Typical antibiotic resistance genes and their association with driving factors in the coastal areas of Yangtze River Estuary.

The massive use of antibiotics has led to the escalation of microbial resistance in aquatic environment, resulting in an increasing concern regarding antibiotic resistance genes (ARGs), posing a serious threat to ecological safety and human health. In this study, surface water samples were collected at eight sampling sites along the Yangtze River Estuary. The seasonal and spatial distribution patterns of 10 antibiotics and target genes in two major classes (sulfonamides and tetracyclines) were analyzed. The findings indicated a high prevalence of sulfonamide and tetracycline resistance genes along the Yangtze River Estuary. Kruskal-Wallis analysis revealed significant seasonal variations in the abundance of all target genes. The accumulation of antibiotic resistance genes in the coastal area of the Yangtze River Estuary can be attributed to the influence of urban instream runoff and the discharge of effluents from wastewater treatment plants. ANISOM analysis indicated significant seasonal differences in the microbial community structure. VPA showed that environmental factors contribute the most to ARG variation. PLS-PM demonstrate that environmental factors and microbial communities pose direct effect to ARG variation. Analysis of driving factors influencing ARGs in this study may shed new insights into the mechanism of the maintenance and propagation of ARGs.

Facile Synthesis of Dual-Network Polymer Hydrogels with Anti-Freezing, Highly Conductive, and Self-Healing Properties.

We report the synthesis of poly(acrylamide-co-acrylic acid)/sodium carboxy methyl cellulose (PAMAA/CMC-Na) hydrogels, and subsequent fabrication of dual-network polymer hydrogels (PAMAA/CMC-Na/Fe) using as-prepared via the salt solution (FeCl3) immersion method. The created dual-network polymer hydrogels exhibit anti-swelling properties, frost resistance, high conductivity, and good mechanical performance. The hydrogel swells sightly when immersed in solution (pH = 2~11). With the increase in nAA:nAM, the modulus of elasticity experiences a rise from 1.1 to 1.6 MPa, while the toughness undergoes an increase from 0.18 to 0.24 MJ/m3. Furthermore, the presence of a high concentration of CMC-Na also contributes to the enhancement of mechanical strength in the resulting hydrogels, ascribing to enhanced physical network of the hydrogels. The minimum freezing point reaches -21.8 °C when the CMC-Na concentration is 2.5%, owing to the dissipated hydrogen bonds by the coordination of Fe3+ with carboxyl (-COO-) in CMC-Na and PAMAA. It is found that the conductivity of the PAMAA/CMC-Na/Fe hydrogels gradually decreased from 2.62 to 0.6 S/m as the concentration of CMC-Na rises. The obtained results indicates that the dual-network hydrogels with high mechanical properties, anti-swelling properties, frost resistance, and electrical conductivity can be a competitive substance used in the production of bendable sensors and biosensors.

Schisandrin B from Schisandra chinensis alleviated pain via glycine receptors, Nav1.7 channels and Cav2.2 channels.

ETHNOPHARMACOLOGICAL RELEVANCE: Schisandra chinensis, the dried and ripe fruit of the magnolia family plant Schisandra chinensis (Turcz.) Baill, was commonly used in traditional analgesic prescription. Studies have shown that the extract of Schisandra chinensis (SC) displayed analgesic activity. However, the analgesic active component and the exact mechanisms have yet to be revealed. AIM OF THE STUDY: The present study was to investigate the anti-nociceptive constituent of Schisandra chinensis, assess its analgesic effect, and explore the potential molecular mechanisms. MATERIALS AND METHODS: The effects of a series of well-recognized compounds from SC on glycine receptors were investigated. The analgesic effect of the identified compound was evaluated in three pain models. Mechanistic studies were performed using patch clamp technique on various targets expressed in recombinant cells. These targets included glycine receptors, Nav1.7 sodium channels, Cav2.2 calcium channels et al. Meanwhile, primary cultured spinal dorsal horn (SDH) neurons and dorsal root ganglion (DRG) neurons were also utilized. RESULTS: Schisandrin B (SchB) was a positive allosteric modulator of glycine receptors in spinal dorsal horn neurons. The EC50 of SchB on glycine receptors in spinal dorsal horn neurons was 2.94 ± 0.28 µM. In three pain models, the analgesic effect of SchB was comparable to that of indomethacin at the same dose. Besides, SchB rescued PGE2-induced suppression of α3 GlyR activity and alleviated persistent pain. Notably, SchB could also potently decrease the frequency of action potentials and inhibit sodium and calcium channels in DRG neurons. Consistent with the data from DRG neurons, SchB was also found to significantly block Nav1.7 sodium channels and Cav2.2 channels in recombinant cells. CONCLUSION: Our results demonstrated that, Schisandrin B, the primary lignan component of Schisandra chinensis, may exert its analgesic effect by acting on multiple ion channels, including glycine receptors, Nav1.7 channels, and Cav2.2 channels.

Inhibition of CISD1 attenuates cisplatin-induced hearing loss in mice via the PI3K and MAPK pathways.

Cisplatin is an effective chemotherapeutic drug for different cancers, but it also causes severe and permanent hearing loss. Oxidative stress and mitochondrial dysfunction in cochlear hair cells (HCs) have been shown to be important in the pathogenesis of cisplatin-induced hearing loss (CIHL). CDGSH iron sulfur domain 1 (CISD1, also known as mitoNEET) plays a critical role in mitochondrial oxidative capacity and cellular bioenergetics. Targeting CISD1 may improve mitochondrial function in various diseases. However, the role of CISD1 in cisplatin-induced ototoxicity is unclear. Therefore, this study was performed to assess the role of CISD1 in cisplatin-induced ototoxicity. We found that CISD1 expression was significantly increased after cisplatin treatment in both HEI-OC1 cells and cochlear HCs. Moreover, pharmacological inhibition of CISD1 with NL-1 inhibited cell apoptosis and reduced mitochondrial reactive oxygen species accumulation in HEI-OC1 cells and cochlear explants. Inhibition of CISD1 with small interfering RNA in HEI-OC1 cells had similar protective effects. Furthermore, NL-1 protected against CIHL in adult C57 mice, as evaluated by the auditory brainstem response and immunofluorescent staining. Mechanistically, RNA sequencing revealed that NL-1 attenuated CIHL via the PI3K and MAPK pathways. Most importantly, NL-1 did not interfere with the antitumor efficacy of cisplatin. In conclusion, our study revealed that targeting CISD1 with NL-1 reduced reactive oxygen species accumulation, mitochondrial dysfunction, and apoptosis via the PI3K and MAPK pathways in HEI-OC1 cell lines and mouse cochlear explants in vitro, and it protected against CIHL in adult C57 mice. Our study suggests that CISD1 may serve as a novel target for the prevention of CIHL.

Coupling of Slack and NaV1.6 sensitizes Slack to quinidine blockade and guides anti-seizure strategy development.

Quinidine has been used as an anticonvulsant to treat patients with KCNT1-related epilepsy by targeting gain-of-function KCNT1 pathogenic mutant variants. However, the detailed mechanism underlying quinidine's blockade against KCNT1 (Slack) remains elusive. Here, we report a functional and physical coupling of the voltage-gated sodium channel NaV1.6 and Slack. NaV1.6 binds to and highly sensitizes Slack to quinidine blockade. Homozygous knockout of NaV1.6 reduces the sensitivity of native sodium-activated potassium currents to quinidine blockade. NaV1.6-mediated sensitization requires the involvement of NaV1.6's N- and C-termini binding to Slack's C-terminus and is enhanced by transient sodium influx through NaV1.6. Moreover, disrupting the Slack-NaV1.6 interaction by viral expression of Slack's C-terminus can protect against SlackG269S-induced seizures in mice. These insights about a Slack-NaV1.6 complex challenge the traditional view of 'Slack as an isolated target' for anti-epileptic drug discovery efforts and can guide the development of innovative therapeutic strategies for KCNT1-related epilepsy.

Schisandrin B, a dual positive allosteric modulator of GABAA and glycine receptors, alleviates seizures in multiple mouse models.

Epilepsy is a prevalent and severe neurological disorder and approximately 30% of patients are resistant to existing medications. It is of utmost importance to develop alternative therapies to treat epilepsy. Schisandrin B (SchB) is a major bioactive constituent of Schisandra chinensis (Turcz.) Baill and has multiple neuroprotective effects, sedative and hypnotic activities. In this study, we investigated the antiseizure effect of SchB in various mouse models of seizure and explored the underlying mechanisms. Pentylenetetrazole (PTZ), strychnine (STR), and pilocarpine-induced mouse seizure models were established. We showed that injection of SchB (10, 30, 60 mg/kg, i.p.) dose-dependently delayed the onset of generalized tonic-clonic seizures (GTCS), reduced the incidence of GTCS and mortality in PTZ and STR models. Meanwhile, injection of SchB (30 mg/kg, i.p.) exhibited therapeutic potential in pilocarpine-induced status epilepticus model, which was considered as a drug-resistant model. In whole-cell recording from CHO/HEK-239 cells stably expressing recombinant human GABAA receptors (GABAARs) and glycine receptors (GlyRs) and cultured hippocampal neurons, co-application of SchB dose-dependently enhanced GABA or glycine-induced current with EC50 values at around 5 µM, and application of SchB (10 µM) alone did not activate the channels in the absence of GABA or glycine. Furthermore, SchB (10 µM) eliminated both PTZ-induced inhibition on GABA-induced current (IGABA) and strychnine (STR)-induced inhibition on glycine-induced current (Iglycine). Moreover, SchB (10 µM) efficiently rescued the impaired GABAARs associated with genetic epilepsies. In addition, the homologous mutants in both GlyRs-α1(S267Q) and GABAARs-α1(S297Q)ß2(N289S)γ2L receptors by site-directed mutagenesis tests abolished SchB-induced potentiation of IGABA and Iglycine. In conclusion, we have identified SchB as a natural positive allosteric modulator of GABAARs and GlyRs, supporting its potential as alternative therapies for epilepsy.

TOM40 mediates the effect of TSPO on postpartum depression partially through regulating calcium homeostasis in microglia.

AIMS: To assess the effect of the translocator protein 18 kDa (TSPO) on postpartum depression and explore its mechanism. METHODS: Postpartum depression (PPD) mouse model was established, and flow cytometry, immunofluorescence, Western blot analysis, real-time quantitative PCR, adeno-associated virus (AAV), co-immunoprecipitation-mass spectrometry and immunofluorescence co-staining were used to detect the effect of TSPO ligand ZBD-2 on PPD mice. RESULTS: ZBD-2 inhibits the overactivation of microglia in the hippocampus and amygdala of PPD model mice. ZBD-2 not only inhibited the inflammation but also repressed the burst of reactive oxygen species (ROS) and mitochondrial ROS (mtROS). Meanwhile, ZBD-2 protects mitochondria from LPS-induced damages through inhibiting the influx of calcium. ZBD-2 modulated the calcium influx by increasing the level of translocase of the outer mitochondrial membrane 40 (TOM40) and reducing the interaction of TSPO and TOM40. In addition, the effect of ZBD-2 was partially dependent on anti-oxidative process. Knockdown of TOM40 by adeno-associated virus (AAV) in the hippocampus or amygdala dramatically reduced the effect of ZBD-2 on PPD, indicating that TOM40 mediates the effect of ZBD-2 on PPD. CONCLUSIONS: TOM40 is required for the effect of ZBD-2 on treating anxiety and depression in PPD mice. This study reveals the role of microglia TSPO in PPD development and provides the new therapeutic strategy for PPD.

Occurrence of sulfonamides and tetracyclines in the coastal areas of the Yangtze River (China) Estuary.

Antibiotics have attracted global attention due to the ecological risks to environment. In this paper, solid-phase extraction and ultra-performance liquid chromatography triple quadrupole mass spectrometry (LC-MS/MS) were utilized to analyze the fugitive characteristics of 10 antibiotics of sulfonamides (sulfadiazine, sulfamethazine, sulfadimidine, sulfathiazole, sulfapyridine, sulfamethoxazole) and tetracyclines (tetracycline, oxytetracycline, chlortetracycline and doxycycline) in the coastal waters and surfece sediments of the Yangtze River Estuary and the ecological risks of antibiotics in water were estimated using ecological risk assessment method. The results have showed that 7 of the 10 antibiotics were detected in the water, with total concentrations ranging from 0.652 to 434.47 ng/L. 8 antibiotics were detected in the sediment, with total concentrations ranging from 0.091 to 499.23 ng/g. The main antibiotic species detected in the sediment and water varied seasonally. Higher concentrations in spatially distributed areas where rivers meet and where human activities have a more significant impact. The ecological risks were found to be higher in spring and autumn than those in winter and summer. Spatial variation in individual microbial communities was not evident in the sediments. The relationship between antibiotics and microorganisms in the environment was predominantly positive. Physical and chemical factors were significantly correlated for both antibiotics and microbial communities. This study can provide research ideas for other types of antibiotics and provide a basis for the prevention of antimicrobial resistance (AMR).

[Transmission and Growth Characteristics of Severe PM2.5 Pollution Events from 2013 to 2021 in Xingtai, Hebei].

The correlation between the growth rate of PM2.5 with transport source, atmospheric circulation, and wind field were analyzed, focusing on the severe and above pollution process (SAAP) in Xingtai, Hebei Province from 2013 to 2021. The results showed that from 2013 to 2021, a total of 164 pollution processes and 103 SAAP occurred in Xingtai. In the ground circulation, although the probability occurrence of the inverted trough was low, the probability of pollution was the highest (61.1%), followed by the high-pressure control type (>50.0%). In the 500 hPa, the control of the straight westerly wind belt had the highest probability of severe and above pollution (20.7%), followed by the post-trough type (16.1%), with the highest occurrence frequency. In SAAP, the distribution of the PM2.5 hourly growth rate (ΔPM2.5) was mainly concentrated between ±150 µg·(m3·h)-1, and the PM2.5 hourly growth rate was positive (+ΔPM2.5), contributing 61.7%. Among them, the average proportion of explosive growth was 13.9% (from 2013 to 2021), and the overall trend was decreasing annually. In the full wind speed, in terms of occurrence frequency and pollution probability, north-east (NE) was the wind direction most closely related to air pollution, especially severe and above pollution. The mean value of ΔPM2.5 in SAAP was lower than that of quiet wind in most wind directions. However, in some of the east-north (EN) and south-west (SW) wind direction intervals, the mean ΔPM2.5 in moderate wind speed was significantly higher than that of quiet wind (related to pollution transmission). The impact of larger wind speed on ΔPM2.5 was more complicated. The backward trajectories showed that the backward trajectories of slow, rapid, and explosive growth in SAAP could be divided into three main paths:west-north, east-north, and south. With the acceleration of the growth rate, the proportion of the west-north air mass gradually increased. The humidity (RH) of the slow-growth air mass was relatively large (more than 80% RH>50%), the relative humidity of the rapidly growing air mass was relatively concentrated (mainly distributed in 35%-55%), and the proportion of low-humidity (<50%) air masses increased significantly (by approximately 63%) in the explosive growth. The simulation analysis showed that the types of SAAP pollution could be divided into five categories:local accumulation, east-northern transmission, north-west transmission, mixed transmission, and south transmission. Among them, the proportion of mixed transmission was the highest, followed by that of the north-west transmission. The high and low-altitude configurations with the highest occurrence probability among the southerly transmission, the local accumulation type, and the north-easterly transport type were all high-altitude trough rear type combined with ground equalization field type. Among the north-westerly type, the high-pressure on the ground with the behind trough on high-altitude had the highest probability of occurrence. In mixed transmission, the probabilities of various circulation ratios were relatively balanced.

Benign vs malignant vertebral compression fractures with MRI: a comparison between automatic deep learning network and radiologist's assessment.

OBJECTIVE: To test the diagnostic performance of a deep-learning Two-Stream Compare and Contrast Network (TSCCN) model for differentiating benign and malignant vertebral compression fractures (VCFs) based on MRI. METHODS: We tested a deep-learning system in 123 benign and 86 malignant VCFs. The median sagittal T1-weighted images (T1WI), T2-weighted images with fat suppression (T2WI-FS), and a combination of both (thereafter, T1WI/T2WI-FS) were used to validate TSCCN. The receiver operator characteristic (ROC) curve was analyzed to evaluate the performance of TSCCN. The accuracy, sensitivity, and specificity of TSCCN in differentiating benign and malignant VCFs were calculated and compared with radiologists' assessments. Intraclass correlation coefficients (ICCs) were tested to find intra- and inter-observer agreement of radiologists in differentiating malignant from benign VCFs. RESULTS: The AUC of the ROC plots of TSCCN according to T1WI, T2WI-FS, and T1WI/T2WI-FS images were 99.2%, 91.7%, and 98.2%, respectively. The accuracy of T1W, T2WI-FS, and T1W/T2WI-FS based on TSCCN was 95.2%, 90.4%, and 96.2%, respectively, greater than that achieved by radiologists. Further, the specificity of T1W, T2WI-FS, and T1W/T2WI-FS based on TSCCN was higher at 98.4%, 94.3%, and 99.2% than that achieved by radiologists. The intra- and inter-observer agreements of radiologists were 0.79-0.85 and 0.79-0.80 for T1WI, 0.65-0.72 and 0.70-0.74 for T2WI-FS, and 0.83-0.88 and 0.83-0.84 for T1WI/T2WI-FS. CONCLUSION: The TSCCN model showed better diagnostic performance than radiologists for automatically identifying benign or malignant VCFs, and is a potentially helpful tool for future clinical application. CLINICAL RELEVANCE STATEMENT: TSCCN-assisted MRI has shown superior performance in distinguishing benign and malignant vertebral compression fractures compared to radiologists. This technology has the value to enhance diagnostic accuracy, sensitivity, and specificity. Further integration into clinical practice is required to optimize patient management. KEY POINTS: • The Two-Stream Compare and Contrast Network (TSCCN) model showed better diagnostic performance than radiologists for identifying benign vs malignant vertebral compression fractures. • The processing of TSCCN is fast and stable, better than the subjective evaluation by radiologists in diagnosing vertebral compression fractures. • The TSCCN model provides options for developing a fully automated, streamlined artificial intelligence diagnostic tool.

A worm-inspired robot based on origami structures driven by the magnetic field.

With the rapid development of origami technology, worm-inspired origami robots have attracted tremendous interest owing to their colourful locomotion behaviours, such as creeping, rolling, climbing and obstacle crossing. In the present study, we aim to engineer a worm-inspired robot based on knitting process with paper, which could realize complicated functions associated large deformation and exquisite locomotion patterns. At first we fabricate the backbone of the robot by using the paper-knitting technique. The experiment shows that the backbone of the robot can endure significant deformation during the tension, compression and bending process, and this feature ensures it can achieve the desired targets of motion. Next, the magnetic forces and torques under the actuation of permanent magnets are analysed, which are just the driving forces of the robot. We then consider three formats of motion on the robot, i.e. the inchworm motion, the Omega motion, and the hybrid motion. Typical examples for the robot fulfil desired tasks are given, including sweeping obstacles, climbing the wall and delivering cargoes. Detailed theoretical analyses and numerical simulations are performed to illustrate these experimental phenomena. The results show that the developed origami robot is equipped with such characteristics as lightweight and great flexibility, which is sufficiently robust in various environments. These promising performances shed new light on design and fabrication of bio-inspired robots with good intelligence.

Spatiotemporal distributions of sulfonamide and tetracycline resistance genes and microbial communities in the coastal areas of the Yangtze River Estuary.

In this paper, water and sediments were sampled at eight monitoring stations in the coastal areas of the Yangtze River Estuary in summer and autumn 2021. Two sulfonamide resistance genes (sul1 and sul2), six tetracycline resistance genes (tetM, tetC, tetX, tetA, tetO, and tetQ), one integrase gene (intI1), 16 S rRNA genes, and microbial communities were examined and analyzed. Most resistance genes showed relatively higher abundance in summer and lower abundance in autumn. One-way analysis of variance (ANOVA) showed significant seasonal variation of some ARGs (7 ARGs in water and 6 ARGs in sediment). River runoff and WWTPs are proven to be the major sources of resistance genes along the Yangtze River Estuary. Significant and positive correlations between intI1 and other ARGs were found in water samples (P < 0.05), implying that intI1 may influence the spread and propagation of resistance genes in aquatic environments. Proteobacteria was the dominant phylum along the Yangtze River Estuary, with an average proportion of 41.7%. Redundancy analysis indicated that the ARGs were greatly affected by temperature, dissolved oxygen, and pH in estuarine environments. Network analysis showed that Proteobacteria and Cyanobacteria were the potential host phyla for ARGs in the coastal areas of the Yangtze River Estuary.

The Modified Assessment Tool Based on Scapular Y-View for Global Fatty Infiltration in the Supraspinatus Muscle: Correlation, Predictive Performance, and Reliability Analyses.

BACKGROUND: The accurate evaluation of rotator cuff (RC) fatty degeneration after tears is critical for appropriate surgical decision making and prognosis. However, there is currently no reliable and practical tool to reflect the global fatty infiltration (Global-FI) throughout the 3-dimensional (3D) volumetric RC muscles. PURPOSE: (1) To determine the correlations between 2 modified assessment tools and the Global-FI and their predictive performances and reliabilities for Global-FI prediction, and (2) to compare these predictive parameters with those of the conventional tool using a single scapular Y-view slice. STUDY DESIGN: Cohort study (diagnosis); Level of evidence, 3. METHODS: A total of 49 patients with full-thickness RC tears scheduled to undergo arthroscopic repairs were included, and their surgical shoulders underwent 6-point Dixon magnetic resonance imaging preoperatively. The Global-FI was measured by calculating the 3D-volumetric fat fraction (FF) of the whole supraspinatus muscle through all acquired oblique sagittal slices. As a commonly used radiological landmark, the scapular Y-view was used to evaluate single-plane fatty infiltration (Y-FI) by calculating the FF in 1 slice, defined as the conventional assessment tool. Two modified assessment tools expand the analytic imaging by integrating the FFs from the scapular Y-view slice and its neighboring slices, which were calculated by averaging the FFs of these 3 slices (meanY3-FI) and accumulating local 3D-volumetric FFs from 3 slices (volY3-FI), respectively. The correlations between 3 assessment tools and the Global-FI were analyzed, and the predictive performance for Global-FI prediction using these tools was determined by goodness of fit and agreement. Moreover, the inter- and intraobserver reliabilities of these assessment tools were evaluated. Similar analyses were performed in the small-medium, large, or massive tear subgroups. RESULTS: The Y-FI was significantly higher than the Global-FI in all cases and tear size subgroups, while the 2 modified assessment tools (meanY3-FI and volY3-FI) did not significantly differ from the Global-FI. All assessment tools were significantly correlated with the Global-FI, but the meanY3-FI and volY3-FI showed stronger correlations than the Y-FI, which was also determined in different tear sizes. Moreover, the regression models of the meanY3-FI and volY3-FI showed superior goodness of fit to Y-FI in Global-FI prediction in all cases and subgroups, with larger coefficients of determination (R2) and smaller root mean square errors. The predicted Global-FI using the regression model of volY3-FI had the best agreement with the measured Global-FI, followed by the meanY3-FI, both showing smaller biases and standard deviation of the percentage difference between predicted- and measured Global-FI than the conventional Y-FI. In addition, the 2 modified assessment tools achieved better interobserver and intraobserver reliabilities than the conventional tool in all cases and subgroups. CONCLUSION: Two modified assessment tools (meanY3-FI and volY3-FI) were comparable with the Global-FI of the whole supraspinatus muscle, showing stronger correlations with the Global-FI and better predictive performances and reliabilities than the conventional tool (Y-FI). Moreover, the volY3-FI was slightly superior to meanY3-FI in the predictive performance and reliability.

NAN-190, a 5-HT1A antagonist, alleviates inflammatory pain by targeting Nav1.7 sodium channels.

AIMS: In the present study, NAN-190 [1-(2-methoxyphenyl)-4-[4-(2-phthalimido) butyl] piperazine] was identified as a Nav1.7 blocker. In the meantime, the compound could alleviate the Complete Freund's Adjuvant (CFA)-induced inflammatory pain. To understand the molecular mechanisms of NAN-190 on pain, the effect of NAN-190 on Nav1.7 sodium channels was studied. MAIN METHODS: Inflammatory pain was induced by injection of CFA solution into the plantar side of the left hindpaw. Thermal hyperalgesia and mechanical allodynia were measured. Whole-cell patch clamp methods were used to record sodium channels and other pain-related targets in the cultured recombinant cells and dorsal root ganglion neurons. KEY FINDINGS: Nan-190 was identified as an inhibitor of Nav1.7 sodium channels and animal experiments showed that NAN-190 significantly alleviated CFA-induced inflammatory pain. Mechanism studies demonstrated that NAN-190 was a state-dependent Nav1.7 blocker with IC50 value on the inactivated state ten-fold more potent than that on the rest state. NAN-190 leftward-shifted the fast and slow inactivation curves about 9.07 mV and 38.56 mV, respectively, but had no effects on channel activation. The compound also slowed the recovery from fast and slow inactivation and showed use-dependent properties. Further, the site-directed mutagenesis experiments demonstrated that NAN-190 mainly worked on the open state of Nav1.7 channels by interacting with sites similar as local anesthetics. In DRG neurons, NAN-190 mainly blocks TTX-sensitive currents but is less sensitive to TTX-R sodium currents. SIGNIFICANCE: Taken together, our results indicated that NAN-190 alleviated pain behaviors by blocking sodium channels by interacting with the open state.

Phytol from Faeces Bombycis alleviated migraine pain by inhibiting Nav1.7 sodium channels.

ETHNOPHARMACOLOGICAL RELEVANCE: Faeces Bombycis (silkworm excrement, called Cansha in Chinese), is the dried faeces of the larvae of silkworm. According to the theories of traditional Chinese medicine recorded in "Compendium of Materia Medica", Faeces Bombycis has often been prescribed in traditional Chinese medicine for the treatment of recurrent headache, rheumatalgia, rubella and itching et al. However, the bioactive components and their exact mechanisms underlying the pain-relieving effects remain to be revealed. AIM OF THE STUDY: The present study aimed to evaluate the analgesic effect of Faeces Bombycis extract (FBE) on migraine, explore the main active constituents and investigate the pharmacological mechanisms for its pain relief. MATERIALS AND METHODS: The bioactivity of different extracts from Faeces Bombycis was tracked by the nitroglycerin (NTG)-induced migraine model on rats and identified by NMR spectroscopic data. Whole-cell patch clamp technique, an electrophysiological method, was used to screen the potential targets and study the mechanism of action for the bioactive compound. The following targets have been screened and studied, including Nav1.7 sodium channels, Nav1.8 sodium channels, TRPV1 channels and TRPA1 channels. The trigeminal ganglion neurons were further used to study the effects of the identified compound on neuronal excitability. RESULTS: By testing the bioactivity of the different extracts proceedingly, fraction petroleum ether showed higher anti-migraine activity. Through further step-by-step isolations, 7 compounds were isolated. Among them, phytol was identified with the highest yield and displayed a potent anti-migraine effect. By screening the potential ion channel targets for migraine, phytol was found to preferentially block the inactivated state of Nav1.7 sodium channels with half-inhibition concentration 0.32 ± 0.05 µM. Thus, the effects of phytol on the biophysical properties of Nav1.7 sodium channels were further characterized. Phytol induced a hyperpolarizing shift of voltage-dependent inactivation and slowed the recovery from inactivation. The affinity of phytol became weaker in the inactivation-deficient Nav1.7 channels (Nav1.7-WCW). And such an effect was independent on the local anesthetic site (Nav1.7 F1737A). Consistent with the data from recombinant channels, the compound also displayed state-dependent inhibition on neuronal sodium channels and further decreased the neuronal excitability in trigeminal ganglion neurons. Moreover, besides Nav1.7 channel, phytol also antagonized the activation of TRPV1 and TRPA1 channels at micromolar concentrations with a weaker affinity. CONCLUSION: Our results demonstrated that phytol is the major anti-migraine ingredient of Faeces Bombycis and alleviates migraine behaviors by acting on Nav1.7 sodium channels in the trigeminal ganglion neurons. This study provided evidences for the therapeutic application of Faeces Bombycis and phytol on migraine disease.

[Distributions of Antibiotic Resistance Genes and Microbial Communities in the Nearshore Area of the Yangtze River Estuary].

Due to the extensive use of antibiotics, the resistance of microorganisms to antibiotics in the environment is increasing, and the problem of antibiotic resistance genes (ARGs) is becoming more and more severe, which seriously threatens ecological security and human health. In order to study the distribution characteristics of ARGs and the microbial community in different media in the coastal area of the Yangtze River Estuary, water and sediment samples from eight sites were collected through a field investigation. Two sulfonamide resistance genes ([STBX]sul1, sul2[STBZ]) and six tetracycline resistance genes (tetM, tetC, tetX, tetA, tetO, and tetQ), one integrase gene intI[STBX]1[STBZ], 16S rRNA gene, and the microbial community were detected and analyzed. The results showed that the detection rate of 10 resistance genes in the coastal area of the Yangtze River Estuary was 100%. [STBX]intI1[STBZ] was significantly positively correlated with various ARGs in the water samples. Proteobacteria and Bacteroidota were the dominant bacteria phyla in the water environment of the Yangtze River Estuary. Chloroplast was the main bacteria genus in water, and Chloroplast and Nitrospira were the main bacteria genera in sediment. In water, Nitrospirota was the common potential host of four tetracycline resistance genes (tetX, tetA, tetO, and tetQ). In sediments, Sva0485 was a potential host community shared by [STBX]sul1 and intI1[STBZ]. The distribution of the microbial community was an important factor affecting the migration and transformation of ARGs in the nearshore area of the Yangtze River Estuary.

The rs41291957 polymorphism of miR-143/145 and cancer risk: a case-control study and meta-analysis.

Recently, the rs41291957 polymorphism in the promoter region of miR-143/145 has been repeatedly investigated for its contribution to cancer susceptibility. However, the results remain conflicting rather than conclusive, which calls for further investigations. Therefore, we here conducted a case-control study and meta-analysis to explore the association between rs41291957 and cancer risk. In the case-control study, a total of 2277 cancer patients (lung, liver, gastric and colorectal cancers) and 800 normal controls were recruited, the genotyping of rs41291957 was performed with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and Sanger sequencing. In the meta-analysis, 5 previously published studies and our present study were included, the STATA 14.0 software was applied to conduct all statistical analyses. The results of case-control study showed that rs41291957 was significantly associated with the risk of gastric cancer, colon cancer, rectal cancer, and colorectal cancer in Hubei Han Chinese population. The results of meta-analysis demonstrated that rs41291957 was significantly associated with overall cancer risk, especially colorectal cancer risk and lung cancer risk. Collectively, the rs41291957 polymorphism of miR-143/145 may be a plausible susceptible locus for cancer risk, which should be validated in future studies with larger samples in different ethnic populations.

Development and comparison of 68Ga/18F/64Cu-labeled nanobody tracers probing Claudin18.2.

Claudin 18.2 (CLDN18.2) is an emerging target for the treatment of gastric cancers. We aim to develop tracers to image the expression of CLDN18.2. A humanized nanobody targeting CLDN18.2 (clone hu19V3) was produced and labeled with 68Ga, 64Cu, and 18F. The tracers were investigated in subcutaneous and metastatic models established using two different mouse types (nude and Balb/c mice) and two different cell lines (CHO-CLDN18.2 and CT26-CLDN18.2). Gastric cancer patient-derived xenograft (PDX) models were further established for validation experiments. Three novel CLDN18.2-targeted tracers (i.e., [68Ga]Ga-NOTA-hu19V3, [64Cu]Cu-NOTA-hu19V3, and [18F]F-hu19V3) were developed with good radiochemical yields and excellent radiochemical purities. [68Ga]Ga-NOTA-hu19V3 immuno-positron emission tomography (immunoPET) rapidly delineated subcutaneous CHO-CLDN18.2 lesions and CT26-CLDN18.2 tumors, as well as showing excellent diagnostic value in PDX models naturally expressing CLDN18.2. While [68Ga]Ga-NOTA-hu19V3 had high kidney accumulation, [64Cu]Cu-NOTA-hu19V3 showed reduced kidney accumulation and improved image contrast at late time points. Moreover, [18F]F-hu19V3 was developed via click chemistry reaction under mild conditions and precisely disseminated CHO-CLDN18.2 lesions in the lungs. Furthermore, region of interest analysis, biodistribution study, and histopathological staining results correlated well with the in vivo imaging results. Taken together, immunoPET imaging with the three tracers can reliably visualize CLDN18.2 expression.

Diagnosis and Treatment of Acute Pleural Effusion following Radioiodine Remnant Ablation Post Lobectomy for Thyroid Cancer.

Radioiodine remnant ablation (RRA) was previously demonstrated to be a safe and effective alternative to completion thyroidectomy for patients with differentiated thyroid cancer (DTC). However, its side effects have not been fully investigated, particularly in patients with lobectomy. We reported a young euthyroidal female who underwent RRA post lobectomy and lymph node dissection for papillary thyroid cancer, whose post-ablation 131I-whole-body scan accidentally showed diffuse radioiodine distribution on chest-mimicking pulmonary metastases. Immediately-added single-photon emission computed tomography/computed tomography (SPECT/CT), nevertheless, revealed a 131I-accumulating swollen left thyroid lobe and emerging pleural effusion, which relieved after short-term treatment with prednisone. In summary, acute pleural effusion ascribed to RRA-induced thoracic duct compression was reported for the first time. 131I-lobectomy-induced pleural effusion could be precisely diagnosed by SPECT/CT and efficiently manipulated via treating radiation thyroiditis with the short-term administration of corticosteroid.

ERBB2 as a prognostic biomarker correlates with immune infiltrates in papillary thyroid cancer.

Epidermal growth factor receptor 2 (ERBB2) is commonly over-expressed in advanced or metastatic tissues of papillary thyroid cancer (PTC) with poor prognosis, while it remains unknown whether ERBB2 plays a role in the progression of PTC. Thus, we analyzed the data derived from online repositories, including TCGA, KEGG, GO, GeneMANIA, and STRING, to explore the relationship between ERBB2 expression and prognosis, tumor phenotypes of interest, and immune infiltrates in PTC. Compared to normal thyroid tissue, ERBB2 was up-regulated in PTC samples (p < 0.001); In comparison with the group with low expression of ERBB2, the group with high expression of ERBB2 had poorer progression-free interval in stage III/IV patients (p = 0.008) and patients aged >45 years (p = 0.019). The up-regulated ERBB2 was associated with iodine metabolism dysfunction, proliferation, metastasis, angiogenesis, and drug resistance. The expression of ERBB2 negatively correlated with enrichment scores of B cells (r = -0.176, p < 0.001), CD8+ T cells (r = -0.160, p < 0.001), cytotoxic cells (r = -0.219, p < 0.001), NK CD56dim cells (r = -0.218, p < 0.001), plasmacytoid dendritic cells (r = -0.267, p < 0.001), T cells (r = -0.164, p < 0.001), T follicular helper cells (r = -0.111, p = 0.012), gamma delta T cells (r = -0.105, p = 0.017), and regulatory T cells (r = -0.125, p = 0.005). In conclusion, ERBB2 may serve as a prognostic biomarker and an immunotherapeutic target in PTC, deserving further exploration.