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In this paper, we propose a novel Electroencephalograph (EEG) emotion recognition method inspired by neuroscience with respect to the brain response to different emotions. The proposed method, denoted by R2G-STNN, consists of spatial and temporal neural network models with regional to global hierarchical feature learning process to learn discriminative spatial-temporal EEG features. To learn the spatial features, a bidirectional long short term memory (BiLSTM) network is adopted to capture the intrinsic spatial relationships of EEG electrodes within brain region and between brain regions, respectively. Considering that different brain regions play different roles in the EEG emotion recognition, a region-attention layer into the R2G-STNN model is also introduced to learn a set of weights to strengthen or weaken the contributions of brain regions. Based on the spatial feature sequences, BiLSTM is adopted to learn both regional and global spatial-temporal features and the features are fitted into a classifier layer for learning emotion-discriminative features, in which a domain discriminator working corporately with the classifier is used to decrease the domain shift between training and testing data. Finally, to evaluate the proposed method, we conduct both subject-dependent and subject-independent EEG emotion recognition experiments on SEED database, and the experimental results show that the proposed method achieves state-of-the-art performance.

In this paper, we propose a novel neural network model, called bi-hemisphere domain adversarial neural network (BiDANN) model, for electroencephalograph (EEG) emotion recognition. The BiDANN model is inspired by the neuroscience findings that the left and right hemispheres of human's brain are asymmetric to the emotional response. It contains a global and two local domain discriminators that work adversarially with a classifier to learn discriminative emotional features for each hemisphere. At the same time, it tries to reduce the possible domain differences in each hemisphere between the source and target domains so as to improve the generality of the recognition model. In addition, we also propose an improved version of BiDANN, denoted by BiDANN-S, for subject-independent EEG emotion recognition problem by lowering the influences of the personal information of subjects to the EEG emotion recognition. Extensive experiments on the SEED database are conducted to evaluate the performance of both BiDANN and BiDANN-S. The experimental results have shown that the proposed BiDANN and BiDANN models achieve state-of-the-art performance in the EEG emotion recognition.

Prime editing (PE) applications are limited by low editing efficiency. Here we show that designing prime binding sites with a melting temperature of 30 °C leads to optimal performance in rice and that using two prime editing guide (peg) RNAs in trans encoding the same edits substantially enhances PE efficiency. Together, these approaches boost PE efficiency from 2.9-fold to 17.4-fold. Optimal pegRNAs or pegRNA pairs can be designed with our web application, PlantPegDesigner. Improved guide RNAs enhance the efficiency of prime editing.

To investigate the longitudinal microstructural and microvascular changes in patients with myelin oligodendrocyte glycoprotein antibody-associated optic neuritis (MOG-ON) without new attacks.We included 20 eyes of 12 MOG-ON patients without new attacks during the follow-up and 24 eyes of 12 age- and sex-matched healthy controls.The BCVA, retinal vessels and structure were significantly lower in MOG-ON eyes than in healthy eyes(allOngoing deterioration was observed in RNFL thickness and parafoveal and peripapillary vessel density, but not GCIPL thinning, in MOG-ON eyes without a new attack of ON.

Pearson syndrome (PS), also known as Pearson marrow-pancreas syndrome, is a rare, multi-systemic disorder caused by large-scale deletion of mitochondrial DNA (mtDNA) ranging from 2.3 kb to 9 kb, with 4,977 bp in length as the most common variant. This paper reported a novel mtDNA deletion of 4,734 bp in size, spanning from nucleotide 11,220 to 15,953. The infant suffered from chronic hepatomegaly, liver dysfunction, anemia and lactic acidosis over 1 year. Evidences of any infections were negative. Bone marrow aspiration and whole exome sequencing covering nearly 20,000 nucleus genes were performed when aged 3.3 and 6 months, respectively, but no genetic cause was identified. However, at his age 13 months, multiplex ligation-dependent probe amplification assay of the mtDNA in the patient detected a large deletion of 4,734 bp in size spanning the mitochondrial genes MTND4, MTTH, MTTS2, MTTL2, MTND5, MTND6, MTTE, MTCYB and MTTT which were functionally crucial for the intact oxidative phosphorylation pathway and adenosine triphosphate production, and PS was thus definitely diagnosed. This large deletion was negative in his parents and elder brother. Oral ursodeoxycholic acid, fat-soluble vitamins and blood transfusions were administrated, his clinical and laboratory presentations remained stable so far, but the long-term prognosis needed to be followed up. These findings enriched the variant spectrum of mtDNA, and demonstrated the importance of considering mitochondrial disorder in patient with intractable anemia, liver dysfunction and lactic acidosis as well as the significance of appropriate choosing of relevant genetic tools in the etiology diagnosis of such patients.

The awnless trait is the favorite trait for wheat breeding in Qinghai, China, but the major gene underlying the trait is unknown. This study aimed to analyze a recombinant inbred line (RIL) population containing 112 lines by crossing common wheat varieties GY448 (awnless) and GY115 (awned) using genotyping-by-sequencing analysis. A total of 56.62 Gb of clean sequence data were generated. A major gene was identified for the awnless trait based on 101,275 single-nucleotide polymorphisms by quantitative trait locus (QTL)-seq analysis. The region from 685.50 to 709.77 Mb on chromosome 5AL contained 334 genes and 394 transcripts, including B1, the major gene controlling the awnless trait. A 25-bp indel located in the B1 promoter region has a diagnostic value for the awnless/awned genotypes in the RIL population. A total of 42 spring wheat cultivars in Qinghai, China, were analyzed. Further, 23 awnless cultivars had the alleles of the GY448 parent, while 19 awned cultivars had the alleles of the GY115 parent. B1 is crucial in the awnless trait in wheat cultivars in Qinghai, China. The findings of this study might guide the breeding of new wheat cultivars in this region.

To determine retinal vessel density in patients with myelin oligodendrocyte glycoprotein antibody-associated disease (MOGAD).Twenty-five patients with MOGAD and 20 healthy participants were enrolled. Patients with MOGAD were divided into myelin oligodendrocyte glycoprotein antibody (MOG-Ab)-positive eyes with a history of optic neuritis (ON; MOG-Ab-ON+ group) or without a history of ON (MOG-Ab-ON- group). Visual function, retinal vessel densities, and thickness were measured.The retinal nerve fiber layer, parafoveal ganglion cell and inner plexiform layers, and vessel densities in the peripapillary and parafoveal areas were significantly decreased in the MOG-Ab-ON+ eyes compared with healthy eyes and MOG-Ab-ON- eyes (all P0.05). An increasing number of ON episodes was associated with greater decreases in these variables (all P0.05). Visual field mean deviation was not significantly decreased in patients with a history of 1 or 2 episodes of ON, although the relative decreases in retinal nerve fiber layer thickness, parafoveal ganglion cell and inner plexiform layer thickness, peripapillary vessel density, and parafoveal vessel density reached 33.1%, 23.2%, 17.0%, and 11.5% (all P0.05), respectively, in eyes with 2 episodes of ON. The mean deviation was significantly correlated with peripapillary vessel density (P0.05) after adjustment for other variables. Best-corrected visual acuity was not significantly correlated with optical coherence tomography variables (all P0.05).MOG-Ab-associated ON was associated with significant decreases in retinal structure and vessel density, without significant deteriorations in visual function. The peripapillary vessel density might predict the visual outcomes in patients with MOG-Ab-associated ON.

Purpose To provide the clinical and genetic characteristics of a large cohort of Chinese patients with vitelliform macular dystrophies. Design Cross-sectional study. Methods One hundred and thirty-four unrelated Chinese patients diagnosed with Best vitelliform macular dystrophy (BVMD), autosomal recessive bestrophinopathy (ARB), or adult vitelliform macular dystrophy (AVMD) were enrolled. Detailed ophthalmic examinations and genetic testing on vitelliform macular dystrophy–related genes were performed. Genotype and phenotype association were analyzed among different diagnostic groups. Results In total, 87 BVMD, 30 AVMD, and 17 ARB patients were enrolled in this study. Genetic analysis identified 37 BEST1 mutations in 53 patients with BVMD and ARB. Of these, 5 variants (c.254A>C, c.291C>G, c.722C>G, c.848_850del, c.1740-2A>C) were novel. The variant c.898G>A was a hotspot mutation, which was identified in 13 patients with BVMD and 1 patient with ARB. There were significant differences of ocular biometric parameters among patients with homozygous or compound heterozygous mutations, heterozygous mutations, and those without mutations of BEST1. Homozygous or compound heterozygous patients had shortest axial length (AL), shallowest anterior chamber depth (ACD), and highest intraocular pressure (IOP); patients without mutations had longest AL, deepest ACD, and lowest IOP; and heterozygous patients were in between. Moreover, 7 patients harboring heterozygous mutations in BEST1 and 3 patients without BEST1 mutations showed similar clinical appearance to ARB in our cohort. Conclusions This is the largest sample size study of Chinese vitelliform macular dystrophy patients. Our results indicated that assessment of angle-closure risk is a necessary consideration for all types of BEST1-related vitelliform macular dystrophies. The study expanded both the clinical and genetic findings of 3 common types of vitelliform macular dystrophies in a Chinese population.

OBJECTIVES Acinar cell carcinoma of the pancreas is a rare tumor with limited data. We aim to evaluate the characteristics, treatments, and outcomes of pancreatic acinar cell carcinoma after 2005. METHODS We retrospectively reviewed patients with pancreatic acinar cell carcinoma treated in Peking University Cancer Hospital and Institute (2005-2018) and identified cases from Surveillance, Epidemiology, and End Results database (2005-2015). RESULTS A total of 306 cases in our institute (n = 11) and Surveillance, Epidemiology, and End Results database (n = 295) were identified. The median age was 67 years, and 73.5% were male. The 5-year survival was 36.8% for all patients (median, 27 months). About 37% underwent surgical resection. The 5-year survival was 65.6% for resected patients as compared with 16.9% for unresected ones (P < 0.0001). Among locoregional and metastatic diseases, surgery significantly prolonged survival as well (P = 0.0003). Stage IV patients who received chemotherapy had a better survival than those without it (median, 16 vs 3 months; P = 0.0019). Aging, stage IV, and no surgery were independent predictors of poor overall survival. CONCLUSIONS For pancreatic acinar cell carcinoma, surgery is a potentially curative treatment contributing to long-term survival and suggested even in advanced diseases. Chemotherapy improved survival for metastatic patients.

In barley, D hordein is one of the storage proteins in the grain, which is thought to have a negative effect on malting quality. In this study, the CRISPR/Cas9 genome editing technology was employed to edit the D hordein gene in the spring barley cultivar, ‘Golden Promise’. Three transgenic lines were obtained but only two lines were modified in the coding region of D hordein. Four new D hordein alleles, with five deletions, were discovered through sequencing analysis of the T1 generation. All alleles carried the Del1 and Del2 deletions, while the Mu2, Mu3 and Mu4 alleles also carried the Del3, Del4 and Del5 deletions, respectively. As the frequency of Mu1 far exceeded the frequency of the other alleles, it was inferred that the Mu2, Mu3 and Mu4 alleles were derived from Mu1. The Mu1 allele was created by genome editing, whilst the other alleles were considered to be somatic mutations. Transcriptome analysis showed that the transcript level of D hordein was lower in the mutated lines than in wild type Golden Promise. Protein SDS-PAGE confirmed this result. Wild type D hordein protein could not be detected in the grain of the edited lines. The creation of new D hordein alleles will provide a new germplasm resource for studying the function of D hordein and may allow the breeding of new cultivars with better malt quality.

Prime editors, which are CRISPR-Cas9 nickase (H840A)-reverse transcriptase fusions programmed with prime editing guide RNAs (pegRNAs), can edit bases in mammalian cells without donor DNA or double-strand breaks. We adapted prime editors for use in plants through codon, promoter, and editing-condition optimization. The resulting suite of plant prime editors enable point mutations, insertions and deletions in rice and wheat protoplasts. Regenerated prime-edited rice plants were obtained at frequencies of up to 21.8%.

AIM: To measure the depths of different regions of the anterior chamber (AC) in healthy Chinese adults, and to explore possible correlations with age or gender. METHODS: The AC was imaged by swept-source optical coherence tomography in healthy Chinese adults. The horizontal scan of the right eye was used to measure the anterior chamber depth (ACD) at 199 points. RESULTS: A total of 309 images from 309 subjects were analyzed. The ACD values at nearly all locations were negatively correlated with age (all P

Spotting off-targets from gene editing Unintended genomic modifications limit the potential therapeutic use of gene-editing tools. Available methods to find off-targets generally do not work in vivo or detect single-nucleotide changes. Three papers in this issue report new methods for monitoring gene-editing tools in vivo (see the Perspective by Kempton and Qi). Wienert et al. followed the recruitment of a DNA repair protein to DNA breaks induced by CRISPR-Cas9, enabling unbiased detection of off-target editing in cellular and animal models. Zuo et al. identified off-targets without the interference of natural genetic heterogeneity by injecting base editors into one blastomere of a two-cell mouse embryo and leaving the other genetically identical blastomere unedited. Jin et al. performed whole-genome sequencing on individual, genome-edited rice plants to identify unintended mutations. Cytosine, but not adenine, base editors induced numerous single-nucleotide variants in both mouse and rice. Science , this issue p. 286 , p. 289 , p. 292 ; see also p. 234

To explore human emotions, in this paper, we design and build a multi-modal physiological emotion database, which collects four modal physiological signals, i.e., electroencephalogram (EEG), galvanic skin response, respiration, and electrocardiogram (ECG). To alleviate the influence of culture dependent elicitation materials and evoke desired human emotions, we specifically collect an emotion elicitation material database selected from more than 1500 video clips. By the considerable amount of strict man-made labeling, we elaborately choose 28 videos as standardized elicitation samples, which are assessed by psychological methods. The physiological signals of participants were synchronously recorded when they watched these standardized video clips that described six discrete emotions and neutral emotion. With three types of classification protocols, different feature extraction methods and classifiers (support vector machine and k-NearestNeighbor) were used to recognize the physiological responses of different emotions, which presented the baseline results. Simultaneously, we present a novel attention-long short-term memory (A-LSTM), which strengthens the effectiveness of useful sequences to extract more discriminative features. In addition, correlations between the EEG signals and the participants' ratings are investigated. The database has been made publicly available to encourage other researchers to use it to evaluate their own emotion estimation methods.

To investigate the clinical features and efficacy of lens surgery in patients with lens subluxation misdiagnosed as primary angle-closure glaucoma.In total, 2054 inpatients with primary angle-closure glaucoma were consecutively recruited. Eighty-five of the patients were rediagnosed as lens subluxation following ultrasound biomicroscopy after hospitalization. Lens surgeries were performed in all patients, and 35 had a follow-up of at least 12 months. Risk factors of postoperative intraocular pressure control were identified with multivariate logistic regression analysis.Eighty-five inpatients (4.1%, 85/2054) with lens subluxation were misdiagnosed as primary angle-closure glaucoma, of which 71.8% (2.97% of all cases, 61 out of 2054 subjects) had ocular blunt trauma and 18.8% (0.78% of all cases, 16 out of 2054 subjects) had spontaneous dislocation. Lens surgery significantly decreased the intraocular pressure and improved best-corrected visual acuity, meanwhile increased the anterior chamber depth (all P 0.001). Postoperatively, the intraocular pressure was controlled with antiglaucomatous eye drops administration in 17 eyes, in which greater range of preoperative angle closure and zonular dialysis, and longer operation time delay were detected. Risk factors for poor postoperative IOP control were more quadrants of angle closure (P = 0.038) and operation time delay (P = 0.045).Lens subluxation was the major cause of the misdiagnosed cases as primary angle-closure glaucoma. Misdiagnosed patients could benefit from the lens surgery, while more angle-closure quadrants and operation time delay indicated worse postoperative intraocular pressure control.

BACKGROUND: Several researchers have used commercial microscope-integrated optical coherence tomography (OCT) systems in glaucoma surgery, including ab interno trabeculectomy and canaloplasty. However, the 840 nm wavelength light source of the OCT systems is not ideal for imaging the anterior chamber angle structures because of its limited penetration. We evaluated the potential value of a microscope-integrated swept-source OCT system with a 1,310 nm center-wavelength light in canaloplasty for glaucoma. METHODS: Sixteen porcine eyes were used to simulate canaloplasty. The critical surgical steps were monitored using a prototype microscope-integrated OCT system with a 1,310 nm light source and a high axial scan rate of 100 kHz. The images from swept-source OCT and three-dimensional images from the microscope were projected simultaneously onto a liquid crystal display three-dimensional monitor (LMD-4251TD, Sony, Japan). The changes in the collector vessel (aqueous drainage structure in the porcine eye, similar to Schlemm’s canal in humans) were measured using Image J software. Histological sections stained with hematoxylin and eosin were used to assess surgical efficacy. RESULTS: High-resolution real-time images of the anterior segment were acquired during canaloplasty using the microscope-integrated OCT system. With the real-time OCT images, the position of the collector vessel was identified and the scleral flap could be created at the ideal location. The expansion of the collector vessel after viscoelastic injection was also visualized in real time. Compared with baseline, there was a significant increase in the cross-sectional area (from 14,502.98±9,242.55 to 59,499.04±20,506.41 µm(2), P

Background The impact of right ventricular dysfunction(RVD) on the prognosis of acute respiratory distress syndrome(ARDS) patients is controversial. Objectives The objectives of this systematic review and meta-analysis was to investigate whether RVD or pulmonary vascular dysfunction are associated with increased mortality in patients with ARDS. Methods We searched Pubmed, Embase, Cochrane Library, Wanfang Data, CNKI, and the WHO Clinical Trial Registry for studies of RVD or pulmonary vascular dysfunction in patients with ARDS. Results The presence of RVD or pulmonary vascular dysfunction in patients with ARDS was associated with an increase in mortality (OR = 1.68, 95% CI = 1.21–2.32, P = 0.069, I2 = 40.8%). Subgroup analyses obtained similar results. Funnel plots and the Egger's test indicated no publication bias, and sensitivity analyses determined that the results were stable. Conclusion The prognosis of patients with ARDS and RVD or pulmonary vascular dysfunction is worse than that of ARDS patients without RVD or pulmonary vascular dysfunction.

It is reported that the symptoms of autism spectrum disorder (ASD) could be improved by effective early interventions, which arouses an urgent need for large-scale early identification of ASD. Until now, the screening of ASD has relied on the child psychiatrist to collect medical history and conduct behavioral observations with the help of psychological assessment tools. Such screening measures inevitably have some disadvantages, including strong subjectivity, relying on experts and low-efficiency. With the development of computer science, it is possible to realize a computer-aided screening for ASD and alleviate the disadvantages of manual evaluation. In this study, we propose a behavior-based automated screening method to identify high-risk ASD (HR-ASD) for babies aged 8–24 months. The still-face paradigm (SFP) was used to elicit baby’s spontaneous social behavior through a face-to-face interaction, in which a mother was required to maintain a normal interaction to amuse her baby for 2 minutes (a baseline episode) and then suddenly change to the no-reaction and no-expression status with 1 minute (a still-face episode). Here, multiple cues derived from baby’s social stress response behavior during the latter episode, including head-movements, facial expressions and vocal characteristics, were statistically analyzed between HR-ASD and typical developmental (TD) groups. An automated identification model of HR-ASD was constructed based on these multi-cue features and the support vector machine (SVM) classifier; moreover, its screening performance was satisfied, for all the accuracy, specificity and sensitivity exceeded 90% on the cases included in this study. The experimental results suggest its feasibility in the early screening of HR-ASD.

To investigate the efficacy and safety of nab-paclitaxel plus S-1 (nab-P/S) versus nab-paclitaxel plus gemcitabine (nab-P/G) as first-line chemotherapy in patients with advanced pancreatic ductal adenocarcinoma (PDAC). Treatment-naive patients with advanced PDAC were equally randomized to receive nab-P/S or nab-P/G. The primary endpoint was the objective response rate (ORR). The secondary endpoints were ORR of the primary lesion, disease control rate, progression-free survival (PFS), overall survival (OS) and safety. The trial was registered at https://clinicaltrials.gov as NCT03636308. A total of 110 patients were planned for enrollment, but the trial was prematurely closed because no better ORR was observed with nab-P/S among the first 40 patients assigned between 08/2018 and 06/2019. The ORR was numerically higher with nab-P/S versus nab-P/G (35.0% vs 25.0%, P = 0.49). The ORRs of the primary lesion for both arms were similar (30.0% and 25.0%, P = 0.72). Disease control rate was 70.0% in each arm. There was no significant difference in PFS and OS between the two arms (median PFS, 6.3 vs 5.7 months, P = 0.34; median OS, 10.2 vs 10.2 months, P = 0.92). Risks of hematological toxicity, liver injury and rash were significantly decreased in the nab-P/S arm. A biweekly combination of nab-P/S yielded comparable efficacy with nab-P/G but improved safety profile. It may be a promising and convenient alternative as first-line and neoadjuvant settings for advanced PDAC.

Purpose. To compare the efficacy of passive drainage (PD) and repeated fluid-air (FA) exchange for removing emulsified silicone oil (SO) droplets. Methods. Twenty rhegmatogenous retinal detachment (RRD) patients who underwent primary pars plana vitrectomy (PPV) with SO tamponade were randomly and evenly divided into two groups for PD and FA exchange. Once the bulk of SO was removed, the first 2 mL of the washout fluid was collected, and then, another 2 mL of the washout fluid was collected after PD or FA exchange. The size and number of SO droplets in the washout fluid were measured using a Coulter counter (Multisizer 3, Beckman Coulter, Indianapolis, IN, USA). The efficiencies of FA exchange and PD for removing emulsified SO were compared. Results. The number of SO droplets decreased significantly after FA exchange and PD. The decrease in the number of droplets was statistically significant for larger droplets (>4 µm) with PD and for all droplet sizes with FA exchange. The decrease in the number of SO droplets of μm was significantly greater with FA exchange than with PD. However, at the end of the procedure, the overall number and size distribution of SO droplets were similar for both procedures. Conclusions. PD and FA exchange reduced the number of residual emulsified SO droplets after SO tamponade efficiently. FA exchange may have some advantages over PD in removing small droplets.

Yuan Zong, Zhi Peng, Xicheng Wang, Ming Lu, Lin Shen, Jun Zhou Department of Gastrointestinal Oncology, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Peking University Cancer Hospital & Institute, Beijing 100142, People’s Republic of ChinaCorrespondence: Jun ZhouDepartment of Gastrointestinal Oncology, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Peking University Cancer Hospital & Institute, No. 52 of Fucheng Road, Haidian District, Beijing 100142, People’s Republic of ChinaTel/Fax +86 10 8819 6561Email zhoujun_dr555@163.comObjective: Nab-paclitaxel plus gemcitabine (nab-P/G) has been established as a standard first-line treatment in metastatic pancreatic ductal adenocarcinoma (PDAC). S-1, as an oral fluoropyrimidine derivative, demonstrated effective for PDAC. This study aimed to evaluate the efficacy and safety of first-line chemotherapy with nab-paclitaxel plus S-1 (nab-P/S) versus nab-P/G in patients with advanced PDAC.Methods: Patients with advanced PDAC receiving nab-P/S (n = 65) or nab-P/G (n = 45) as first-line chemotherapy between November 2013 and June 2019 were reviewed.Results: The objective response rate (ORR) and disease control rate were numerically higher with nab-P/S than with nab-P/G (38.5% vs 28.9%, P = 0.30, 73.8% vs 66.7%, P = 0.42, respectively). ORRs of the primary lesion were similar for both groups (30.8% and 22.2%, P = 0.32). The median progression-free survival and overall survival were comparable between the two groups (5.5 vs 5.7 months, P = 0.34, 10.2 vs 11.3 months, P = 0.74, respectively). Nab-P/S was associated with a numerically lower risk of adverse events, especially hematologic adverse events.Conclusion: Nab-P/S could be a convenient alternative with similar efficacy and a favorable safety profile compared with nab-P/G as first-line chemotherapy for advanced PDAC, as well as an option for neoadjuvant therapy.Keywords: advanced pancreatic ductal adenocarcinoma, nab-paclitaxel, S-1, gemcitabine, objective response rate

Background Jerusalem artichoke (Helianthus tuberosus) is a fructan-accumulating plant, and an industrial source of raw material for fructan production, but the crucial enzymes involved in fructan biosynthesis remain poorly understood in this plant. Results In this study, a fructan: fructan 1-fructosyl-transferase (1-FFT) gene, Ht1-FFT, was isolated from Jerusalem artichoke. The coding sequence of Ht1-FFT was 2025 bp in length, encoding 641 amino acids. Ht1-FFT had the type domain of the 1-FFT protein family, to which it belonged, according to phylogenetic tree analysis, which implied that Ht1-FFT had the function of catalyzing the formation and extension of beta-(2,1)-linked fructans. Overexpression of Ht1-FFT in the leaves of transgenic tobacco increased fructan concentration. Moreover, the soluble sugar and proline concentrations increased, and the malondialdehyde (MDA) concentration was reduced in the transgenic lines. The changes in these parameters were associated with increased stress tolerance exhibited by the transgenic tobacco plants. A PEG-simulated drought stress experiment confirmed that the transgenic lines exhibited increased PEG-simulated drought stress tolerance. Conclusions The 1-FFT gene from Helianthus tuberosus was a functional fructan: fructan 1-fructosyl-transferase and played a positive role in PEG-simulated drought stress tolerance. This transgene could be used to increase fructan concentration and PEG-simulated drought stress tolerance in plants by genetic transformation.

We describe here a CRISPR simultaneous and wide-editing induced by a single system (SWISS), in which RNA aptamers engineered in crRNA scaffold recruit their cognate binding proteins fused with cytidine deaminase and adenosine deaminase to Cas9 nickase target sites to generate multiplexed base editing. By using paired sgRNAs, SWISS can produce insertions/deletions in addition to base editing. Rice mutants are generated using the SWISS system with efficiencies of cytosine conversion of 25.5%, adenine conversion of 16.4%, indels of 52.7%, and simultaneous triple mutations of 7.3%. The SWISS system provides a powerful tool for multi-functional genome editing in plants.

Short insertions and deletions can be produced in plant genomes using CRISPR-Cas editors, but reliable production of larger deletions in specific target sites has proven difficult to achieve. We report the development of a series of APOBEC-Cas9 fusion-induced deletion systems (AFIDs) that combine Cas9 with human APOBEC3A (A3A), uracil DNA-glucosidase and apurinic or apyrimidinic site lyase. In rice and wheat, AFID-3 generated deletions from 5'-deaminated C bases to the Cas9-cleavage site. Approximately one-third of deletions produced using AFID-3 in rice and wheat protoplasts (30.2%) and regenerated plants (34.8%) were predictable. We show that eAFID-3, in which the A3A in AFID-3 is replaced with truncated APOBEC3B (A3Bctd), produced more uniform deletions from the preferred TC motif to the double-strand break. AFIDs could be applied to study regulatory regions and protein domains to improve crop plants.

Purpose. To compare the efficacy of passive drainage (PD) and repeated fluid-air (FA) exchange for removing emulsified silicone oil (SO) droplets. Methods. Twenty rhegmatogenous retinal detachment (RRD) patients who underwent primary pars plana vitrectomy (PPV) with SO tamponade were randomly and evenly divided into two groups for PD and FA exchange. Once the bulk of SO was removed, the first 2 mL of the washout fluid was collected, and then, another 2 mL of the washout fluid was collected after PD or FA exchange. The size and number of SO droplets in the washout fluid were measured using a Coulter counter (Multisizer 3, Beckman Coulter, Indianapolis, IN, USA). The efficiencies of FA exchange and PD for removing emulsified SO were compared. Results. The number of SO droplets decreased significantly after FA exchange and PD. The decrease in the number of droplets was statistically significant for larger droplets (>4 µm) with PD and for all droplet sizes with FA exchange. The decrease in the number of SO droplets of

Purpose. To investigate the characteristics of silicone oil (SO) emulsification after vitrectomy for rhegmatogenous retinal detachment and their possible correlations with clinical factors. Methods. The first 2 mL of washing out fluid after SO removal was collected, and used for the measurement of the size and number of SO droplets using a Multisizer® 3 Coulter counter (Beckman Coulter, USA). The correlations between SO droplets and clinical factors were analyzed. Results. A total of 38 patients (23 males, 15 females) who underwent primary PPV with SO injection for RRD and whose retina stayed attached for ≥3 months after SO removal were included in the study. The average number of oil droplets was 1.96 × 106 ± 3.95 × 106/mL (range 0.17 × 106 to 21.71 × 106/ml), and 80.8% (range 64.23%–99.07%) of the droplets were 1-2 μm in diameter. The total number of emulsified SO droplets was not correlated with any clinical factor (all P>0.05). When the emulsified SO droplets were divided into groups by their diameter, multiple linear regression revealed that age was negatively correlated with the numbers of 5–7-μm-diameter and 7–12-μm-diameter droplets (both P<0.05). Patients using antiglaucoma medications had more 5–7-μm-diameter and 7–12-μm-diameter droplets than those not using (all P<0.05). Conclusion. Using a Multisizer® Coulter counter, we successfully determined the number and size of SO droplets after emulsification. We found that the number of 5–12-μm-diameter droplets was higher in younger-age patients and was higher in patients using antiglaucoma eyedrops.

Base editors (BEs) have been used to create C-to-T substitutions in various organisms. However, editing with rat APOBEC1-based BE3 is limited to a 5-nt sequence editing window and is inefficient in GC contexts. Here, we show that a base editor fusion protein composed of Cas9 nickase and human APOBEC3A (A3A-PBE) converts cytidine to thymidine efficiently in wheat, rice and potato with a 17-nucleotide editing window at all examined sites, independent of sequence context.

Purpose To evaluate the potential value of microscope-integrated optical coherence tomography (MI-OCT) in anterior segment surgical maneuvers. Methods Twenty-four ophthalmology residents, who were randomly and evenly divided into two groups, performed four anterior segment surgical maneuvers (corneal tunnel, scleral tunnel, simple corneal suture, and corneal laceration repair) on porcine eyes with (group B) or without (group A) real-time MI-OCT feedback. All residents performed the maneuvers again without MI-OCT. Results Compared with group A, group B (with MI-OCT) showed better accuracy in the length/depth of the corneal tunnel and the length of the scleral tunnel. However, both groups showed similar performances in the depth of both the simple corneal suture and the corneal laceration suture. When both groups performed the maneuvers again without MI-OCT, group B still showed better results than group A for the length of both the corneal and scleral tunnels. Conclusions Primary results suggest that real-time MI-OCT images are valuable for some anterior segment surgical maneuvers and could be helpful in surgical training. Translational relevance MI-OCT systems can be valuable in improving accuracy and decision making during anterior segment surgery and will be useful in surgical training.

The basic helix-loop helix (bHLH) transcription factor has been inferred to play an important role in blue and purple grain traits in common wheat, but to date, its overexpression has not been reported. In this study, the bHLH transcription factor ThMYC4E, the candidate gene controlling the blue grain trait from Th. Ponticum, was transferred to the common wheat JW1. The positive transgenic lines displayed higher levels of purple anthocyanin pigments in their grains, leaves and glumes. Stripping the glumes (light treatment) caused white grains to become purple in transgenic lines. RNA-Seq and qRT-PCR analysis demonstrated that the transcript levels of structural genes associated with anthocyanin biosynthesis were higher in transgenic wheat than the wild-type (WT), which indicated that ThMYC4E activated anthocyanin biosynthesis in the transgenic lines. Correspondingly, the anthocyanin contents in grains, roots, stems, leaves and glumes of transgenic lines were higher than those in the WT. Metabolome analysis demonstrated that the anthocyanins were composed of cyanidin and delphinidin in the grains of the transgenic lines. Moreover, the transgenic lines showed higher antioxidant activity, in terms of scavenging DPPH radicals, in the ethanol extracts of their grains. The overexpression of ThMYC4E sheds light on the traits related to anthocyanin biosynthesis in common wheat and provide a new way to improve anthocyanin content.

Sodium taurocholate cotransporting polypeptide (NTCP) is encoded by the gene SLC10A1 and expressed in the basolateral membrane of the hepatocyte, functioning to uptake bile acids from plasma. Although SLC10A1 has been cloned and NTCP function studied intensively for years, clinical description of NTCP deficiency remains rather limited. This study reported the genotypic and phenotypic features of two neonatal patients with NTCP deficiency. They both presented with neonatal indirect hyperbilirubinemia and remarkable hypercholanemia, and harbored the SLC10A1 variants c.800C>T (p.S267F) and c.263T>C (p.I88T). On genetic analysis of the two family trios, the latter missense variant was detected in trans with the former, a reported loss-of-function variant. Having not been reported in any databases, the c.263T>C (p.I88T) variant demonstrated an allele frequency of 0.67% (1/150) in healthy controls. Moreover, this variant involved a relatively conservative amino acid, and was predicted to be pathogenic or deleterious by changing the conformation of the NTCP molecule. In conclusion, the novel variant c.263T>C (p.I88T) in this study enriched the SLC10A1 mutation spectrum; the clinical findings lent support to the primary role of NTCP in hepatic bile acid clearance, and suggested that NTCP deficiency might be a contributing factor for the development of neonatal indirect hyperbilirubinemia.

Background/Aims: Cannabinoids are vasoactive substances that act as key regulators of arterial tone in the blood vessels supplying peripheral tissues and the central nervous system. We therefore investigated the effect of cannabinoids on retinal capillaries and pericytes. Methods: The effects of cannabinoids on capillary diameters were determined using an ex vivo whole-mount rat retinal model. Western blotting, quantitative PCR, and immunohistochemistry were performed to explore the underlying mechanism. Results: Endogenous cannabinoid 2-arachidonoylglycerol and anandamide and exogenous cannabinoid (R-(+)-WIN55212-2) dilated the noradrenaline-precontracted capillaries in a concentration-dependent manner (1 µM to 0.1 mM). The extent of vasorelaxation was positively correlated with changes in pericyte width. The effects of R-(+)-WIN55212-2 on vasorelaxation and pericyte width were inhibited by a cannabinoid receptor type-1 (CB1) antagonist, AM251 or rimonabant (SR141716A), the nitric oxide synthase inhibitor l-NAME, and the guanylate cyclase inhibitor ODQ. They were also abolished by the removal of the endothelium, but not by the cannabinoid receptor-2 antagonist SR144528, the endothelial cannabinoid receptor antagonist O-1918, or the cyclooxygenase inhibitor indomethacin. Conclusion: The exogenous cannabinoid R-(+)-WIN55212-2 promotes the vasorelaxation of pericyte-containing rat retinal capillaries. This effect of R-(+)-WIN55212-2 is dependent on CB1 and the nitric oxide–cyclic guanosine monophosphate pathway, and requires an intact endothelium.

Rc1 (red coleoptile) encode a R2R3-MYB transcription factor TaMYB-A1 in common wheat, but the homologous genes have not been reported in Triticum urartu or Triticum monococcum, both species being sources of the ancient A genome. In this study, four alleles were discovered in 120 T. urartu accessions, and two alleles from nine T. monococcum accessions. The six alleles encoded five proteins because TuMYB-A1-F and TuMYB-A1-C encoded the same protein. All mutated sites in the amino acid sequence of these alleles were out of the R2R3 and transcript activator domains, and did not affect the conserved tryptophan (W) residues nor the “D/ELX2R/KX3LX6LX3R” motif of the MYB proteins. The alleles from T. monococcum were more similar in sequence to TaMYB-B1 and TaMYB-D1 than were those from T. urartu in the phylogenetic tree, while the alleles from T. urartu resembled TaMYB-A1 more closely than did the T. monococcum alleles. TuMYB-A1-C was the most widely distributed allele, and Turkey contained the greatest genetic variation compared with the five other countries in the geographical distribution analysis. Transient expression of TuMYB-A1-D or TuMYB-A1-F induced anthocyanin biosynthesis in white coleoptiles of wheat cultivar ‘Opata’ with the help of ZmR. RT-PCR analysis showed that the transcript level of Rc1 allele in white coleoptiles was clearly lower than in red coleoptiles, suggesting that Rc1 allele plays an important role in anthocyanin biosynthesis in coleoptiles of T. urartu. The allele isolation, phylogenetic analysis, geographical distribution and functional verification should be helpful in understanding the functions of Rc1 alleles and the molecular mechanism underlying the coleoptile color trait in Triticum.

Purple pericarp is an interesting and useful trait in Triticum aestivum, but the molecular mechanism behind this phenotype remains unclear. The allelic variation in the MYB transcriptors is associated with the phenotype of pigmented organs in many plants. In this study, a MYB transcription factor gene, TaMYB3, was isolated using homology-based cloning and a differentially expressed gene mining approach, to verify the function of the MYB transcriptor in the purple pericarp. The coding sequence of TaMYB3 in cultivar Gy115 was the same as that in cultivar Opata. TaMYB3 was localized to FL0.62–0.95 on chromosome 4BL. The TaMYB3 protein contains DNA-binding and transcription-activation domains, and clustered on a phylogenetic tree with the MYB proteins that regulates anthocyanin and proanthocyanin biosynthesis. TaMYB3 localized in the nuclei of Arabidopsis thaliana and wheat protoplasts after it was transiently expressed with PEG transformation. TaMYB3 induced anthocyanin synthesis in the pericarp cells of Opata in the dark in collaboration with the basic helix–loop–helix protein ZmR, which is also the function of ZmC1. However, TaMYB3 alone did not induce anthocyanin biosynthesis in the pericarp cells of the white grain wheat cultivar Opata in the light after bombardment, whereas the single protein ZmR did. Light increased the expression of TaMYB3 in the pericarp of Gy115 and Opata, but only induced anthocyanin biosynthesis in the grains of Gy115. Our results extend our understanding of the molecular mechanism of the purple pericarp trait in T. aestivum.

// Zhan-Hui Zhang 1, * , Wei-Xia Lin 2, * , Qi-Qi Zheng 2 , Li Guo 2 and Yuan-Zong Song 2 1 Clinical Medicine Research Institute, The First Affiliated Hospital, Jinan University, Guangzhou 510630, China 2 Department of Pediatrics, The First Affiliated Hospital, Jinan University, Guangzhou 510630, China * These authors have contributed equally to this study Correspondence to: Yuan-Zong Song, email: songyuanzong@hotmail.com, songyuanzong@vip.tom.com Keywords: citrin deficiency, intrahepatic cholestasis, SLC25A13 , mutation, Western blotting Received: December 06, 2016 Accepted: July 18, 2017 Published: August 03, 2017 ABSTRACT Neonatal Intrahepatic Cholestasis caused by Citrin Deficiency (NICCD) arises from biallelic SLC25A13 mutations, and SLC25A13 analysis provides reliable evidences for NICCD definite diagnosis. However, novel large insertions/deletions in this gene could not be detected just by conventional DNA analysis. This study aimed to explore definite diagnostic evidences for an infant highly-suspected to have NICCD. Prevalent mutation screening and Sanger sequencing of SLC25A1 3 gene just revealed a paternally-inherited mutation c.851_854del4. Nevertheless, neither citrin protein nor SLC25A13 transcripts of maternal origin could be detected on Western blotting and cDNA cloning analysis, respectively. On this basis, the hidden maternal mutation was precisely positioned using SNP analysis and semi-quantitative PCR, and finally identified as a novel large deletion c.-3251_c.15+18443del21709bp, which involved the SLC25A13 promoter region and the entire exon 1 where locates the translation initiation codon. Hence, NICCD was definitely diagnosed in the infant. To the best of our knowledge, the novel gross deletion, which silenced the transcriptional and translational expression of the affected SLC25A13 allele, is the hitherto largest deletion in SLC25A13 mutation spectrum. The Western blotting approach using mitochondrial protein extracted from expanded peripheral blood lymphocytes, of particular note, might be a new minimally-invasive and more-feasible molecular tool for NICCD diagnosis.

AIM: To evaluate the long-term outcome of viscocanalostomy combined with trabeculotomy and mitomycin C in the treatment of primary congenital glaucoma. METHODS: This is a retrospective study. Forty-two eyes of 26 patients with primary congenital glaucoma were enrolled. Intraocular pressure (IOP), corneal diameter (mm) and cup/disc (C/D) were measured before and after the surgery respectively. Follow-up period was 30mo. RESULTS: The mean preoperative IOP was 30.6±7.35 mm Hg. Of the 42 eyes, 2 eyes were required conversion to trabeculectomy for the absence of Schlemm’s canal. Of remained 40 eyes, 38 eyes (95%) achieved successful IOP control. The average postoperative IOP was 11.69±4.18 mm Hg at 12mo. The mean reduction was 18.91 mm Hg (P

Purpose The aims of this study were to evaluate how the corneal deformation parameters provided by the Corvis ST tonometer (CST) were influenced by pressure-lowering ocular surgery, and to determine the correlations of intraocular pressure (IOP) and axial length (AL) with CST corneal deformation parameters. Methods This prospective 1-month study enrolled 22 subjects (22 eyes) who underwent trabeculectomy combined with mitomycin C. The corneal deformation parameters were measured using the CST. IOP was measured before and after surgery by a Goldmann applanation tonometer and the CST. The central corneal thickness and AL were also recorded. The correlations of the corneal deformation parameters with central corneal thickness, AL, and IOP changes were determined by linear regression analysis. Results IOP decreased significantly after surgery. AL was significantly shorter at 1 week after surgery. There were significant changes in the CST parameters time 1, velocity 1, velocity 2, peak distance highest concavity, and deformation amplitude highest concavity at 1 week and 1 month after surgery and in time highest concavity at 1 week after surgery. The change in time 1 was significantly correlated with preoperative IOP and the IOP reductions, and was positively correlated with the decrease in AL at 1 week. The increase in velocity 1 was negatively correlated with preoperative IOP, and IOP reductions. The change in time highest concavity was negatively correlated with IOP before surgery and the decrease in IOP at 1 week. Conclusion CST is a good choice for measuring IOP, especially when aiming for normalization of IOP after glaucoma surgery.

Sodium taurocholate cotransporting polypeptide (NTCP), a carrier protein encoded by the gene SLC10A1, is expressed in the basolateral membrane of the hepatocyte to uptake bile acids from plasma. As a new inborn error of bile acid metabolism, NTCP deficiency remains far from being well understood in terms of the clinical and molecular features. Citrin deficiency is a well-known autosomal recessive disease arising from SLC25A13 mutations, and in neonates or infants, this condition presents as transient intrahepatic cholestasis which usually resolves before 1 year of age. All the three patients in this paper exhibited cholestatic jaundice and elevated total bile acids in their early infancy, which were attributed to citrin deficiency by SLC25A13 genetic analysis. In response to feeding with lactose-free and medium-chain triglycerides-enrich formula, their clinical and laboratory presentations disappeared gradually while the hypercholanemia persisted, even beyond 1 year of age. On subsequent SLC10A1 analysis, they were all homozygous for the well-known pathogenic variant c.800C > T (p.Ser267Phe), and NTCP deficiency was thus definitely diagnosed. The findings in this paper indicated that NTCP deficiency could be covered up by citrin deficiency during early infancy; however, in citrin-deficient patients with intractable hypercholanemia following resolved cholestatic jaundice, NTCP deficiency should be taken into consideration.

Sodium taurocholate cotransporting polypeptide (NTCP), a carrier protein encoded by solute carrier family 10 member 1 (SLC10A1), is expressed in the basolateral membrane of hepatocytes, where it is responsible for the uptake of bile acids from plasma into hepatocytes. The first patient with NTCP deficiency was described in 2015. A limited number of such patients have been reported in the literature and their genotypic and phenotypic features require further investigation. The current study investigated 4 patients with NTCP deficiency from two unrelated families. The patients were subjected to SLC10A1 genetic analysis and it was revealed that all patients were compound heterozygous for the c.800C>T (p.Ser267Phe) and c.595A>C (p.Ser199Arg) SLC10A1 variants. To the best of the authors' knowledge, the latter variant had not been previously reported. Further analysis in 50 healthy individuals did not identify carriers. The c.595A>C (p.Ser199Arg) variant exhibited co-segregation with hypercholanemia and exhibited a relatively conserved amino acid when compared with homologous peptides. Moreover, SWISS-MODEL prediction revealed that the mutation affected the conformation of the NTCP molecule. The 4 patients demonstrated varying degrees of hypercholanemia while a downward trend in the plasma levels of total bile acids (TBA) in 2 pediatric patients and occasionally normal TBA level in an adult case were observed. The results indicated an autosomal recessive trait for NTCP deficiency, supported the primary role of NTCP in the uptake of bile acids from plasma and suggested that hepatic uptake of bile acids may occur by means other than NTCP uptake. Moreover, the novel missense variant c.595A>C(p.Ser199Arg) enriched the SLC10A1 mutation spectrum and may serve as a new genetic marker for the molecular diagnosis and genetic counseling of NTCP deficiency.

Background Overexpression of MYB transcription factors can induce the expression of structural genes for anthocyanin biosynthesis and increase the anthocyanin content of plant tissues. However, it remains unclear whether MYB transcription factor overexpression effects the activation of other genes and the concomitant accumulation of chemical compounds. Results Overexpression of LrAN2 promoted anthocyanin accumulation in a variety of tissues in tobacco cultivar Samsun. Only 185 unigenes, from total of 160,965, were expressed differently in leaves and 241 chemical compounds exhibited differences in accumulation. Four anthocyanins, including apigeninidin chloride, cyanidin 3-O-malonylhexoside, pelargonidin 3-O-beta-D-glucoside, and cyanidin 3,5-O-diglucoside were detected only in transgenic lines, which could explain the purple leaf phenotype. Beside anthocyanins, the phenylpropanoids, polyphenols (catechins), flavonoids, flavones, and flavonols were also upregulated. Overexpression of LrAN2 activated the basic helix-loop-helix transcription factor AN1b, and the MYB transcription factor MYB3. Additionally, structural genes associated with the phenylpropanoid biosynthetic pathway were activated, which lead to the upregulated accumulation of phenylpropanoid, polyphenol (catechin), flavonoid, flavone, flavonol, and anthocyanin. The MYB transcription factor CPC, a negative regulator of anthocyanin biosynthesis, was also expressed at increased levels in transgenic lines, which implie that a negative regulation mechanism existed in the anthocyanin biosynthesis pathway. The relative contents of all 19 differently accumulated amino groups and derivatives were decreased in transgenic lines, which meant that the phenylalanine biosynthesis pathway used other amino acids as substrates. Interestingly, the expression of acetylalkylglycerol acetylhydrolase was suppressed in transgenic lines, which caused the accumulation of 19 lyso-phosphatidylcholine derivatives and a decrease in production of eight octodecane derivatives. Conclusions Overexpression of LrAN2 activates the pathway of anthocyanin synthesis and metabolism in tobacco. Four anthocyanins lead to the purple leaf phenotype The main pathways of flavonoid biosynthesis were up-regulated. This research provides more information about the function of MYB transcription factors in anthocyanin biosynthesis and the production of other chemical compounds. This work will help breeders to obtain new plant cultivars with high anthocyanin contents using biotechnology.

Purpose To develop a software package for automated classification of anterior chamber angle of the eye by using ultrasound biomicroscopy. Methods Ultrasound biomicroscopy images were collected, and the trabecular-iris angle was manually measured and classified into three categories: open angle, narrow angle, and angle closure. Inception v3 was used as the classifying convolutional neural network and the algorithm was trained. Results With a recall rate of 97% in the test set, the neural network's classification accuracy can reach 97.2% and the overall area under the curve was 0.988. The sensitivity and specificity were 98.04% and 99.09% for the open angle, 96.30% and 98.13% for the narrow angle, and 98.21% and 99.05% for the angle closure categories, respectively. Conclusions Preliminary results show that an automated classification of the anterior chamber angle achieved satisfying sensitivity and specificity and could be helpful in clinical practice. Translational relevance The present work suggests that the algorithm described here could be useful in the categorizing of anterior chamber angle and screening for subjects who are at high risk of angle closure.

Targeted saturation mutagenesis of crop genes could be applied to produce genetic variants with improved agronomic performance. However, tools for directed evolution of plant genes, such as error-prone PCR or DNA shuffling, are limited1. We engineered five saturated targeted endogenous mutagenesis editors (STEMEs) that can generate de novo mutations and facilitate directed evolution of plant genes. In rice protoplasts, STEME-1 edited cytosine and adenine at the same target site with C > T efficiency up to 61.61% and simultaneous C > T and A > G efficiency up to 15.10%. STEME-NG, which incorporates the nickase Cas9-NG protospacer-adjacent motif variant, was used with 20 individual single guide RNAs in rice protoplasts to produce near-saturated mutagenesis (73.21%) for a 56-amino-acid portion of the rice acetyl-coenzyme A carboxylase (OsACC). We also applied STEME-1 and STEME-NG for directed evolution of the OsACC gene in rice and obtained herbicide resistance mutations. This set of two STEMEs will accelerate trait development and should work in any plants amenable to CRISPR-based editing. Saturation mutagenesis using dual base editors improves the herbicide resistance of rice.

Intrahepatic cholestasis of pregnancy (ICP) is the most common pregnancy-related liver disorder. Although the etiology of ICP is not fully understood thus far, some genetic factors might contribute to the development of this condition. Sodium-taurocholate cotransporting polypeptide (NTCP), the protein encoded by the gene Solute Carrier Family 10, Member 1 (SLC10A1), is the primary transporter expressed in the basolateral membrane of the hepatocyte to uptake conjugated bile salts from the plasma. NTCP deficiency arises from biallelic SLC10A1 mutations which impair the NTCP function and cause intractably elevated levels of total bile acids (TBA) in the plasma (hypercholanemia). In this study, all the SLC10A1 exons and their flanking sequences were analyzed by Sanger sequencing to investigate the etiology for hypercholanemia in two male infants aged 2 and 20 months, respectively, from two unrelated families. As a result, both patients are homozygous for the reported pathogenic variant c.800C>T (p.Ser267Phe) that could impair the NTCP function to uptake bile acids, and the diagnosis of NTCP deficiency was thus made. Their mothers are also homozygotes of the same variant and both had been diagnosed to have ICP in the third trimester, with one of them undergoing cesarean section. The father of the first patient in this paper has the same SLC10A1 genotype c.800C>T/c.800C>T, also exhibiting slight hypercholanemia with a plasma TBA level of 21.5 μmol/L. In conclusion, we suggest that with hypercholanemia being a common laboratory change, NTCP deficiency may be a genetic factor leading to ICP and even cesarean section in clinical practice.

PURPOSE: To investigate the diagnostic values of intereye or intraeye asymmetry of retinal perfused vessel density and neural structure parameters for detection of glaucoma. METHODS: In total, 152 healthy subjects and 72 bilateral primary open-angle glaucoma (POAG) patients were enrolled. Total POAG group contains all glaucoma patients. Early to moderate POAG group contains patients whose binocular mean defect values were larger than −12 dB. The retinal perfused vessel densities were acquired using optic coherence tomography angiography. The neural structure parameters include RNFL, GCC thickness and its derivative indices like focal loss volume percentage (FLV%) and global loss volume percentage (GLV%). Intereye asymmetry equaled to the absolute difference of parameters between paired eyes. Intraeye asymmetry was defined as absolute difference between the inferior and superior hemisphere values from one random selected eye. The areas under the receiver operating characteristic curves (AUROCs) were calculated to evaluate diagnostic ability. RESULTS: From pairwise comparison analysis of ROC curves, the intereye asymmetric parameters with the largest diagnostic accuracy were FLV and GLV% (AUROC = 0.944), which were significantly superior to the intereye asymmetry of perfused vessel density in peripapillary area and parafoveal area (P

The red flesh trait gives red pitayas more healthful components and a higher price, while the genetic mechanism behind this trait is unknown. In this manuscript, transcriptome analysis was employed to discover the genetic differences between white and red flesh in pitayas. A total of 27.99 Gb clean data were obtained for four samples. Unigenes, 79,049 in number, were generated with an average length of 1333 bp, and 52,618 Unigenes were annotated. Compared with white flesh, the expression of 10,215 Unigenes was up-regulated, and 4853 Unigenes were down-regulated in red flesh. The metabolic pathways accounted for 64.6% of all differentially expressed Unigenes in KEGG pathways. The group with high betalain content in red flesh and all structural genes, related to betalain biosynthesis, had a higher expression in red flesh than white flesh. The expression of the key gene, tyrosine hydroxylase CYP76AD1, was up-regulated 245.08 times, while 4,5-DOPA dioxygenase DODA was up-regulated 6.46 times. Moreover, the special isomers CYP76AD1&alpha, and DODA&alpha, were only expressed in red flesh. The competitive anthocyanin biosynthesis pathway had a lower expression in red flesh. Two MYB transcription factors were of the same branch as BvMYB1, regulating betalain biosynthesis in beet, and those transcription factors had expression differences in two kinds of pitayas, which indicated that they should be candidate genes controlling betalain accumulation in red pitayas. This research would benefit from identifying the major gene controlling red flesh trait and breed new cultivars with the red flesh trait. Future research should aim to prove the role of each candidate gene in betalain biosynthesis in red pitayas.

Objectives: To evaluate the efficacy and safety of erlotinib for the treatment of advanced hepatocellular carcinoma (HCC). Methods: A systematic literature search was undertaken in June 2015. Phase II/III trials of erlotinib for the treatment of advanced HCC were included. A descriptive analysis was applied. The study was conducted in College of Medicine, Honghui Hospital, Xi’an Jiaotong University, Xi’an, China, between June 2015 and January 2016. Results: Ten trials, comprising 9 phase II and one phase III trial, were included in the systematic review. The tumor response rate was 0% in 4 of the phase II trials, less than 10% in 3 of the phase II trials and the phase III trial, and greater than 20% in 2 of the phase II trials. The disease control rate was 42.5-79.6% in most studies. Three studies reported a median progression-free survival (PFS) of 6.5-9.0 months, although PFS was less than 3.5 months in most studies. Most trials reported a median overall survival of 6.25-15.65 months. The most frequent grade 3/4 toxicities were fatigue (11.9%), diarrhea (10%), increased alanine and aspartate transaminases (7.3%), and rash/desquamation (6.9%). Conclusion: Erlotinib provides efficacious and well-tolerated treatment for advanced HCC. However, more detailed investigations of HCC pathogenesis and evaluation of sensitive patient subsets are needed to improve outcomes of patients with advanced HCC. Additional well-designed, randomized, controlled trials are needed to evaluate the efficacy and safety of erlotinib as monotherapy or combination with other drugs for advanced HCC. Saudi Med J 2016; Vol. 37 (11): 1184-1190 doi: 10.15537/smj.2016.11.16267 How to cite this article: Zhang J, Zong Y, Xu GZ, Xing K. Erlotinib for advanced hepatocellular carcinoma. A systematic review of phase II/III clinical trials. Saudi Med J . 2016 Nov;37(11):1184-1190. doi: 10.15537/smj. 2016 .11.16267.

Idiopathic eruptive macular pigmentation (IEMP) is a rare dermatological disorder with generally unclear etiology and pathogenesis. A 5½-year-old girl was referred to hospital with a 10 month history of brown skin rashes. In early infancy, citrin deficiency had been diagnosed with the SLC25A13 genotype c.851_854del4/c.998G > A, but all clinical and laboratory abnormalities recovered following the introduction of a lactose-free and medium-chain triglyceride-enriched formula. Physical examination at referral indicated symmetric, multiple and non-scaly brown macules on the neck, trunk, buttocks and proximal parts of the extremities. Histopathology indicated epidermal basal layer hyperpigmentation with an irregular distribution, along with a large number of melanophages in the upper dermis. The diagnosis of IEMP was thus made. Within 2 years of follow up, the rashes disappeared spontaneously and gradually. To our knowledge, this is the first description of IEMP in a patient with silent citrin deficiency.

In this letter, a novel cross-corpus speech emotion recognition (SER) method using domain-adaptive least-squares regression (DaLSR) model is proposed. In this method, an additional unlabeled data set from target speech corpus is used to serve as an auxiliary data set and combined with the labeled training data set from source speech corpus for jointly training the DaLSR model. In contrast to the traditional least-squares regression (LSR) method, the major novelty of DaLSR is that it is able to handle the mismatch problem between source and target speech corpora. Hence, the proposed DaLSR method is very suitable for coping with cross-corpus SER problem. For evaluating the performance of the proposed method in dealing with the cross-corpus SER problem, we conduct extensive experiments on three emotional speech corpora and compare the results with several state-of-the-art transfer learning methods that are widely used for cross-corpus SER problem. The experimental results show that the proposed method achieves better recognition accuracies than the state-of-the-art methods.

Precise genome modification using artificial nucleases is a powerful tool for in-depth understanding of gene functions and for creating new varieties. The CRISPR/Cas system, derived from an adaptive immunity system in bacteria and archaea, can introduce DNA double-strand breaks (DSBs) into pre-selected genomic loci and lead to loss of gene function due to error-prone non-homologous end joining (NHEJ). RNA-guided nucleases have been widely used in several eukaryotic organisms. In this article, we provide a detailed protocol for designing and constructing gRNA targets, detecting nuclease activity in transient protoplast assays, and identifying mutations in transgenic plants (including rice, wheat and maize). Targeted mutations in T0 plants can be generated in 14 to 18 weeks. © 2016 by John Wiley & Sons, Inc.

Background.Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) is a Mendelian disorder arising from biallelicSLC25A13mutations, andSLC25A13genetic analysis was indispensable for its definite diagnosis. However, conventionalSLC25A13analysis could not detect all mutations, especially obscure large insertions/deletions. This paper aimed to explore the obscureSLC25A13mutation in an NICCD infant.Methods.Genomic DNA was extracted to screen for 4 high-frequencySLC25A13mutations, and then all 18 exons and their flanking sequences were analyzed by Sanger sequencing. Subsequently, cDNA cloning, SNP analyses, and semiquantitative PCR were performed to identify the obscure mutation.Results.A maternally inherited mutation IVS16ins3kb was screened out, and then cDNA cloning unveiled paternally inherited alternative splicing variants (ASVs) featuring exon 5 skipping. Ultimately, a large deletion c.329-1687_c.468+3865del5692bp, which has never been described in any other references, was identified via intensive study on the genomic DNA around exon 5 ofSLC25A13gene.Conclusions.An NICCD patient was definitely diagnosed as a compound heterozygote of IVS16ins3kb and c.329-1687_c.468+3865del5692bp. The large deletion enriched theSLC25A13mutation spectrum, and its identification supported the concept that cDNA cloning analysis, along with other molecular tools such as semiquantitative PCR, could provide valuable clues, facilitating the identification of obscureSLC25A13deletions.