Patch-seq: Advances and Biological Applications.

Multimodal analysis of gene-expression patterns, electrophysiological properties, and morphological phenotypes at the single-cell/single-nucleus level has been arduous because of the diversity and complexity of neurons. The emergence of Patch-sequencing (Patch-seq) directly links transcriptomics, morphology, and electrophysiology, taking neuroscience research to a multimodal era. In this review, we summarized the development of Patch-seq and recent applications in the cortex, hippocampus, and other nervous systems. Through generating multimodal cell type atlases, targeting specific cell populations, and correlating transcriptomic data with phenotypic information, Patch-seq has provided new insight into outstanding questions in neuroscience. We highlight the challenges and opportunities of Patch-seq in neuroscience and hope to shed new light on future neuroscience research.

The necessity of transperineal ultrasound view in urethra malacoplakia.

This case illustrates the untypical presentation of primary bladder malacoplakia. The patient was in her mid-50s have impaired immunity by the long-term hyperglycemic condition. She presented with symptoms of urinary tract infection and dysuria, and had multiple nodulars in bladder and significantly mass in urethra. Although the diagnosis of bladder malacoplakia was established on bladder biopsy, transperineal ultrasound examination can find its distinct clinical presentation.

Neurotransmitters and Motoneuron Contacts of Multifunctional and Behaviorally Specialized Turtle Spinal Cord Interneurons.

The spinal cord can appropriately generate diverse movements, even without brain input and movement-related sensory feedback, using a combination of multifunctional and behaviorally specialized interneurons. The adult turtle spinal cord can generate motor patterns underlying forward swimming, three forms of scratching, and limb withdrawal (flexion reflex). We previously described turtle spinal interneurons activated during both scratching and swimming (multifunctional interneurons), interneurons activated during scratching but not swimming (scratch-specialized interneurons), and interneurons activated during flexion reflex but not scratching or swimming (flexion reflex-selective interneurons). How multifunctional and behaviorally specialized turtle spinal interneurons affect downstream neurons was unknown. Here, we recorded intracellularly from spinal interneurons activated during these motor patterns in turtles of both sexes in vivo and filled each with dyes. We labeled motoneurons using choline acetyltransferase antibodies or earlier intraperitoneal FluoroGold injection and used immunocytochemistry of interneuron axon terminals to identify their neurotransmitter(s) and putative synaptic contacts with motoneurons. We found that multifunctional interneurons are heterogeneous with respect to neurotransmitter, with some glutamatergic and others GABAergic or glycinergic, and can directly contact motoneurons. Also, scratch-specialized interneurons are heterogeneous with respect to neurotransmitter and some directly contact motoneurons. Thus, scratch-specialized interneurons might directly excite motoneurons that are more strongly activated during scratching than forward swimming, such as hip-flexor motoneurons. Finally, and surprisingly, we found that some motoneurons are behaviorally specialized, for scratching or flexion reflex. Thus, either some limb muscles are only used for a subset of limb behaviors or some limb motoneurons are only recruited during certain limb behaviors.SIGNIFICANCE STATEMENT Both multifunctional and behaviorally specialized spinal cord interneurons have been described in turtles, but their outputs are unknown. We studied responses of multifunctional interneurons (activated during swimming and scratching) and scratch-specialized interneurons, filled each with dyes, and used immunocytochemistry to determine their neurotransmitters and contacts with motoneurons. We found that both multifunctional and scratch-specialized interneurons are heterogeneous with respect to neurotransmitter, with some excitatory and others inhibitory. We found that some multifunctional and some scratch-specialized interneurons directly contact motoneurons. Scratch-specialized interneurons may excite motoneurons that are more strongly activated during scratching than swimming, such as hip-flexor motoneurons, or inhibit their antagonists, hip-extensor motoneurons. Surprisingly, we also found that some motoneurons are behaviorally specialized, for scratching or for flexion reflex.

Rapid construction of cyclopenta[b]naphthalene frameworks from propargylic alcohol tethered methylenecyclopropanes.

We have developed a new synthetic methodology for the rapid construction of cyclopenta[b]naphthalene frameworks from the reaction of propargylic alcohol tethered methylenecyclopropanes with mesyl chloride in the presence of triethylamine through cascade cyclization. The reaction can be performed under mild conditions without the use of transition metals, affording the target products in moderate to good yields, and this cyclization reaction process can be scaled up to a gram-scale synthesis.

[Network Meta-analysis of Huoxue Huayu Chinese medicine injections on hypertensive nephropathy].

To assess the clinical efficacy of Huoxue Huayu Chinese medicine injections on hypertensive nephropathy by using Network Meta-analysis method. The relative randomized controlled trials(RCTs) of Huoxue Huayu Chinese medicine injections on hypertensive nephropathy were retrieved from CNKI, WanFang, VIP, SinoMed, PubMed, Cochrane Library and Web of Science in November 2019. Two researchers independently conducted literature screening, data extraction and quality evaluation. The data was analyzed by RevMan 5.3, Stata 15.1, ADDIS 1.16.8 software. Finally, 4 518 patients in 50 RCTs were included, involving 5 kinds of Huoxue Huayu Chinese medicine injections. The five Chinese medicine injections were Breviscapine Injection, Danhong Injection, Sodium Tanshinone Ⅱ_A Sulfonate Injection, Ginkgo Dipyridamole Injection, and Salvianolate Injection. Meta-analysis showed that, in terms of renal function protection, urine protein quantification and serum creatinine of each Chinese medicine injections combined with conventional Western medicine group, except Salvianolate Injection combined with conventional Western medicine group, was significantly lower than those of the conventional Western medicine treatment group. Ginkgo Dipyridamole Injection combined with conventional Western medicine had the best effect in reducing urine protein, and Danhong Injection combined with conventional Western medicine had the best effect in reducing serum creatinine. In terms of blood pressure control, the blood pressure of each Chinese medicine injections combined with conventional Western medicine group, except Breviscapine Injection combined with conventional Western medicine group, was significantly lower than that of conventional Western medicine treatment group. Salvianolate Injection combined with conventional Western medicine had the best effect in reducing blood pressure. In terms of safety, no serious adverse reactions were found in all studies. The five kinds of Huoxue Huayu Chinese medicine injections combined with conventional Western medicine had significantly advantages in the treatment of hypertensive nephropathy. Ginkgo Dipyridamole Injection combined with conventional Western medicine is most likely to be the optimal therapy. However, limited by the quantity and quality of the documents included, the conclusion of this study still needs to be verified by more high-quality, multi-center and large-sample RCT.

Lewis or Brønsted acid-catalysed reaction of propargylic alcohol-tethered alkylidenecyclopropanes with indoles and pyrroles for the preparation of polycyclic compounds tethered with indole or pyrrole motif.

We developed a facile synthetic method to access cyclopenta[b]naphthalene derivatives via the Lewis or Brønsted acid catalysed cascade nucleophilic addition, electronic cyclization, ring-opening rearrangement of propargylic alcohol-tethered alkylidenecyclopropanes with indole and pyrrole derivatives. The reaction exhibited a broad substrate scope and good functional group tolerance under metal-free conditions, affording the desired products in moderate to good yields.

Direct observation of a widely tunable bandgap in bilayer graphene.

The electronic bandgap is an intrinsic property of semiconductors and insulators that largely determines their transport and optical properties. As such, it has a central role in modern device physics and technology and governs the operation of semiconductor devices such as p-n junctions, transistors, photodiodes and lasers. A tunable bandgap would be highly desirable because it would allow great flexibility in design and optimization of such devices, in particular if it could be tuned by applying a variable external electric field. However, in conventional materials, the bandgap is fixed by their crystalline structure, preventing such bandgap control. Here we demonstrate the realization of a widely tunable electronic bandgap in electrically gated bilayer graphene. Using a dual-gate bilayer graphene field-effect transistor (FET) and infrared microspectroscopy, we demonstrate a gate-controlled, continuously tunable bandgap of up to 250 meV. Our technique avoids uncontrolled chemical doping and provides direct evidence of a widely tunable bandgap-spanning a spectral range from zero to mid-infrared-that has eluded previous attempts. Combined with the remarkable electrical transport properties of such systems, this electrostatic bandgap control suggests novel nanoelectronic and nanophotonic device applications based on graphene.

Rostral spinal cord segments are sufficient to generate a rhythm for both locomotion and scratching but affect their hip extensor phases differently.

Rostral segments of the spinal cord hindlimb enlargement are more important than caudal segments for generating locomotion and scratching rhythms in limbed vertebrates, but the adequacy of rostral segments has not been directly compared between locomotion and scratching. We separated caudal segments from immobilized low-spinal turtles by sequential spinal cord transections. After separation of the caudal four segments of the five-segment hindlimb enlargement, the remaining enlargement segment and five preenlargement segments still produced rhythms for forward swimming and both rostral and pocket scratching. The swimming rhythm frequency was usually maintained. Some animals continued to generate swimming and scratching rhythms even with no enlargement segments remaining, using only preenlargement segments. The preenlargement segments and rostral-most enlargement segment were also sufficient to maintain hip flexor (HF) motoneuron quiescence between HF bursts [which normally occurs during each hip extensor (HE) phase] during swimming. In contrast, the HF-quiescent phase was increasingly absent (i.e., HE-phase deletions) during rostral and pocket scratching. Moreover, respiratory motoneurons that normally burst during HE bursts continued to burst during the HF quiescence of swimming even with the caudal segments separated. Thus the same segments are sufficient to generate the basic rhythms for both locomotion and scratching. These segments are also sufficient to produce a reliable HE phase during locomotion but not during rostral or pocket scratching. We hypothesize that the rostral HE-phase interneurons that rhythmically inhibit HF motoneurons and interneurons are sufficient to generate HF quiescence during HE-biased swimming but not during the more HF-biased rostral and pocket scratching.

Toward Low-Voltage and High-Sensitivity Direct X-ray Detectors Based on Thick Bulk Heterojunction Organic Device.

Organic semiconducting materials are promising for the fabrication of flexible ionizing radiation detectors for imaging because of their tissue equivalence, simple large-scale processing, and mass production. However, it is challenging to achieve high-sensitivity detection for organic direct detectors prepared by low-cost solution processing because of the compromise between thickness and carrier transport. In this study, high-performance organic direct X-ray detectors were fabricated by building a micrometer-thick bulk heterojunction (BHJ) using poly(3-hexylthiophene-2,5-diyl) (P3HT):(6,6)-phenyl c71 butyric acid methyl ester. A 5 µm BHJ film was fabricated by drop-casting and enhanced crystallization of P3HT using binary solvents and high-boiling-point additives to improve the charge carrier mobility. Furthermore, this organic direct X-ray detector has a sensitivity of >654.26 µC Gyair s-1 and a self-powered response. Because of the architecture of the thick active layer and the energy cascade in this diode detector, it has a very low dark current of 46.26 pA at -2 V. A fast and efficient approach was developed for fabricating thick, highly mobile organic BHJ films for high-performance direct X-ray detectors. It has great potential for application in a new generation of flexible and portable large-area flat-panel detectors.

Multimodal Nature of the Single-cell Primate Brain Atlas: Morphology, Transcriptome, Electrophysiology, and Connectivity.

Primates exhibit complex brain structures that augment cognitive function. The neocortex fulfills high-cognitive functions through billions of connected neurons. These neurons have distinct transcriptomic, morphological, and electrophysiological properties, and their connectivity principles vary. These features endow the primate brain atlas with a multimodal nature. The recent integration of next-generation sequencing with modified patch-clamp techniques is revolutionizing the way to census the primate neocortex, enabling a multimodal neuronal atlas to be established in great detail: (1) single-cell/single-nucleus RNA-seq technology establishes high-throughput transcriptomic references, covering all major transcriptomic cell types; (2) patch-seq links the morphological and electrophysiological features to the transcriptomic reference; (3) multicell patch-clamp delineates the principles of local connectivity. Here, we review the applications of these technologies in the primate neocortex and discuss the current advances and tentative gaps for a comprehensive understanding of the primate neocortex.

Adult neurogenesis in the primate hippocampus.

Adult hippocampal neurogenesis (AHN) is crucial for learning, memory, and emotion. Deficits of AHN may lead to reduced cognitive abilities and neurodegenerative disorders, such as Alzheimer's disease. Extensive studies on rodent AHN have clarified the developmental and maturation processes of adult neural stem/progenitor cells. However, to what extent these findings apply to primates remains controversial. Recent advances in next-generation sequencing technologies have enabled in-depth investigation of the transcriptome of AHN-related populations at single-cell resolution. Here, we summarize studies of AHN in primates. Results suggest that neurogenesis is largely shared across species, but substantial differences also exist. Marker gene expression patterns in primates differ from those of rodents. Compared with rodents, the primate hippocampus has a higher proportion of immature dentate granule cells and a longer maturation period of newly generated granule cells. Future research on species divergence may deepen our understanding of the mechanisms underlying adult neurogenesis in primates.

Visible-light-induced reactions of methylenecyclopropanes (MCPs).

Diverse, visible-light-induced transformations of methylenecyclopropanes (MCPs) have been reported in recent years, attracting significant attention from synthetic chemists. As readily accessible strained molecules, MCPs have sufficient reactivity to selectively generate different target products, through reactions with various radical species upon visible-light irradiation under regulated reaction conditions. These transformations can be classified into three subcategories of reaction pathway, forming ring-opened products, cyclopropane derivatives, and alkynes. These products include pharmaceutical intermediates and polycyclic/heterocyclic compounds that are challenging to obtain using traditional methods. This review summarizes the recent advancements in this field.

Synthesis and Characterization of Photoswitchable Dithienylethene-Based Chiral Bisoxazoline Compounds with Bidirectional Visible-Light Control.

In this paper, we have successfully synthesized dithienylethene-based chiral bisoxazoline ligands with bidirectional photoswitching capabilities under visible light irradiation and proposed a strategy for adjusting the conjugation system length in sensitizer groups. The detailed experimental procedures and the characterization data are presented in the main text and the Supporting Information. Despite their moderate photoswitching rates, these ligands provide a promising approach towards developing fully visible light-responsive chiral catalysts.

A primate nigrostriatal atlas of neuronal vulnerability and resilience in a model of Parkinson's disease.

The degenerative process in Parkinson's disease (PD) causes a progressive loss of dopaminergic neurons (DaNs) in the nigrostriatal system. Resolving the differences in neuronal susceptibility warrants an amenable PD model that, in comparison to post-mortem human specimens, controls for environmental and genetic differences in PD pathogenesis. Here we generated high-quality profiles for 250,173 cells from the substantia nigra (SN) and putamen (PT) of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced parkinsonian macaques and matched controls. Our primate model of parkinsonism recapitulates important pathologic features in nature PD and provides an unbiased view of the axis of neuronal vulnerability and resistance. We identified seven molecularly defined subtypes of nigral DaNs which manifested a gradient of vulnerability and were confirmed by fluorescence-activated nuclei sorting. Neuronal resilience was associated with a FOXP2-centered regulatory pathway shared between PD-resistant DaNs and glutamatergic excitatory neurons, as well as between humans and nonhuman primates. We also discovered activation of immune response common to glial cells of SN and PT, indicating concurrently activated pathways in the nigrostriatal system. Our study provides a unique resource to understand the mechanistic connections between neuronal susceptibility and PD pathophysiology, and to facilitate future biomarker discovery and targeted cell therapy.

Synchrotron infrared measurements of protein phosphorylation in living single PC12 cells during neuronal differentiation.

Protein phosphorylation is a post-translational modification that is essential for the regulation of many important cellular activities, including proliferation and differentiation. Current techniques for detecting protein phosphorylation in single cells often involve the use of fluorescence markers, such as antibodies or genetically expressed proteins. In contrast, infrared spectroscopy is a label-free and noninvasive analytical technique that can monitor the intrinsic vibrational signatures of chemical bonds. Here, we provide direct evidence that protein phosphorylation in individual living mammalian cells can be measured with synchrotron radiation-based Fourier transform-infrared (SR-FT-IR) spectromicroscopy. We show that PC12 cells stimulated with nerve growth factor (NGF) exhibit statistically significant temporal variations in specific spectral features, correlating with changes in protein phosphorylation levels and the subsequent development of neuron-like phenotypes in the cells. The spectral phosphorylation markers were confirmed by bimodal (FT-IR/fluorescence) imaging of fluorescently marked PC12 cells with sustained protein phosphorylation activity. Our results open up new possibilities for the label-free real-time monitoring of protein phosphorylation inside cells. Furthermore, the multimolecule sensitivity of this technique will be useful for unraveling the associated molecular changes during cellular signaling and response processes.

Distributions of active spinal cord neurons during swimming and scratching motor patterns.

The spinal cord can generate motor patterns underlying several kinds of limb movements. Many spinal interneurons are multifunctional, contributing to multiple limb movements, but others are specialized. It is unclear whether anatomical distributions of activated neurons differ for different limb movements. We examined distributions of activated neurons for locomotion and scratching using an activity-dependent dye. Adult turtles were stimulated to generate repeatedly forward swimming, rostral scratching, pocket scratching, or caudal scratching motor patterns, while sulforhodamine 101 was applied to the spinal cord. Sulforhodamine-labeled neurons were widely distributed rostrocaudally, dorsoventrally, and mediolaterally after each motor pattern, concentrated bilaterally in the deep dorsal horn, the lateral intermediate zone, and the dorsal to middle ventral horn. Labeled neurons were common in all hindlimb enlargement segments and the pre-enlargement segment following swimming and scratching, but a significantly higher percentage were in the rostral segments following swimming than rostral scratching. These findings suggest that largely the same spinal regions are activated during swimming and scratching, but there are some differences that may indicate locations of behaviorally specialized neurons. Finally, the substantial inter-animal variability following a single kind of motor pattern may indicate that essentially the same motor output is generated by anatomically variable networks.

Intrinsic optical properties of vanadium dioxide near the insulator-metal transition.

We studied the insulator-metal transition (IMT) in single-domain, single crystalline vanadium dioxide (VO(2)) microbeams with infrared microspectroscopy. The unique nature of such samples allowed us to probe the intrinsic behavior of both insulating and metallic phases in the close vicinity of IMT, and investigate the IMT driven by either strain or temperature independently. We found that the VO(2) insulating band gap narrows rapidly upon heating, and the infrared response undergoes an abrupt transition at both strain- and temperature-induced IMT. The results are consistent with recent studies attributing the opening of VO(2) insulating band gap to a correlation-assisted Peierls transition.

Deciphering the core seed endo-bacteriome of the highland barley in Tibet plateau.

Highland barley (Hordeum vulgare var. nudum (L.) Hook.f., qingke) has unique physical and chemical properties and good potential for industrial applications. As the only crop that can be grown at high altitudes of 4200-4500 m, qingke is well adapted to extreme habitats at high altitudes. In this study, we analysed the seed bacterial community of 58 genotypes of qingke grown in different regions of Tibet, including qingke landraces, modern cultivars, and winter barley varieties, and characterised endophytic bacterial communities in seeds from different sources and the core endo-bacteriome of qingke. This study aim to provide a reference for the application of seed endophytes as biological inoculants for sustainable agricultural production and for considering microbe-plant interactions in breeding strategies. A total of 174 qingke seed samples from five main agricultural regions in Tibet were collected and subjected to investigation of endophytic endo-bacteriome using high-throughput sequencing and bioinformatics approaches. The phyla of endophytic bacteria in qingke seeds from different sources were similar; however, the relative proportions of each phylum were different. Different environmental conditions, growth strategies, and modern breeding processes have significantly changed the community structure of endophytic bacteria in seeds, among which the growth strategy has a greater impact on the diversity of endophytic bacteria in seeds. Seeds from different sources have conserved beneficial core endo-bacteriome. The core endo-bacteriome of qingke seeds dominated by Enterobacteriaceae may maintain qingke growth by promoting plant growth and assisting plants in resisting pests and diseases. This study reveals the core endo-bacteriome of qingke seeds and provides a basis for exploiting the endophytic endo-bacteriome of qingke seeds.

1,1-Dimethylhydrazine adsorption on intrinsic, vacancy, and N-doped graphene: a first-principle study.

Graphene and its derivatives have been widely used in the area of high-sensitivity sensing materials for small-molecule gases. Based on first-principle calculations, the present study systematically investigated the adsorption of 1,1-dimethylhydrazine on intrinsic graphene, graphene with vacancy defects, and nitrogen-doped graphene. The adsorption effects, when 1,1-dimethylhydrazine takes on different orientations, were determined separately, and the specific adsorption energy and charge transfer were calculated accordingly. The results reveal that the 1,1-dimethylhydrazine adsorption on intrinsic graphene, graphene with vacancy defects, and nitrogen-doped graphene falls into physical adsorption. Besides, both vacancy defect and nitrogen doping help enhance the adsorption but the effect of vacancy defect is superior to that of nitrogen doping.

Routes and rates of bacterial dispersal impact surface soil microbiome composition and functioning.

Recent evidence suggests that, similar to larger organisms, dispersal is a key driver of microbiome assembly; however, our understanding of the rates and taxonomic composition of microbial dispersal in natural environments is limited. Here, we characterized the rate and composition of bacteria dispersing into surface soil via three dispersal routes (from the air above the vegetation, from nearby vegetation and leaf litter near the soil surface, and from the bulk soil and litter below the top layer). We then quantified the impact of those routes on microbial community composition and functioning in the topmost litter layer. The bacterial dispersal rate onto the surface layer was low (7900 cells/cm2/day) relative to the abundance of the resident community. While bacteria dispersed through all three routes at the same rate, only dispersal from above and near the soil surface impacted microbiome composition, suggesting that the composition, not rate, of dispersal influenced community assembly. Dispersal also impacted microbiome functioning. When exposed to dispersal, leaf litter decomposed faster than when dispersal was excluded, although neither decomposition rate nor litter chemistry differed by route. Overall, we conclude that the dispersal routes transport distinct bacterial communities that differentially influence the composition of the surface soil microbiome.