Gene editing by SSB/CRISPR-Cas9 ribonucleoprotein in bacteria.

The application of CRISPR-Cas9 ribonucleoprotein (RNP) for gene editing is commonly used in plants and animals, but its application in bacteria has not been reported. In this study, we employed DNA single-strand binding protein (SSB) to construct an SSB/CRISPR-Cas9 RNP-editing system for non-homologous recombination and homologous recombination gene editing of the upp gene in bacteria. The RNP targeting the upp gene, along with SSB, was introduced into the protoplasts of Escherichia coli, Pseudomonas, and Bacillus subtilis. Transformants were obtained on plates containing 5-fluorouracil (5-FU) with gene editing efficiencies (percentage of transformants relative to the number of protoplasts) of 9.75 %, 5.02 %, and 8.37 %, respectively, and sequencing analysis confirmed 100 % non-homologous recombination. When RNP, SSB, and a 100-nucleotide single-stranded oligodeoxynucleotide (ssODN) donor were introduced into the protoplasts of these bacteria, transformants were obtained with editing efficiencies of 45.11 %, 30.13 %, and 27.18 %, respectively, and sequencing confirmed 100 % homologous recombination knockout of the upp gene. Additionally, introducing RNP, SSB, and a 100 base-pair double-stranded oligodeoxynucleotide (dsODN) donor containing a tetracycline resistance gene (tetR-dsODN) resulted in transformants on 5-FU plates with editing efficiencies of 35.94 %, 22.46 %, and 19.08 %, respectively, with sequencing confirming 100 % homologous recombination replacement of the upp gene with tetR. These results demonstrate that the SSB/CRISPR-Cas9 RNP system can efficiently, simply, and rapidly edit bacterial genomes without the need for plasmids. This study is the first to report the use of RNP-based gene editing in bacteria.

The Influence of Homologous Arm Length on Homologous Recombination Gene Editing Efficiency Mediated by SSB/CRISPR-Cas9 in Escherichia coli.

The precise editing of genes mediated by CRISPR-Cas9 necessitates the application of donor DNA with appropriate lengths of homologous arms and fragment sizes. Our previous development, SSB/CRISPR-Cas9, has demonstrated high efficiency in homologous recombination and non-homologous end joining gene editing within bacteria. In this study, we optimized the lengths and sizes of homologous arms of the donor DNA within this system. Two sets of donor DNA constructs were generated: one set comprised donors with only 10-100 bp homologous arms, while the other set included donors with homologous arms ranging from 10-100 bp, between which was a tetracycline resistance expression cassette (1439 bp). These donor constructs were transformed into Escherichia coli MG1655 cells alongside pCas-SSB/pTargetF-lacZ. Notably, when the homologous arms ranged from 10 to 70 bp, the transformation efficiency of non-selectable donors was significantly higher than that of selectable donors. However, within the range of 10-100 bp homologous arm lengths, the homologous recombination rate of selectable donors was significantly higher than that of non-selectable donors, with the gap narrowing as the homologous arm length increased. For selectable donor DNA with homologous arm lengths of 10-60 bp, the homologous recombination rate increased linearly, reaching a plateau when the homologous arm length was between 60-100 bp. Conversely, for non-selectable donor DNA, the homologous recombination rate increased linearly with homologous arm lengths of 10-90 bp, plateauing at 90-100 bp. Editing two loci simultaneously with 100 bp homologous arms, whether selectable or non-selectable, showed no difference in transformation or homologous recombination rates. Editing three loci simultaneously with 100 bp non-selectable homologous arms resulted in a 45% homologous recombination rate. These results suggest that efficient homologous recombination gene editing mediated by SSB/CRISPR-Cas9 can be achieved using donor DNA with 90-100 bp non-selectable homologous arms or 60-100 bp selectable homologous arms.

Low-Dimensional-Materials-Based Photodetectors for Next-Generation Polarized Detection and Imaging.

The vector characteristics of light and the vectorial transformations during its transmission lay a foundation for polarized photodetection of objects, which broadens the applications of related detectors in complex environments. With the breakthrough of low-dimensional materials (LDMs) in optics and electronics over the past few years, the combination of these novel LDMs and traditional working modes is expected to bring new development opportunities in this field. Here, the state-of-the-art progress of LDMs, as polarization-sensitive components in polarized photodetection and even the imaging, is the main focus, with emphasis on the relationship between traditional working principle of polarized photodetectors (PPs) and photoresponse mechanisms of LDMs. Particularly, from the view of constitutive equations, the existing works are reorganized, reclassified, and reviewed. Perspectives on the opportunities and challenges are also discussed. It is hoped that this work can provide a more general overview in the use of LDMs in this field, sorting out the way of related devices for "more than Moore" or even the "beyond Moore" research.

Antimicrobial films fabricated with myricetin nanoparticles and chitosan derivation microgels for killing pathogenic bacteria in drinking water.

Pathogenic bacteria in drinking water threaten human health and life. In the work, antimicrobial films composed of myricetin@tannic acid (My@TA) nanoparticles (NPs) and chitosan derivation microgels were developed to kill pathogenic bacteria in drinking water. Hydrophobic My was first made into water soluble My@TA NPs using a solvent exchange method with TA as stabilizer. Polymeric microgels of carboxymethyl chitosan (CMCS)/hydroxypropyltrimethyl ammonium chloride chitosan (HACC) were then fabricated with a blending method. CMCS&HACC/My@TA multilayer films were further deposited on the internal surface of PET bottles by using a layer-by-layer (LbL) assembly technique. The PET bottles coated with the films could effectively kill pathogenic bacteria in water such as S. aureus, E. coli, Staphylococcus epidermidis, Pseudomonas fluorescens, Listeria monocytogenes and methicillin resistant Staphylococcus aureus (MRSA). In addition, CMCS&HACC/My@TA films displayed good antioxidant activity, water resistance, and in vivo biocompatibility with heart, liver, spleen, lung and kidney organs. We believe that the container coated with CMCS&HACC/My@TA films can be applied to prevent microbial contamination of drinking water.

Peripheral neuropathy associated with chronic lymphoproliferative disorders of natural killer cells (CLPD-NK): a case report and literature review.

BACKGROUND: Chronic lymphoproliferative disorders of natural killer cells (CLPD-NK) is a rare lymphoproliferative disease. Peripheral neuropathy is an unusual symptom of CLPD-NK. We report a case of peripheral neuropathy associated with CLPD-NK and perform a review of literatures. CASE PRESENTATION: a 62-year-old woman presented with progressive numbness and weakness in both extremities. Electrophysiological examinations indicated a sensorimotor polyneuropathy. Peripheral blood examination revealed that the number of white blood cells (WBC) and lymphocytes were significantly increased. Flow cytometry analysis identified that 84% of the lymphocytes are NK cells that mainly expressed CD56, combined with variable expression of CD16, CD2, CD7, CD94, granzyme B, perforin, and CD158 but negative for CD3. Sural nerve biopsy revealed that a plethora of NK cells infiltrated into nerve fascicles. On treatment with combined cyclophosphamide and corticosteroids, her symptoms rapidly improved. Moreover, the absolute lymphocyte count and its proportion recovered to normal range after 3 months' treatment. CONCLUSION: To the best of our knowledge, this is the first case report of peripheral neuropathy associated with CLPD-NK from Chinese. This rare lymphoproliferative disease should be considered if peripheral neuropathy combines with increased WBC or lymphocytes. Immunosuppressive drugs are the major treatment and most patients can achieve a good prognosis.

Bandage modified with antibacterial films of quaternized chitosan & sodium carboxymethyl cellulose microgels/baicalein nanoparticles for accelerating infected wound healing.

Wound dressings capable of sterilizing pathogenic bacteria and scavenging free radicals are important to inhibit bacterial invasion and accelerate wound healing. The target of this work is to develop an antibacterial dressing by modifying bandages with films composed of biological macromolecule microgels and baicalein@tannic acid (Bai@TA) nanoparticles (NPs). Firstly, hydrophobic Bai was made into water soluble Bai@TA NPs using a solvent exchange method with TA as stabilizer. Polymeric microgels of sodium carboxymethyl cellulose (CMC)&hydroxypropyltrimethyl ammonium chloride chitosan (HACC) were then prepared by a simple blending method. Further, CMC&HACC/Bai@TA multilayer films were deposited on medical bandages by using a layer-by-layer assembly technique to obtain an antibacterial dressing. The as-prepared dressings showed great antibacterial ability against E. coli, S. aureus and methicillin resistant Staphylococcus aureus (MRSA), excellent antioxidant activity and good biological safety. In addition, compared to conventional medical bandages, the dressings could efficaciously diminish inflammation in the wound, accelerate skin regeneration and functional restoration, and promote the in vivo healing speed of full-thickness skin wounds infected by MRSA. We believe that as a low-cost but effective wound dressing, the antibacterial bandage modified with CMC&HACC/Bai@TA films has potentials to replace traditional dressings in the clinical management of infected wounds.

Photonic Modulation Enabled by Controlling the Edge Structures of Boron-Doped Molecular Carbons.

Control over topological edges of molecular carbons (MCs) is of importance for achieving diverse molecular topologies and desirable physical properties. However, it remains very challenging for heteroatom-doped MCs due to the synthetic difficulty. Herein, we report control over the edge structures of boron-doped MCs (BMCs) via the sequential cyclization strategy. Three BMC molecules that feature the C56 B2 or C84 B2 polycyclic π-skeletons with selective cove/fjord or cove/bay edges, respectively, were synthesized through the rational combination of Mallory photoreaction and Scholl reaction. We not only obtain the largest boron-doped π-system reported so far, but also disclose that fine control of their edges and length greatly affects electronic structures and thereby photonic properties of BMCs, such as tunable aromaticity, decreased band gaps, as well as redshifted absorptions and fluorescence. Remarkably, the C56 B2 molecule exhibits stimulated emission behavior and amplified spontaneous emission property, both of which have never been reported for pristine boron-doped π-systems, thus demonstrating the potential of BMCs as optical gain materials for laser cavities.

Long-term effect of transurethral partial cystectomy with a 2-micrometer continuous-wave laser for non-muscle-invasive bladder cancer.

Purpose: We have reported the efficacy and safety of 2-micrometer continuous-wave laser cystectomy of non-muscle invasive bladder tumor (NMIBC) (J Urol. 2009;182:66-9). In this study, we evaluated the long-term outcomes of patients with NMIBC who underwent transurethral partial cystectomy with a 2-micrometer continuous-wave laser, and explored the risk factors for tumor recurrence. Methods: This was a retrospective study of patients with NMIBC planned to undergo transurethral partial cystectomy with a 2-micrometer continuous-wave laser at the Fourth Medical Center of the PLA General Hospital between January 2012 and December 2014. The primary outcome was bladder cancer recurrence. Results: A total of 75 patients were enrolled. Sixty-two (82.7%) were male. The patients were 59.8 ± 12.9 years of age. The mean operation time was 38.7 ± 20.4 min. No Clavien grade >2 complications occurred. The duration of catheter indwelling was 3.6 ± 1.8 days. The hospital stay was 6.0 ± 2.3 days. The median follow-up was 80 months. A total of 17 patients had a recurrence during follow-up, and the recurrence-free survival (RFS) rate was 77.3%. In the multivariable analysis, the tumor risk group were independently associated with the recurrence of NMIBC (p = 0.026). Conclusions: After TURBT with a 2-micrometer continuous-wave laser, RFS was 77.3% at the median follow-up of 80 months. All complications were mild. Only tumor risk group was independently associated with the recurrence of NMIBC.

Synergistic effects of extrinsic photoconduction and photogating in a short-wavelength ZrS3 infrared photodetector.

Two-dimensional (2D) material-based photodetectors, especially those working in the infrared band, have shown great application potential in the thermal imaging, optical communication, and medicine fields. Designing 2D material photodetectors with broadened detection band and enhanced responsivity has become an attractive but challenging research direction. To solve this issue, we report a zirconium trisulfide (ZrS3) infrared photodetector with enhanced and broadened response with the assistance of the synergistic effects of extrinsic photoconduction and photogating effect. The ZrS3 photodetectors can detect infrared light up to 2 µm by extrinsic photoconduction and exhibit a responsivity of 100 mA W-1 under 1550 nm illumination. Furthermore, the ZrS3 infrared photodetectors with an oxide layer show a triple enhanced responsivity due to the photogating effect. Additionally, the infrared imaging capability of the ZrS3 infrared photodetectors is also demonstrated. This work provides a potential way to extend the response range and improve the responsivity for nanomaterial-based photodetectors at the same time.

Substrate engineering for wafer-scale two-dimensional material growth: strategies, mechanisms, and perspectives.

The fabrication of wafer-scale two-dimensional (2D) materials is a prerequisite and important step for their industrial applications. Chemical vapor deposition (CVD) is the most promising approach to produce high-quality films in a scalable way. Recent breakthroughs in the epitaxy of wafer-scale single-crystalline graphene, hexagonal boron nitride, and transition-metal dichalcogenides highlight the pivotal roles of substrate engineering by lattice orientation, surface steps, and energy considerations. This review focuses on the existing strategies and underlying mechanisms, and discusses future directions in epitaxial substrate engineering to deliver wafer-scale 2D materials for integrated electronics and photonics.

Distorted B/O-containing nanographenes with tunable optical properties.

We report the synthesis of two B/O-containing nanographenes, which feature the fusion of three or six planar B/O-heterocycles onto one hexa-peri-hexabenzocoronene π-framework. Incorporation of the B/O-heterocycles not only leads to distorted geometries, but also modulates the electronic structures and results in gradually red-shifted absorptions and fluorescence.

Diradical B/N-Doped Polycyclic Hydrocarbons.

Heterocyclic diradicaloids with atom-precise control over open-shell nature are promising materials for organic electronics and spintronics. Herein, we disclose quinoidal π-extension of a B/N-heterocycle for generating B/N-type organic diradicaloids. Two quinoidal π-extended B/N-doped polycyclic hydrocarbons that feature fusion of the B/N-heterocycle motif with the antiaromatic s-indacene or dicyclopenta[b,g]naphthalene core were synthesized. This quinoidal π-extension and B/N-heterocycle leads to their open-shell electronic nature, which stands in contrast to the multiple-resonance effect of conventional B/N-type emitters. These B/N-type diradicaloids have modulated (anti)aromaticity and enhanced diradical characters comparing with the all-carbon analogues, as well as intriguing properties, such as magnetic activities, narrow energy gaps and highly red-shifted absorptions. This study thus opens the new space for both of B/N-doped polycyclic π-systems and heterocyclic diradicaloids.

Differential diagnosis of Crohn's disease and intestinal tuberculosis: development and assessment of a nomogram prediction model.

BACKGROUND: China is a region with a high incidence of tuberculosis, and the incidence of IBD has also been rising rapidly in recent years. Differentiating Crohn's disease(CD) from intestinal tuberculosis (ITB) has become a very challenging issue. We aimed to develop and assess a diagnostic nomogram to differentiate between CD and ITB to improve the accuracy and practicability of the model. METHODS: A total of 133 patients (CD 90 and ITB 43) were analyzed retrospectively. Univariate and multivariate logistic regression analysis was included to determine the independent predictive factors and establish the regression equation. On this basis, the nomogram prediction model was constructed. The discrimination, calibration and clinical efficiency of the nomogram were assessed using area under the curve(AUC), C-index, calibration curve, decision curve analysis (DCA) and clinical impact curve. RESULTS: T-SPOT positive, cobblestone appearance, comb sign and granuloma were significant predictors in differentiating CD from ITB. Base on the above independent predictors, a diagnostic nomogram was successfully established. The sensitivity, specificity, accuracy of the prediction model are 94.4%, 93.0%, 94.0% respectively. The AUC and the C-index of the prediction model are both 0.988, which suggest that the model had a good discrimination power. The calibration curve indicated a high calibration degree of the prediction model. The DCA and clinical impact curve indicated a good clinical efficiency of the prediction model which could bring clinical benefits. CONCLUSION: A nomogram prediction model for distinguishing CD from ITB was developed and assessed, with high discrimination, calibration and clinical efficiency. It can be used as an accurate and convenient diagnostic tool to distinguish CD from ITB, facilitating clinical decision-making.

SRH suppressed P-G-I design for very long-wavelength infrared HgCdTe photodiodes.

The very long wavelength infrared (VLWIR, >14 µm) spectral band is an indispensable part of new-generation infrared remote sensing. Mercury cadmium telluride (HgCdTe or MCT) has shown excellent potential across the entire infrared band. However, the dark current, which is extremely sensitive to the technological level and small Cd composition, severely limits the performance of VLWIR HgCdTe photodiodes. In this study, cut-off wavelengths of up to 15 µm for HgCdTe devices with novel P-G-I (including wide bandgap p-type cap layer, grading layer and intrinsic absorption layer) designs have been reported. Compared with a device with a double-layer heterojunction (DLHJ) structure, the designed P-G-I structure successfully reduced dark current by suppressing the Shockley-Read-Hall process. Considering the balance of quantum efficiency and dark current, with the introduction of an approximately 0.8 µm thickness Cd composition grading layer, the device can achieve a high detectivity of up to 2.5×1011 cm Hz1/2 W-1. Experiments show that the P-G-I-T device has a lower dark current and a better SRH process suppressing ability than DLHJ devices, the measured detectivity achieved 8.7×1010 cm Hz1/2 W-1. According to additional research, the trap-assisted tunneling current is the primary component of the dark current. Controlling the trap concentration to as low as 1×1013 cm-3 will be continuous and meaningful work. The proposed study provides guidance for VLWIR HgCdTe photodetectors.

Ribbon-Type Boron-Doped Polycyclic Aromatic Hydrocarbons: Conformations, Dynamic Complexation and Electronic Properties.

Molecular ribbons (MRs), one-dimentional topological polycyclic aromatic hydrocarbons (PAHs), are of importance for synthetic chemistry and material sciences. Herein, we disclose an effective strategy to develop boron-doped MRs, i.e. photochemical cyclization on conjugated organoboranes for rapid π-extension. A series of ribbon-type boron-doped PAHs that own multiple cove edges were synthesized using Mallory photoreaction in solution. Two of them feature isomeric C68 B2 π-skeletons with 2.2 nm in length, thus representing a new kind of boron-doped MRs. The boron atoms endow them with sufficient Lewis acidity, and notably, the formed Lewis acid-base adducts based on boron-doped MRs display the photo-induced dual-dissociation behavior in the excited state and thus photochromism property. Moreover, despite of the highly contorted topological conformations, their potential utility as organic semiconductor was demonstrated by fabrication of organic field-effect transistors.

Tuning Diradical Properties of Boron-Containing π-Systems by Structural Isomerism.

Tuning diradical character is an important topic for organic diradicaloids. Herein, we report the precise borylation enabling structural isomerism as an effective strategy to modulate diradical character and thereby properties of organic diradicaloids. We synthesized a new B-containing polycyclic hydrocarbon that has the indeno[1,2-b]fluorene π-skeleton with the ß-carbons bonding to two boron atoms. Detailed theoretical and experimental results show that this bonding pattern leads to its distinctive electronic structures and properties in comparison to that of its isomeric molecule. This molecule has the efficient conjugation between boron atoms and π-skeleton, resulting in downshifted LUMO and HOMO levels. Moreover, it exhibits smaller diradical character and thereby inhibited diradical properties, such as significantly blue-shifted light absorption, larger energy bandgap and weak para-magnetic resonance. Notably, this B-containing polycyclic hydrocarbon possesses much stronger Lewis acidity and its Lewis acid-base adducts display enhanced diradical character, demonstrating the positive effects of Lewis coordination on modulating diradical performance.

Emerging Single-Photon Detectors Based on Low-Dimensional Materials.

Single-photon detectors (SPDs) that can sense individual photons are the most sensitive instruments for photodetection. Established SPDs such as conventional silicon or III-V compound semiconductor avalanche diodes and photomultiplier tubes have been used in a wide range of time-correlated photon-counting applications, including quantum information technologies, in vivo biomedical imaging, time-of-flight 3D scanners, and deep-space optical communications. However, further development of these fields requires more sophisticated detectors with high detection efficiency, fast response, and photon-number-resolving ability, etc. Thereby, significant efforts have been made to improve the performance of conventional SPDs and to develop new photon-counting technologies. In this review, the working mechanisms and key performance metrics of conventional SPDs are first summarized. Then emerging photon-counting detectors (in the visible to infrared range) based on 0D quantum dots, 1D quantum nanowires, and 2D layered materials are discussed. These low-dimensional materials exhibit many exotic properties due to the quantum confinement effect. And photodetectors built from these nD-materials (n = 0, 1, 2) can potentially be used for ultra-weak light detection. By reviewing the status and discussing the challenges faced by SPDs, this review aims to provide future perspectives on the research directions of emerging photon-counting technologies.

Antibacterial, antioxidant and biocompatible nanosized quercetin-PVA xerogel films for wound dressing.

Topical use of antimicrobial agents to treat wounds to inhibit bacterial invasion and facilitate wound healing is an effective strategy. In this work, an antibacterial xerogel film for potential applications in wound dressings was developed. First, a natural antibacterial agent, quercetin (Qu), was made into water-soluble quercetin-borate (QuB) nanoparticles by merging a solvent exchange method with the borate esterification reaction. QuB nanoparticles were then employed as the cross-linking agent to achieve gelation of poly(vinyl alcohol) (PVA) to obtain antimicrobial QuB-PVA composite microgels. Furthermore, QuB-PVA microgels were utilized as raw materials to produce xerogel films via an electrospray technique. The as-prepared QuB-PVA xerogel films exhibited excellent bacteriostasis, antioxidation, biocompatibility, self-healing, accelerated skin regeneration and functional restoration, and promoted skin wound healing. The QuB-PVA films significantly facilitated the in vivo healing speed of full-thickness skin wounds compared to commercial dressings. We believe that the present multifunctional QuB-PVA xerogel film is an excellent candidate for the wound dressings.

Boron-Containing Organic Diradicaloids: Dynamically Modulating Singlet Diradical Character by Lewis Acid-Base Coordination.

Organic diradicaloids have unique open-shell structures and properties and promising applications in organic electronics and spintronics. Incorporation of heteroatoms is an effective strategy to alter the electronic structures of organic diradicaloids. However, B-containing organic diradicaloids are very challenging due to their high reactivities, which are caused by not only diradical nature but also the B atom. In this article, we report a new kind of organic diradicaloids containing boron atoms. Our strategy is to incorporate planarized triarylboranes to antiaromatic polycyclic hydrocarbons (PHs). We synthesized two isomeric B-containing PHs composed of indenofluorene π-skeletons and two dioxa-bridged triphenylborane moieties. As proved by theoretical and experimental results, both of them have excellent ambient stability and open-shell singlet diradical structures, as well as intriguing magnetic and optoelectronic properties, such as thermally accessible triplet species, reversible multiredox ability, and narrow energy gaps. Notably, they possess sufficient Lewis acidity, which has never been observed for organic diradicaloids. In addition, they can coordinate with Lewis bases to form Lewis adducts, achieving unprecedented dynamic modulations of (anti)aromaticity and thus diradical character of organic diradicaloids.