Rhizosphere Mortierella strain of alfalfa exerted weed growth inhibition by inducing expression of plant hormone-related genes.

Introduction: Weeds are significant factors that detrimentally affect crop health and hinder optimal herbage yield. Rhizosphere microorganisms play crucial roles in plant growth, development, and nutrient uptake. Therefore, research focusing on weed control through the lens of microorganisms has emerged as a prominent area of study. The oil-producing fungus Mortierella, which is known for its numerous agricultural benefits, has garnered significant attention in recent years. Methods: In this study, we conducted inoculation experiments in a controlled artificial culture climate chamber to investigate the effects of differential hormones and differentially expressed genes in the stems and leaves of Digitaria sanguinalis using Liquid Chromatography Tandem Mass Spectrometry and RNA-seq techniques, respectively. Additionally, Pearson's correlation analysis was used to establish correlations between differential hormones and growth indicators of Digitaria sanguinalis. Results and discussion: The results demonstrated that inoculation with Mortierella sp. MXBP304 effectively suppressed aboveground biomass and plant height in Digitaria sanguinalis. Furthermore, there was significant upregulation and downregulation in the expression of genes involved in the synthesis and metabolism of phenylalanine and L-phenylalanine. Conversely, the expression of genes related to tryptophan, L-tryptophan, and indole was significantly downregulated. The addition of Mortierella sp. MXBP304 can influence the gene expression associated with phenylalanine and tryptophan synthesis and metabolism during Digitaria sanguinalis growth, subsequently reducing the relative contents of phenylalanine and tryptophan, thereby directly inhibiting Digitaria sanguinalis growth.

Are recyclable plastics eco-friendly? Recycled PVC microplastics show higher toxicity than pristine PVC on Vallisneria natans, regardless of Cadmium.

As environmental awareness increases, the use of recyclable plastics has risen. However, it is currently unclear whether recycled microplastics (MPs) pose a lesser or greater environmental risk than pristine MPs. Cadmium (Cd), known for its toxicity to most organisms, can bind with MPs and accumulate in sediments. Few studies have explored the environmental risks posed by the coexistence of recycled MPs and pristine MPs with Cd to rooted macrophytes. We investigated the effects of recycled PVC MPs (R-PVC-MPs) and pristine PVC MPs (PVC-MPs) on Vallisneria natans in the presence and absence of Cd. Results showed that at moderate and high Cd levels, R-PVC-MPs reduced plant Cd enrichment. Despite this, the fresh weight of V. natans exposed to R-PVC-MPs was significantly lower than those exposed to PVC-MPs. Furthermore, R-PVC-MPs had more negative impacts on the physiological traits of V. natans than PVC-MPs, as chlorophyll was significantly reduced across all Cd levels. At high Cd levels, both R-PVC-MPs and PVC-MPs caused significantly high oxidative stress, with no significant differences observed. The PCoA plot showed that different MPs cause noticeable variations within the same Cd concentration. The trait network diagrams illustrated strong interactions among traits, with R-PVC-MPs showing the highest complexity. Lower average degree and decreased edge density indicate that traits of plants with R-PVC-MPs addition are more independent of each other. Our findings suggest that recycled PVC MPs pose a greater environmental risk than pristine PVC MPs, offering reference for assessing the risks of recycled plastics in freshwater ecosystems.

The antibodies against the A137R protein drive antibody-dependent enhancement of African swine fever virus infection in porcine alveolar macrophages.

African swine fever virus (ASFV) is the causative agent of African swine fever (ASF), a highly contagious disease that can kill up to 100% of domestic pigs and wild boars. It has been shown that the pigs inoculated with some ASF vaccine candidates display more severe clinical signs and die earlier than do pigs not immunized. We hypothesize that antibody-dependent enhancement (ADE) of ASFV infection may be caused by the presence of some unidentified antibodies. In this study, we found that the ASFV-encoded structural protein A137R (pA137R) can be recognized by the anti-ASFV positive sera, indicating that the anti-pA137R antibodies are induced in the ASFV-infected pigs. Interestingly, our results demonstrated that the anti-pA137R antibodies produced in rabbits or pigs enhanced viral replication of different ASFV strains in primary porcine alveolar macrophages (PAMs), the target cells of ASFV. Mechanistic investigations revealed that anti-pA137R antibodies were able to promote the attachment of ASFV to PAMs and two types of Fc gamma receptors (FcγRs), FcγRII and FcγRIII, mediated the ADE of ASFV infection. Taken together, anti-pA137R antibodies are able to drive ASFV ADE in PAMs. These findings shed new light on the roles of anti-ASFV antibodies and have implications for the pathophysiology of the disease and the development of ASF vaccines.

Construction and Evaluation of an AI-based CBCT Resolution Optimization Technique for Extracted Teeth.

INTRODUCTION: In dental clinical practice, cone-beam computed tomography (CBCT) is commonly used to assist practitioners to recognize the complex morphology of root canal systems; however, because of its resolution limitations, certain small anatomical structures still cannot be accurately recognized on CBCT. The purpose of this study was to perform image super-resolution (SR) processing on CBCT images of extracted human teeth with the help of a deep learning model, and to compare the differences among CBCT, super-resolution computed tomography (SRCT), and micro-computed tomography (Micro-CT) images through three-dimensional reconstruction. METHODS: The deep learning model (Basicvsr++) was selected and modified. The dataset consisted of 171 extracted teeth that met inclusion criteria, with 40 maxillary first molars as the training set and 40 maxillary first molars as well as 91 teeth from other tooth positions as the external test set. The corresponding CBCT, SRCT, and Micro-CT images of each tooth in test sets were reconstructed using Mimics Research 17.0, and the root canal recognition rates in the 3 groups were recorded. The following parameters were measured: volume of hard tissue (V1), volume of pulp chamber and root canal system (V2), length of visible root canals under orifice (VL-X, where X represents the specific root canal), and intersection angle between coronal axis of canal and long axis of tooth (∠X, where X represents the specific root canal). Data were statistically analyzed between CBCT and SRCT images using paired sample t-test and Wilcoxon test analysis, with the measurement from Micro-CT images as the gold standard. RESULTS: Images from all tested teeth were successfully processed with the SR program. In 4-canal maxillary first molar, identification of MB2 was 72% (18/25) in CBCT group, 92% (23/25) in SRCT group, and 100% (25/25) in Micro-CT group. The difference of hard tissue volume between SRCT and Micro-CT was significantly smaller than that between CBCT and Micro-CT in all tested teeth except 4-canal mandibular first molar (P < .05). Similar results were obtained in volume of pulp chamber and root canal system in all tested teeth (P < .05). As for length of visible root canals under orifice, the difference between SRCT and Micro-CT was significantly smaller than that between CBCT and Micro-CT (P < .05) in most root canals. CONCLUSIONS: The deep learning model developed in this study helps to optimize the root canal morphology of extracted teeth in CBCT. And it may be helpful for the identification of MB2 in the maxillary first molar.

Appropriate supply of sulfur alleviates lead toxicity and stimulates its accumulation in hyperaccumulator Arabis alpina.

Widespread lead (Pb) contamination of agricultural soils is a global issue stemming from human activities. The remediation of Pb-contaminated soils used for agricultural purposes is critically important to safeguard food crop safety. Despite the modulating effects of sulfur (S) on plant responses to toxic heavy metals, the ecological, physiological, and molecular mechanisms driving such modulation in the Pb hyperaccumulator Arabis alpina L. remain unclear. Here, we investigated the effects of five S concentrations (0, 50, 100, 150, and 200 mg kg-1) on A. alpina grown in Pb-contaminated soil from a lead-zinc mining area. Under S50 (i.e., 50 mg kg-1) and S100 treatments, the Pb concentration in both shoots and roots of A. alpina significantly decreased compared to the control (S0). Specifically, the S50 treatment significantly enhanced Pb accumulation, plant biomass, and plant height, indicating that low S applications facilitate Pb accumulation from the soil and alleviate Pb toxicity. Additionally, S50, S100, and S150 treatments significantly improved photosynthetic rate, stomatal conductance, and intercellular CO2 concentration in A. alpina. Transcriptomic analysis showed that S50 and S100 treatments increased the expression of the LHCA, LHCB, psa, and psb genes, which had a significant impact on photosynthetic efficiency. S50 and S100 boosted glutathione (GSH) levels in A. alpina roots, and the increased expression of GST gene enhanced tolerance to environmental stress. In summary, these results suggest that an appropriate supply of S (S50 and S100) not only alleviates Pb toxicity by enhancing plant biomass, height, photosynthetic features, and sulfur metabolites but also stimulates Pb accumulation in the hyperaccumulator A. alpina. Our study elucidated the specific concentrations of sulfur that optimally enhance both Pb accumulation and stress tolerance in the hyperaccumulator A. alpina, providing novel insights into the practical application of sulfur in phytoremediation strategies and advancing our understanding of the underlying molecular mechanisms.

The features of male breast cancer in China: A real-world study.

BACKGROUND: Male breast cancer (MBC) is a rare disease. Although several large-scale studies have investigated MBC patients in other countries, the features of MBC patients in China have not been fully explored. This study aims to explore the features of Chinese MBC patients comprehensively. METHODS: We retrospectively collected data of MBC patients from 36 centers in China. Overall survival (OS) was evaluated by the Kaplan-Meier method, log-rank test, and Cox regression analyses. Multivariate Cox analyses were used to identify independent prognostic factors of the patients. RESULTS: In total, 1119 patients were included. The mean age at diagnosis was 60.9 years, and a significant extension over time was observed (P < 0.001). The majority of the patients (89.1 %) received mastectomy. Sentinel lymph node biopsy was performed in 7.8 % of the patients diagnosed in 2009 or earlier, and this percentage increased significantly to 38.8 % in 2020 or later (P < 0.001). The five-year OS rate for the population was 85.5 % [95 % confidence interval (CI), 82.8 %-88.4 %]. Multivariate Cox analysis identified taxane-based [T-based, hazard ratio (HR) = 0.32, 95 % CI, 0.13 to 0.78, P = 0.012] and anthracycline plus taxane-based (A + T-based, HR = 0.47, 95 % CI, 0.23 to 0.96, P = 0.037) regimens as independent protective factors for OS. However, the anthracycline-based regimen showed no significance in outcome (P = 0.175). CONCLUSION: As the most extensive MBC study in China, we described the characteristics, treatment and prognosis of Chinese MBC population comprehensively. T-based and A + T-based regimens were protective factors for OS in these patients. More research is required for this population.

Single-cell transcriptome reveals a novel mechanism of C-Kit+-liver sinusoidal endothelial cells in NASH.

AIM: To understand how liver sinusoidal endothelial cells (LSECs) respond to nonalcoholic steatohepatitis (NASH). METHODS: We profiled single-LSEC from livers of control and MCD-fed mice. The functions of C-Kit+-LSECs were determined using coculture and bone marrow transplantation (BMT) methods. RESULTS: Three special clusters of single-LSEC were differentiated. C-Kit+-LSECs of cluster 0, Msr1+-LSECs of cluster 1 and Bmp4+Selp+-VECs of cluster 2 were revealed, and these cells with diverse ectopic expressions of genes participated in regulation of endothelial, fibrosis and lipid metabolism in NASH. The number of C-Kit+-primary LSECs isolated from MCD mice was lower than control mice. Immunofluorescence co-staining of CD31 and C-KIT showed C-Kit+-LSECs located in hepatic sinusoid were also reduced in NASH patients and MCD mice, compared to AIH patients and control mice respectively. Interestingly, lipotoxic hepatocytes/HSCs cocultured with C-Kit+-LSECs or the livers of MCD mice receipting of C-Kit+-BMCs (bone marrow cells) showed less steatosis, inflammation and fibrosis, higher expression of prolipolytic FXR and PPAR-α, lower expression of TNF-α and α-SMA. Furthermore, coculturing or BMT of C-Kit+-endothelial derived cells could increase the levels of hepatic mitochondrial LC3B, decrease the degree of mitochondrial damage and ROS production through activating Pink1-mediated mitophagy pathway in NASH. CONCLUSIONS: Hence, a novel transcriptomic view of LSECs was revealed to have heterogeneity and complexity in NASH. Importantly, a cluster of C-Kit+-LSECs was confirmed to recovery Pink1-related mitophagy and NASH progression.

Si/Ge phototransistor with responsivity >1000A/W on a silicon photonics platform.

In this article, we report a Si/Ge waveguide phototransistor with high responsivity and low dark current under low bias voltages, due to an engineered electric field distribution. The photodetector consists of n-i-p-i-n doping regions and shows a responsivity of 606 A/W at 1 V bias, and 1032 A/W at 2.8V bias with an input optical power of -50 dBm, and dark current of 4 µA and 42 µA respectively. This is achieved by placing two p+-doped regions in the silicon slab region beneath the Ge epitaxial layer. A measured small signal -3 dB bandwidth of 1.5 GHz with a -80 dBc/Hz phase noise response at 1 KHz frequency offset were demonstrated experimentally.

Design of pH-responsive antimicrobial peptide melittin analog-camptothecin conjugates for tumor therapy.

Melittin, a classical antimicrobial peptide, is a highly potent antitumor agent. However, its significant toxicity seriously hampers its application in tumor therapy. In this study, we developed novel melittin analogs with pH-responsive, cell-penetrating and membrane-lytic activities by replacing arginine and lysine with histidine. After conjugation with camptothecin (CPT), CPT-AAM-1 and CPT-AAM-2 were capable of killing tumor cells by releasing CPT at low concentrations and disrupting cell membranes at high concentrations under acidic conditions. Notably, we found that the C-terminus of the melittin analogs was more suitable for drug conjugation than the N-terminus. CPT-AAM-1 significantly suppressed melanoma growth in vivo with relatively low toxicity. Collectively, the present study demonstrates that the development of antitumor drugs based on pH-responsive antimicrobial peptide-drug conjugates is a promising strategy.

Revealing How Polyvinyl Chloride Microplastic Physicochemically Affect the Anaerobic Digestion of Waste Activated Sludge.

Recent studies have shown that the microplastics in waste activated sludge (WAS) can directly reduce the microbial activity and influence the performance of anaerobic digestion. Unfortunately, few studies paid attention on the interactions between WAS and MPs, since MPs could impact the contact between sludge flocs and microorganisms. We found that PVC-MP changed the interfacial energy properties of the WAS surface and affected methane production. Low concentration (40 mg/L) of PVC-MP changed the water affinity and greatly reduced the energy barrier of interfacial reaction. Simultaneously, WAS surface charge characteristics changed with increasing MPs concentration, which made the sludge difficult to contact with microorganisms. The change process of WAS surface functional groups also indicated that PVC-MP first cover the sludge surface to prevent from being utilized by microorganisms, and then affect the surface protein structure before toxic substances leaching. Our study provides new insights into how MPs affect anaerobic digestion.

Evaluating the cytotoxicity mechanism of the cell-penetrating peptide TP10 on Jurkat cells.

TP10, a classic cell-penetrating peptide, shows a high degree of similarity to AMPs in structure. Although TP10 has been widely used in drug delivery, the mechanism underlying its cytotoxicity is yet to be elucidated. Herein, we explored the cell-killing mechanism of TP10 against human leukemia Jurkat cells. TP10 induced necrosis in Jurkat cells via rapid disruption of cell membranes, particularly at high concentrations. Although mitochondria in Jurkat cells were damaged by TP10, mitochondria-mediated apoptosis did not occur, possibly due to intracellular ATP depletion. Necroptosis in TP10-treated Jurkat cells became an alternative route of apoptosis. Our results demonstrate that necrosis and necroptosis rather than apoptosis are involved in the cell-killing mechanism of TP10, which contributes to the understanding of its toxicity.

Altered brain network centrality in patients with orbital fracture: A resting­state functional MRI study.

The present study aimed to investigate potential functional network brain-activity abnormalities in individuals with orbital fracture (OF) using the voxel-wise degree centrality (DC) technique. The present study included 20 patients with OF (12 males and 8 females) and 20 healthy controls (HC; 12 males and 8 females), who were matched for gender, age and educational attainment. Functional magnetic resonance imaging (fMRI) in the resting state has been widely applied in several fields. Receiver operating characteristic (ROC) curves were calculated to distinguish between patients with OF and HCs. In addition, correlation analyses were performed between behavioral performance and average DC values in various locations. The DC technique was used to assess unprompted brain activity. Right cerebellum 9 region (Cerebelum_9_R) and left cerebellar peduncle 2 area (Cerebelum_Crus2_L) DC values of patients with OF were increased compared with those in HCs. Cerebelum_9_R and Cerebelum_Crus2_L had area under the ROC curve values of 0.983 and 1.000, respectively. Patients with OF appear to have several brain regions that exhibited aberrant brain network characteristics, which raises the possibility of neuropathic causes and offers novel therapeutic options.

Morphological and transcriptomic responses/acclimations of erect-type submerged macrophyte Hydrilla verticillata both at low-light exposure and light recovery phases.

Light intensity is a determinant for submerged macrophytes. Little is known about their molecular responses to low-light exposure, despite more informative and responsive than morphological traits. For erect-type submerged macrophytes, the stem is more crucial relative to the leaf in acclimation to low-light stress, but receives less attention. We determined morphological and stem transcriptomic responses/acclimations of Hydrilla verticillata to extremely and mildly low light (7.2 and 36 µmol photons m-2 s-1, respectively), that is, EL and ML, with the radiation intensity of 180 µmol photons m-2 s-1 as the control. Low-light exposure continued for 9 days, followed by a 7-day recovery phase (180 µmol photons m-2 s-1). At the exposure phase, the low-light treatments, in particular the EL, decreased the relative growth ratio, but induced greater height and longer stem internode distance and epidermal cell. Such responses/acclimations continued into the recovery phase, despite more or less changes in the magnitude. Transcriptome showed that the photosynthetic system was inhibited at the exposure phase, but the macrophyte adjusted hormone synthesis relating to cell division and elongation. Moreover, the EL activated cell stress responses such as DNA repair. Following light recovery, the macrophyte exhibited a strong-light response, although energy metabolism enhanced. Especially, the EL enriched the pathways relating to anthocyanin synthesis at such phase, indicating an activation of photoprotective mechanism. Our findings suggest that negative influences of low light occur at both low-light exposure and recovery phases, but submerged macrophytes would acclimate to light environments. Transcriptome can show molecular basis of plant responses/acclimations, including but not limited to morphology. This study establishes a bridge connecting morphological and molecular responses/acclimations.

Event-Triggered Adaptive Antidisturbance Switching Control for Switched Systems With Dynamic Neural Network Disturbance Modeling.

In this article, a dynamic event-triggered adaptive antidisturbance (ETAAD) switching control strategy is proposed for switched systems subject to multisource disturbances. The disturbances are divided into two categories: the available unmodeled disturbance and the unavailable dynamic neural network modeled disturbance. First, a dynamic ET criterion is set based on the system state. Then, a novel dynamic ETA disturbance estimator is introduced to observe the modeled disturbance. Furthermore, according to the ET rule and adaptive disturbance observer, a switched controller is designed. Next, under the controller and switching criterion with the average dwell time limitation, sufficient conditions are given to force the switched systems to realize multisource disturbance suppression (DS), trajectory tracking, and communication resource (CR) saving simultaneously. Meanwhile, the Zeno phenomenon may be caused by the ET rule being excluded. In addition, the presented ETAAD approach is also applicable to the nonswitched systems case. Finally, a simulation case is given to validate the effectiveness of the dynamic ETAAD switching control method.

Mechanism of microbial regulation on methane metabolism in saline-alkali soils based on metagenomics analysis.

Saline-alkali soils constitute a globally important carbon pool that plays a critical role in soil carbon dioxide (CO2) and methane (CH4) fluxes. However, the relative importance of microorganisms in the regulation of CH4 emissions under elevated salinity remains unclear. Here, we report the composition of CH4 production and oxidation microbial communities under five different salinity levels in the Yellow River Delta, China. This study also obtained the gene number of microbial CH4 metabolism via testing the soil metagenomes, and further investigated the key soil factors to determine the regulation mechanism. Spearman correlation analysis showed that the soil electrical conductivity, salt content, and Na+, and SO42- concentrations showed significantly negative correlations with the CO2 and CH4 emission rates, while the NO2--N concentration and NO2-/NO3- ratio showed significantly positive correlations with the CO2 and CH4 emission rates. Metabolic pathway analysis showed that the mcrA gene for CH4 production was highest in low-salinity soils. By contrast, the relative abundances of the fwdA, ftr, mch, and mer genes related to the CO2 pathway increased significantly with rising salinity. Regarding CH4 oxidation processes, the relative abundances of the pmoA, mmoB, and mdh1 genes transferred from CH4 to formaldehyde decreased significantly from the control to the extreme-salinity plot. The greater abundance and rapid increase of methanotrophic bacteria compared with the lower abundance and slow increase in methanogenic archaea communities in saline-alkali soils may have increased CH4 oxidation and reduced CH4 production in this study. Only CO2 emissions positively affected CH4 emissions from low- to medium-salinity soils, while the diversities of CH4 production and oxidation jointly influenced CH4 emissions from medium- to extreme-salinity plots. Hence, future investigations will also explore more metabolic pathways for CH4 emissions from different types of saline-alkali lands and combine the key soil enzymes and regulated biotic or abiotic factors to enrich the CH4 metabolism pathway in saline-alkali soils.

PiRNA hsa_piR_019914 Promoted Chondrocyte Anabolic Metabolism By Inhibiting LDHA-Dependent ROS Production.

OBJECTIVES: Osteoarthritis (OA) is the most common joint disease. The occurrence and progression of OA are regulated by epigenetics. A large number of studies have shown the important regulatory role of noncoding RNAs in joint diseases. As the largest class of noncoding small RNAs, the importance of piRNAs in many diseases, especially cancer, has been increasingly recognized. However, few studies have explored the role of piRNAs in OA. Our study showed that hsa_piR_019914 decreased significantly in OA. This study aimed to demonstrate the role of hsa_piR_019914 as a potential biological target of OA in chondrocytes. DESIGN: The GEO database and bioinformatics analysis were used for a series of screenings, and the OA model using human articular chondrocytes (C28/I2 cells), SW1353 cells under inflammatory factor stimulation was used to determine that hsa_piR_019914 was significantly downregulated in OA. Overexpression or inhibition of hsa_piR_019914 in C28/I2 cells was achieved by transfecting mimics or inhibitors. The effect of hsa_piR_019914 on the biological function of chondrocytes was verified by qPCR, flow cytometry, and colony formation assays in vitro. The target gene of hsa_piR_019914, lactate dehydrogenase A (LDHA), was screened by small RNA sequencing and quantitative polymerase chain reaction (qPCR), LDHA was knocked out in C28/I2 cells by the transfection of siRNA LDHA, and the relationship between hsa_piR_019914, LDHA, and reactive oxygen species (ROS) production was verified by flow cytometry. RESULTS: The piRNA hsa-piR-019914 was significantly downregulated in osteoarthritis (OA). Hsa-piR-019914 reduced inflammation-mediated chondrocyte apoptosis and maintained cell proliferation and clone formation in vitro. Hsa-piR-019914 reduced the production of LDHA-dependent ROS through targeted regulation of LDHA expression, maintained chondrocyte-specific gene expression of ACAN and COL2, and inhibited the gene expression of MMP3 and MMP13. CONCLUSIONS: Collectively, this study showed that hsa_piR_019914 was negatively correlated with the expression of LDHA, which mediates ROS production. Under the stimulation of inflammatory factors, overexpression of hsa_piR_019914 had a protective effect on chondrocytes in vitro, and the absence of hsa_piR_019914 exacerbated the negative effect of inflammation on chondrocytes. Studies on piRNAs provide new therapeutic interventions for OA.

The emerging role of snoRNAs in human disease.

Small nucleolar RNAs (snoRNAs) play critical roles in various biological processes. The aberrant expression or depletion of snoRNAs is related to various diseases. In previous research, most of the snoRNAs were categorized as C/D box snoRNAs and H/ACA box snoRNAs, whose typical functions were thought of as regulation of 2'-O-ribose methylation and pseudouridylation of ribosome RNAs, respectively. However, in the past two decades, studies have revealed an increasing number of snoRNAs without specific targets or determined cell functions. These findings indicated that some potential roles of snoRNAs are still unknown. Numerous studies have indicated the correlation of snoRNAs with human diseases. SnoRNAs play various roles in abundant biological processes, and they have great potential in controlling human diseases. This new and rising field could benefit from investigations of the disease pathogenesis, biomarker identification, and the determination of novel therapeutic targets. This review summarized the reports on snoRNAs and the regulation of different diseases in recent years.

Assessment of the coronal root canal morphology of permanent maxillary first molars using digital 3D-reconstruction technology based on micro-computed tomography data.

Background: The design of minimally invasive access has become a hotspot. This study aimed to evaluate the coronal root canal morphology of permanent maxillary first molars to facilitate the design of endodontic access cavities for minimally invasive linear access. Materials and methods: A selection of 91 permanent maxillary first molars was evaluated. Three-dimensional tooth models were reconstructed using micro-computed tomography data. Root canal prevalence and coronal root canal landmarks were recorded. The positional coordinates of landmarks in the horizontal plane and the angles and directions of coronal root canal curvature in the horizontal and axial planes were also assessed. Results: The detection rates of the mesiobuccal (MB), distobuccal (DB), and palatal (P) canals were 100%, whereas that of the second mesiobuccal (MB2) canals was 68.1%. All landmarks were located near the central fossa. In the axial plane, the average angles of coronal root canal curvature were DB (27.05°) > MB (25.43°) > P (20.71°) in teeth with three canals, and MB2 (33.20°) > MB (29.61°) > DB (28.40°) > P (23.69°) in teeth with four canals. In the horizontal plane, the average angles were P (78.15°) > DB (42.34°) > MB (32.41°) in teeth with three canals, and P (81.26°) > DB (43.44°) > MB (41.22°) > MB2 (9.41°) in teeth with four canals. Conclusion: In maxillary first molars, coronal root canals tend to converge towards the occlusal surface. The results of this study could be applied to improve the precision of endodontic access cavity designs of minimally invasive access.

Piperlonguminine attenuates renal fibrosis by inhibiting TRPC6.

ETHNOPHARMACOLOGICAL RELEVANCE: Liuwei Dihuang (LWDH) is a classic prescription that has been used to the treatment of "Kidney-Yin" deficiency syndrome for more than 1000 years in China. Recent studies have confirmed that LWDH can prevent the progression of renal fibrosis. Numerous studies have demonstrated the critical role that TRPC6 plays in the development of renal fibrosis. Due to the complex composition of LWDH and its remarkable therapeutic effect on renal fibrosis, it is possible to discover new active ingredients targeting TRPC6 for the treatment of renal fibrosis. AIM OF STUDY: This study aimed to identify selective TRPC6 inhibitors from LWDH and evaluate their therapeutical effects on renal fibrosis. MATERIALS AND METHODS: Computer-aided drug design was used to screen the biologically active ingredients of LWDH, and their affinities to human TRPC6 protein were detected by microcalorimetry. TRPC6, TRPC3, and TRPC7 over-expressed HEK293 cells were constructed, and the selective activities of the compounds on TRPC6 were determined by measuring [Ca2+]i in these cells. To establish an in vitro model of renal fibrosis, human renal proximal tubular epithelial HK-2 cells were stimulated with TGF-ß1. The therapeutic effects of LWDH compounds on renal fibrosis were then tested by detecting the related proteins. TRPC6 was knocked-down in HK-2 cells to investigate the effects of LWDH active ingredients on TRPC6. Finally, a unilateral ureteral obstruction model of renal fibrosis was established to test the therapeutic effect. RESULTS: From hundreds of LWDH ingredients, 64 active components with oral bioavailability ≥30% and drug-likeness index ≥0.18 were acquired. A total of 10 active components were obtained by molecular docking with TRPC6 protein. Among them, 4 components had an affinity with TRPC6. Piperlonguminine (PLG) had the most potent affinity with TRPC6 and blocking effect on TRPC6-mediated Ca2+ entry. A 100 µM of PLG showed no detectable inhibition on TRPC1, TRPC3, TRPC4, TRPC5, or TRPC7-mediated Ca2+ influx into cells. In vitro results indicated that PLG concentration-dependently inhibited the abnormally high expression of α-smooth muscle actin (α-SMA), collagen I, vimentin, and TRPC6 in TGF-ß1-induced HK-2 cells. Consistently, PLG also could not further inhibit TGF-ß1-induced expressions of these protein biomarkers in TRPC6 knocked-down HK-2 cells. In vivo, PLG dose-dependently reduced urinary protein, serum creatinine, and blood urea nitrogen levels in renal fibrosis mice and markedly alleviated fibrosis and the expressions of α-SMA, collagen I, vimentin, and TRPC6 in kidney tissues. CONCLUSION: Our results showed that PLG had anti-renal fibrosis effects by selectively inhibiting TRPC6. PLG might be a promising therapeutic agent for the treatment of renal fibrosis.

Development of Neuropeptide Hemokinin-1 Analogues with Antimicrobial and Wound-Healing Activity.

Wound healing is a complex process that can be delayed in some pathological conditions, such as infection and diabetes. Following skin injury, the neuropeptide substance P (SP) is released from peripheral neurons to promote wound healing by multiple mechanisms. Human hemokinin-1 (hHK-1) has been identified as an SP-like tachykinin peptide. Surprisingly, hHK-1 shares similar structural features with antimicrobial peptides (AMPs), but it does not display efficient antimicrobial activity. Therefore, a series of hHK-1 analogues were designed and synthesized. Among these analogues, AH-4 was found to display the greatest antimicrobial activity against a broad spectrum of bacteria. Furthermore, AH-4 rapidly killed bacteria by membrane disruption, similar to most AMPs. More importantly, AH-4 showed favorable healing activity in all tested mouse full-thickness excisional wound models. Overall, this study suggests that the neuropeptide hHK-1 can be used as a desirable template for developing promising therapeutics with multiple functions for wound healing.