Single-neuron analysis of axon arbors reveals distinct presynaptic organizations between feedforward and feedback projections.

The morphology and spatial distribution of axon arbors and boutons are crucial for neuron presynaptic functions. However, the principles governing their whole-brain organization at the single-neuron level remain unclear. We developed a machine-learning method to separate axon arbors from passing axons in single-neuron reconstruction from fluorescence micro-optical sectioning tomography imaging data and obtained 62,374 axon arbors that displayed distinct morphology, spatial patterns, and scaling laws dependent on neuron types and targeted brain areas. Focusing on the axon arbors in the thalamus and cortex, we revealed the segregated spatial distributions and distinct morphology but shared topographic gradients between feedforward and feedback projections. Furthermore, we uncovered an association between arbor complexity and microglia density. Finally, we found that the boutons on terminal arbors show branch-specific clustering with a log-normal distribution that again differed between feedforward and feedback terminal arbors. Together, our study revealed distinct presynaptic structural organizations underlying diverse functional innervation of single projection neurons.

[Plant biomass degradation by filamentous fungi and production of renewable chemicals: a review].

Plant biomass represents a vast resource of carbon. In China, it is estimated that 1 billion tons of biomass is available each year. The conversion of these biomass resources into bioethanol or other bio-based chemicals, if fully commercialized, may reduce at least 200 million tons of crude oil import. Therefore, bioethanol and bulk chemicals are the core components of the biomanufacturing using plant biomass as carbon sources. Since the foundation of Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences (TIB, CAS), we have proposed a strategy of "two replacements and one upgrade". Utilizing renewable carbon resources instead of non-renewable petrochemical resources to produce bulk chemicals is included in our strategy. It is a long-term effort for TIB to develop plant biomass biomanufacturing to produce renewable chemicals. Continuous and systematic research was carried out in these two fields, and significant progress has been made in the past 10 years since the foundation of TIB. Here we review the progress of TIB in this field, mainly focusing on fungal system, including the mechanism of cellulose degradation by filamentous fungi and the strategy of consolidated bioprocessing of biomass. Based on this, malic acid, fuel ethanol and other bulk chemicals were produced through one-step conversion of biomass. Besides, the commercial processes for production of bulk chemicals such as succinic and lactic acid from renewable carbon resources, which were developed by TIB, were also be discussed. These examples clearly demonstrated that bulk chemicals can be obtained from biomass instead of from petroleum. Research on plant biomass biotransformation and renewable chemicals production in TIB has provided an alternative route for the development of low-carbon bioeconomy in China, and will contribute to the goal of carbon neutralization of China.

Analyses and identification of the chemical constituents of the Uighur formulation Baixuan Xiatare using three-stage infrared spectroscopy.

OBJECTIVE: To identify, rapidly and accurately, the chemical composition of the traditional Uighur formulation Baixuan Xiatare (BXXTR-FU). METHODS: We investigated if application of three-stage infrared (IR) spectroscopy enabled identification of the main chemical constituents (and their origins) in BXXTR-FU. RESULTS: The characteristic peaks of herbal material(s) and BXXTR-FU were assigned. In Fourier transform-IR (FT-IR) spectroscopy of BXXTR-FU, peaks at 1616 and 1605 cm-1 of BXXTR-FU were considered to denote anthraquinones and their derivatives; 1066 cm-1 was regarded as the characteristic absorption peak of resin glycosides. In second-derivative IR (SD-IR) spectroscopy, the main carbonyl types of BXXTR-FU in the range 1743-1636 cm-1 were assigned: 1651 cm-1 belonged to the carbonyl stretching vibrations of flavonoids and chromones; 1717 cm-1 belonged to tannins; 1699 cm-1 belonged to carboxylic acids; 1636 cm-1 belonged to anthraquinones and their derivatives. SD-IR spectroscopy further confirmed that the characteristic absorption peaks at 1636, 1618 and 1603 cm-1 could be used as markers that BXXTR-FU contained anthraquinones and their derivatives. Synchronous 2D-IR correlation spectra of chemical groups further confirmed the results of FT-IR and SD-IR spectroscopy. CONCLUSION: Our study strongly supported the necessity and importance of three-stage IR spectroscopy owing to its rapid and accurate identification of herbal formulations.

Screening and Identification of the Main Metabolites of Schisantherin a In Vivo and In Vitro by Using UHPLC-Q-TOF-MS/MS.

Schisantherin A is an active ingredient originating from Schisandra chinensis (Turcz.) which has hepatoprotective and anti-oxidation activities. In this study, in vitro metabolisms investigated on rat liver microsomes (RLMs) and in vivo metabolisms explored on male Sprague Dawley rats of Schisantherin A were tested, respectively. The metabolites of Schisantherin A were identified using ultra-high-performance liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS). Based on the method, 60 metabolites were successfully identified and structurally characterized including 48 phase-I and 12 phase-II metabolites. Among the metabolites, 45 metabolites were reported for the first time. Moreover, 56 and eight metabolites were detected in urine and bile and 19 metabolites were identified in rats' plasma. It demonstrated that hepatic and extra-hepatic metabolic pathways were both involved in Schisantherin A biotransformation in rats. Five in vitro metabolites were structurally characterized for the first time. The results indicated that the metabolic pathways mainly include oxidation, reduction, methylation, and conjugation with glucuronide, taurine, glucose, and glutathione groups. This study provides a practical strategy for rapidly screening and identifying metabolites, and the results provide basic data for future pharmacological and toxicology studies of Schisantherin A and other lignin ingredients.

Comparative Proteomic Analysis of the Hevea brasiliensis Latex under Ethylene and Calcium Stimulation.

BACKGROUND: Calcium ions usually act as a second messenger in the signal transmission process and a major element required by plants. In Hevea, calcium ion could alleviate the negative effects of long-term ethylene application to a certain extent. However, the molecular mechanisms remain unclear. METHODS: Two-dimensional electrophoresis was used to determine the pattern of protein changes in latex after treatments with calcium and/or ethylene. Quantitative real-time polymerase chain reaction and Western blotting were used to determine the expression levels of some proteins and genes. STRING software was used to determine the protein-protein interaction network of the identified proteins. RESULTS: Comparative proteomics identified 145 differentially expressed proteins, which represented 103 unique proteins. The abundance change patterns of some proteins involved in signal transduction, rubber particle aggregation, and natural rubber biosynthesis were altered upon calcium stimulation. Quantitative real-time polymerase chain reaction analysis of 29 proteins showed that gene expression did not always maintain the same trend as protein expression. The increased enzyme activities of superoxide dismutase, ascorbate peroxidase, and glutathione reductase suggested that calcium can enhance the antistress ability of plants by increasing the activity of their antioxidant enzyme systems. CONCLUSION: These results supplement the rubber latex proteome, and provide evidence for investigating the molecular mechanisms by which calcium alleviates the negative effects of ethylene stimulation.

Mechanism Underlying Antitumor Effects of Sinomenine.

Sinomenine (SIN) is a bioactive alkaloid compound extracted from a Chinese medicinal plant Sinomenium acutum. It is a multitarget antitumor natural substance. Various mechanisms have been proposed for the antitumor effects of SIN, such as direct cytotoxicity, induction of apoptosis, sensitization attenuating radiotherapy and chemotherapy, reversal of drug resistance, resistance to distant metastasis, and antiangiogenesis. SIN can be used as a tumor cell killer and an adjuvant to radiotherapy and chemotherapy. However, recent studies are mostly limited to the basic experimental stage; no systematic clinical studies have yet been reported. Therefore, this paper aimed to review the mechanism underlying the antitumor effects of SIN by consulting relevant domestic and foreign studies and to provide a relevant reference for further development, use, and exploration of SIN.

Commonalities and characteristics of aqueous extracts from three Uighur medicines were analyzed by using three-stage infrared spectroscopy combined with ultra-performance liquid chromatography-time of flight-mass spectra.

OBJECTIVE: To establish a method to analyze the commonalities and characteristics of the aqueous extracts from three Uighur medicines. METHODS: In this study, a combination method of three-stage infrared spectroscopy and ultra-performance liquid chromatography-time of flight-mass spectra (UPLC-TOF-MS) method was used to analyze the commonalities and characteristics of the aqueous extracts from Hezi (Fructus Chebulae), Maohezi (Terminalia Belliricae Fructus) and Xiqingguo (Chebulae Fructus Immaturus). RESULTS: In Fourier transform-infrared spectroscopy spectra, all three samples showed the characteristic absorption peaks of tannins similarly. According to UPLC-TOF-MS data analysis, the difference of the position and intensity of the peaks at 1713- 1707 cm－1 in the three samples were related to the difference in the relative content of tannin and pentacyclic triterpenoids. In second derivative infrared spectroscopy spectra, further analysis of the differences in the infrared spectra of the three samples was performed by increasing the apparent resolution. In combination with UPLC-TOF-MS data, some features infrared absorption peaks were assigned. The absorption peaks at 1032-1030 cm－1 assigned to the stretching vibration of C-O-C (ether bond) groups of tannins; 1164-1163, 1063-1062, 1009- 1005, 904 cm－1 attributed to the stretching vibration of C-O (ether bond) and C-O-C groups of glycosidic compounds. 1385-1383 cm－1 attributed to the bending vibration of C-H (alkyl hydrogen) of methoxyl group of pentacyclic triterpenoids. Peak 835 cm－1 was assigned to the characteristic absorption peak of bending vibration of = C-H groups of pentacyclic triterpenoids. The two dimensional correlation infrared spectroscopy could further quickly distinguish three samples through the dynamic structural information of their chemical components and discrepancy of auto-peaks and cross- peaks intuitively in the range of 1720-419 cm－1. CONCLUSION: By comparing the intensity of the infrared characteristic absorption peaks, the main chemical components were assigned, which significantly enriched and perfected the data analysis of the infrared spectra of three Uighur Medicines. It provided a rationale for the identification of medicinal materials with complex and similar chemical components using the three-stage infrared spectroscopy and UPLC-TOF-MS.

Ma Huang Tang ameliorates bronchial asthma symptoms through the TLR9 pathway.

CONTEXT: Ma Huang Tang (MHT) has been used to treat influenza, fever, bronchial asthma, etc. as a traditional Chinese medication. However, the anti-inflammation mechanism of MHT remains unclear. OBJECTIVE: The study identifies the possible mechanisms of MHT on ovalbumin (OVA)-induced acute bronchial asthma in mice. MATERIALS AND METHODS: First, an asthma-related protein-protein interaction (PPI) network was constructed. And then, the acute bronchial asthma mice models were established by exposing to aerosolized 1% ovalbumin for 30 min/day for 1 week, and the mice were administered 2.0, 4.0, or 8.0 g/kg of MHT daily. To evaluate therapeutic effect, sensitization time, abdominal breathing time, eosinophils in bronchoalveolar lavage fluid, and tissue and trachea pathology were examined. Related genes were measured using RNA sequencing (RNA-seq). The expression levels of TLR9 in lung and trachea tissues were determined by immunohistochemical staining. RESULTS: MHT had a LD50 = 19.2 g/kg against asthma, while MHT at high doses (8 g/kg) effectively extended the sensitization time and abdominal breathing time and alleviated OVA-induced eosinophilic airway inflammation and mitigated pathological changes. The RNA-seq assay showed that the high-dose MHT resulted in a significant decrease in the levels of TLR9, TRAF6, TAB2, etc. in the lung tissue. Immunohistochemical assay confirmed the down-regulated of TLR9. Molecular docking revealed that six MHT compounds potentially mediated the TLR9 signaling pathway. DISCUSSION AND CONCLUSIONS: MHT could mitigate the pathological changes of acute asthma-like syndrome through inhibition of the TLR9 pathway. Results of this study may provide a reference for the development of a novel therapy for patients with allergic asthma.

Decryption of Active Constituents and Action Mechanism of the Traditional Uighur Prescription (BXXTR) Alleviating IMQ-Induced Psoriasis-Like Skin Inflammation in BALB/c Mice.

Bai Xuan Xia Ta Re Pian (BXXTR) is a traditional Uighur medicine ancient prescription in China widely used in the treatment of psoriasis, presenting a high curative rate and few side effects. Given that the active constituents and action mechanism still remain unclear, the aim of this study is to explore the potential active constituents and mechanism of antipsoriasis of BXXTR. Psoriasis-like lesions model in BALB/c mice was induced by Imiquimod (IMQ), including five treatment groups: control group, IMQ-treated group, IMQ-ACITRETIN group (Positive control group), IMQ-BXXTR low dose group, IMQ-BXXTR medium dose group and IMQ-BXXTR high dose group. The Psoriasis Area and Severity Index (PASI) score, skin and ear thickness, and histologic section were collected. The differentially expressed genes were determined by using RNAseq technology and the relevant pathways were analyzed by KEGG database. The ELISA kit and western blot assays were used to detect the related protein expression levels. In addition, the chemical constituents of BXXTR were determined by UPLC-TOF-MS analysis and the potential active constituents were predicted by SEA DOCK and Gene Ontology (GO). The data demonstrated that BXXTR significantly alleviated IMQ-induced psoriasis. RNA-seq analysis showed that BXXTR induced the expression levels of 31 genes; the KEGG analysis suggested that BXXTR could significantly change IL-17-related inflammatory pathways. The ELISA kit confirmed that the expression level of IL-17A protein was significantly reduced. 75 compounds of BXXTR were determined by UPLC-TOF-MS analysis, 11 of 75 compounds were identified as potential active compounds by similarity ensemble approach docking (SEA DOCK) and Gene Ontology (GO). BXXTR reduced the severity of skin lesions by inhibiting IL-17-related inflammatory pathways. The results indicated that BXXTR could suppress psoriasis inflammation by multiple-constituents-regulated multiple targets synergistically. Collectively, this study could provide important guidance for the elucidation of the active constituents and action mechanism of BXXTR for the treatment of psoriasis.

Ecdysterones from Rhaponticum carthamoides (Willd.) Iljin reduce hippocampal excitotoxic cell loss and upregulate mTOR signaling in rats.

Glutamate-induced excitotoxicity is a key pathological mechanism in many neurological disease states. Ecdysterones derived from Rhaponticum carthamoides (Willd.) Iljin (RCI) have been shown to alleviate glutamate-induced neuronal damage; although their mechanism of action is unclear, some data suggest that they enhance signaling in the mechanistic target of rapamycin (mTOR) signaling pathway. This study sought to elucidate the mechanisms underlying ecdysterone-mediated neuroprotection. We used in silico target prediction and simulation methods to identify putative ecdysterone binding targets, and to specifically identify those that represent nodes where several neurodegenerative diseases converge. We then used histological analyses in a rat hippocampal excitotoxicity model to test the effectiveness of ecdysterones in vivo. We found that RCI-derived ecdysterones should bind to glutamatergic NMDA-type receptors (NMDARs); specifically, in vivo modeling showed binding to the GRIN2B subunit of NMDARs, which was found also to be a node of convergence in several neurodegenerative disease pathways. Computerized network construction by using pathway information from the Kyoto Encyclopedia of Genes and Genomes (KEGG) database showed putative links between GRIN2B and mTOR pathway elements including phosphoinositide-3kinase (PI3K), mTOR, and protein kinase C (PKC); these elements are associated with neuronal survival. Brain tissue western blots of ecdysterone-treated rats showed upregulated PI3K, Akt, mTOR, and phosphorylated Akt and mTOR, and down regulated GRIN2B and the apoptotic enzyme cleaved caspase-3. Ecdysterone treatment also prevented glutamate-induced rat hippocampal cell loss. In summary, RCI-derived ecdysterones appear to prevent glutamatergic excitotoxicity by increasing mTOR/Akt/PI3K signaling activity.

Chondromalacia patellae treated by warming needle and rehabilitation training.

OBJECTIVE: To observe the effect of warming needle combined with rehabilitation training on chondromalacia patellae in a randomized controlled trial. METHODS: The 92 cases were randomly divided into a treatment group treated by warming needle plus rehabilitation training (47 cases) and a control group treated by medication plus rehabilitation training (45 cases), and the therapeutic effect was compared after 20 sessions. RESULTS: The pain was relieved more obviously in the treatment group than in the control group (P < 0.05), and the total effective rate was 91.8% and 71.1% respectively (P < 0.01). CONCLUSION: Warming needle plus rehabilitation training was superior in the therapeutic effect and duration of producing relief of pain to medication plus rehabilitation training in treating chondromalacia patellae.