they could not identify whether the effect was determined by the size of the reward or the probability of receiving it (in that Dabrafenib study the effect related to a strong reduction in uncertainty when observing whether a reward materialized over a specific time interval). We suspect that the changing reward probabilities drove the observed effect and, therefore, having an action was not critical in their paradigm. Here, in our money paradigm, the task was set up to measure the strength of the urge, manipulated solely by the size of the monetary reward ($5 or $0.1), while keeping the probability of seeing $5 or $0.1 on any trial exactly the same. In these experiments, the MEPs likely reflect multiple contributing factors,

including not only the urge (determined by the value of the stimulus), but also action preparation. Hence, we caution the reader that comparing MEPs (raw or normalized) across different experiments might be misleading. It is only the relative difference between MEPs observed for different levels of urge (within an experiment, when all other factors are controlled) that can be reliably interpreted. Even a comparison of MEPs between baseline and non-baseline trials within

the same experiment is difficult to interpret because the probability of seeing a baseline trial was lower than the probability of seeing a non-baseline trial in all three experiments and, therefore, differences in the probabilities could confound such a comparison. We used baseline trials only for normalizing the MEPs within each participant (to reduce variance between participants). Thus, we have shown that the strength of an urge can be indexed via ‘spill over’ into motor system excitability, at one time-point and not another, and only when a response is needed Selleck Regorafenib for satisfying the urge. Moreover, unlike prior studies, we have separated the preparation to make a response from response execution itself. Further, by recording motor excitability before the participant knew which response to prepare, we also show that the effect on motor excitability is not purely one of motor preparation but must also reflect a motivational component. Further, by manipulating the response-requirement in the money task, we show that the effect is also not purely related to general brain arousal but must also include an action-relevant component.

The number of visible viral lesions was

counted and the diameter and area of lesions were measured 6 days after inoculation. For each treatment, six plants were used and each experiment Daporinad was repeated three times. The production of superoxide anion radical (O2−) and peroxide (H2O2) in the leaves of the 8–10-leaf stage tobacco plants treated with Trichokonins or control solution were examined using the procedure of Fitzgerald et al. (2004). For detection of systemic responses, seedlings were cultured in MS-medium containing 100 nM Trichokonins or control solution for 4 days, after which the top leaf was harvested. For detection of local responses, Trichokonins (100 nM) or 2 μL control solution were placed on the adaxial surface PLX4032 order of scratched leaves. Leaves were harvested and analyzed immediately. The leaves treated with Trichokonins or control solution were vacuum-infiltrated with nitrotetrazolium blue chloride (NBT) or 3,3-diaminobenzidine (DAB), incubated overnight at 28 °C, fixed and cleared in alcoholic lactophenol solution and examined for the formation of precipitates. Microscopic analysis was performed using an Olympus Stereoscope SZX-9 (Olympus America Inc., Melville, NY)

at × 40 magnification. To test autofluorescence, 2 μL of 100 nM Trichokonins or 2 μL control solution were placed on the adaxial surface of scratched leaves. After 24 h incubation at 25±1 °C, the autofluorescence in the leaves was assessed (Fitzgerald et al., 2004). Microscopic analysis was performed using Olympus BX-51 fluorescent microscope (Olympus America Inc.) at × 200 Thiamet G magnification. The excitation wavelengths were 470–490 nm and the emission wavelengths were 510–550 nm. Each tobacco plant at the 8–10-leaf stage was sprayed with 1 mL of 100 nM Trichokonins or 1 mL control solution. After 0, 1, 2, 3, 4, 5 or 6 days, the leaves of tobacco plants were harvested, ground to a fine powder in liquid

nitrogen and stored at −80 °C until analysis. Phenylalanine Ammonia-Lyase (PAL, E.C.4.3.1.5) activity was determined as described by González-Aguilar et al. (2004). Peroxidases (POD, E.C.1.11.1.7) activity was determined as described by Rathmell & Sequeira (1974). Polyphenol oxidases (PPO, E.C. 1.14.18.1) activity was assayed using Flurkey’s method (Flurkey, 1985). For each treatment, three tobacco plants were used and each experiment was repeated three times. RT-PCR analysis was conducted to determine the expression of selected defense-related genes in Trichokonins-treated tobacco plants. Each tobacco plant at the 8–10-leaf stage was sprayed with 1 mL of 100 nM Trichokonins. Total RNA was extracted from treated tobacco leaves after 0, 1, 2, 4, 6, 9, 12, 24 or 48 h treatment. The quality of extracted RNA was tested by 1.0% agarose electrophoresis. The transcription levels of genes were detected by RT-PCR using a One Step RNA PCR Kit (TaKaRa, Japan). RT-PCR products were loaded on 1.