To evaluate quality of life (QoL) and psychological score in adults with allergic rhinitis (AR) and to assess the possible risk factors.A total of 110 adult patients with a define diagnosis of AR, who were referred to an outpatient clinic of allergy and immunology, were included in this study. The modified Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ) was applied for the evaluation of QoL in these patients. The psychological score was also assessed, using the Esprint-15 questionnaire.In a total of 110 patients  with the mean±SEM age of 31.92±1.03 years, nasal congestion was the most common symptom (89.1%). Fifty-one (51%) out of 110 AR patients were found to have olfactory dysfunction . The relationship between nasal congestion and psychological score was significant (P= 0.005). OD had a significant relationship with QoL, as well as the psychological score . Furthermore, psychological score had a significant relationship with reduction in QoL and sleep impairment . Statistical analysis showed that 54.5% of patients had a high psychological score, which was more common in females, although gender differences were not significant.Considering the increasing prevalence of AR and the significant relationship of OD and sleep impairment with QoL and psychological score, early diagnosis and treatment of AR may be important in improving QoL. Allergic rhinitis (AR), which is the most common form of noninfectious rhinitis, is caused by airborne allergens and is characterized by nasal mucosal inflammation, rhinorrhea, congestion, itching, sneezing, and daily changes in the sense of smell for more than 1 hour as a result of immunoglobuin E (IgE)-mediated nasal inflammation .Additionally, AR may cause significant morbidity, creating major economic burdens, increasing sedative consumption, and disrupting sleep patterns , 4. The Several conditions, such as sinonasal diseases, upper respiratory tract infection, trauma, and aging, may cause olfactory dysfunction (OD), which is more frequent in adults with AR, compared to healthy people . OD is aIn the current study, we aimed to evaluate QoL and psychological score in adult patients with AR and to assess the possible risk factors.In this cross sectional survey, adult patients, aged 18–60 years, with reversible symptoms of AR, who were referred to the outpatient clinic of allergy and immunology, affiliated to Immunology, Asthma, and Allergy Research Institute (IAARI) of Tehran University of Medical Sciences  were consecutively recruited. Considering a 95% confidential interval and precision of 3%, a total of 110 individuals were enrolled in this study. The study design was approved by the IAARI local ethics committee and research committee, and written informed consents were obtained from all the patients.In this study, patients with 1 or more AR symptoms, such as itchy nose, rhinorrhea, repeated sneezing, and nasal congestion, were included. The Allergic Rhinitis and its Impact on Asthma (ARIA) guidelines were applied for AR diagnosis . In thisThe demographic data were collected, and comorbidities were also recorded. Individuals with symptoms persisting for less than 4 days or less than 4 consecutive weeks were categorized in the intermittent AR group, while those with AR symptoms for more than 4 days per week or more than 4 consecutive weeks were classified in the persistent AR group. Finally, a total of 110 patients, who met the mentioned criteria, were included in this study.QoL was evaluated by Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ), developed by Juniper under her permission for the application of the English version . Four quA trained professional physician  completed the modified RQLQ in face-to-face interviews. The mean score of QoL was measured, based on the responses to the modified RQLQ for each individual. A mean score ≥ 3 was considered as high QoL impairment, while a mean score ≤ 3 was considered as low QoL impairment. Moreover, the integrated questions were related to the “management of rhinitis”, “irritability or bad-temperedness because of rhinitis”, “feeling unwell because of rhinitis”, and “general health status with respect to rhinitis”. A mean score of ≥ 3 was considered as a high psychological score, while a mean score of ≤ 3 was considered as a low psychological score.P-value < 0.05 was considered to be significant.SPSS for Windows 18.0  was used for data analysis. The results are presented as frequency and mean±standard deviation. Pearson’s Chi square test was used for the analysis of categorical data. P> 0.05).A total of 110 consecutive patients, including 68 (61.8%) women and 42 (38.2%) men with the mean±SEM age of 31.92±1.03 years , met the inclusion criteria. The clinical features of the patients are outlined in P> 0.05).Overall, 7 (6.4%) and 41 (37.3%) patients suffered from mild and severe intermittent AR, respectively. Mild and severe persistent AR were reported in 20 (18.2%) and 39 (35.5%) patients, respectively. However, no significant relationship was found between gender and AR grade. Assessment of comorbidities in this subgroup of patients revealed that the prevalence of sinusitis, asthma, conjunctivitis, reflux, dermatitis, and urticaria was 57.3% (n=63), 28.2% (n=31), 26.4% (n=29), 20% (n=22), 19.1% (n=21), and 18.2% (n=20), respectively. Additionally, the frequency of impaired taste, hyposmia, and anosmia was 10.9% (n=12), 34.5% (n=38), and 11.8% (n=13) among patients, respectively, and no significant relationship was found between gender and different comorbidities . However, no significant relationship was found between OD (hyposmia and anosmia) and gender . Although no significant relationship was detected between anosmia and nasal congestion (P= 0.69), the statistical analysis revealed a significant relationship between hyposmia and nasal congestion (P= 0.008). The relationship between impaired taste and QoL was also significant (P= 0.034). However, the statistical analysis showed no significant association between impaired taste and nasal congestion or impaired taste and gender .The relationship between QOL score and different variables is summarized in P= 0.72). Also, the results revealed a significant relationship between psychological score and QoL, as well as morning symptoms (P< 0.0001). The association of psychological score with nasal congestion (P= 0.005) and sleep impairment (P< 0.0001) was also significant.P= 0.005). In fact, the prevalence of OD was higher in the persistent AR group, compared to the intermittent AR group. The statistical analysis showed that AR class had a significant relationship with QoL (P< 0.0001). Similarly, patients with persistent AR were more prone to psychological problems (P= 0.001).Based on the findings, the relationship between AR class and OD was significant . In addition, the relationships between OD and QOL, OD and psychological score, and psychological score and QOL were found to be significant. Although the frequency of OD in healthy adult males was significantly higher than females , 16, no The frequency of OD seems to increase with the duration and severity of AR, especially when accompanied by rhinosinusitis . Rhee etThe results of another study revealed that OD occurs in patients with seasonal AR only at the beginning of the pollen season . HoweverPrevious studies have suggested an association between mood and allergic disorders. Additionally, immune-mediated clinical symptoms are amplified by psychological stress in individuals with allergic disorders. In a study by Bedolla-Barajas et al., over one-third of individuals with rhinitis had anxiety or depression , which iIn another study, the frequency of anxiety disorders and mood swings was higher in subjects with allergic diseases, compared to patients without allergies . AdditioOverall, the risk of depression is greater in women with a history of allergies . In a reInteraction of genetic and environmental factors, such as weather conditions and local/cultural aspects, is correlated with allergic sensitization . UnderstIn the current study, qualitative ratings were reported by the patients. As a result, quality of the data is limited considering the difficulty in distinguishing nasal patency and olfactory function. In addition, patients were not compared with healthy controls, which is one of the important limitations of the current study. It is important to carry out a population-based study in this subgroup of patients in order to determine the potential risk factors, such as the building age, type of flooring, presence of carpets or rugs, type of materials used in pillows and blankets, presence of furry pets or cockroaches, exposure to second-hand smoking and nonspecific irritants, presence of plants in the household, air-conditioning equipment, and external pollution. Moreover, actions should be taken to eliminate these risk factors and to increase QoL and general health.In the current study, the relationship between psychological score and nasal congestion, sleep impairment, morning symptoms, and QoL was well described in 110 adult patients with AR. The association of QoL with OD and impaired taste was also documented. These findings highlight the importance of controlling AR symptoms, as they may cause severe QoL and psychological disturbances.


This study aimed to correlate male age with embryo morphokinetic parameterson D3 considering the timing and the exact moment of embryo cleavage.Time-lapse imaging was used to produce an ideal cleavage curve for theembryos analyzed. The percentage of embryos under the curve was analyzed andcorrelated with male age.32.6% of the embryos from patients aged 28-33 years were under the curve;36.2% of the embryos from patients aged 34-39 years were under the curve;41.3% of the embryos from patients aged 40-45 years were under the curve;and 26.3% of the embryos fro patients aged 46-57 years were under thecurve.a statistically non-significant decrease was observed in the percentage ofembryos under the optimal cleavage curve on D3 in the group of men agedbetween 40 and 45 years. Further studies looking into embryos in theblastocyst stage (D5 or D6) are required. Male age has been associated with decreased semen quality .Time-lapse imaging has been used to monitor embryo development and help pick the bestembryo based on cleavage kinetics (Considering the timing and exact moment of embryo cleavage described by Two hundred and ninety-six embryos obtained from intracytoplasmic sperm injection(ICSI) procedures were included in the study.The embryos were analyzed using time-lapse imaging (10/10 min), and the exact time ofoccurrence of significant embryo development events was noted.®Eclipse TE 2000-S microscope at 250x magnification was used. Temperature wascontrolled in the central vinyl surface of the micro-handler table with aGreisinger® GMH 3230 surface thermometer (Germany) with validatedcalibration. After ICSI, the embryos were rinsed with the same culture medium inwhich they developed. Rinsing was carried out with at least three drops (~ 50mL)of pre-equilibrated medium. Then the oocytes were placed in micro-wells from thespecial time-lapse board and taken to an incubator.ICSI was performed on culture medium containing HEPES. A NikonThe same culture medium was used for all embryos included in this study -., 2013.Culture plates with nine or sixteen wells were prepared and pre-equilibrated inthe incubator. After pre-equilibration, all micro-bubbles were carefullyremoved.The images were captured with a microscope camera placed inside a "big box"incubator type. Photos were taken every 10 minutes for the composition of a timeline. The system used a green homogeneous LED light source.A software program was used to retrospectively analyze the images depicting theevents that occurred after ICSI, and identify the precise moments at whichpronuclei and cell walls disappeared and abnormalities arose.An ideal cleavage curve plotted with the aid of analysis software was considered.The chance of an individual aged 34-39 having an embryo under of thecurve was 1.17  times the chance of an individual aged 28-33years, but this difference was not statistically significant.The chance of an individual aged 40-45 having an embryo under of thecurve was 1.46  times the chance of an individual aged 28-33years, but this difference was not statistically significant.The chance of an individual aged 46-57 having an embryo inside of thecurve was 0.76  times the chance of an individual aged 28-33years, but this difference was not statistically significant.The chance of an individual aged 40-45 having an embryo under of thecurve was 1.24  times the chance of an individual aged 34-39years, but this difference was not statistically significant.The chance of an individual aged 46-57 having an embryo inside of thecurve was 0.63  times the chance of an individual aged 34-39years, but this difference was not statistically significant.The chance of an individual aged 46-57 having an embryo inside of thecurve was 0.51  times the chance of an individual aged 40-45years, but this difference was not statistically significant.


Oryza sativa) LNC. Spectral reflectance and biochemical composition were determined in rice leaves of different cultivars (Yongyou 4949 and Yangliangyou 6) throughout two growing seasons (2014–2015). Results demonstrated that HSL provided the best indicator for predicting rice LNC, yielding a coefficient of determination (R2) of 0.74 and a root mean square error of 2.80 mg/g with a support vector machine, similar to the performance of ASD (R2 = 0.73). Estimation of rice LNC could be significantly improved with the finer spectral resolution of HSL compared with MSL (R2 = 0.56).Fast and nondestructive assessment of leaf nitrogen concentration (LNC) is critical for crop growth diagnosis and nitrogen management guidance. In the last decade, multispectral LiDAR (MSL) systems have promoted developments in the earth and ecological sciences with the additional spectral information. With more wavelengths than MSL, the hyperspectral LiDAR (HSL) system provides greater possibilities for remote sensing crop physiological conditions. This study compared the performance of ASD FieldSpec Pro FR, MSL, and HSL for estimating rice ( Oryza sativa) is a daily necessity among people. The expansion of cities has resulted in a decrease in available land for paddies. In addition, immoderate application of fertilizers has given rise to serious environmental consequences, such as water eutrophication and soil hardeningRice  information, light detection and ranging (LiDAR) has undergone advanced developments in quantifying the 3D surface properties and processes in earth and ecological sciences9Recently, MSL/HSL systems have been applied to estimate leaf moisture contents under laboratory conditions1819With regard to pattern recognition algorithms, multiple linear regression methods such as partial least-squares regression2224The development of HSL offers a way to address the limitations of traditional optical remote sensing and LiDAR. However, the performance of HSL in quantifying LNC has yet to be directly compared with that of other remote sensing technologies. The current study compared the performance of an HSL system in discriminating rice LNC with that of the passive sensor ASD FieldSpec Pro FR  and an MSL system using SVM. The experiments were based on rice samples of different cultivars and growth states, grown in different places for two consecutive years (2014–2015). The objectives of this study were to (1) estimate rice LNC in different situations  with data collected by three sensors  and (2) compare their capability to predict rice LNC with SVM. This study focused mainly on the novel use of the LiDAR intensities of HSL and MSL to reflect foliar biochemistry.Located on the Jianghan Plain, Hubei Province is one of the largest provinces for rice plantations in China. The yield of Hubei Province ranked first in the nation’s yields in 2014Yongyou 4949 was grown in Junchuan County, Suizhou during the growing season of 2014. The crops were seeded on April 27, and transplanted on June 1. Six levels of urea fertilizer  were implemented. For every urea fertilizer level, three fields with identical cultivation conditions were used as replicates. The fields were designed as randomized blocks. Each plot was separated completely from the others by setting plastic films on the ridges of adjacent fields to avoid water leakage, thereby ensuring the precise application of N. The paddy rice samples were collected on July 15 and August 1, 2014.Yangliangyou 6 was grown in the experimental station of the Huazhong Agricultural University in Wuhan in the growing season of 2015. The crops were seeded on April 30, and transplanted on May 27. Four levels of urea fertilizer  were implemented. Replications and separation between fields were similar to the procedures used in 2014. The paddy rice samples were gathered on July 20, 22, 24, and 26, 2015.In each experimental field, at least six fully expanded second leaves from the top were selected randomly. The fresh leaf samples were sealed in plastic bags, kept in ice chests, and then transported to the laboratory for spectral measurementset al.The passive sensor ASD FieldSpec Pro FR  was used for spectrum measurements. The measurement was performed as described by Song, R2 for LNC is concentrated in the red-edge bands (700–760 nm) paired with the red-edge to near-infrared (NIR) bands (700–1100 nm), followed by blue to green bands (450–520 nm) paired with red-edge to NIR bands (740–1000 nm)32Previous rice LNC studies showed that the highest et al.One of the investigated active sensors was an MSL system developed by Wuhan University, operating at four wavelengths  covering VIS and NIR wavelengths. A detailed description of the system is provided in the study by Wei, et al.The HSL system employed in this study was developed by Wuhan University, and a detailed description can be found in the study by Du, All of the rice leaves were measured perpendicular at a distance of 4.2 m. Thus, the effects of incidence angle and distance were eliminated. The influence of certain factors, such as the dark current of the instrument, can be weakened by calculating the reflectance from the spectral measurements of a reference white panelFoliar reflectance in the VIS and NIR regions (including the red edge) has often been considered a good candidate for representing the biochemical or biophysical parameters of vegetationThe exact relationship between LNC and the reflectance spectra remains unclear. With this consideration, a SVM, capable of constructing both linear and nonlinear inversion, was employed in this study. Different from an artificial neural network, a SVM has excellent generalization performance with a strong theoretical foundation in statistical learning theoryWe consider a set of training points, The dual problem isAfter solving problem (2), the approximate function isα*−α is the result. Using MATLAB , where relevant inherent functions and the library LIBSVMThe model output R2), root mean square error (RMSE), and relative error (RE) were calculated as shown below to evaluate the performance of the estimation models:Altogether 220 rice samples were collected in 2014 and 2015. They were divided randomly into two datasets: 80% (176) as the training dataset and the remaining 20% (44) as the validation dataset for predicting LNC. The coefficient of determination  was similar to that of Eitel, et al.R2 =  0.71–0.76)..82 mg/g . Compareet al.W in t ha−1) estimates in winter wheat decreased when the actual W was large. The saturation effect described above with increasing LNC still exists even when using the complete range of reflectance instead of red light-dependent or NIR-based indicesThe scatter of the predicted and observed rice LNC compared with the 1:1 line  suggesteet al.R2 ranging from 0.45 to 0.69 for narrow bands). Chen, et al.R2 of 0.72 for corn and 0.44 for wheat. Erdle, et al.et al.1510) −log (1/R1680)]/[log (1/R1510) + log (1/R1680)]) correlated with the foliar N concentration of native shrub vegetation Spectral indices have been widely applied detect crop N status. For example, Eitel, Based on a systematic analysis of the quantitative relationships between LNC and reflectance characteristics, the capability of an HSL system with 32 channels to estimate rice LNC was evaluated through comparisons with the passive hyperspectral sensor ASD FieldSpec Pro FR (using the reflectance spectra from 400 to 1000 nm) and an MSL system with four bands. Through the regression results of SVM, rice LNC was best predicted by HSL. ASD provided comparable results with HSL in this study. Limited by the number of wavelengths, the MSL provided a moderate regression.HSL demonstrated its potential as a rapid and non-destructive tool for assessing rice LNC, which can facilitate real-time N management decisions. Additional studies should be carried out to test the monitoring relationships further using independent datasets, which can help to test the reliability under a range of conditions. In addition, the HSL system shows promise for other agronomic applications, such as examining other crop biochemical properties and soil parameters, which merits exploration in future research.How to cite this article: Sun, J. et al. Evaluation of hyperspectral LiDAR for monitoring rice leaf nitrogen by comparison with multispectral LiDAR and passive spectrometer. Sci. Rep.7, 40362; doi: 10.1038/srep40362 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


The advancement and refinement in prostate cancer detection and treatment modalities have contributed to a younger patient population undergoing radical prostatectomy (RP) . AlthougThe discovery of the neurovascular bundle sparing technique by Dr. Patrick Walsh enabled urologists to provide hope of regaining erectile function after radical prostatectomy . HoweverAfter understanding the mechanisms that promote ED after radical prostatectomy, multiple studies have been focused on evaluating ways to increase oxygenation to the cavernosal bodies, decrease tissue fibrosis and apoptosis, and consequentially improve erectile function. In theory, the role of penile rehabilitation is to maintain tissue oxygenation and prevent tissue fibrosis until the cavernosal nerves recover from neuropraxia with the return of spontaneous nocturnal tumescence. However, evidence from our daily clinical practice demonstrates that penile rehabilitation does not necessarily guarantee the return of unassisted spontaneous erections.By preventing the breakdown of cGMP, PDE-5 inhibitors may exert a protective effect on cavernosal smooth muscle after prostatectomy . HoweverIn 2008, Montorsi et al.  publishePavlovich et al.  pursued The REACTT study conducted by Montorsi et al.  aimed toAll these studies evaluated the use of PDE5is by relying on self-reported outcomes to determine efficacy of therapy which could lead to response bias. Kim et al.  conducteAlthough there is not enough evidence to create an algorithm for penile rehabilitation, the use of PDE5-inhibitors has been well-tolerated and no significant harm of rehabilitation has been demonstrated provided the patients understand the side-effects and costs. This has driven urologists to include penile rehabilitation programs in their practices . Most ha


NPR1, an evolutionary conserved gene, plays a central role in SAR, and PR-1 is widely used as a marker for effective SA signaling.Systematic acquired resistance (SAR) is an effective broad-spectrum defense mechanism that confers long-lasting protection against biotrophic pathogens trough defense related salicylic acid (SA) signaling. Gene(s) involved in SAR have been extensively studied in dicot plants; however, remains largely unresolved in monocot plants. NPR1 and PR-1 homologous genes, PhaNPR1 and PhaPR1, from an economically important orchid, Phalaenopsis aphrodite, and characterized their roles in SA signaling and Cymbidium mosaic virus (CymMV) resistance. A phylogenetic analysis of NPR1 homologs showed that these genes appear to have evolved before angiospermy. Similar to Arabidopsis NPR1, PhaNPR1 was only moderately induced upon SA treatment and CymMV infection. Although PhaPR1 shows only 36% identity with AtPR1, its promoter shared conserved elements with those of other PR-1 genes, and it was induced upon SA treatment and CymMV infection. After CymMV infection, silencing on PhaNPR1 also reduced PhaPR1 expression; however, CymMV accumulation was not affected.We identified PhaNPR1 is required for PhaPR1 induction, but plays little role in defense against CymMV.In conclusion, after virus infection, The online version of this article (doi:10.1186/1999-3110-54-31) contains supplementary material, which is available to authorized users. Orchidaceae is a widespread monocot family that undergoes rapid speciation. More than 20,000 species in 850 genera have been recorded, and it is believed to be the largest family of angiosperms. Due to adaptive radiation, orchids have evolved an array of strategies to successfully colonize diverse terrestrial ecosystems. Therefore, they provide rich resources to study evolution and the mechanisms of plant-environment interactions. Their diverse flower colors and shapes make them attractive in the floral industry. Orchids are economically important in countries worldwide; however, their production is harmed by pathogen attacks. Our knowledge concerning orchid defense is still limited.PR genes) are induced locally at the site of infection and systemically in distal plant tissues , jasmonic acid (JA), and ethylene , is induced by SA, and it has been suggested to be the lone PR-1 protein for induced resistance , which is a conserved central positive regulator of SA signaling , Theobroma cacao (TcNPR1), or Vitis vinifera (VvNPR1.1) were able to complement an npr1 mutation in Arabidopsis . The plants were maintained in an insect-proof controlled greenhouse with a 12 h photoperiod (200 μmole m-2s-1) at 25°C. Upon receipt, reverse transcription-polymerase chain reaction (RT-PCR) was conducted to ensure that the orchid plants used were free of virus. The primer pairs, ORSV-CP-F/ORSV-CP-R and CymMV-CP-F/CymMV-CP-R, targeting the coating proteins of two prevalent orchid viruses, ORSV and CymMV, were used  according to the manufacturer’s instructions. The primers used for this study are described in Additional file NPR1 conserved region to amplify partial PhaNPR1 cDNA using RNA isolated from P. aphrodite subsp. formosana treated with salicylic acid. The amplified fragment was cloned, and the complete sequences were determined. NPR1R was used as a gene-specific primer (GSP) for 1st RACE PCR and NPR1 5' NGSP was used as a 2nd GSP for nested PCR to obtain the 5′ end of PhaNPR. The 3′ end of PhaNPR1 was amplified using NPR1 F and NPR13′ NGSP as 1st and 2nd GSPs for 3′RACE to amplify the 3′ end of PhaNPR1. Both RACE amplified 5′ and 3′ ends of PhaNPR1 were cloned, and the complete sequences were determined. The primer pairs NPR1 ORFF/NPR1 ORFR were designed from the obtained 5′- and 3′- RACE amplified fragments and used in a PCR reaction to obtain the full-length open reading frame (ORF) of PhaNPR1. The full-length ORF of PhaNPR1 was cloned, and the complete sequence was determined. The cloning of PhaPR1 was performed using essentially the same method as that used to clone PhaNPR1, except the primer pair PR1F/PR1R were used in the PCR reaction to obtain partial PhaPR1 cDNA, PR1 5′ GSP and PR1 5′ NGSP primers were used as1st and 2nd GSPs in the 5′-RACE reaction, PR1 3′ GSP and PR1 3′ NGSP primers were used as 1st and 2nd GSPs in the 3′-RACE reaction, and the entire PR1 ORF was amplified using the primer pair PR1 ORFF/PR1 ORFR.The rapid amplification of cDNA ends (RACE) was performed to obtain 5′ and 3′-end cDNAs of the NPR1 and PR-1 homologs P. aphrodite was purified as previously described . The genomic DNA from P. aphrodite was digested with restriction enzymes, Dra I, EcoR V, Pvu II and Stu I, to generate blunt-end fragments, and subsequently the GenomeWalker Adaptors were ligated to the DNA fragments to generated four DNA libraries. The ligated products were used as templates, and the primer pair, Adaptor Primer 1/PR1GSP1  and manually rubbed into carborundum-dusted Phalaenopsis leaves with hands wearing latex gloves. The inoculated leaves were washed with excess distilled water.A total of 0.5 g of CymMV-infected plant tissue was ground in 300 μl of inoculation buffer  followed by an LR Gateway cloning reaction (Invitrogen) to transfer the fragments into pB7GWIWG2(I) to obtain pB7G-NPR1-1 and pB7G-NPR1- 2. The constructs were transformed into Agrobacterium tumefaciens LBA4404 by electroporation. The verified A. tumefaciens strains were cultured in 2 ml of YEB medium  containing 100 mg/L kanamycin and 100 μM acetosyringone and grown at 28°C for overnight. The next day, 1 ml of the bacterial culture was transferred into 10 ml of YEB medium containing 100 mg/L kanamycin and 100 μM acetosyringone and further incubated at 28°C until reaching an OD600 of 1.0~1.2. The A. tumefaciens cultures were centrifuged at 3,000 × g for 10 min, and the cells were resuspended in 1 ml of infiltration medium  and incubated at room temperature for 3 h. Each Phalaenopsis plant was infiltrated with 100 μl of A. tumefaciens suspension.The oligonucleotide pairs PhaNPR1-hpRNA-F1/PhaNPR1-hpRNA-R1 and PhaNPR1-hpRNA-F2/PhaNPR1-hpRNA-R2  method. The branch support was estimated using bootstrapping with 1,000 replicates. The sequences used in this study were obtained from NCBI GenBank and are listed in the supplementary information.Phylogenetic analysis of the NPR1 and PR1 homologs in Phalaenopsis aphrodite subsp. formosana, primers were designed based on the conserved domain sequences to amplify partial sequences of NPR1 and PR1. Subsequently, RACE-PCR was used to obtain full-length transcripts of both genes. The NPR1 homolog PhaNPR1 (GenBank accession no. JN630802) was identified, which comprises a full-length 1938 bp sequence encoding a 546 amino acid protein with a predicted molecular weight of 60.9 kDa and pI of 5.4. The protein shows 50% sequence identity with the Arabidopsis NPR1 (AT1G64280) and 61% identity with the rice NPR1 (Os01g0194300). The protein also shared conserved domains with other known NPR homologs: a BTB/POZ domain (amino acids 56 to 177), ankyrin repeat domains (amino acids 238 to 369), and a NPR1/NIM1 like defense protein C terminal motif (amino acids 353 to 546) , is 767 bp sequence encoding a 169 amino acid protein containing a 21 amino acid signal peptide, as predicted by SignalP, and a SCP domain, which is a conserved domain present in most PR1-like proteins. The predicted molecular weight and pI of the mature protein are 16.2 kDa and 6.95, respectively. The conserved features, which include four α-helices, four β-strands and six conserved cysteine residues, were present in PhaPR1 , rice PR-1a (Os07g0129200), and tobacco PR-1a (CAA31233) are 40%, 36%, 35%, and 35%, respectively. PhaPR1 is closely related to STS14-like proteins (~47% identity), which were first identified in the potato and highly expressed in the pistil , with 43% identity. Although detailed characterizations of these proteins have not been attempted, a search for microarray studies on these genes using Genevestigator revealed that AT5G66590 was induced upon infection of the fungal pathogen Alternaria brassicicola , and a zinc-finger motif (LS4). The LS10 and LS7 motifs are essential for SAR-mediated AtPR1 regulation , at position -440 to -430, a LS9 motif at position -1125 to -1117, and two LS4 motifs at positions -647 to -641 and -1011 to -1005 were identified; however, the LS7 motif was not located within the promoter region  , whose function has not been characterized, than to Arabidopsis PR1 (AT2G14610), which is the only SA inducible PR1-like protein in Arabidopsis. However, sequence similarity is not a reliable method for the prediction of protein function; therefore, we identified the promoter region of PhaPR1 and analyzed whether the PhaPR1 can be induced upon SA treatment and virus infection. The promoter region contains many conserved motifs, including an ASF1 motif , an LS10-like motif, and many W-box motifs , which are found in promoter of other PR-1 genes . The role of NPR1 in viral resistance is often contradicted among different plant species. NPR1-dependent pathway is required for proper N-mediated resistance to Tobacco mosaic virus (TMV) in tobacco  seems required SA but independent to NPR1 because resistance to TCV was not compromised in both npr1-1 and npr1-5 mutant backgrounds  was increased without alteration on virus concentration . Table S2. NPR1 homologs used for phylogenetic analysis. Table S3. Primers used in this study. (DOC 144 KB)Additional file 1: Table S1: Differences in the expression of Authors’ original file for figure 1Authors’ original file for figure 2Authors’ original file for figure 3Authors’ original file for figure 4Authors’ original file for figure 5Below are the links to the authors’ original submitted files for images.


Genotyping was performed using Taqman probe technique (rs5787 and rs5911) and gene sequencing technology (rs3842788). By using binary logistic regression analysis, the impact of clinical and genetic determinants on AR was evaluated. The prevalence of AR and aspirin semiresistance (ASR) was 3.86% and 20.76%, respectively, in Chinese chronic stable angina patients. rs5911 A/C and C/C versus A/A genotype , rs3842788 A/G versus G/G genotype , and blood stasis syndrome  were associated with AR, but rs5787 variants were all homozygous of G/G genotype. Plasma TXB2 and hs-CRP increased significantly in AR and ASR group, while 6-keto-PGF1α showed no difference, and TXB2 level was significantly higher in carriers of the rs3842788 A/G genotype. According to our results, rs5911 and rs3842788 are proved to be specific genetic markers of AR in Chinese chronic stable angina patients for the first time, and BSS was also proved to be a remarkable determinant for AR. The AR and ASR patients were with increased plasma TXB2 and hs-CRP levels, and the TXB2 level was influenced by the variation of rs3842788 genotype.The identification of single nucleotide polymorphisms (SNPs) related to aspirin resistance (AR) is of great significance for the explanation why some individuals demonstrate an incomplete response to aspirin and for optimizing the antiplatelet therapy strategy. The study was designed to investigate the possible associated genetic markers and clinical factors of AR for Chinese patients with chronic stable angina after PCI and to analyze the association between TXA2, PGI2, hs-CRP level, AR, and gene polymorphisms. Totally 207 chronic stable angina patients who received 100 mg maintenance dose daily of aspirin for more than 7 days were enrolled. The inhibition of platelets was assessed using light transmittance aggregometry. TXB2, 6-keto-PGF1 Despite disease . Accordihibitors . Kranzhohibitors . As comphibitors . Since gProstaglandin I2 (PGI2) counteracts the vasoconstrictor and platelet aggregation effects of thromboxane A2 (TXA2), which maintains an important balance in cardiovascular homeostasis . InactivStudies have shown that platelet activation, thrombosis, and restenosis after coronary stenting are closely related to blood stasis syndrome , 9. HoweChiCTR-OCC-12002415 . All subjects enrolled were chronic stable angina patients with coronary angiography and showed stenosis ≥50% in at least one main coronary artery or previous myocardial infarction [9/L, or >450 × 109/L, (3) hemoglobin < 90 g/L, (4) taking other antiplatelet, anticoagulant drugs or nonsteroidal antiinflammatory drugs within 30 days before test, (5) history of trauma or surgery within 2 weeks before enrollment, (6) severe primary diseases like renal insufficiency, liver dysfunction, hematopoietic system diseases, mental disorder, or malignant tumor, and (7) female in pregnancy or lactation period [This study was approved by the institutional Ethics Committee of China Academy of Chinese Medical Sciences, and all subjects gave written informed consent before participating. The trial is registered with farction , who wern period . Classifn period . For stan period , 20  and arachidonic acid  as inducers. AR is defined as ADP-induced platelet aggregation rate ≥70% and AA-induced platelet aggregation rate ≥ 20%, and aspirin semiresistance (ASR) is defined when one of the results was observed [Platelet aggregation rate was assayed using light transmittance aggregometry  with diphosphate adenosine  [The TXA2- and PGI2-stable metabolites  , as wellhttp://hapmap.ncbi.nlm.nih.gov/) and previous investigations [rs5787, rs3842788, and rs5911 were selected based on results from the International HapMap Project , and one-way analysis of variance (ANOVA) was carried out for the comparison of means. Categorical data were described by frequency tables, percentage, or constituent ratio and compared using Chi-square test. Chi-square test or Fisher's exact test was used for testing whether the possible deviation of genotype distribution was consistent with Hardy-Weinberg equilibrium. A binary logistic regression model was used to identify independently associated clinical and genetic factors which significantly determine early AR in chronic stable angina patients.All statistical analyses were performed using SPSS version 15.0, and P > 0.05) (P < 0.05), whereas there were more males in the ASS group . AR + ASR group also had more patients with hypertension  and longer duration of CAD  when compared to ASS group.Among 207 patients recruited in the present study, 8 (3.86%) patients were classified as AR, 43 (20.77%) as ASR, and the remaining 156 (75.36%) as aspirin sensitive (ASS) patients. AA-induced ASR occurred in 2.90% (6/207) patients and ADP-induced ASR in 17.87% (37/207) patients. The data of AR patients in combination with ASR patients were compared with that of ASS patients. There was no significance between the AR + ASR group and ASS group regarding differences in age, body mass index (BMI), underlying diseases , being current smoker, drinking history, statins medication history, and serum lipid levels ( > 0.05) . NotablyP < 0.01). Concerning rs3842788 gene polymorphism, 188 were homozygous with G/G genotype (90.8%) and 19 were heterozygous with A/G genotype (9.2%), but no patients with A/A genotype were observed in the present study. Patients with A/G genotype of rs3842788 were more susceptible to AR or ASR compared to G/G genotype . All patients enrolled in the study were homozygous of G/G genotype in regard to rs5787 gene polymorphism. The genotype frequency distributions of rs5911 and rs3842788 in AR + ASR and ASS groups were presented in Tables Among the 207 enrolled patients, 76 patients were A/A homozygous (36.71%), 98 were A/C heterozygous 47.34%), and 33 were C/C homozygous 15.94%) of rs5911 gene polymorphism, which was in accordance with Hardy-Weinberg equilibrium. More patients carrying at least one C allele of rs5911 gene polymorphism were observed in AR + ASR group compared to those in ASS group . For the rs3842788 polymorphism, the A/G genotype significantly increased odds for AR development compared to G/G homozygous carriers .To explore the impact of risk factors on ASS and AR/ASR patients, binary logistic regression analysis adjusted by age, gender, and BMI is shown in P < 0.001), which may contribute to a higher proportion of postmenopausal females in AR + ASR patients than that in ASS patients . Patients with BSS had significantly increased odds for AR , which indicated that BSS was an important independent risk factor for AR.Female gender was found with an increased risk compared to male gender by binary logistic regression , while no significant difference was present in plasma level of 6-keto-PGF1α . TXB2 level was significantly higher in carriers of the rs3842788 A/G genotype than in G/G carriers (P < 0.05). However, there was no difference in 6-keto-PGF1α and hs-CRP levels with different genotypes of rs3842788 (P > 0.05).Compared to ASS group, plasma TXB2 and hs-CRP levels increased significantly in AR + ASR group .Individualized medicine promises to take individual genome variations into account to optimize disease treatment . Since ars5911, with higher mutation rate in Chinese population than the other SNPs in GP IIb, was proved to be a specific marker of AR for the first time. The patients with mutant genotype of rs5911 carrying at least one C allele were proved to be more susceptible to AR or ASR. More chronic stable angina patients with A/G genotype in rs3842788 were AR or ASR when compared to G/G genotype. C allele carriers of rs5911 and A/G genotype carriers of rs3842788 were independently associated with 4.5-fold and 8.3-fold higher incidence of AR when analyzed by binary logistic regression. Since the genetic variations of rs5911 in GP IIb may not directly modulate aspirin pharmacokinetics and pharmacodynamics as the SNPs in COX-1, its underlining mechanisms need further study.Previous studies confirmed that BSS is closely related to platelet activation and thrombosis, which is the most common TCM syndrome in coronary heart disease , 9. In tα (the PGI2-stable metabolite), indicating TXB2 could be used as a specific marker to evaluate COX-1 inhibition and patients' response to low-dose aspirin. And plasma TXB2 generation was also proved to be related to the variation of rs3842788 genotype in COX-1, which in part explains the heterogeneity in the way patients respond to aspirin. Plasma hs-CRP level also increased significantly in AR and ASR group, which was in accordance with the results from Larsen et al. [TXB2, mainly derived from platelets COX-1 pathway, is a potent stimulus for platelet aggregation reflecting the inhibition of COX-1 and COX-2 enzymes by aspirin upon a maximal platelet activation during clotting, while PGI2, mainly derived from endothelial cells, serves as a potent inhibitor of platelet aggregation , 30. In n et al. , who repIt is proved for the first time that rs5911 and rs3842788 are independently associated with AR in Chinese chronic stable angina patients, with an increased risk as high as 4.5-fold and 8.3-fold, respectively. Blood stasis syndrome was also proved to be a remarkable determinant for AR, with as high as 10.2-fold incidence of AR compared to non-blood stasis syndrome. Significantly increased TXB2 and hs-CRP levels were demonstrated in AR and ASR patients and the TXB2 level was also proved to be related to the variation of rs3842788 genotype. Therefore, rs5911 and rs3842788 could be used as specific genetic markers of AR in Chinese population for optimizing the antiplatelet therapy strategy in clinic, and BSS could also be used as potential predictor for AR. More rigorous randomized controlled trials should be considered to further clarify the association between the polymorphisms and AR because of the relatively small study size in our present study.


It is rapidly absorbed and extensively metabolized. AR-C124910XX, the major active metabolite, antagonizes the P2Y12 receptor at approximately equal potency. The metabolism of ticagrelor to AR-C124910XX involves CYP3A4 and CYP3A5. CYP3A polymorphisms have been well documented, and ∗1GCYP3A4  and ∗3CYP3A5  are the most important single nucleotide polymorphisms in Chinese. Genetic differences in CYP3A4 and CYP3A5 expression in human volunteers and patients might affect the clearance of ticagrelor or AR-C124910XX in vivo resulting in subsequent variable patient response. Thus, this study is designed to explore the effects of ∗1GCYP3A4 and ∗3CYP3A5 polymorphisms on the pharmacokinetics and pharmcodynamics of ticagrelor in healthy Chinese subjects. The results indicated that the ∗1GCYP3A4 polymorphism significantly influenced the pharmacokinetics of AR-C124910XX, and it may be more important than ∗3CYP3A5 with respect to influencing ticagrelor pharmacokinetics by increasing CYP3A4 activity. However, the significant effect of ∗1GCYP3A4 polymorphism on AR-C124910XX plasma levels did not translate into detectable effect on inhibition of platelet aggregation. Therefore, it seems not necessary to adjust the dosage of ticagrelor according to the CYP3A4 or 3A5 genotype.Ticagrelor is the first reversible, direct-acting, potent P2Y The frequencies of ∗1GCYP3A4 and ∗3CYP3A5 alleles were 22.7–38.99%  or ∗3CYP3A5  variant or both were recruited from 136 male healthy Chinese volunteers. Each subject was physically healthy with no prior history of significant medical illness according to a medical history, physical examinations, vital signs, routine clinical laboratory tests  and ECGs. Subjects were required to have the following laboratory values within normal ranges: body mass index (BMI) of 19–24 kg⋅m-2, baseline maximal platelet aggregation (MPA) response to 5 μM ADP of ≥70%. Volunteers who had donated blood within the past 2 months, who had a history of conditions affecting drug disposition ; who had a personal or family history of bleeding disorders/events; who smoked ≥5 cigarettes per week ; who consumed grapefruit products 1 week before the study, or who had taken any prescription medication within 2 weeks of the start of the study were excluded.Fourteen healthy male Chinese volunteers who had the All subjects provided written informed consent before participation and the Ethics Committee of the First Affiliated Hospital of Soochow University approved the study protocol. Studies were conducted in accordance with the International Conference on Harmonisation Guideline for Good Clinical Practice and the Declaration of Helsinki.∗1GCYP3A4 or ∗3CYP3A5 alleles by direct sequencing. Briefly, the genomic DNA was amplified by use of two primers for each genotype. The sequences of PCR primers are shown in Table 1. PCR was performed by using Taq DNA polymerase (1.25 U/50 μL), 10× Taq Buffer, 25 mM MgCl2, dNTP and genomic DNA (50–100 ng/50 μL) with a pair of primers (0.3 μM) on T100TM Thermal Cycler. The PCR conditions for ∗1GCYP3A4 were 95°C for 5 min, followed by 35 cycles of denaturizing at 95°C for 30 s, annealing at 59°C for 30 s, and extension at 72°C for 30 s, and a final extension at 72°C for 10 min. The PCR conditions for ∗3CYP3A5 is almost the same as ∗1GCYP3A4 except for the annealing temperature that was 56°C. Then, the genotyping was carried out by direct sequencing on an ABI 3730 DNA Analyzer.Genomic DNA from EDTA-treated blood samples was extracted using a Wizard Genomic DNA Purification kit  according to the manufacturer’s instructions. Genomic DNA extracted from subjects’ blood was genotyped for AM on the subsequent morning, each subject received one dose  with 200 mL of water after an overnight fast. Blood samples for pharmacokinetics (4 mL each) were collected via an indwelling catheter or direct venipuncture into tubes containing sodium heparin immediately before and 0.5, 1, 1.5, 2, 3, 4, 5, 6, 8, 12, 16, 24, 36, and 48 h after drug administration. Blood samples were processed by centrifugation , and plasma was harvested and stored at -80°C until analysis. Pharmacodynamics samples were collected via direct venipuncture into tubes containing 0.109 mmol⋅L-1 sodium citrate before and 1, 2, 4, 12, 24, and 48 h after drug administration.All subjects were admitted to the clinical trial center the night before ticagrelor dosing. At 7:00 -1. The linear calibration ranges were 0.5–2,000 ng⋅mL-1 for ticagrelor and AR-C124910XX (r2 ≥ 0.99) and the lower limits of quantification for ticagrelor and AR-C124910XX were both 0.5 ng⋅mL-1. Intraday precision values [relative standard deviations (RSD)] ranged from 6.0 to 13.2% for ticagrelor and 4.2–11.5% for AR-C124910XX at three quality control levels. Interday precision values ranged from 8.7 to 11.4% for ticagrelor and 6.5 to 10.2% for AR-C124910XX.Plasma concentrations of ticagrelor and the metabolite, AR-C124910XX, were analyzed by validated UPLC-MS-MS methods as previously described . ProteinThe inhibition of ADP-induced platelet aggregation of platelet-rich plasma (PRP) was measured in response to 5 μM ADP as described elsewhere . The meat represents the IPA at time t, MPAt represents the MPA at time t, and MPAbaseline represents the MPA value at baseline. In essence, a higher IPA indicated greater antiplatelet effect.where IPAmax was estimated directly from observed plasma concentration–time data. AUC0-t was calculated using the linear trapezoidal rule, AUC0-∞ was calculated as AUC0-∞ = AUC0-48 + Ct/ke, where Ct was the last measured concentration and ke was calculated using linear regression analysis of the log-linear part of the plasma concentration–time curve. t1/2 was calculated as ln2/ke.Pharmacokinetics was analyzed using non-compartmental methods and WinNonlin Pro 5.2 . Concentration–time curves were generated for plasma ticagrelor and AR-C124910XX, and CThe pharmacodynamics effect (IPA) of ticagrelor was expressed as the percentage change from baseline platelet aggregation to 48 h after initial administration of the study drugs. The area under the time-effect curve (AUEC) for the IPA of ticagrelor was calculated from the time vs. IPA value curve, using the linear trapezoidal rule.post hoc test for multiple comparisons and an unpaired Student’s t-test for two groups as appropriate. Comparison of data not normally distributed was performed with a Mann–Whitney U test for comparison of two groups, while a Kruskal–Wallis H test was used for comparison of multiple groups. Two-sided P values ≤ 0.05 were considered statistically significant.Statistical analyses were performed using SPSS 16.0. Continuous variables were presented as means ± SD, and a Kolmogorov–Smirnov test was used to confirm normal distribution of continuous data. Data normally distributed were analyzed using a one-way analysis of variance (ANOVA) with the least significant difference (LSD) All participants given the medication evaluated here were observed closely to preserve their safety. All adverse events (AEs), such as symptoms and their severity, duration, time of onset and disappearance, and relationships to the study drug, were recorded. Physical examinations were performed and vital signs were evaluated, including systolic and diastolic blood pressures. Laboratory tests included urinalysis and hematologic, coagulation, and chemical analysis were performed.∗1GCYP3A4 and ∗3CYP3A5. All enrolled subjects completed the study with no major protocol violations. Subjects’ demographics and SNP characteristics appear in Table 2, and there were no significant differences in age, weight, height, or BMI among the six genotypes.Fourteen healthy Chinese male subjects were recruited from 136 individuals genotyped for CYP3A4 and CYP3A5 genotypes are presented in Figures 2. Pharmacokinetic comparisons for both genotypes are summarized in Tables 4. Because no significant differences exist between the CYP3A4: g.20230GA heterozygotes and CYP3A4: g.20230AA homozygotes, both were combined and compared to CYP3A4: g.20230GG homozygotes.Mean ± SD plasma concentration–time profiles for ticagrelor and AR-C124910XX for 0-last, AUC0-∞, t1/2, tmax, or Cmax between the two CYP3A4 genotypes (P > 0.05). No statistically significant differences were found in pharmacokinetics for ticagrelor among the three CYP3A5 genotypes.No statistically significant differences were observed in the AUC0-last, AUC0-∞, and Cmax of AR-C124910XX in ∗1GCYP3A4 carriers were 1.38 (P = 0.001), 1.43 (P = 0.001), and 1.42 (P = 0.003)-fold higher than in those with the CYP3A4: g.20230GG homozygotes, respectively. The t1/2 of AR-C124910XX in the ∗1GCYP3A4 group was also significantly longer than the CYP3A4: g.20230GG homozygotes (P = 0.015). The CYP3A4 genotype had no statistically significant effect on the tmax of AR-C124910XX. The CYP3A5 genotype had no statistically significant effect on pharmacokinetics parameters of AR-C124910XX.The mean AUCCYP3A4 and CYP3A5 genotypes are presented in Figures 4. Pharmacodynamics comparisons for both genotypes are summarized in Table 5. Strong inhibitory effects on platelet aggregation were observed. A single oral dose of ticagrelor (180 mg) induced a maximum inhibition of 75% at an average of 4 h after dosing. No statistically significant difference in IPA was observed for either CYP3A4 or CYP3A5 genotype.Pharmacodynamics properties of ticagrelor for Ticagrelor was well tolerated, and no study volunteers withdrew from the protocol, and no clinically significant bleeding event or thrombocytopenia was documented.in vivo resulting in subsequent variable patient response. However, influence of ∗1GCYP3A4 and ∗3CYP3A5 on ticagrelor pharmacokinetics and pharmacodynamics is still not quite clear. The present study was thus designed to extensively investigate the effects of ∗1GCYP3A4 and ∗3CYP3A5 genetic polymorphisms on the pharmacokinetics and pharmacodynamics of ticagrelor and AR-C124910XX in Chinese healthy subjects.Effective antiplatelet treatment is particularly important in patients with a high risk of cardiovascular events. As an orally active antiplatelet agent, ticagrelor has been demonstrated to exert a faster and more powerful IPA in comparison to clopidogrel in coronary artery disease patients. Although ticagrelor does not require metabolic activation for antiplatelet activity, it is still extensively metabolized to an active metabolite, AR-C124910XX . Signifi∗1GCYP3A4 allele had greater AR-C124910XX AUC0-last, AUC0-∞, and Cmax and longer t1/2 compared with those subjects with the ∗1/∗1CYP3A4 genotypes. Thus, the ∗1GCYP3A4 allele may enhance CYP3A4 expression and increase biotransformation of ticagrelor. This finding is consistent with previously published studies  polymorphism decreased CYP3A4 activity and modified fentanyl pharmacokinetics. So, patients with the ∗1G/∗1GCYP3A4 genotype required significantly less fentanyl for postoperative pain control compared to those with wild type or the ∗1/∗1GCYP3A4 genotype.Indeed, our results suggest that subjects with the  studies  which shs groups  found th∗1GCYP3A4 on plasma AR-C124910XX levels. Considering the fact that CYP3A4 and CYP3A5 convert ticagrelor into its active metabolite, AR-C124910XX, whose potency is almost the same as ticagrelor in antagonizing P2Y12 receptor, it became not so surprised. However, it’s important to note that a strong linkage disequilibrium exists between the ∗1GCYP3A4 and ∗1CYP3A5 polymorphisms, the presence of ∗1G might be only a marker of yet unstudied allele of greater influence of the variability on ticagrelor therapy. Therefore, to further examine the associations of the ∗3CYP3A5 and ∗1GCYP3A4 polymorphisms on CYP3A activity and to assess the combined influence of these two polymorphisms, a larger study should be on the way.However, statistically significant difference in IPA was not observed as well as the effects of SLCO1B1, CYP3A4, and UGT2B7) with plasma ticagrelor. Because the CYP3A4: is infrequent in the Chinese, we selected ∗1GCYP3A4 for the study, and our data showed that ∗1GCYP3A4 significantly affected AR-C124910XX pharmacokinetics but not the parent drug or platelet reactivity, which is in agreement with data published by Christophe’s group. Meanwhile, considering the fact that, there was no direct evidence whether SLCO1B1 or UGT2B7 participated in the metabolism of ticagrelor, and the effect of genetic polymorphisms of the two proteins on ticagrelor weren’t assessed in our study.∗1GCYP3A4 polymorphism significantly influenced the pharmacokinetics of AR-C124910XX, and it may be more important than ∗3CYP3A5 with respect to influencing ticagrelor pharmacokinetics by increasing CYP3A4 activity. Dosage adjustment according to the CYP3A4 or 3A5 genotype seems unnecessary considering their minor impact on platelet reactivity.In summary, the SL, LM, and ZS contributed to the study design and protocol. SL and XS conducted the study. SL, XZ, and XT performed the pharmacokinetic and pharmacodynamics measurements. All authors reviewed, interpreted the data and agreed on the content. LM and ZS were the lead authors, directed the manuscript content of each draft, supervising the medical writer. All authors approved the final version for submission.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


KLK3 gene which encodes the prostate specific antigen (PSA) and promoted cell growth of PC cells. The cell growth inhibition by CNPY2 knockdown was rescued by AR overexpression. Furthermore, positive correlation of expression levels between CNPY2 and AR/AR target genes was observed in tissue samples from human prostate cancer patients. Together, these results suggested that CNPY2 promoted cell growth of PC cells by inhibition of AR protein degradation through MYLIP-mediated AR ubiquitination.The androgen receptor (AR) is a ligand-dependent transcription factor that promotes prostate cancer (PC) cell growth through control of target gene expression. This report suggests that Canopy FGF signaling regulator 2 (CNPY2) controls AR protein levels in PC cells. We found that AR was ubiquitinated by an E3 ubiquitin ligase, myosin regulatory light chain interacting protein (MYLIP) and then degraded through the ubiquitin-proteasome pathway. CNPY2 decreased the ubiquitination activity of MYLIP by inhibition of interaction between MYLIP and UBE2D1, an E2 ubiquitin ligase. CNPY2 up-regulated gene expression of AR target genes such as  Signaling by the androgen-induced androgen receptor (AR) promotes cell growth of prostate cancer (PC) cells. AR is a ligand-dependent transcription factor. Androgen-bound ARs exerts both genomic action and non-genomic action in PC cells. In genomic pathway, AR transported into the cellular nucleus, bind to chromatin and control the expression of target genes . AlternaAR protein levels are controlled by both transcriptional regulation , 5 and pDrosophila PC model to identify novel regulators of PC cell growth and invasion . In experiments to detect auto-ubiquitination of MYLIP, 300 ng of GST-RING and 2 μg of GST-CNPY2 were used for the reaction. Reactions were stopped by addition of SDS-PAGE loading buffer and subjected to immunoblotting.ications , 15. 1.5Recombinant GST-RING (8 μg) plus Glutathione Sepharose beads were incubated with recombinant UBE2D-His (5 μg) or immunoprecipitated FLAG-CNPY2 (5 or 15 μg) at 4° C for 30 min in binding buffer , washed with binding buffer and subjected to immunoblotting.Human prostate tumor samples were obtained from University Hospital, Kyoto Prefectural University of Medicine, in accordance with protocols approved by the Hospital’s Institutional Ethical Committee. Written informed consent was obtained from each patient. Tumor samples were taken from freshly isolated surgical resections or needle biopsy.Prostate cancer patient samples were embedded in paraffin. Immunostaining and analyses were carried out as previously described , 40. Sert-test as appropriate. All data are reported as means ± SD. A P-value of < 0.05 was considered significant.Statistical analyses were carried out by


AR) gene underlie complete androgen insensitivity syndrome (CAIS), the most common cause of 46,XY sex reversal. Molecular genetic diagnosis of CAIS, however, remains uncertain in patients who show normal coding region of AR. Here, we describe a novel mechanism of AR disruption leading to CAIS in two 46,XY sisters. We analyzed whole-genome sequencing data of the patients for pathogenic variants outside the AR coding region. Patient fibroblasts from the genital area were used for AR cDNA analysis and protein quantification. Analysis of the cDNA revealed aberrant splicing of the mRNA caused by a deep intronic mutation (c.2450-118A>G) in the intron 6 of AR. The mutation creates a de novo 5′ splice site and a putative exonic splicing enhancer motif, which leads to the preferential formation of two aberrantly spliced mRNAs (predicted to include a premature stop codon). Patient fibroblasts contained no detectable AR protein. Our results show that patients with CAIS and normal AR coding region need to be examined for deep intronic mutations that can lead to pseudoexon activation.Mutations in the X-linked androgen receptor ( AR, the X-linked gene encoding the androgen receptor, cause the different forms of AIS, which total more than 500 mutations reported by 2012 in the Androgen Receptor Gene Mutations DatabaseAR mutationAR confirms the diagnosis of AIS, especially in the milder forms which have some phenotypic overlap with other disorders of sex developmentAndrogen insensitivity syndrome (AIS) is the most common known cause of 46,XY disorders of sex development, ranging from mild (MAIS) and partial (PAIS) to complete (CAIS) forms of androgen resistance. CAIS is characterized by a female phenotype in a genetically male  individual, whereas PAIS ranges from predominantly female to predominantly male phenotype, and MAIS patients have normal external male genitalia but they may suffer from infertility due to defective spermatogenesisAR has not even been identified in all patients with the complete form of the disorder raises the possibility of mutations in necessary AR cofactor(s) as the underlying cause in some casesAndrogen receptor mutations are mostly missense and enriched in the ligand-binding domain (LBD) of the proteinAR, detected in two siblings with CAIS with normal AR coding region and conserved splice sites, leads to aberrant splicing of the AR mRNA and insufficient AR protein production.Here, we demonstrate a new mechanism for CAIS. We show that a deep intronic pseudoexon-activating mutation in the intron between exons 6 and 7 of AR coding sequence from genomic DNA of the two siblings with CAIS revealed only one synonymous polymorphism . No rare potentially pathogenic variants in the coding region or in the conserved splice sites of the gene were found. PCR-amplification of the AR cDNA from the patients’ genital fibroblasts with primers situated on exon 5 and the 3′UTR, however, revealed abnormally long products (approximately 800 and 900 bp) in comparison to controls  for each DNA sample. Consistent with the result from AR cDNA PCR-amplification, the sequencing data revealed a point mutation in intron 6  that was not reported in variation databases [dbSNP (http://www.ncbi.nlm.nih.gov/projects/SNP/index.html) and 1000 Genomes]. This mutation was confirmed by Sanger-sequencing to be present as hemizygotic in both affected sisters and heterozygotic in the mother, whereas it was absent in the father and the healthy sister  in the y sister . No otheIn silico analysis with Human Splicing Finder suggested that the mutation leads to the formation of potential splice sites (ss), either a cryptic acceptor site  or a donor site . Additionally, the analysis suggested that the mutation may also create a new exonic splicing enhancer (ESE) motif specific for the SR-family protein SRSF1 (SF2/ASF) [mutant motif value 75.19 with ESE Finder algorithm ].in silico predictions and indicate that the mutation leads to the formation of a novel 5′ss (shorter product) and an ESE (longer product), which both subsequently promote the use of the upstream 3′ss shared between the cryptic transcripts.Sequencing of the two aberrantly spliced RT-PCR products  revealedAR mRNA   followed by a premature stop codon (PTC) after amino acid 816. Because of an intron downstream of the PTC, both aberrantly spliced mRNA isoforms are potential targets for the nonsense-mediated decay (NMD) pathway that would lead to the degradation of the AR mRNAression) . In accoaP cells .AR coding sequence or conserved splice sites. In our knowledge, although several splicing mutations of AR have already been reported in AIS patients, most are situated in the conserved splice sites close to the exon-intron junctions. Only one deep intronic mutation (in intron 6 located 44 bases from the exon-intron border) in AR has been previously reported in AISet al., who showed that a mutation in the AR 5′ untranslated region (UTR) created an upstream translation start site and subsequently led to the diminished expression of the full-length AR proteinAR coding region in AIS.In the current work, by combining whole-genome sequencing and cDNA analysis we were able to pinpoint the molecular genetic cause of CAIS in two 46,XY sisters with no mutations in the AR mRNA, creating two abnormally long products and a significantly reduced amount of the normal-sized mRNA. Computational analysis and sequencing of the 800 bp product suggested that the c.2450-118A>G mutation creates a de novo 5′ss facilitating the binding of U1 small nuclear ribonucleoprotein (snRNP) to this sequenceThe deep intronic mutation causes the aberrant splicing of AR mRNA by the NMD pathway, as is predicted to occur when a premature stop codon is located at least 50–55 nt upstream of the last exon-exon boundaryAR mRNA in the patient fibroblasts revealed only approximately 10% levels compared to the control fibroblasts, the overall abundance of AR mRNA was not clearly lower in the patient fibroblasts than in the controls. The destruction of the aberrant mRNA products by the NMD pathway cannot be ruled out, however, especially since the half-life of the PTC-containing mRNA products is not known; it is for example possible that some of the aberrant product is destroyed quickly, while the rest has a half-life similar to the normal AR mRNAThe inclusion of both the shorter and the longer intronic sequences is predicted to cause a premature stop codon before exon 7, which may result in degradation of the AR cDNA together with intronic sequences should be examined in CAIS patients with normal coding region and conserved splice sites of AR. The possibility of mutations in the 5′UTR or other regulatory regions should also be taken into consideration. Other deep intronic or regulatory region mutations, often missed in routine genetic screenings, may underlie CAIS in patients who still lack a molecular genetic diagnosis. The precise molecular genetic diagnosis is of crucial importance for genetic counseling of the patients and their family members. In addition, the molecular genetic diagnosis in DSD patients is valuable in predicting the clinical course of the disease, and in decision-making related to the possible need of gonadectomy.In conclusion, AR gene. At the age of 1.6 years, in an hCG test, Pregnyl 50 IU/kg i.m. was given on days 0, 3, and 6, and serum samples were collected before the injections and eight days after the last injection. The testosterone and the dihydrotestosterone (DHT) levels before stimulation were 1.1 nmol/l and 0.6 nmol/l, respectively, and SHBG concentration was 100 nmol/l. Serum estradiol concentration was under detection limit (0.025 nmol/l). After the stimulation, the testosterone level increased to 17 nmol/l, DHT to 1.1 nmol/l. SHBG level remained at 101 nmol/l and estradiol became measurable, 0.028 nmol/l. At the age of 10, she entered puberty with a growth spurt, breast development, and increased levels of LH (9.2 U/l), FSH (3.7 U/l), testosterone (30 nmol/l), and estradiol (0.078 nmol/l). Lanugo-type pubic hair appeared at the age of 11, and at 12 years of age she was at Tanner stage M4 P3, with scarce lanugo-type pubic hair. In annual check-ups gonads have not shown any signs of malignancy and serum hCG and alpha fetoprotein (AFP) levels have been low. Her current serum hormone levels at the age of 12.9 years are: LH 19 U/L, FSH 4.1 U/L, testosterone 29 nmol/L, estradiol 0.149 nmol/L, and inhibin B 426 ng/L, verifying functional testicular tissue.The two cases are the first (A) and the third (B) child from a non-consanguineous couple. The second child in the family is a 46,XX girl. The birth and the pregnancy of A were normal. Her birth weight was 3690 g, length 50 cm, head circumference 39.5 cm, Apgar score 9/10/10. Her growth started to follow +1.1−1.6 SDS curve. At the age of three months, she presented with an inguinal hernia on the right side. During the operation, 10 × 7 × 7 mm-size gonad was found in the hernia sac and was returned to the abdominal cavity. The vagina was short, only 0.7 cm . At that time, all girls with hernias were tested for the karyotypeChild B was karyotyped from the amniotic fluid and the 46,XY result was verified after the birth. Her phenotype was identical to A’s. No uterine structures were found in mri. The pregnancy and the birth were normal, size at birth 3860 g, 53 cm, head circumference 35.5 cm, and Apgar score 9/10/10. At 5 days age, the serum testosterone level was 1.2 nmol/l, DHT under 0.2 nmol/l, LH less than 0.1 U/l, FSH 0.24 U/l. At 2.5 months, testosterone level was surprisingly less than 0.5 nmol/l, but DHT level was 0.2 nmol/l; LH 0.15 U/l and FSH 0.76 U/l. The low testosterone level is consistent with a previous report of absent postnatal T surge in CAIS patientsThe parents and the children have given their informed consent and assent, respectively, to the study that was approved by the Ethics committee of Helsinki University Central Hospital. The study was carried out in accordance with the approved guidelines.AR (RefSeq ID NM_000044.3) were PCR-amplified from the genomic DNA of Child A and Child B and the PCR products were purified with ExoSAP-IT treatment , and bi-directionally sequenced by using the ABI BigDyeTerminator Cycle Sequencing Kit (v3.0) and ABI 3730xl 96-capillary DNA Analyzer automated sequencer . The sequences were aligned and read with Sequencher 4.9 software . All primer sequences and PCR conditions are available upon request.Genomic DNA was extracted from the peripheral blood leukocytes of the subjects. The entire coding region (exons 1–8) and the exon-intron junctions of AR variant found in the genome-sequencing data was confirmed by Sanger-sequencing from all family members.The whole-genome sequencing of the two patients and their father, who served as a healthy control, was performed in the Beijing Genomic Institute  by using Illumina’s HiSeq 2000 technology. The intronic Human control and patient fibroblasts (obtained from the labia majora) were grown in DMEM  supplemented with 1% (vol/vol) penicillin and streptomycin and 10% fetal calf serum . LNCaP prostatic cancer cells were used as a control for AR expression and androgen response and were grown in RPMI  supplemented with 1% (vol/vol) penicillin and streptomycin and 10% FCS. For immunoprecipitations and RNA extractions, the cells were split onto 6-well plates (250 000 cells/well) and allowed to grow for 24 h. Medium was changed to steroid-depleted medium (for fibroblasts: DMEM supplemented with 2.5% stripped serum and for LNCaP: RPMI supplemented with 10% stripped serum) for 6 h and a half of the wells were treated with vehicle (0.1% EtOH) and a half with 1 nM AR agonist methyltrienolone (R1881)  for the next 18 h.Immunoprecipitation was performed with Magna ChIP™ Protein A magnetic beads  from protein extract of 2 million cells. Beads needed for preclearing were equilibrated in lysis buffer  Triton X-100, 10% glycerol, 10 mM Na-phosphate, 50 mM NaF, 10 mM N-ethylmaleimide (NEM), 1x protease inhibitor cocktail (PIC) ) which contained 0.5% bovine serum albumin (BSA). Immunoprecipitating polyclonal antibody (rabbit α-AR)Cells were harvested (three wells in one sample) in PBS containing 10 mM NEM and suspended in lysis buffer. After 20 min incubation on ice, the samples were sonicated 10 sec and centrifuged 13000 × g for 20 min at +4 °C. Supernatants were transferred into clean tubes and precleared with equilibrated magnetic beads in rotation at +4 °C for 1 h. The beads and the supernatant were separated with a magnet, and an aliquot was taken into a separate tube as an input. The rest of the supernatants were immunoprecipitated with antibody-coupled beads in rotation at +4 °C overnight. The magnetic beads were washed three times with lysis buffer. Immunoprecipitated proteins were eluted from the beads by 1 × SDS sample buffer containing 10 mM NEM and 1 × PIC. The inputs and the immunoprecipitated samples were run on PAGE gels and transferred onto nitrocellulose membranes. The amount loaded into each lane corresponded to 40% of each immunoprecipitate. AR was recognized by mouse monoclonal α-AR antibody 441  and HRP-conjucated anti-mouse secondary antibody . Membranes were detected also by rabbit polyclonal α-GAPDH antibody  to determine the loading of the input samples.2O and 1 μg was used for cDNA synthesis in 20 μl volume by using Transcriptor First Strand cDNA Synthesis Kit (Roche Diagnostics). cDNA samples were diluted with sterile H2O into 200 μl. An aliquot (4 μl) was used in a qPCR reaction containing 0.4 μM each primer and 1 × LightCycler® 480 SYBR Green I Master (Roche) in 10 μl reaction volume by using the following PCR procedure: denaturation at 95 °C for 10 min, 40 cycles of 95 °C, 20 sec; 58 °C, 20 sec and 72 °C, 20 sec, and final extension 72 °C for 5 min. Primer sequences were for AR: forward 5′-TTGGAGACTGCCAGGGAC-3′ and reverse 5′-TCAGGGGCGAAGTAGAGC-3′, for FKBP5: forward 5′-AAAAGGCCAAGGAGCACAAC-3′ and reverse 5′-TTGAGGAGGGGCCGAGTTC-3′, for GAPDH: forward 5′-TGGGGAAGGTGAAGGTCGG-3′ and reverse 5′-TCTCAGCCTTGACGGTGCC-3′. Quantification of the normally-spliced AR mRNA was performed with primers targeting specifically the normal variant: forward 5′-CAGTGTGTCCGAATGAGGCA-3′ and reverse (located at the junction of exons 6 and 7) 5′-CCCATCCACTGGAATAATGCTGA-3′. The results were calculated for each gene of interest in relation to GAPDH expression. Further, the results of all samples were expressed in relation to control sample XX31B vehicle (0.1% ethanol) treatment  using the formula 2−(ΔΔCt), where ΔΔCt is ΔCt(sample)−ΔCt(XX31B vehicle), ΔCt is Ct(gene of interest)−Ct(GAPDH) and Ct is the cycle where the threshold value is crossed.Total RNA was extracted by using TriPure Isolation Reagent (Roche) according to the manufacturer’s instructions. RNA pellets were suspended in 30 μl of sterile HAR was PCR-amplified from the cDNA with cDNA-specific primers in overlapping fragments. The PCR products were visualized on a 1.5% agarose gel. In lanes with multiple-sized PCR products, the products were extracted from the gel with QIAquick gel extraction kit  for sequencing. All PCR products were sequenced as described above.The entire coding region of The effects of the identified intronic mutation were predicted with the web-based analysis tool Human Splicing Finder v.3.0How to cite this article: Känsäkoski, J. et al. Complete androgen insensitivity syndrome caused by a deep intronic pseudoexon-activating mutation in the androgen receptor gene. Sci. Rep.6, 32819; doi: 10.1038/srep32819 (2016).


Vestibular rehabilitation is a treatment option for the management of vertigo and unsteadiness, which are very common in head trauma patients and more challenging in the early months after trauma. This study evaluated the effectiveness of a vestibular rehabilitation program in the recovery of acute and sub-acute head trauma patients. The goal of this study was evaluation of the effect of early vestibular rehabilitation on patients with acute and sub-acute head trauma.This study was performed in 20 head trauma patients with vertigo and unsteadiness. The patients were randomly divided into two groups: one group received medical therapy (Betaserc) and the other received rehabilitation and medical therapy. An individualized vestibular rehabilitation program was designed that was then revised and verified by a joint committee of vestibular rehabilitation groups. The effectiveness of interventions was measured using the Dizziness Handicap Inventory (DHI) by comparing the results before and after therapy.The physical conditions and DHI scores of patients in both groups were similar at baseline. After 1 month of rehabilitation, patients receiving vestibular rehabilitation and medication showed greater progress than patients receiving medication only (P=0.000).Vestibular rehabilitation can aid in the recovery from vertigo and increase the stability of head trauma patients. Simultaneous treatment with medicine and vestibular rehabilitation exercises can result in quicker and better therapeutic effects. Head trauma can be caused by any incident such as falling, an accident at work or a quarrel. It is a common event, and the associated symptoms of vertigo and unsteadiness are prevalent in head trauma patients ,2. The vVestibular rehabilitation is a treatment option for the management of vertigo and unsteadiness that could result in the recovery or improvement of symptoms. In most cases, because of the patient’s physical and emotional condition, vestibular rehabilitation is initiated in the chronic stage of the disease, and it can be very challenging in the acute stage. Despite the large number of patients and the damaging consequences of vertigo and unsteadiness, only a few studies have assessed the effect of vestibular rehabilitation in acute and sub-acute head trauma patients (up to 6 months after head trauma). These few studies have suggested that early rehabilitation can obviate vertigo associated with head trauma and be beneficial in head trauma patients . The mosFor this reason, a questionnaire is commonly used for the assessment of patients before and after therapy.Vestibular rehabilitation is usually performed at the chronic stage of head trauma, although it would actually be helpful in the acute stage. Therefore, the objective of the present study was to evaluate the effect of early vestibular rehabilitation in patients with acute and sub-acute head trauma using DHI.ParticipantsThis study was performed from March 2013 to January 2014 in 20 adult patients (aged 18–60 years) referred to a vestibular rehabilitation center by ear-nose-throat or neurosurgery specialists. All patients had undergone complete neurologic and otologic evaluations and hospital discharge, and had normal computed tomography (CT) scans and an absence of physical abnormalities including benign paroxysmal positional vertigo, rupture of the round window membrane, perilymphatic fistula and endolymphatic hydrops. Patients showed no remarkable pathology .A hearing assessment revealed five patients with normal hearing, eight with bilateral and slight sensorineural hearing loss (SNHL) at high frequencies that could be related to their age, two with general bilateral and slight SNHL, three with bilateral moderate SNHL, one with bilateral profound SNHL and one with unilateral moderate SNHL. No patients with conductive or mixed hearing loss were enrolled into the study. The SNHL was not permitted to affect the vestibular tests including cervical vestibular evoked myogenic potential (cVEMP), but was required to show damage to the auditory system. The inclusion criteria were vertigo and unsteadiness due to head trauma that was caused by falling or an accident in the past 6-months; lack of cognitive, cervical, visual and neck problems in the neurologic and otologic evaluations; Glasgow coma score higher than 9 ; and abnPatients who met the study criteria were randomly classified into two groups. All patients were informed about the study in detail and their consent was obtained for participation in the study. This study was approved by the Ethics Committee of our local university (91d1303430).ProceduresVestibular assessment: Vestibular assessment of the patients was performed at the beginning of the program, including a case history, bedside examination, assessment of spontaneous nystagmus, horizontal and vertical gaze and saccade, Romberg’s test, Dix–Hallpike maneuver, cVEMP  and Dynamic Posturography . These tests were used to determine the site of the lesion of the vestibular disorder and to confirm inclusion criteria. The Persian version of the DHI was used to measure the physical, emotional and functional outcomes of head trauma and the progress during and after 4 weeks of vestibular rehabilitation .cVEMP: cVEMPs were recorded ipsilaterally from the sternocleidomastoid (SCM) muscle using acoustic stimulation. Tone bursts stimuli  were presented monaurally through headphones. The active, reference and ground electrodes were located over the middle of the SCM, upper sternum and forehead, respectively. The electrode impedance was maintained under 3 kΩ. Subjects turned their head to the contralateral shoulder before starting the measurement and held this position constantly to achieve a constant tonic activation of the SCM during the recording period. The tonic activation also was monitored.Dynamic posturography: This test was performed under the usual six conditions of the sensory organization test, and the results of positions 5 and 6 were used for detecting vestibular abnormalities as inclusion criteria.DHI: DHI is a valuable tool for assessing the patient’s recovery during the follow-up period. The DHI is a valid and reliable questionnaire that measures the effect of vertigo and unsteadiness on different physical , emotionInterventionAfter the vestibular assessment, participants were randomly divided into two groups. The first group received the usual medical therapy , and the second group received medical therapy and vestibular rehabilitation.The vestibular rehabilitation program was designed for daily training for a 4-week period and included mostly gaze stability and adaptation exercises, and some substitution exercises . These exercises were selected for each patient based on their function and vestibular assessment test results. Different gaze stability and adaptation exercises were used in all patients, although substitution exercises including standing and walking exercise were used only in patients with unsteadiness. The vestibular rehabilitation program was revised and verified by the joint committee of vestibular rehabilitation groups at our local university. These exercises were performed by therapists with specialized training in the assessment and treatment of vestibular disorders.Data analysisData were analyzed using SPSS version 19.0 computer software. A p-value below 0.05 was considered statistically significant. Descriptive analysis (mean and standard deviation) included demographic factors such as age, sex and duration of symptoms. The DHI was the only assessment that was repeated every week, and its scores were compared with repeated measures analysis of variance (ANOVA) at baseline and during and after vestibular rehabilitation in the two groups.Patients had a mean age of 44.2 (standard deviation [SD]: 12.6) years. Vestibular rehabilitation is a procedure that can be used for the treatment of vertigo and unsteadiness . Owing iIn the present study, we used an individualized vestibular rehabilitation program for head trauma patients with vertigo and unsteadiness. The results show that an early vestibular rehabilitation program within a time period of 1 month can decrease vertigo symptoms and increase stability and balance performance, thereby improving the patient’s functional and physical conditions.Our results show that early vestibular rehabilitation could resolve the patient’s problem, or at least decrease it. Similar studies have also reported the ability of vestibular rehabilitation to decrease the harmful effect of vertigo and unsteadiness ,5,16–19.Although clinical experience shows that medications can help some patients to resolve their symptoms, the control group in our study showed only a modest change during the 1-month follow-up. This could be due to the fact that this follow-up period is a short time for observing the therapeutic effects. Therefore, simultaneous treatment with medicine and vestibular rehabilitation exercises can have a quicker and better therapeutic effect, at least during the early treatment phase.Most of our study participants who underwent rehabilitation had high DHI scores; however, after completion of the program, most were symptom free or experiencing a mild level of disability. Decreasing the severity of disability leads to an independent life and increases the patient’s quality of life . AccordiVestibular rehabilitation can decrease the functional, emotional and physical consequences of vertigo and unsteadiness and improve the patients’ quality of life. In our study, the functional and physical subtests of DHI showed a higher change than the emotional subtest. It is possible that changing the emotional status requires more time and psychological support.The strength of the present study lies in using a control group to better describe the vestibular rehabilitation effect and begin the rehabilitation program in the short period after the head trauma event. It has been reported that a long period between the head trauma and onset of rehabilitation can decrease the effectiveness of the procedure , and usiWe used an individualized vestibular rehabilitation program that is based on the patient’s problem, site of lesion and the patient’s functional status. Different treatment strategies would be necessary for different types of abnormalities in vestibular mechanisms . IndividVestibular rehabilitation can aid in the recovery from vertigo and increase the stability of head trauma patients. Simultaneous treatment with medicine and vestibular rehabilitation exercises can result in quicker and better therapeutic effects.