Advances in CRISPR/Cas9-based research related to soybean [Glycine max (Linn.) Merr] molecular breeding.

Soybean [Glycine max (Linn.) Merr] is a source of plant-based proteins and an essential oilseed crop and industrial raw material. The increase in the demand for soybeans due to societal changes has coincided with the increase in the breeding of soybean varieties with enhanced traits. Earlier gene editing technologies involved zinc finger nucleases and transcription activator-like effector nucleases, but the third-generation gene editing technology uses clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9). The rapid development of CRISPR/Cas9 technology has made it one of the most effective, straightforward, affordable, and user-friendly technologies for targeted gene editing. This review summarizes the application of CRISPR/Cas9 technology in soybean molecular breeding. More specifically, it provides an overview of the genes that have been targeted, the type of editing that occurs, the mechanism of action, and the efficiency of gene editing. Furthermore, suggestions for enhancing and accelerating the molecular breeding of novel soybean varieties with ideal traits (e.g., high yield, high quality, and durable disease resistance) are included.

α-Difluoroalkylation of Benzyl Amines with Trifluoromethylarenes.

An α-difluoroalkylation of benzyl amines with trifluoromethylarenes is disclosed herein. This protocol is characterized by its operational simplicity, excellent chemoselectivity and broad scope-even with advanced synthetic intermediates-, thus offering a new entry point to medicinally-relevant α-difluoroalkylated amines from simple, yet readily accessible, precursors.

The Structural Changes of Frontal Subregions and Their Correlations with Cognitive Impairment in Patients with Alzheimer's Disease.

BACKGROUND: The frontal lobe is affected by Alzheimer's disease (AD) and mild cognitive impairment (MCI). However, we still lack sufficient understanding of subregion atrophy in the frontal cortex, and the relationship between subregions volume and cognitive decline in AD or MCI remains unclear. METHODS: This study enrolled 434 participants from the Alzheimer's Disease Neuroimaging Initiative (ADNI), including 150 cognitively normals (CN), 187 subjects with MCI, and 97 patients with AD. The gray matter of frontal regions and subregions was divided based on the BNA-246 atlas and its volume was measured by voxel-based morphometry (VBM). Analysis of covariance was performed to compare the differences in frontal regions and subregions volume. Then, receiver operating characteristic (ROC) curve and the area under the curve (AUC) were used to analyze the discriminative ability of subregion volume to distinguish the three groups. In addition, we investigated the association of subregion volume with Mini-Mental State Examination (MMSE) score and Alzheimer's Disease Assessment Scale-Cognitive Behavior section (ADAS-cog) scores with age, gender, education, and the estimated total intracranial volume (eTIV) as covariates. RESULTS: In addition to the regions of frontal lobe atrophy found in previous studies, atrophy of the precentral gyrus (PrG) and some of its subregions were found in MCI. The volume of the right dorsal area 9/46 (MFG_7_1) was the best index to differentiate AD from CN, with an AUC value of 0.7. Moreover, we found that some subregions are associated with cognition in patients with MCI and AD. CONCLUSIONS: Frontal lobe atrophy in MCI is more extensive than we assumed. In addition, the volume of right MFG_7_1 has the potential to distinguish AD from CN.

Comparison between pre- and post-contrast cardiac MRI cine images: the impact on ventricular volume and strain measurement.

To explore whether contrast agent administration will affect ventricular volume and strain parameters measured on cardiac magnetic resonance cine images. This prospective study enrolled 88 patients, including 32 patients with cardiac amyloidosis (CA), 32 patients with hypertrophic cardiomyopathy (HCM), and 24 control participants, to perform steady-state free precession (SSFP)-cine imaging twice, respectively before and after contrast agent injection. Indexed left and right ventricular (LV and RV) volume and LV strain parameters (peak radial strain [PRS], peak circumferential strain [PCS], peak longitudinal strain [PLS]) were analyzed and compared between the pre- and post-contrast cine groups. Compared to the group of pre-contrast cine, the end-diastolic volume index (EDVi) and end-systolic volume index (ESVi) significantly increased in the group using post-contrast cine images (all p < 0.05), especially in the right ventricle. After contrast injection, the right ventricular ejection fraction (RVEF) decreased significantly (p < 0.05), while the left ventricular ejection fraction (LVEF) only reduced for patients with HCM (p < 0.05). The PRS (37.1 ± 15.2 vs. 32.0 ± 15.4, p < 0.001) and PCS (- 14.9 ± 4.3 vs. - 14.0 ± 4.1, p < 0.001) derived from post-contrast cine images reduced significantly in all patients and this tendency remained in subgroup analysis except for PCS in the control group. The administration of a contrast agent may influence the measurements of ventricular volume and strain. Acquiring pre-contrast cine images were suggested for patients who required more accurate right ventricle evaluation or precise strain assessment.

Comprehensive mapping of alternative polyadenylation site usage and its dynamics at single-cell resolution.

Alternative polyadenylation (APA) plays an important role in posttranscriptional gene regulation such as transcript stability and translation efficiency. However, our knowledge about APA dynamics at the single-cell level is largely unexplored. Here, we developed single-cell polyadenylation sequencing, a strand-specific approach for sequencing the 3' end of transcripts, to investigate the landscape of APA at the single-cell level. By analyzing several cell lines, we found many genes using multiple polyA sites in bulk data are prone to use only one polyA site in each single cell. Interestingly, cell cycle genes were significantly enriched in genes with high variation in polyA site usages. Furthermore, the 414 genes showing a polyA site usage switch after cell synchronization enriched cell cycle genes, while the differentially expressed genes after cell synchronization did not enrich cell cycle genes. We further identified 812 genes showing polyA site usage changes between neighboring cell cycles, which were grouped into six clusters, with cell phase-specific functional categories enriched in each cluster. Deletion of one polyA site in MSL1 and SCCPDH results in slower and faster cell cycle progression, respectively, supporting polyA site usage switch played an important role in cell cycle. These results indicate that APA is an important layer for cell cycle regulation.

Catalytic Hydrodifluoroalkylation of Unactivated Olefins.

Herein, we report a modular catalytic technique that streamlines the preparation of gem-difluoroalkanes from unactivated sp3 precursors. The method is characterized by its simplicity, generality, and site selectivity, including the functionalization of advanced intermediates and olefin feedstocks. Our approach is enabled by a cooperative interplay of halogen- and hydrogen-atom transfer, thus offering a new entry point to difluorinated alkyl bioisosteres of interest in drug discovery.

[Feasibility of Single-Breath-Hold Compressed Sensing Real-Time Cine Imaging for Assessment of Ventricular Function and Left Ventricular Strain in Cardiac Magnetic Resonance].

Objective: To explore the feasibility of single-breath-hold compressed sensing real-time cine imaging (CS-cine) in the assessment of ventricular function and left ventricular (LV) strain. Methods: A total of 70 subjects were enrolled prospectively, and all subjects underwent cardiac magnetic resonance imaging (cardiac MRI) using both the standard steady-state free procession cine (sta-cine) acquisition and a prototype CS-cine sequence. For both CS-cine and sta-cine imaging, continuous short-axis cine images were acquired from the base to the apex to cover the entire left ventricle, and long-axis cine images including two-, three-, and four-chamber views were also acquired. The scanning range, number of slices, slice thickness and intervals were kept identical for the two cine images of the same participant. Subjective evaluation of the image quality was performed on all cine images. For both sequences, the conventional function parameters of the left and the right ventricles and LV strain values were assessed with post-processing software analysis. The cine image quality, conventional ventricular function parameters, and LV strain values were compared between the two cine groups and the differences were examined. Inter- and intraobserver agreements for CS-cine images were measured using intraclass correlation coefficient ( ICC). Bland-Altman analysis was performed to assess reproducibility between the two cine methods. Results: The median scanning time of CS-cine was 21 s versus 272 s for sta-cine ( P<0.001). The median image quality scores of two groups were significantly different, 4 points for sta-cine and 2 points for CS-cine ( P<0.001). Bi-ventricular end-diastolic volumes (EDV), stroke volume (SV) and ejection fraction (EF) were significantly smaller in CS-cine ( P<0.001). Nevertheless, no significant differences between the two groups in bi-ventricular ESV or LV mass were observed ( P>0.05). LV strain parameters, including the peak radial strain, peak circumferential strain and peak longitudinal strain derived from LV mid-ventricular slice, were significantly different in the two sequences ( P<0.001). Moreover, CS-cine-derived functional parameters and strain measurements have a good correlation with those of sta-cine (for RV function parameters, and left ventricular PLS, PCS values, more than 95% points fell within the limits of agreement [ LoA]; meanwhile, more than 91% points fell within the LoA for other parameters) and inter- and intraobserver agreements were strong ( ICC=0.88 to 0.99) for CS-cine. Conclusion: CS-cine can well realize the rapid acquisition of cine images for quantitative analysis of cardiac function, and the conventional ventricular function parameters and LV globalized strain values obtained from CS-cine imaging have good reproducibility.

Optimize the irrigation and fertilizer schedules by combining DSSAT and genetic algorithm.

As one of the main food crops in the world, the yield of maize directly affects the food security of the world. The optimization of irrigation and fertilizer schedules is also one of the hot issues in the world. However, the traditional optimization methods are mainly based on field experiment or crop model. The research on combining crop model with optimization algorithm to optimize irrigation and fertilizer schedule is rare. In this paper, the genetic algorithm (GA) and DSSAT crop model were combined to provide theoretical basis for the optimization of irrigation and fertilizer schedules of maize in China. On the basis of field experimental data in previous references, the model was calibrated and verified, and get a well simulation result with RMSE ranged from 0.262 to 0.580 Mg/ha. After that, GA and DSSAT were run to obtain the optimized irrigation and fertilizer schedules. Compared with the results of previous references, the new optimization schedules can improve the yield (1.9 ~ 2.6%) and economic benefits (7.3 ~ 8.9%). It is proved that this method has a good optimization effect, and the method also has a wide range of research prospects.

Copper(I)-Catalyzed Asymmetric Synthesis of Unnatural α-Amino Acid Derivatives and Related Peptides Containing γ-(aza)Aryls.

Chiral α-amino acids are indispensable compounds in organic chemistry, biochemistry, and medicinal chemistry. Herein, by means of copper(I)-catalyzed asymmetric conjugate addition of derivatives of glycine, serine, cysteine, and ß-amino-alanine to electron-deficient vinyl(aza)arenes, an array of novel unnatural chiral α-amino acid derivatives bearing a γ-(aza)aryl is prepared in moderate to high yields with high enantioselectivity. Various azaarenes, such as pyrimidine, 1,3,5-triazine, pyridine, pyridine-N-oxide, quinoline, quinoxaline, purine, benzo[d]imidazole, benzothiazole, and 1,2,4-oxadiazole, are well tolerated. Moreover, the electrophiles are nicely extended to (Z)/(E) mixtures of electron-deficient butadienylpyridine and benzene, which are transformed to the corresponding chiral α-amino acid derivatives in high (E)/(Z) ratio and high enantioselectivity. More importantly, the present methodology is successfully applied in the catalytic asymmetric functionalization of Schiff bases derived from peptides, which finally afforded a new chiral tripeptide bearing two electron-deficient azaaryls and one electron-deficient aryl in high total yield with high diastereo- and excellent enantioselectivities.

Site-Selective Defluorinative sp3 C-H Alkylation of Secondary Amides.

A site-selective defluorinative sp3 C-H alkylation of secondary amides that rapidly and reliably incorporates gem-difluoroalkene motifs into previously unfunctionalized sp3 sites is disclosed. This protocol is distinguished by its mild conditions, wide scope, and exquisite site-selectivity, thus unlocking a new platform to introduce carbonyl isosteres at saturated hydrocarbon sites.

Copper(I)-Catalyzed Asymmetric Vinylogous Aldol-Type Reaction of Allylazaarenes.

A vinylogous aldol-type reaction of allylazaarenes and aldehydes is disclosed that affords a series of chiral γ-hydroxyl-α,ß-unsaturated azaarenes in moderate to excellent yields with high to excellent regio- and enantioselectivities. With (R,RP )-TANIAPHOS and (R,R)-QUINOXP* as the ligand, the carbon-carbon double bond in the products is generated in (E)-form. With (R)-DTBM-SEGPHOS as the ligand, (Z)-form carbon-carbon double bond is formed in the major product. In this vinylogous reaction, aromatic, α,ß-unsaturated, and aliphatic aldehydes are competent substrates. Moreover, a variety of azaarenes, such as pyrimidine, pyridine, pyrazine, quinoline, quinoxaline, quinazoline, and benzo[d]imidazole are well-tolerated. At last, the chiral vinylogous product is demonstrated as a suitable Michael acceptor towards CuI-catalyzed nucleophilic addition with organomagnesium reagents.

Discrepant antitumor efficacies of three CpG oligodeoxynucleotide classes in monotherapy and co-therapy with PD-1 blockade.

Unmethylated CpG oligodeoxynucleotides (ODNs) activate plasmacytoid dendritic cells (pDCs) and B cells to induce humoral and cellular immunity, and are under development for the treatment of multiple cancers. However, the specific differences in antitumor effects among the three CpG ODN classes when administered as a monotherapy or in co-therapy with the anti-PD-1 antibody are unclear. We compared the immunostimulatory effects in vitro and antitumor effects in a CT26 subcutaneous mouse tumor model among the three CpG ODN classes. We found that CpG-A slightly suppressed tumor growth but possessed no synergistic antitumor effects with the anti-PD-1 antibody. CpG-B at low doses significantly inhibited tumor growth and possessed synergistic antitumor effects with the anti-PD-1 antibody. A high dose of CpG-C was required to achieve antitumor effects comparable to those of CpG-B, which was consistent with the immunostimulatory effects in B-cell proliferation and TLR9-NF-κB activation. Importantly, CpG-C in combination with anti-PD-1 antibody inhibited tumor growth more quickly and effectively than CpG-B because CpG-B significantly upregulated PD-L1 expression on multiple host immune cells to promote tumor immune escape. Moreover, co-therapy increased the infiltration of effector memory T cells. In summary, CpG-B and CpG-C with different optimal concentrations possessed strong antitumor effects, while CpG-C was more rapid and effective for co-therapy with the anti-PD-1 antibody.

A Novel C Type CpG Oligodeoxynucleotide Exhibits Immunostimulatory Activity In Vitro and Enhances Antitumor Effect In Vivo.

BACKGROUND: C type CpG oligodeoxynucleotides (CpG-C ODNs), possessing the features of both A type and B type CpG ODNs, exert a variety of immunostimulatory activities and have been demonstrated as an effective antitumor immunotherapy. Based on the structural characteristics, we designed 20 potential ODNs with the aim of synthesizing an optimal, novel CpG-C ODN specific to human and murine Toll-like receptor 9 (TLR9). We also sought to investigate the in vitro immunostimulatory and in vivo antitumor effects of the novel CpG-C ODN. METHODS: Twenty potential CpG-C ODNs were screened for their ability to secrete interferon (IFN)-α, and interleukin (IL)-6 and tumor necrosis factor (TNF)-α production for the three most promising sequences were assayed in human peripheral blood mononuclear cells (PBMCs) by enzyme-linked immunosorbent assay (ELISA) or cytometric bead array assay. The functions of human and mouse B cells, and cytokine production in mice induced by the most promising sequence, HP06T07, were determined by flow cytometry and ELISA. Growth and morphology of tumor tissues in in vivo murine models inoculated with CT26 cells were analyzed by a growth inhibition assay and immunohistochemistry, respectively. RESULTS: Among the 20 designed ODNs, HP06T07 significantly induced IFN-α, IL-6, and TNF-α secretion, and promoted B-cell activation and proliferation in a dose-dependent manner in human PBMCs and mouse splenocytes in vitro. Intratumoral injection of HP06T07 notably suppressed tumor growth and prolonged survival in the CT26 subcutaneous mouse model in a dose-dependent manner. HP06T07 administered nine times at 2-day intervals (I2) eradicated tumor growth at both primary and distant sites of CT26 tumors. HP06T07 restrained tumor growth by increasing the infiltration of T cells, NK cells, and plasmacytoid dendritic cells (pDCs). CONCLUSIONS: HP06T07, a novel CpG-C ODN, shows potent immunostimulatory activity in vitro and suppresses tumor growth in the CT26 subcutaneous mouse model.

Rapid Synthesis of Chiral 1,2-Bisphosphine Derivatives through Copper(I)-Catalyzed Asymmetric Conjugate Hydrophosphination.

1,2-Bisphosphines have been identified as one class of important and powerful chiral ligands in asymmetric catalysis with transition metals. Herein, a copper(I)-catalyzed asymmetric hydrophosphination of α,ß-unsaturated phosphine sulfides was developed with the assistance of "soft-soft" interaction between copper(I)-catalyst and the phosphine sulfide moiety, which afforded 1,2-bisphosphine derivatives with diversified electronic nature and steric hindrance in high to excellent yields with high to excellent enantioselectivity. Moreover, the challenging catalytic asymmetric hydrophosphination/protonation reaction was achieved with excellent enantioselectivity. Strikingly, the dynamic kinetic resolution of racemic diarylphosphines was also successfully carried out with high to excellent diastereo- and enantioselectivities. Interestingly, the nucleophilic copper(I)-diphenylphosphide species was characterized by 31 P NMR spectrum and mass spectrum. At last, three products were transformed to chiral 1,2-bisphosphines, which were employed as ligands in Rh-catalyzed asymmetric hydrogenation of α-amino-α,ß-unsaturated ester. The α-amino acid derivative was produced in high enantioselectivity, which demonstrated the utility of the present methodology.

The role of NF-κB signaling pathway in diffuse large B cell lymphoma and its application in targeted therapy / 中国肿瘤生物治疗杂志

@#弥漫大B细胞淋巴瘤（diffuse large B-cell lymphoma，DLBCL）是一种起源于B淋巴细胞的侵袭性恶性肿瘤，是世界上 最常见的非霍奇金淋巴瘤。多种基因的遗传损伤引发B细胞内NF-κB信号通路持续性激活，促进B细胞恶性增殖，从而导致肿 瘤生成。NF-κB信号通路的组成性激活在DLBCL的发病机制中起着重要作用，因此抑制NF-κB信号通路的过度激活成为DLBCL治疗研究的热点。近年来，多种NF-κB信号通路相关靶向抑制剂相继开发并进行临床试验。本文主要阐述DLBCL中B细胞 受体和Toll样受体依赖性NF-κB信号通路异常激活的遗传学机制，及其在DLBCL发生和发展过程中的作用，并总结该信号通路 靶向抑制剂的临床研究进展，旨在为DLBCL治疗方案的选择和靶向药物的开发提供参考依据。

Asymmetric Vinylogous Aldol-type Reactions of Aldehydes with Allyl Phosphonate and Sulfone.

Two catalytic asymmetric vinylogous aldol-type reactions of aldehydes with allyl phosphonate and allyl sulfone have been uncovered in good to high yields for the first time. The bulky ligand-(R)-DTBM-SEGPHOS-was found to be the key to perfectly control both regio- and enantioselectivities. Transformations of the vinylogous products (including Horner-Wadsworth-Emmons and Julia olefinations) were successfully realized by virtue of the phosphonate and sulfone moieties. Moreover, the present methodology was successfully applied in the asymmetric synthesis of natural products.

Combining the Arterial Phase of Contrast-Enhanced Ultrasonography, Gadoxetic Acid-Enhanced Magnetic Resonance Imaging and Diffusion-Weighted Imaging in the Diagnosis of Hepatic Nodules ≤20 mm in Patients with Cirrhosis.

Contrast-enhanced ultrasonography (CEUS) and gadoxetic acid-enhanced magnetic resonance imaging (EOB-MRI) were compared with respect to diagnostic efficacy in the detection of small hepatocellular carcinoma. A new diagnostic strategy that combines the arterial phase of CEUS, the hepatobiliary phase of EOB-MRI and diffusion-weighted MR imaging (DWI) is described. One hundred sixteen nodules were enrolled to validate the performance of the strategy. For lesions ≤20 mm in size, the areas under the receiver operating characteristic curves (Az) of CEUS and EOB-MRI were 0.930 (95% confidence interval [CI]: 0.867-0.969) and 0.920 (95% CI: 0.855-0.962) (p = 0.796), respectively. The Az value of the new diagnostic strategy was 0.985 (95% CI: 0.942-0.999) (vs. CEUS, p = 0.026; vs. EOB-MRI, p = 0.014). The sensitivity, specificity and diagnostic accuracy of the new strategy were 95.5% (95% CI: 88.9%-98.8%), 96.3% (95% CI: 81.0%-99.9%) and 95.7% (95% CI: 91.9%-99.4%), respectively. The new diagnostic strategy based on the arterial phase of CEUS, hepatobiliary phase of EOB-MRI and DWI represents an appealing solution for distinguishing small hepatocellular carcinomas from benign lesions, especially when the nodules present atypical enhancement patterns.

Catalytic Asymmetric Construction of Halogenated Stereogenic Carbon Centers by Direct Vinylogous Mannich-Type Reaction.

A catalytic asymmetric vinylogous Mannich-type reaction of γ-halo-α,ß-unsaturated N-acylpyrazoles and N-Boc-aldimines was disclosed, which afforded an array of halogenated (F-, Cl-, and Br-) allylic stereogenic carbon centers in high yields with good to high regio-, diastereo-, and enantioselectivities. The brominated product served as a suitable electrophile for common SN2 nucleophilic substitution and copper-mediated SN2' allylic alkylation with metal reagents. The utility of present methodology was demonstrated by the asymmetric synthesis of a common intermediate toward the synthesis of two chiral 2,3-disubstituted piperidine pharmaceuticals.