Distribution of the active components from Xianglian Pill in tissues of healthy and antibiotic-associated diarrhea model mice and the mechanism study.

Antibiotic-associated diarrhea (AAD) is a common side effect of antibiotic therapy, characterized by intestinal inflammation which reduces the quality of life of patients. Xianglian Pill (XLP) has long been used to treat abdominal pain, diarrhea, bacillary dysentery and enteritis. Studies found that XLP has curative effect on AAD; however, the chemical constituents and mechanism of XLP have not been fully elucidated because of the lack of in vitro and in vivo studies. In this study, ultra-high performance liquid chromatography mass spectrometry method (UPLC-Q-Exactive-Orbitrap-HRMS) was used to examine the components of the XLP. Then, the binding between active compounds and the key targets was studied using network pharmacology and molecular docking. A comparative tissue distribution study was established for the simultaneous determination of the 10 active components in healthy and AAD mouse models. Forty-six components were characterized from XLP. According to the network pharmacology degree value, a prediction was made that encompassed 42 components and 14 core targets, which were intricately involved in crucial biological pathways, such as the AGE-RAGE signaling, cellular senescence, and MAPK signaling. Tissue distribution analysis showed that the 10 components were widely distributed in the heart, liver, spleen, lungs, kidneys, small intestine, and large intestine of mice, with varying concentrations in healthy and AAD mice. Molecular docking analysis also indicated that the active compounds in the tissue distribution could bind tightly to key targets of network pharmacological studies. This study provides a reference for further investigations of the relationships between the chemical components and pharmacological activities of XLP.

Comparative transcriptomic insights into molecular mechanisms of the susceptibility wheat variety MX169 response to Puccinia striiformis f. sp. tritici (Pst) infection.

Stripe rust of wheat is caused by the fungal pathogen Puccinia striiformis f. sp. tritici (Pst). Breeding durably resistant wheat varieties by disrupting the susceptibility (S) gene has an important impact on the control of wheat stripe rust. Mingxian169 (MX169) showed strong stripe rust susceptibility to all the races of Pst. However, molecular mechanisms and responsive genes underlying susceptibility of the wheat variety MX169 to Pst have not been elucidated. Here, we utilized next-generation sequencing technology to analyze transcriptomics data of "MX169" and high-resistance wheat "Zhong4" at 24, 48, and 120 h post-inoculation (hpi) with Pst. Comparative transcriptome analysis revealed 3,494, 2,831, and 2,700 differentially expressed genes (DEGs) at different time points. We observed an upregulation of DEGs involved in photosynthesis, flavonoid biosynthesis, pyruvate metabolism, thiamine metabolism, and other biological processes, suggesting their involvement in MX169's response to Pst. DEGs encoding transcription factors were also identified. Our study suggested the potential susceptibility gene resources in MX169 related to stripe rust response could be valuable for understanding the mechanisms involved in stripe rust susceptibility and for improving wheat resistance to Pst. IMPORTANCE: Our study suggests the potential susceptibility gene resources in MX169 related to stripe rust response could be valuable for understanding the mechanisms involved in stripe rust susceptibility and for improving wheat resistance to Pst.

Structural Optimization of Marine Natural Product Pretrichodermamide B for the Treatment of Colon Cancer by Targeting the JAK/STAT3 Signaling Pathway.

Marine natural product (MNP) pretrichodermamide B (Pre B, 9) was identified as a novel STAT3 inhibitor in our previous work, while its metabolic instability hindered its further development. To address this drawback, ligand structure-based drug design was adopted leading to a series of Pre B derivatives. Among them, MNP trichodermamide B (tri B, 24) obtained by skeletal rearrangement exhibited more potent antiproliferative activity with an IC50 value of 0.12 µM against HCT116. Notably, 24 stood out with improved metabolic stability (T1/2 = 31 min) and more favorable oral bioavailability (F = 37.5%). Further studies indicated that 24 blocked JAK/STAT3 signaling in dose- and time-dependent manner. In vivo, 24 suppressed tumor growth (TGI = 65%) at a dose of 20 mg/kg in a HCT116-derived xenograft mouse model. Overall, 24 might be a promising lead compound for colon cancer and is worthy of further investigation.

Resolving syntactic-semantic conflicts: comprehension and processing patterns by deaf Chinese readers.

The current study combined sentence plausibility judgment and self-paced reading tasks to examine the comprehension strategies and processing patterns of Chinese deaf individuals when comprehending written Chinese sentences with syntactic-semantic cue conflicts. Similar to findings from previous crosslinguistic studies on deaf readers, the Chinese deaf readers showed great variability in their comprehension strategies, with only 38% robustly relying on syntactic cues. Regardless of their overall comprehension preferences, the deaf readers all showed additional processing efforts as reflected by longer reading time at the verb regions when they relied on the syntactic cues. Those with less robust reliance on syntactic cues also showed longer reading time at the verb regions even when they relied on the semantic cues, suggesting sensitivity to the syntactic cues regardless of the comprehension strategy. These findings suggest that deaf readers in general endure more processing burden while resolving conflicting syntactic and semantic cues, likely due to their overall high reliance on semantic information during sentence comprehension. Increased processing burden thus may contribute to an overall tendency of over-reliance on semantic cues when comprehending sentences with cue conflicts.

Prelimbic cortical pyramidal neurons to ventral tegmental area projections promotes arousal from sevoflurane anesthesia.

AIMS: General anesthesia has been used in surgical procedures for approximately 180 years, yet the precise mechanism of anesthetic drugs remains elusive. There is significant anatomical connectivity between the ventral tegmental area (VTA) and the prelimbic cortex (PrL). Projections from VTA dopaminergic neurons (VTADA ) to the PrL play a role in the transition from sevoflurane anesthesia to arousal. It is still uncertain whether the prelimbic cortex pyramidal neuron (PrLPyr ) and its projections to VTA (PrLPyr -VTA) are involved in anesthesia-arousal regulation. METHODS: We employed chemogenetics and optogenetics to selectively manipulate neuronal activity in the PrLPyr -VTA pathway. Electroencephalography spectra and burst-suppression ratios (BSR) were used to assess the depth of anesthesia. Furthermore, the loss or recovery of the righting reflex was monitored to indicate the induction or emergence time of general anesthesia. To elucidate the receptor mechanisms in the PrLPyr -VTA projection's impact on anesthesia and arousal, we microinjected NMDA receptor antagonists (MK-801) or AMPA receptor antagonists (NBQX) into the VTA. RESULTS: Our findings show that chemogenetic or optogenetic activation of PrLPyr neurons prolonged anesthesia induction and promoted emergence. Additionally, chemogenetic activation of the PrLPyr -VTA neural pathway delayed anesthesia induction and promoted anesthesia emergence. Likewise, optogenetic activation of the PrLPyr -VTA projections extended the induction time and facilitated emergence from sevoflurane anesthesia. Moreover, antagonizing NMDA receptors in the VTA attenuates the delayed anesthesia induction and promotes emergence caused by activating the PrLPyr -VTA projections. CONCLUSION: This study demonstrates that PrLPyr neurons and their projections to the VTA are involved in facilitating emergence from sevoflurane anesthesia, with the PrLPyr -VTA pathway exerting its effects through the activation of NMDA receptors within the VTA.

Sevoflurane exerts antidepressant-like effects via the BDNF-TrkB pathway.

Depression has emerged as the predominant psychiatric affliction affecting individuals. Prior research has substantiated the antidepressant properties exhibited by numerous anesthetics. Sevoflurane, a widely utilized inhalant anesthetic in clinical practice, remains relatively uncharted in terms of its specific antidepressant effects. In this study, we used open field test, forced swimming test and novelty-suppressed feeding test to investigate the anxiety and depression-like behaviors in C57BL/6 mice following the inhalation of sevoflurane. We then used western blotting to scrutinized the expression levels of proteins associated with the brain-derived neurotrophic factor (BDNF)-tryosine receptor kinase B (TrkB) pathway in the hippocampus and prefrontal cortex. To further investigate whether sevoflurane exerts antidepressant-like effects via the BDNF-TrkB pathway, we downregulated TrkB expression by administering siRNA into the lateral ventricle. We found that the inhalation of 2.5 % sevoflurane exerted a significant antidepressant-like effect, accompanied by an elevation in p-TrkB expression levels in the hippocampus and prefrontal cortex. Intriguingly, this antidepressant-like effect was abrogated following the downregulation of TrkB expression through the microinjection of siRNA into the lateral ventricle. In conclusion, this study provides evidence supporting the notion that sevoflurane exerts its antidepressant-like effect via the BDNF-TrkB signaling pathway.

A flame-retardant and conductive fabric-based triboelectric nanogenerator: Application in fire alarm and emergency evacuation.

The fabrics commonly used in architectural decorative materials pose significant fire hazards due to their flammability and rapid fire spread. Moreover, the traditional fire-alarm systems may fail to function properly in complex fire environments owing to power supply disruptions. In this study, we developed a low-cost and eco-friendly flame-retardant conductive fabric-based triboelectric nanogenerator (FCF-TENG) by integrating flame-retardant conductive nylon fabric and polytetrafluoroethylene soaked cotton fabric. This nanogenerator exhibits excellent flame-retardant properties and remarkable energy-harvesting capabilities. The nylon fabric, treated with layer-by-layer self-assembly method, possesses outstanding self-extinguishing capability and melt-dripping resistance. Additionally, the electrical performance of FCF-TENG significantly improves, with a 10-fold boost in conductivity, and the open-circuit voltage increases by 84% to 92 V. Besides, by incorporating the rectifier circuit, the FCF-TENG is capable of completely charging a 1 µF capacitor within 30 s. Furthermore, the FCF-TENG was successfully applied as a self-powered sensor in the fire-alarm system and served as a safety exit indicator for evacuees and fire rescue. This work presents an effective and innovative application of multifunctional smart textiles for energy harvesting and self-powered sensing.

A deep lung cell atlas reveals cytokine-mediated lineage switching of a rare cell progenitor of the human airway epithelium.

The human airway contains specialized rare epithelial cells whose roles in respiratory disease are not well understood. Ionocytes express the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR), while chemosensory tuft cells express asthma-associated alarmins. However, surprisingly, exceedingly few mature tuft cells have been identified in human lung cell atlases despite the ready identification of rare ionocytes and neuroendocrine cells. To identify human rare cell progenitors and define their lineage relationship to mature tuft cells, we generated a deep lung cell atlas containing 311,748 single cell RNA-Seq (scRNA-seq) profiles from discrete anatomic sites along the large and small airways and lung lobes of explanted donor lungs that could not be used for organ transplantation. Of 154,222 airway epithelial cells, we identified 687 ionocytes (0.45%) that are present in similar proportions in both large and small airways, suggesting that they may contribute to both large and small airways pathologies in CF. In stark contrast, we recovered only 3 mature tuft cells (0.002%). Instead, we identified rare bipotent progenitor cells that can give rise to both ionocytes and tuft cells, which we termed tuft-ionocyte progenitor cells (TIP cells). Remarkably, the cycling fraction of these TIP cells was comparable to that of basal stem cells. We used scRNA-seq and scATAC-seq to predict transcription factors that mark this novel rare cell progenitor population and define intermediate states during TIP cell lineage transitions en route to the differentiation of mature ionocytes and tuft cells. The default lineage of TIP cell descendants is skewed towards ionocytes, explaining the paucity of mature tuft cells in the human airway. However, Type 2 and Type 17 cytokines, associated with asthma and CF, diverted the lineage of TIP cell descendants in vitro , resulting in the differentiation of mature tuft cells at the expense of ionocytes. Consistent with this model of mature tuft cell differentiation, we identify mature tuft cells in a patient who died from an asthma flare. Overall, our findings suggest that the immune signaling pathways active in asthma and CF may skew the composition of disease-relevant rare cells and illustrate how deep atlases are required for identifying physiologically-relevant scarce cell populations.

Host-parasite interactions: a study on the pathogenicity of different Mesanophrys sp. densities and hemocytes-mediated parasitic resistance of swimming crabs (Portunus trituberculatus).

Mesanophrys sp. is a parasitic ciliate that invades and destroys the hemocytes of the swimming crab (Portunus trituberculatus). In the present study, we employed an in vitro model to elucidate how Mesanophrys sp. destroys crab hemocytes. We also evaluated the relationship between the parasite's density, the destruction rate of the hemocytes, and the rapid proliferation pattern of parasites in host crabs. We found that the survival rate and cell integrity of crab hemocytes decreased with an increase in Mesanophrys sp. density, depicting a negative correlation between hemocyte viability and parasite density. Further analyses revealed that crab hemocytes could resist destruction by a low density (10 ind/mL) of Mesanophrys sp. for a long time (60 h). Mesanophrys sp. and its culture medium (containing the ciliate secretions) destroy the host hemocytes. The natural population growth rate of Mesanophrys sp. decreased with an increase in the parasite density, but the Mesanophrys sp. density did not affect the generation time of the parasites. In summary, Mesanophrys sp. can destroy crab hemocytes, and the degree of destruction is directly proportional to the parasite density. The resistance of crab hemocytes to Mesanophrys sp. decreased gradually with an increase in the parasite density.

Economic and Humanistic Burden of Osteoarthritis: An Updated Systematic Review of Large Sample Studies.

OBJECTIVE: A previous systematic literature review demonstrated a significant economic and humanistic burden on patients with osteoarthritis (OA). The aim of this study was to systematically review and update the burden of OA reported by large sample studies since 2016. METHODS: We searched Medline (via Ovid) and Embase using the updated search strategy based on the previous review. Those studies with a sample size ≥ 1000 and measuring the cost (direct or indirect) or health-related quality of life (HRQL) of OA were included. Pairs of reviewers worked independently and in duplicate. An arbitrator was consulted to resolve discrepancies between reviewers. The Kappa value was calculated to examine the agreement between reviewers. All costs were converted to 2021 US dollars according to inflation rates and exchange rates. RESULTS: A total of 1230 studies were screened by title and abstract and 159 by full text, and 54 studies were included in the review. The Kappa value for the full-text screening was 0.71. Total annual OA-related direct costs ranged from US$326 in Japan to US$19,530 in the US. Total annual all-cause direct costs varied from US$173 in Italy to US$41,433 in the US. The annual indirect costs ranged from US$736 in the US to US$18,884 in the Netherlands. Thirty-four studies reported HRQL, with EQ-5D (13, 38%) and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) (6, 18%) being the most frequently used instruments. The EQ-VAS and utility scores ranged from 41.5 to 81.7 and 0.3 to 0.9, respectively. The ranges of WOMAC pain (range 0-20, higher score for worse health), stiffness (range 0-8), and physical functioning (range 0-68) were 2.0-3.0, 1.0-5.0, and 5.8-42.8, respectively. CONCLUSION: Since 2016, the ranges of direct costs of OA became wider, while the HRQL of patients remained poor. More countries outside the US have published OA-related disease burden using registry databases.

Semi-rational design of an imine reductase for asymmetric synthesis of alkylated S-4-azepanamines.

Although imine reductase (IRED)-catalyzed reductive amination is promising for the synthesis of alkylated chiral amines, precisely regulating the stereoselectivity of IRED remains a great challenge. Herein, focusing on the residues directly in contact with the ketone moiety, we applied structure-guided semi-rational design to obtain the triple-mutant I149Y/L200H/W234K. This mutant showed high stereoselectivity, of up to >99% (S), toward reductive amination of N-Boc-4-oxo-azepane and different amines, and to the best of our knowledge is the first biocatalyst developed for asymmetric synthesis of chiral azepane-4-amines.

Identification of marine natural product Pretrichodermamide B as a STAT3 inhibitor for efficient anticancer therapy.

The Janus kinase (JAK)/signal transducer and activator of transcription 3 (STAT3) regulates the expression of various critical mediators of cancer and is considered as one of the central communication nodes in cell growth and survival. Marine natural products (MNP) represent great resources for discovery of bioactive lead compounds, especially anti-cancer agents. Through the medium-throughput screening of our in-house MNP library, Pretrichodermamide B, an epidithiodiketopiperazine, was identified as a JAK/STAT3 signaling inhibitor. Further studies identified that Pretrichodermamide B directly binds to STAT3, preventing phosphorylation and thus inhibiting JAK/STAT3 signaling. Moreover, it suppressed cancer cell growth, in vitro, at low micromolar concentrations and demonstrated efficacy in vivo by decreasing tumor growth in a xenograft mouse model. In addition, it was shown that Pretrichodermamide B was able to induce cell cycle arrest and promote cell apoptosis. This study demonstrated that Pretrichodermamide B is a novel STAT3 inhibitor, which should be considered for further exploration as a promising anti-cancer therapy. Supplementary Information: The online version contains supplementary material available at 10.1007/s42995-022-00162-x.

Single-Case Experimental Design in Rehabilitation: Basic Concepts, Advantages, and Challenges.

ABSTRACT: Single-case experimental design is a family of experimental methods that can be used to examine the efficacy of interventions by testing a small number of patients or cases. This article provides an overview of single-case experimental design research for use in rehabilitation as another option along with traditional group-based research when studying rare cases and rehabilitation interventions of unknown efficacy. Basic concepts related to single-case experimental design and the characteristics of common subtypes ( N-of-1 randomized controlled trial, withdrawal design, multiple-baseline design, multiple-treatment design, changing criterion/intensity design, and alternating treatment design) are introduced. The advantages and disadvantages of each subtype are discussed along with challenges in data analysis and interpretation. Criteria and caveats for interpreting single-case experimental design results and their use in evidence-based practice decisions are discussed. Recommendations are provided for appraising single-case experimental design articles as well as using single-case experimental design principles to improve real-world clinical evaluation.

Effects of solvent polarity on the novel excited-state intramolecular thiol proton transfer and photophysical property compared with the oxygen proton transfer.

Recently, the dual-fluorescent phenomena of excited state intramolecular thiol proton transfer (ESIPT) for 3-thiolflavone derivative (3NTF) were reported by Chou and coworkers for the first time [J. Am. Chem. Soc. 143 (2021) 12715-12724], which opened a new chapter in the field of ESIPT. Based on density functional theory (DFT) and time-dependent density functional theory (TDDFT), the proton transfer processes of 3NTF in toluene, dichloromethane and acetonitrile were studied. By optimizing the structure of the ground (S0) state and first excited (S1) state of 3NTF in different solvents, the hydrogen-bond parameters and proton-transfer potential energy curves were calculated. It was shown that although photo-excitation enhanced the intramolecular hydrogen bonding strength and thus promoted the occurrence of ESIPT, the solvent polarities inhibited the enhancement of the hydrogen bond of S1 state, which was not conducive to ESIPT. The electron spectra analyses were consistent with experimental data, which confirmed the rationality of molecular configurations. The time-evolved excited state dynamics simulation was performed based on the optimized structure of 3NTF, indicating that the ESIPT was an ultrafast photochemical reaction less than 180 fs. Moreover, we compared the potential energy surfaces of ESIPT, electronic structures based on natural transition orbitals (NTOs) method and electron-hole isosurfaces for the 3NTF and the traditional flavone molecule (3NHF), concluded that the unusually large Stokes shift fluorescence of 3NTF was mainly caused by the coupling of ESIPT and twisting intramolecular charge transfer (TICT), and the 3NTF isomer had the more nπ* character in the electron transition process. The nπ* ICT significantly increased with the decrease of solvent polarities, affecting the molecular photophysical properties, this made it more widely used in biomedical, photochemical, materials science and other fields.

Influence of Ambivalent Sexism on Intimate Partner Violence Tolerance and Mental Violence in a Chinese Female Sample: Relationship Causality Orientation as a Moderator.

The current study examined how the general tolerance of women's intimate partner violence and mental violence perpetration are affected by women's ambivalent sexism and relationship causality orientation. One hundred and forty-nine of 221 Chinese female participants recruited on an online platform were included in the final data analysis. The results showed that causality orientation plays a moderating role. Specifically, as controlled orientation increased, the relationship between hostile sexism and intimate partner violence tolerance became stronger. As the autonomous orientation increased, the relationship between benevolent sexism and intimate partner violence tolerance became weaker. Hostile sexism and controlled orientation positively predict women's mental violence perpetration.

Activation of the ERK1/2 pathway mediates the neuroprotective effect provided by calycosin treatment.

Calycosin is a natural product extracted from some plant families and exhibits various biological properties. But the effect of calycosin on cerebral ischemia-reperfusion injury has not been fully elucidated. In this study, the neuroprotective effect of calycosin treatment on the differentiated SH-SY5Y cells exposed to OGD was evaluated using MTT and flow cytometry. Rats that were pretreatment with calycosin were subjected to MCAO, neurological behavior scores and brain infarct volume were evaluated. The protein expression of pERK/ERK were assessed using Western blot. siRNA-pERK and U0126 were administered to investigate the impact of the ERK pathway on calycosin preconditioning. The results demonstrated the neuronal viability in the calycosin-treated SH-SY5Y cells increased significantly, and the rate of apoptosis decreased compared with the Oxygen-glucose deprivation only SH-SY5Y cells. Calycosin pretreatment reduced infarct volume and improved neurological outcome in rats subjected to MCAO. Administration of calycosin increased the ratio of pERK/ERK expression, which was down-regulated in ischemia-reperfusion group. Down-regulation of pERK/ERK significantly attenuated the neuroprotective effect induced by calycosin pretreatment in vitro and in vivo. We concluded calycosin treatment could induce a neuroprotective effect against ischemia, which was related to the regulation of the ERK1/2 pathway.

Assessing Macro Disease Index of Wheat Stripe Rust Based on Segformer with Complex Background in the Field.

Wheat stripe rust (WSR) is a foliar disease that causes destructive damage in the wheat production context. Accurately estimating the severity of WSR in the autumn growing stage can help to objectively monitor the disease incidence level of WSR and predict the nationwide disease incidence in the following year, which have great significance for controlling its nationwide spread and ensuring the safety of grain production. In this study, to address the low accuracy and the efficiency of disease index estimation by traditional methods, WSR-diseased areas are segmented based on Segformer, and the macro disease index (MDI) is automatically calculated for the measurement of canopy-scale disease incidence. The results obtained with different semantic segmentation algorithms, loss functions, and data sets are compared for the segmentation effect, in order to address the severe class imbalance in disease region segmentation. We find that: (1) The results of the various models differed significantly, with Segformer being the best algorithm for WSR segmentation (rust class F1 score = 72.60%), based on the original data set; (2) the imbalanced nature of the data has a significant impact on the identification of the minority class (i.e., the rust class), for which solutions based on loss functions and re-weighting of the minority class are ineffective; (3) data augmentation of the minority class or under-sampling of the original data set to increase the proportion of the rust class greatly improved the F1-score of the model (rust class F1 score = 86.6%), revealing that re-sampling is a simple and effective approach to alleviating the class imbalance problem. Finally, the MDI was used to evaluate the models based on the different data sets, where the model based on the augmented data set presented the best performance (R2 = 0.992, RMSE = 0.008). In conclusion, the deep-learning-based semantic segmentation method, and the corresponding optimization measures, applied in this study allow us to achieve pixel-level accurate segmentation of WSR regions on wheat leaves, thus enabling accurate assessment of the degree of WSR disease under complex backgrounds in the field, consequently providing technical support for field surveys and calculation of the disease level.

Dynamic changes of inducible nitric oxide synthase expression in rat's retina and its role on blood-retinal barrier injury after acute high intraocular pressure.

AIM: To clarify the role of inducible nitric oxide synthase (iNOS) in blood-retinal barrier (BRB) injury after acute high intraocular pressure (IOP) in rats. METHODS: Forty-two Sprague-Dawley (SD) rats were randomized into 7 groups [control (Cont), 3, 6, 12, 24, 48, and 72h, n=6]. Except Cont group, other groups' retina tissue was obtained at corresponding time points after a model of acute high IOP have been established in rats. The expression of iNOS and tight junction protein zonula occludens (ZO)-1 was detected by Western blotting. Evans blue (EB; 3% ) was injected into the great saphenous vein to detect the leakage of EB by spectrophotometer. Nine rats were divided into Cont, 6h, 12h groups, the expression of iNOS was localized by immunofluorescence. In order to verify the role of iNOS in the damage to BRB, thirty-six rats were randomly divided into 4 groups [Cont, Cont+inhibitor (Inh), 6h and 6h+Inh, n=9]. After treatment with the iNOS-specific inhibitor 1400W, the expression of iNOS and ZO-1 and the leakage of BRB were detected again. RESULTS: The immunofluorescence results showed that the expression of iNOS was observed in the Cont group and 6h group, but not in the 12h group. iNOS was mainly expressed in the retinal nerve fiber layer, ganglion cell layer and inner nuclear layer and that it did not colocalize with the retinal ganglion cell marker NeuN but was co-expressed with the vascular endothelial cell marker CD31. Western blotting showed that in the early period (3h, 6h) after acute high IOP, the expression of iNOS was upregulated, then the down-regulation of iNOS were tested in the follow-up timing spots. ZO-1 expression showed a continuous down-regulation after 6h. The quantitative results for EB showed that the amount of EB leakage began to increase at 3h after acute high IOP. At 6h, the leakage of EB was lower, but at 12h, the leakage of EB was highest, after which it gradually recovered but remained higher than that in the Cont group. The expression of iNOS was down-regulated after 1400W treatment. ZO-1 expression was not significantly changed in the Cont+Inh group and the 6h group, and significantly down-regulated in the 6h+Inh group, and the leakage of EB was significantly increased after 1400W treatment. CONCLUSION: These results suggest that the upregulation of iNOS expression in the early stage after acute high IOP may have a protective effect on BRB injury.

Evaluation of the effect of perioperative administration of S(+)-ketamine hydrochloride injection for postoperative acute pain in children: study protocol for a prospective, multicenter, randomized, open-label, parallel-group, pragmatic clinical trial.

BACKGROUND: Inadequate postoperative pain management increases the risk of adverse events after the surgery and aggressive perioperative pain prevention has both short-term and long-term benefits. S(+)-ketamine is an N-methyl-D-aspartic acid (NMDA) receptor antagonist with a strong analgesic effect and can significantly relieve postoperative acute pain and reduce opioid consumption. However, for children, it still needs to be confirmed by large sample clinical studies. METHODS: This is a pragmatic, randomized controlled trial which will evaluate the effect of perioperative administration of S(+)-ketamine hydrochloride injection for postoperative acute pain in children in a pragmatic clinical setting. A total of 3000 children (≤17 years old) undergoing surgery will be included in this protocol. Subjects will be randomized 2:1 to either receive S(+)-ketamine hydrochloride injection or conventional therapy without S(+)-ketamine during the entire perioperative period. The primary endpoints are the area under the receiver operating characteristic (ROC) curve of Face Legs Activity Cry and Consolability (FLACC, 0-7 years old) scale score or Numerical Rating Scale (NRS, 8-17 years old) score within 48 h after surgery, and the consumption of opioids within 48 h after surgery. The secondary endpoints include the time of first use of rescue analgesics after surgery, rescue analgesia rate within 48 h after surgery, anesthesia recovery time, incidence of emergency delirium (for 0-7 years old), changes of anxiety and depression scale scores at 48 h after surgery (for 8-17 years old), incidence of intraoperative adverse events (AEs), and incidence of postoperative AEs and pharmacoeconomic indicators. AEs and serious AEs were recorded to evaluate safety. DISCUSSION: This trial will be the first pragmatic clinical trial to prospectively assess the effect of perioperative administration of S(+)-ketamine hydrochloride injection for postoperative acute pain in children, which is of great significance to the continuous optimization of clinical anesthesia and analgesia programs for children. TRIAL REGISTRATION: This trial was registered in the U.S. National Institutes of Health ClinicalTrials.gov database ( http://clinicaltrials.gov ; Registration number: NCT04834427). Registered on 8 April 2021.

Actinomycetes-derived imine reductases with a preference towards bulky amine substrates.

Since imine reductases (IREDs) were reported to catalyze the reductive amination reactions, they became particularly attractive for producing chiral amines. Though diverse ketones and aldehydes have been proved to be excellent substrates of IREDs, bulky amines have been rarely transformed. Here we report the usage of an Increasing-Molecule-Volume-Screening to identify a group of IREDs (IR-G02, 21, and 35) competent for accepting bulky amine substrates. IR-G02 shows an excellent substrate scope, which is applied to synthesize over 135 amine molecules as well as a range of APIs' substructures. The crystal structure of IR-G02 reveals the determinants for altering the substrate preference. Finally, we demonstrate a gram-scale synthesis of an analogue of the API sensipar via a kinetic resolution approach, which displays ee >99%, total turnover numbers of up to 2087, and space time yield up to 18.10 g L-1 d-1.