Selective Hydrofunctionalization of Alkenyl Fluorides Enabled by Nickel-Catalyzed Hydrogen Atoms and Group Transfer: Reaction Development and Mechanistic Study.

Due to the unique effect of fluorine atoms, the efficient construction of high-value alkyl fluorides has attracted significant interest in modern drug development. However, enantioselective catalytic strategies for the efficient assembly of highly functionalized chiral C(sp3)-F scaffolds from simple starting materials have been underutilized. Herein, we demonstrate a nickel-catalyzed radical transfer strategy for the efficient, modular, asymmetric hydrogenation and hydroalkylation of alkenyl fluorides with primary, secondary, and tertiary alkyl halides under mild conditions. The transformation provides facile access to various structurally complex secondary and tertiary α-fluoro amide products from readily available starting materials with excellent substrate compatibility and distinct selectivity. Furthermore, the utility of this method is demonstrated by late-stage modifications and product derivatizations. Detailed mechanistic studies and DFT calculations have been conducted, showing that the rate-determining step for asymmetric hydrogenation reaction is NiH-HAT toward alkenyl fluorides and the stereo-determining step is alcohol coordination to Ni-enolates followed by a barrierless protonation. The mechanism for the asymmetric hydroalkylation reaction is also delivered in this investigation.

Investigating relationships among regulated learning, teaching presence and student engagement in blended learning: An experience sampling analysis.

This study explored the relationships among regulated learning, teaching presence and student engagement in blended learning. A two-level model was designed based on contextual factors (teaching presence) and individual factors (regulated learning), and experience sampling method was employed to collect intensive longitudinal data on 139 participants across three universities over 13 weeks in a blended course. Furthermore, multilevel regression analysis were conducted to examine the effects of teaching presence, self-regulated learning (SRL), co-regulated learning (CoRL) on intra- and interindividual variance in student engagement. The findings were as follows. 1) Perceived teacher support and instructional design fit had a significant positive effect on cognitive and emotional engagement and were crucial contextual factors that influenced intraindividual variance in learning engagement. 2) SRL and CoRL were copredictors of student engagement in blended learning. CoRL was more related to emotional engagement, while SRL was more related to cognitive engagement. 3) Modality had a significant effect on cognitive engagement but not on emotional engagement. 4) SRL and CoRL positively moderated the relationship between perceived teaching presence and cognitive engagement, while they negatively moderated the relationship between teacher support and emotional engagement, i.e., the relation between teacher support and emotional engagement was stronger in situations of low SRL or CoRL. Implications for teaching practice on blended learning were also discussed. Supplementary Information: The online version contains supplementary material available at 10.1007/s10639-023-11717-5.

Alternative deep learning method for fast spatial-frequency shift imaging microscopy.

Spatial-frequency shift (SFS) imaging microscopy can break the diffraction limit of fluorescently labeled and label-free samples by transferring the high spatial-frequency information into the passband of microscope. However, the resolution improvement is at the cost of decreasing temporal resolution since dozens of raw SFS images are needed to expand the frequency spectrum. Although some deep learning methods have been proposed to solve this problem, no neural network that is compatible to both labeled and label-free SFS imaging has been proposed. Here, we propose the joint spatial-Fourier channel attention network (JSFCAN), which learns the general connection between the spatial domain and Fourier frequency domain from complex samples. We demonstrate that JSFCAN can achieve a resolution similar to the traditional algorithm using nearly 1/4 raw images and increase the reconstruction speed by two orders of magnitude. Subsequently, we prove that JSFCAN can be applied to both fluorescently labeled and label-free samples without architecture changes. We also demonstrate that compared with the typical spatial domain optimization network U-net, JSFCAN is more robust to deal with deep-SFS images and noisy images. The proposed JSFCAN provides an alternative route for fast SFS imaging reconstruction, enabling future applications for real-time living cell research.

Fate of carotenoids in yeasts: synthesis and cleavage.

Carotenoids and their cleavage products (norisoprenoids) have excellent functional properties with diverse applications in foods, medicaments, cosmetics, etc. Carotenoids can be oxidatively cleaved through nonspecific reactions or by carotenoid cleavage oxygenases (CCOs), the product of which could further modify food flavor. This review provides comprehensive information on both carotenoid synthesis and cleavage processes with emphasis on enzyme characterization and biosynthetic pathway optimization. The use of interdisciplinary approaches of bioengineering and computer-aided experimental technology for key enzyme modification and systematic pathway design is beneficial to monitor metabolic pathways and assess pathway bottlenecks, which could efficiently lead to accumulation of carotenoids in microorganisms. The identification of CCOs spatial structures isolated from different species has made a significant contribution to the current state of knowledge. Current trends in carotenoid-related flavor modification are also discussed. In particular, we propose the carotenoid-synthesizing yeast Rhodotorula spp. for the production of food bioactive compounds. Understanding the behavior underlying the formation of norisoprenoids from carotenoids using interdisciplinary approaches may point toward other areas of investigation that could lead to better exploiting the potential use of autochthonous yeast in flavor enhancement.

The increased prevalence of depression and anxiety in T2DM patients associated with blood glucose fluctuation and sleep quality.

BACKGROUND: Current evidence demonstrates that blood glucose fluctuation can be associated with depression and anxiety. The association among blood glucose fluctuation, traditional risk factors and emotional disorders in T2DM should be studied and clarified. METHODS: A total of 182 diabetic patients including 81 patients with depression or anxiety and 101 patients without emotional disorder were enrolled into this study. Data were obtained through medical history and questionnaire survey. Data were analyzed using appropriate statistical methods. RESULTS: The comparison results of basic information between the two groups showed that the differences of the proportion of female were statistically significant (p = 0.002). There was no statistical difference in laboratory examination indexes between the two groups, however, standard deviation of blood glucose (SDBG) and postprandial glucose excursion (PPGE) of the comorbidity group were significantly higher than that of control group (p = 0.032 and p = 0.037). The results of questionnaire survey showed that there were statistically significant differences in sleep quality, PSQI and dietary habit between the two groups (p < 0.001, p < 0.001 and p < 0.001). Stratified analysis results according to gender showed that the percentage of cognitive disorder, anxiety and depression in female group was significantly higher than that in male group (p = 0.001, p < 0.001 and p < 0.001). Mini-mental state examination (MMSE), self-rating anxiety scale (SAS) and patient health questionnaire (PHQ-9) score in female group were also higher than male group (p = 0.001, p < 0.001 and p < 0.001). Logistic regression analysis results showed that SDBG and sleep quality were associated with emotional disorders in T2DM (p = 0.040 and p < 0.001) and the OR values of these factors were 7.588 (1.097-52.069) and 4.428 (2.649-7.401). CONCLUSIONS: Blood glucose fluctuation and sleep quality are associated with the increased prevalence of depression and anxiety disorders in T2DM.

Optimization of Cockpit Ventilation for Polar Cruise Ships in Combination with Windscreen Defogging and Cabin Comfort Considerations.

Polar cruise ships are exposed to extreme external conditions during voyages, resulting in cockpit windscreens that are prone to fogging and frosting, seriously affecting the driver's vision and even threatening navigation safety. However, the current research mainly focuses on cabin comfort, ignoring the coupling of defogging and comfort. Accordingly, this paper combines cockpit-windshield-defogging design and cockpit comfort considerations, and proposes 108 orthogonal-ventilation design parameters based on the four basic ventilation methods. The effects of different air supply parameters on comfort and anti-fog characteristics are investigated by using fluid dynamics simulation methods. The entropy weight-TOPSIS algorithm is employed to find the optimal ventilation parameters. The results show that the "Down-supply up-return type vertical jet" air supply method corresponding to an air supply velocity of 1 m/s, an air supply temperature of 297 K, and an air supply relative humidity of 30% has the smallest Euclidean distance di+ from the positive ideal solution, and the largest Euclidean distance di- from the negative ideal solution; thus, it obtains a higher ci and the highest priority. This air supply method provides the best thermal comfort for the drivers, as well as the best anti-fogging and defogging effect. The results can be useful to provide suggestions for the future design of the air-conditioning systems in polar cruise ships.

Genome-Wide Identification of the TIFY Gene Family in Brassiceae and Its Potential Association with Heavy Metal Stress in Rapeseed.

The TIFY gene family plays important roles in various plant biological processes and responses to stress and hormones. The chromosome-level genome of the Brassiceae species has been released, but knowledge concerning the TIFY family is lacking in the Brassiceae species. The current study performed a bioinformatics analysis on the TIFY family comparing three diploid (B. rapa, B. nigra, and B. oleracea) and two derived allotetraploid species (B. juncea, and B. napus). A total of 237 putative TIFY proteins were identified from five Brassiceae species, and classified into ten subfamilies (six JAZ types, one PPD type, two TIFY types, and one ZML type) based on their phylogenetic relationships with TIFY proteins in A. thaliana and Brassiceae species. Duplication and synteny analysis revealed that segmental and tandem duplications led to the expansion of the TIFY family genes during the process of polyploidization, and most of these TIFY family genes (TIFYs) were subjected to purifying selection after duplication based on Ka/Ks values. The spatial and temporal expression patterns indicated that different groups of BnaTIFYs have distinct spatiotemporal expression patterns under normal conditions and heavy metal stresses. Most of the JAZIII subfamily members were highest in all tissues, but JAZ subfamily members were strongly induced by heavy metal stresses. BnaTIFY34, BnaTIFY59, BnaTIFY21 and BnaTIFY68 were significantly upregulated mostly under As3+ and Cd2+ treatment, indicating that they could be actively induced by heavy metal stress. Our results may contribute to further exploration of TIFYs, and provided valuable information for further studies of TIFYs in plant tolerance to heavy metal stress.

Metabolite Characteristics Analysis of Siliques and Effects of Lights on the Accumulation of Glucosinolates in Siliques of Rapeseed.

Glucosinolates (GSLs) are naturally occurring secondary metabolites found in the Brassicaceae family, which mainly synthesize in the siliques with a wide range of functions. In this study, we investigated the effects of lights on metabolites in siliques of rapeseed through ultra high-performance liquid chromatography (UPLC)-heated electrospray ionization (HESI)-tandem mass spectrometry (MS/MS). A total of 249 metabolites, including 29 phenolic acids, 38 flavonoids, 22 GSLs, 93 uncalculated and 67 unknown compounds, were identified in siliques of rapeseed. Meanwhile, 62 metabolites showed significant differences after shading treatment, which were mainly GSLs and unknown compounds. Interestingly, the amounts of 10 GSLs had high accumulation levels in siliques, while the expression levels of their corresponding biosynthetic genes (AOP, GSL-OH, IGMT, and ST5a) were obviously reduced after shading treatment. Further evidence showed that the amounts of GSLs were significantly reduced in seeds, in accordance with the expression profiles of transporter genes (BnaGTRs). Our findings indicated that lights could affect the accumulation and transportation of GSLs from siliques to seeds in rapeseed. Therefore, this study facilitates a better understanding of metabolic characteristics of siliques and provides insight into the importance of light for GSLs accumulation and transportation in siliques and seeds of rapeseed.

High-Refractive-Index Chip with Periodically Fine-Tuning Gratings for Tunable Virtual-Wavevector Spatial Frequency Shift Universal Super-Resolution Imaging.

Continued research in fields such as materials science and biomedicine requires the development of a super-resolution imaging technique with a large field of view (FOV) and deep subwavelength resolution that is compatible with both fluorescent and nonfluorescent samples. Existing on-chip super-resolution methods exclusively focus on either fluorescent or nonfluorescent imaging, and, as such, there is an urgent requirement for a more general technique that is capable of both modes of imaging. In this study, to realize labeled and label-free super-resolution imaging on a single scalable photonic chip, a universal super-resolution imaging method based on the tunable virtual-wavevector spatial frequency shift (TVSFS) principle is introduced. Using this principle, imaging resolution can be improved more than threefold over the diffraction limit of a linear optical system. Here, diffractive units are fabricated on the chip's surface to provide wavevector-variable evanescent wave illumination, enabling tunable spatial frequency shifts in the Fourier space. A large FOV and resolutions of λ/4.7 and λ/7.1 were achieved for label-free and fluorescently labeled samples using a gallium phosphide (GaP) chip. With its large FOV, compatibility with different imaging modes, and monolithic integration, the proposed TVSFS chip may advance fields such as cell engineering, precision industry inspection, and chemical research.

Highly efficient genome editing in Xanthomonas oryzae pv. oryzae through repurposing the endogenous type I-C CRISPR-Cas system.

Efficient and modular genome editing technologies that manipulate the genome of bacterial pathogens will facilitate the study of pathogenesis mechanisms. However, such methods are yet to be established for Xanthomonas oryzae pv. oryzae (Xoo), the causal agent of rice bacterial blight. We identified a single type I-C CRISPR-Cas system in the Xoo genome and leveraged this endogenous defence system for high-efficiency genome editing in Xoo. Specifically, we developed plasmid components carrying a mini-CRISPR array, donor DNA, and a phage-derived recombination system to enable the efficient and programmable genome editing of precise deletions, insertions, base substitutions, and gene replacements. Furthermore, the type I-C CRISPR-Cas system of Xoo cleaves target DNA unidirectionally, and this can be harnessed to generate large genomic deletions up to 212 kb efficiently. Therefore, the genome-editing strategy we have developed can serve as an excellent tool for functional genomics of Xoo, and should also be applicable to other CRISPR-harbouring bacterial plant pathogens.

A toolkit for haptic force feedback in a telerobotic ultrasound system.

OBJECTIVE: To develop a collision engine (haptic force feedback simulator) compatible with a 5-degrees-of-freedom (DOF) haptic wand. This has broad applications such as telerobotic ultrasound systems. Integrating force feedback into systems is critical to optimize remote scanning. A collision engine compatible with a 5-DOF haptic wand was developed based on the Gilbert-Johnson-Keerthi algorithm. The collision engine calculated force during collision between the wand and a virtual object based on code developed using MATLAB. A proportional force was subsequently returned to a user via the haptic wand, thereby simulating the collision force for the user. Three experiments were conducted to assess the accuracy of the collision engine on curved and flat surfaces. RESULTS: The average errors in calculation of distances between the wand and virtual object were 2.1 cm, 3.4 cm, and 4.2 cm for the model of the human hand, cylinder, and cuboid, respectively. The collision engine accurately simulated forces on a flat surface, though was less accurate on curved surfaces. Future work will incorporate haptic force feedback into a telerobotic ultrasound system. The haptic force simulator presented here may also be used in the development of ultrasound simulators for training and education.

Development and assessment of a telesonography system for musculoskeletal imaging.

BACKGROUND: Telesonography systems have been developed to overcome barriers to accessing diagnostic ultrasound for patients in rural and remote communities. However, most previous telesonography systems have been designed for performing only abdominal and obstetrical exams. In this paper, we describe the development and assessment of a musculoskeletal (MSK) telesonography system. METHODS: We developed a 4-degrees-of-freedom (DOF) robot to manipulate an ultrasound probe. The robot was remotely controlled by a radiologist operating a joystick at the master site. The telesonography system was used to scan participants' forearms, and all participants were conventionally scanned for comparison. Participants and radiologists were surveyed regarding their experience. Images from both scanning methods were independently assessed by an MSK radiologist. RESULTS: All ten ultrasound exams were successfully performed using our developed MSK telesonography system, with no significant delay in movement. The duration (mean ± standard deviation) of telerobotic and conventional exams was 4.6 ± 0.9 and 1.4 ± 0.5 min, respectively (p = 0.039). An MSK radiologist rated quality of real-time ultrasound images transmitted over an internet connection as "very good" for all telesonography exams, and participants rated communication with the radiologist as "very good" or "good" for all exams. Visualisation of anatomic structures was similar between telerobotic and conventional methods, with no statistically significant differences. CONCLUSIONS: The MSK telesonography system developed in this study is feasible for performing soft tissue ultrasound exams. The advancement of this system may allow MSK ultrasound exams to be performed over long distances, increasing access to ultrasound for patients in rural and remote communities.

[Association of different stages of histological chorioamnionitis with respiratory distress syndrome in preterm infants with a gestational age of < 32 weeks].

OBJECTIVE: To study the association of different stages of histological chorioamnionitis (HCA) with the incidence rate and severity of respiratory distress syndrome (RDS) in preterm infants. METHODS: Related data were collected from the infants and their mothers who were treated in the Neonatal Intensive Care Unit of Qingdao Women and Children's Hospital, Qingdao University, from January 2018 to June 2020. According to the presence or absence of HCA and its stage, the infants were divided into four groups: control (n=109), early-stage HCA (n=126), middle-stage HCA (n=105), and late-stage HCA (n=36). The four groups were compared in terms of gestational age, birth weight, sex, maternal age, placental abruption, prenatal use of antibiotics, and incidence rate of RDS. The correlation between HCA stage and RDS severity was analyzed. RESULTS: Compared with the control and late-stage HCA groups, the early-stage HCA group had a significantly lower incidence rate of placental abruption and a significantly higher rate of prenatal use of antibiotics (P < 0.05), and the early-stage HCA group had a significantly lower incidence rate of RDS than the control group (P < 0.05). The multivariate logistic regression analysis showed that early-, middle-, and late-stage HCA were protective factors against RDS (P < 0.05). The Spearman test showed that the severity of RDS in preterm infants was not correlated with the HCA stage (P > 0.05). CONCLUSIONS: Early-, middle-, and late-stage HCA can reduce the incidence rate of RDS in preterm infants. HCA stage may not be correlated with RDS severity in preterm infants, which needs to be verified by further research.

LiCl induces apoptosis via CHOP/NOXA/Mcl-1 axis in human choroidal melanoma cells.

BACKGROUND: Choroidal melanoma is the most common primary intraocular malignancy that occurs in adults. Lithium Chloride Promotes Apoptosis in Human Leukemia NB4 Cells by Inhibiting Glycogen Synthase Kinase-3 Beta. In this study, we aimed to understand whether LiCl exerts anticancer effects on choroidal melanoma cells and elucidate the underlying molecular mechanisms. METHODS: Human choroidal melanoma cells were treated with LiCl, and cell survival was assessed with MTT assays. Cell reproductive viability was measured by plate colony formation assays. Cell apoptosis was evaluated using flow cytometry, and proteins were detected using western blotting. A human choroidal melanoma xenograft model was established to demonstrate the effect of LiCl on human choroidal melanoma in vivo. RESULTS: We found that LiCl inhibited cell survival and clonogenic potential and induced apoptosis in human choroidal melanoma cells. LiCl also reduced the proliferation of choroidal melanoma cells in vivo. Moreover, the upregulation of NOXA and downregulation of Mcl-1 were responsible for LiCl-induced apoptosis. Mcl-1 overexpression obviously impaired LiCl-induced apoptosis and cleavage of caspase8, caspase9, caspase3 and PARP. Moreover, the protein expression of endoplasmic reticulum stress markers, including IRE1α, Bip, p-eIF2α, ATF4 and CHOP, were upregulated following treatment with LiCl. When CHOP expression was knocked down and cells were treated with LiCl, the protein level of NOXA was partially increased, and Mcl-1 expression was increased, while the cleavage of caspase8, caspase9, caspase3 and PARP that was induced by the LiCl was reduced compared with the vehicle treated group. Prolonged ER stress results in the activation of the apoptotic pathway. CONCLUSIONS: In summary, LiCl induced an endoplasmic reticulum stress response while activating intrinsic apoptosis. Furthermore, the CHOP/NOXA/Mcl-1 axis contributed to LiCl-induced apoptosis both in vitro and in vivo. The present study provides important mechanistic insight into potential cancer treatments involving LiCl and enhances the understanding of human choroidal melanoma.

The relationship between platelet indices and patent ductus arteriosus in preterm infants: a systematic review and meta-analysis.

Patent ductus arteriosus (PDA), one of the most common disorders in newborns, is associated with many complications in premature infants such as respiratory distress syndrome (RDS) and bronchopulmonary dysplasia (BPD). However, the diagnosis of hemodynamically significant patent ductus arteriosus (hsPDA) is still an ongoing debate. The relationship between platelet parameters and hsPDA has been explored in many studies over the last decade, but there is still no definite conclusion. We aim to explain the relationship between platelet parameters and hsPDA through this meta-analysis. Therefore, we used PubMed, Embase, the Cochrane Library, and Web of Science databases as well as the Google Scholar to search for studies up to May 2020. Three reviewers independently screened the articles, evaluated the quality of the articles, and collected the data. The random-effects model and fixed-effects model were used to evaluate pooled results. We used the I-square (I2) test to examine heterogeneity and the funnel plot; Egger's test and meta-regression analysis were used to test for publication bias. Influence analysis was also carried out in this study. Stata version 12.0 software was used for data analysis. Fourteen studies, which included 3330 newborns, were extracted from 986 studies. The weighted mean difference (WMD) of the platelet count was - 17.98 (p < 0.001), the platelet distribution width (PDW) was 0.27 (p = 0.266), the mean platelet volume (MPV) was 0.01 (p = 0.958), the plateletcrit (PCT) was - 0.03 (p < 0.001), and the platelet mass was - 150.10 (p = 0.001).Conclusion: Platelet count, PCT, and platelet mass of the first 3 days of life are potentially helpful in identifying premature infants at risk of hsPDA. More prospective studies on the relationship between different degrees of thrombocytopenia and platelet function and hsPDA should be conducted. What is Known: • Platelets are involved in the formation of thrombi during closure of the arterial duct. • The diagnosis of hsPDA by Doppler echocardiography and clinical signs is not precise enough. What is New: • Preterm newborns with hsPDA in the first week of life demonstrated a significant reduction in platelet count, platelet mass, and plateletcrit in the first 3 days of life. • No significant difference was shown between hsPDA and non-hsPDA infants in platelet distribution width and mean platelet volume in the first 3 days of life.

Genome-wide identification and comparative analysis of diacylglycerol kinase (DGK) gene family and their expression profiling in Brassica napus under abiotic stress.

BACKGROUND: Diacylglycerol kinases (DGKs) are signaling enzymes that play pivotal roles in response to abiotic and biotic stresses by phosphorylating diacylglycerol (DAG) to form phosphatidic acid (PA). However, no comprehensive analysis of the DGK gene family had previously been reported in B. napus and its diploid progenitors (B. rapa and B. oleracea). RESULTS: In present study, we identified 21, 10, and 11 DGK genes from B. napus, B. rapa, and B. oleracea, respectively, which all contained conserved catalytic domain and were further divided into three clusters. Molecular evolutionary analysis showed that speciation and whole-genome triplication (WGT) was critical for the divergence of duplicated DGK genes. RNA-seq transcriptome data revealed that, with the exception of BnaDGK4 and BnaDGK6, BnaDGK genes have divergent expression patterns in most tissues. Furthermore, some DGK genes were upregulated or downregulated in response to hormone treatment and metal ion (arsenic and cadmium) stress. Quantitative real-time PCR analysis revealed that different BnaDGK genes contribute to seed oil content. CONCLUSIONS: Together, our results indicate that DGK genes have diverse roles in plant growth and development, hormone response, and metal ion stress, and in determining seed oil content, and lay a foundation for further elucidating the roles of DGKs in Brassica species.

Fusing T5 exonuclease with Cas9 and Cas12a increases the frequency and size of deletion at target sites.

CRISPR/Cas systems, especially CRISPR/Cas9, generally result in small insertions/deletions, which are unlikely to eliminate the functions of regulatory and other non-coding sequences. To generate larger genomic deletions usually requires the use of pairs of guide RNAs. Here we show that it is possible to create such deletions with a single guide RNA by fusing Cas9 or Cas12a with T5 exonuclease (T5exo). These fusion constructs were found to increase both the frequency and size of deletions at target loci in rice protoplasts and seedlings. Moreover, the genome editing efficiencies of Cas9 and Cas12a were also enhanced by fusion with T5 exonuclease. These T5exo-Cas fusions expand the CRISPR toolbox, and facilitate knockout of regulatory and non-coding DNA sequences. From a wider standpoint, our results suggest a general strategy for producing larger deletions using other Cas nucleases.

Knockdown of miR-124 Reduces Depression-like Behavior by Targeting CREB1 and BDNF.

OBJECTIVE: As a brain-specific microRNA, the mechanism of miR-124 in depression has not been clarified so far. The present study aimed to explore the role of miR-124 in depression and its potential targets. METHODS: The depression model was first replicated by the chronic unpredictable mild stress (CUMS) method. miR-124 antagomir was injected into the hippocampus of CUMS rats. Sucrose preference test (SPT), open-field test (OFT), elevated-plus maze (EPM), and forced swimming test (FST) were used to analyze the depression-like behavior. The content of norepinephrine (NE), dopamine (DA) and 5-hydroxytryptamine (5-HT) in the hypothalamus was analyzed by ELISA. qRT-PCR and western blot assay were used for functional analysis. RESULTS: miR-124 expression was up-regulated in the hippocampus of CUMS -induced depression model rats, while CREB1 and BDNF were down-regulated. Administration of miR-124 antagomir in the hippocampus inhibited miR-124 expression in the hippocampus of CUMS rats. Additionally, SPT, OFT, EPM, and FST also showed that miR-124 antagomir can reduce the depression-like behavior of CUMS rats. Furthermore, miR-124 antagomir injection increased the levels of NE, DA and 5-HT in the hypothalamus of CUMS rats. Moreover, miR-124 antagomir injection increased the expression of cyclic AMP-responsive element-binding protein1 (CREB1) and brain-derived neurotrophic factor (BDNF) in the hippocampus. Using the dual-luciferase reporter assay, it was confirmed that miR-124 directly targets 3'UTR of CREB1 and BDNF genes. CONCLUSION: Knockdown of miR-124 can improve depression-like behavior in CUMS-induced depressive rats, which may be related at least in part to the up-regulation of CREB1 and BDNF expression in the hippocampus.

Modulating chromatin accessibility by transactivation and targeting proximal dsgRNAs enhances Cas9 editing efficiency in vivo.

The CRISPR/Cas9 system is unable to edit all targetable genomic sites with full efficiency in vivo. We show that Cas9-mediated editing is more efficient in open chromatin regions than in closed chromatin regions in rice. A construct (Cas9-TV) formed by fusing a synthetic transcription activation domain to Cas9 edits target sites more efficiently, even in closed chromatin regions. Moreover, combining Cas9-TV with a proximally binding dead sgRNA (dsgRNA) further improves editing efficiency up to several folds. The use of Cas9-TV/dsgRNA thus provides a novel strategy for obtaining efficient genome editing in vivo, especially at nuclease-refractory target sites.

Enantioselective Hydrolysis of Styrene Oxide and Benzyl Glycidyl Ether by a Variant of Epoxide Hydrolase from Agromyces mediolanus.

Enantiopure epoxides are versatile synthetic intermediates for producing optically active pharmaceuticals. In an effort to provide more options for the preparation of enantiopure epoxides, a variant of the epoxide hydrolase (vEH-Am) gene from a marine microorganism Agromyces mediolanus was synthesized and expressed in Escherichia coli. Recombiant vEH-Am displayed a molecular weight of 43 kDa and showed high stability with a half-life of 51.1 h at 30 °C. The purified vEH-Am exhibited high enantioselectivity towards styrene oxide (SO) and benzyl glycidyl ether (BGE). The vEH-Am preferentially converted (S)-SO, leaving (R)-SO with the enantiomeric excess (ee) >99%. However, (R)-BGE was preferentially hydrolyzed by vEH-Am, resulting in (S)-BGE with >99% ee. To investigate the origin of regioselectivity, the interactions between vEH-Am and enantiomers of SO and BGE were analyzed by molecular docking simulation. In addition, it was observed that the yields of (R)-SO and (S)-BGE decreased with the increase of substrate concentrations. The yield of (R)-SO was significantly increased by adding 2% (v/v) Tween-20 or intermittent supplementation of the substrate. To our knowledge, vEH-Am displayed the highest enantioselectivity for the kinetic resolution of racemic BGE among the known EHs, suggesting promising applications of vEH-Am in the preparation of optically active BGE.