Integrating single-cell and multi-omic approaches reveals Euphorbiae Humifusae Herba-dependent mitochondrial dysfunction in non-small-cell lung cancer.

Euphorbiae Humifusae Herba (EHH) is a pivotal therapeutic agent with diverse pharmacological effects. However, a substantial gap exists in understanding its pharmacological properties and anti-tumour mechanisms. This study aimed to address this gap by exploring EHH's pharmacological properties, identifying NSCLC therapy-associated protein targets, and elucidating how EHH induces mitochondrial disruption in NSCLC cells, offering insights into novel NSCLC treatment strategies. String database was utilized to explore protein-protein interactions. Subsequently, single-cell analysis and multi-omics further unveiled the impact of EHH-targeted genes on the immune microenvironment of NSCLC, as well as their influence on immunotherapeutic responses. Finally, both in vivo and in vitro experiments elucidated the anti-tumour mechanisms of EHH, specifically through the assessment of mitochondrial ROS levels and alterations in mitochondrial membrane potential. EHH exerts its influence through engagement with a cluster of 10 genes, including the apoptotic gene CASP3. This regulatory impact on the immune milieu within NSCLC holds promise as an indicator for predicting responses to immunotherapy. Besides, EHH demonstrated the capability to induce mitochondrial ROS generation and perturbations in mitochondrial membrane potential in NSCLC cells, ultimately leading to mitochondrial dysfunction and consequent apoptosis of tumour cells. EHH induces mitochondrial disruption in NSCLC cells, leading to cell apoptosis to inhibit the progress of NSCLC.

Genome-wide identification of polyamine metabolism and ethylene synthesis genes in Chenopodium quinoa Willd. and their responses to low-temperature stress.

BACKGROUND: Quinoa (Chenopodium quinoa Willd.) is valued for its nutritional richness. However, pre-harvest sprouting poses a significant threat to yield and grain quality. This study aims to enhance our understanding of pre-harvest sprouting mitigation strategies, specifically through delayed sowing and avoiding rainy seasons during quinoa maturation. The overarching goal is to identify cold-resistant varieties and unravel the molecular mechanisms behind the low-temperature response of quinoa. We employed bioinformatics and genomics tools for a comprehensive genome-wide analysis of polyamines (PAs) and ethylene synthesis gene families in quinoa under low-temperature stress. RESULTS: This involved the identification of 37 PA biosynthesis and 30 PA catabolism genes, alongside 227 ethylene synthesis. Structural and phylogenetic analyses showcased conserved patterns, and subcellular localization predictions indicated diverse cellular distributions. The results indicate that the PA metabolism of quinoa is closely linked to ethylene synthesis, with multiple genes showing an upregulation in response to cold stress. However, differential expression within gene families suggests a nuanced regulatory network. CONCLUSIONS: Overall, this study contributes valuable insights for the functional characterization of the PA metabolism and ethylene synthesis of quinoa, which emphasize their roles in plant low-temperature tolerance and providing a foundation for future research in this domain.

The process of solid-state fermentation of soybean meal: antimicrobial activity, fermentation heat generation and nitrogen solubility index.

BACKGROUND: Bacillus amyloliquefaciens has excellent protease production ability and holds great prospects for application in the solid-state fermentation of soybean meal (SBM). RESULTS: Among eight strains of bacteria, Bacillus amyloliquefaciens subsp. plantarum CICC 10265, which exhibited higher protease production, was selected as the fermentation strain. The protease activity secreted by this strain reached 106.41 U mL-1 . The microbial community structure differed significantly between natural fermentation and inoculation-enhanced fermented soybean meal (FSBM), with the latter showing greater stability and inhibition of miscellaneous bacterial growth. During fermentation, the temperature inside the soybean meal increased, and the optimal environmental temperature for FSBM was found to be between 35 and 40 °C. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and nitrogen solubility index (NSI) results demonstrated that solid-state fermentation had a degrading effect on highly denatured proteins in SBM, resulting in an NSI of 67.1%. CONCLUSION: Bacillus amyloliquefaciens subsp. plantarum CICC 10265 can enhance the NSI of SBM in solid-state fermentation and inhibit the growth of miscellaneous bacteria. © 2023 Society of Chemical Industry.

Anion-modulated Ion Conductor with Chain Conformational Transformation for stabilizing Interfacial Phase of High-Voltage Lithium Metal Batteries.

In solid-state lithium metal batteries (SSLMBs), the inhomogeneous electrolyte-electrode interphase layer aggravates the interfacial stability, leading to discontinuous interfacial ion/charge transport and continuous degradation of the electrolyte. Herein, we constructed an anion-modulated ionic conductor (AMIC) that enables in situ construction of electrolyte/electrode interphases for high-voltage SSLMBs by exploiting conformational transitions under multiple interactions between polymer and lithium salt anions. Anions modulate the decomposition behavior of supramolecular poly (vinylene carbonate) (PVC) at the electrode interface by changing the spatial conformation of the polymer chains, which further enhances ion transport and stabilizes the interfacial morphology. In addition, the AMIC weakens the "Li+-solvation" and increases Li+ vehicle sites, thereby enhancing the lithium-ion transport number (tLi +=~0.67). Consequently, Li || LiNi0.8Co0.1Mn0.1O2 cell maintains about 85 % capacity retention and Coulombic efficiency >99.8 % in 200 cycles at a charge cut-off voltage of 4.5 V. This study provides a new understanding of lithium salt anions regulating polymer chain segment behavior in the solid-state polymer electrolyte (SPE) and highlights the importance of the ion environment in the construction of interfacial phases and ionic conduction.

BSA-stabilized silver nanoclusters for efficient photoresponsive colorimetric detection of chromium(VI).

Hexavalent chromium is a highly toxic substance, which will pose a serious threat to human life and health and the entire ecosystem. Therefore, it is crucial to establish a simple and rapid detection method for hexavalent chromium. In this work, we fabricated bovine serum albumin-stabilized silver nanocluster (BSA-Ag13 NC) which exhibited photoresponsive oxidase-like activity, catalyzing the oxidation of colorless 3,3',5,5'-tetramethylbenzidine (TMB) to the blue oxidized state TMB (oxTMB) in a short time. Interestingly, 8-hydroxyquinoline (8-HQ) can significantly inhibit the color reaction of TMB oxidation while Cr(VI) can interact specifically with 8-HQ to restore this chromogenic reaction. Based on the above facts, a colorimetric sensing system for detecting Cr(VI) was developed. The sensing system shows a wide linear range, and good selectivity, with a low detection limit of 2.32 nM. Moreover, this sensing system could be successfully applied to the detection of Cr(VI) in lake water, tap water, and sewage with satisfactory results.

Assessment of DNA mutagenicity induced by He-Ne laser using Salmonella typhimurium strains.

Helium-neon (He-Ne) laser mutagenesis is widely used in microbiology and plant breeding. In this study, two frameshift mutant representative strains of Salmonella typhimurium TA97a and TA98 and two base pair substitution types TA100 and TA102 were employed as model microorganisms to assess DNA mutagenicity induced by He-Ne laser (3 J·cm-2·s-1, 632.8 nm) for 10, 20, and 30 min. The results revealed that the optimal laser application was 6 h in the mid-logarithmic growth stage. Low-power He-Ne laser for short treatment inhibited cell growth, and continued treatment stimulated the metabolism. The effects of the laser on TA98 and TA100 were the most prominent. Sequencing results from 1500 TA98 revertants showed that there were 88 insertion and deletion (InDel) types in the hisD3052 gene, of which the InDels unique to laser were 21 more than that of the control. Sequencing results from 760 TA100 revertants indicated that laser treatment created Pro (CCC) in the product of the hisG46 gene more likely to be replaced by His (CAC) or Ser (TCC) than by Leu (CTC). Two unique non-classical base substitutions, CCC → TAC and CCC → CAA, also appeared in the laser group. These findings will provide a theoretical basis for further exploration of laser mutagenesis breeding. KEY POINTS: • Salmonella typhimurium served as model organism for laser mutagenesis study. • Laser promoted the occurrence of InDels in the hisD3052 gene of TA98. • Laser promoted the occurrence of base substitution in the hisG46 gene of TA100.

Respiratory fabric sensor based on the side luminescence and photosensitivity mechanism of polymer optical fibers.

It is significant to monitor respiration conveniently and in real time for people suffering from respiratory diseases. Polymer optical fibers (POFs) have the advantages of flexibility and light weight, which is highly desirable for wearable respiratory monitoring. However, in most current applications, the POFs are stitched on the textile substrates in the form of macro-bending. This method is complex to fix the bending with certain curvatures and uncomfortable compared with the POF sensors woven into the textile. In this paper, a respiratory fabric sensor based on the side luminescence and photosensitivity mechanism of POF is proposed and demonstrated. The 750µm-diameter POFs were woven into a fabric as warp and laser marking was performed at their designed positions to make them release or couple light. The spacing change between the POFs caused by the respiratory movement accordingly makes the light intensity change in the photosensitive fiber. We chose four fabric widths (10cm, 8cm, 6cm and 4cm) and four fabric weaves (plain weave, honeycomb weave, 1/3 right twill weave and 8/3 warp satin weave) to implement the full-factor experiment for exploring the measurement effect of the respiratory fabric sensor. The result is that the fabric with width of 4cm and weave of 8/3 warp satin is optimal. The calm and deep respiratory tests of the human chest and abdomen in sitting and standing posture were carried out and the test performance of the fabric sensor is almost comparable to that of the medical monitor. The proposed respiratory fabric sensor is comfortable, easily woven and high in precision, which is expected to realize industrialized scale production.

Pd Nanosheet-Decorated 2D/2D g-C3N4/WO3·H2O S-Scheme Photocatalyst for High Selective Photoreduction of CO2 to CO.

The development of the global economy in recent years, environmental problems, greenhouse effect, and so forth have been of concern for countries all over the world. The key for solving the greenhouse effect is the reduction of CO2. With the development of photocatalytic reduction of CO2, hybrid photocatalytic nanostructures composed of noble metals and plasmonic semiconductors are being widely studied. In this work, S-scheme photocatalysts with a g-C3N4/WO3·H2O/Pd heterostructure was constructed by introducing ultrathin Pd nanosheets into the optimized 2D/2D g-C3N4/WO3·H2O binary system. The S-scheme charge transfer generated by the matched band gap of g-C3N4 and WO3·H2O can effectually improve the electron transfer rate and the redox ability of photogenerated carriers. The introduction of Pd nanosheets can inject a large number of hot electrons into the semiconductor on the basis of the S-scheme heterojunction to participate in the reaction. The S-scheme electron transfer method is used to improve the utilization rate of thermionic electrons and achieve the effect of widening the near-infrared-light absorption area of the composite material. Moreover, the reaction was carried out in water without the addition of any sacrificial agent, which can better reflect the green environmental protection of the experiment. This investigation will promote the broad-spectrum application of new and environment-friendly thermoelectron-assisted S-scheme photocatalysts, and on this basis, the possible reaction mechanism is discussed.

Tailored Linker Defects in UiO-67 with High Ligand-to-Metal Charge Transfer toward Efficient Photoreduction of CO2.

Defect engineering can be used as a potential tool to activate metal-organic frameworks by regulating the pore structure, electronic properties, and catalytic activity. Herein, linker defects were effectively controlled by adjusting the amount of formic acid, and UiO-67 with different CO2 reduction capabilities was obtained. Among them, UiO-67-200 had the highest ability to selectively reduce CO2 to CO (12.29 µmol g-1 h-1). On the one hand, the results based on time-resolved photoluminescence decay curves and photochemical experiments revealed that UiO-67-200 had the highest charge separation efficiency. On the other hand, the linker defects affected the band structure of UiO-67 by changing the lowest unoccupied molecular orbital (LUMO) based on the density functional theory and UV-vis spectra. Hence, the proper linker defects enhanced the ligand-to-metal charge transfer process by promoting the transfer of electrons between the highest occupied molecular orbital and LUMO. Additionally, in situ Fourier transform infrared spectra and 13CO2 labeling experiments also indicated that COOH* was an important intermediate for CO formation and that CO originated from the photoreduction of CO2.

Construction of a ZnIn2S4/Au/CdS Tandem Heterojunction for Highly Efficient CO2 Photoreduction.

Photocatalytic CO2 reduction technology is of great importance to alleviate energy crisis and environmental pollution; however, it remains a serious challenge due to the fast recombination of carriers. In this study, we report a three-dimensional structure of a ZnIn2S4/Au/CdS composite photocatalyst for the CO2 reduction reaction, where Au nanoparticles (NPs) are evenly anchored on the surface of ZnIn2S4 by photodeposition and Au NPs are wrapped around by CdS. In ZnIn2S4/Au/CdS composite photocatalysts, Au NPs act as a bridge to construct a "semiconductor-metal-semiconductor" tandem electron transfer mechanism (ZnIn2S4 → Au → CdS) heterojunction, which greatly promotes the transfer of photogenerated electrons. It is worth noting that Au NPs, as a local surface plasmon resonance (LSPR) effect excited source to generate excited-state electrons, further improve the photoreduction CO2 activity. Under UV-vis light irradiation, the CO yield of ZnIn2S4/Au/CdS can reach 63.07 µmol·g-1·h-1, which is higher than that of 6.37 µmol·g-1·h-1 for pure ZnIn2S4, 0.93 µmol·g-1·h-1 for CdS, 8.9 µmol·g-1·h-1 for ZnIn2S4/CdS, 31.04 µmol·g-1·h-1 for ZnIn2S4/Au, and 5.37 µmol·g-1·h-1 for CdS/Au. In addition, the ternary ZnIn2S4/Au/CdS composite photocatalyst has good cyclic stability. This study broadens the idea of designing photocatalysts with good carrier separation efficiency.

A novel missense mutation of LRP6 identified by whole-exome sequencing in a Chinese family with non-syndromic tooth agenesis.

OBJECTIVE: The aim of this study was to explore the genetic basis of non-syndromic tooth agenesis (TA) in a Chinese family of five individuals using whole-exome sequencing (WES) analysis. SETTINGS AND SAMPLE POPULATION: Five participants/Family-based study of a non-syndromic TA proband. METHODS: The proband, proband's mother and grandmother displayed congenital tooth deficiency. Genomic DNA was extracted from the peripheral blood or saliva samples of the proband, her parents and her grandmother, and WES was utilized to identify the causal genetic mutation. The identified mutation was further verified by Sanger sequencing and analysed using bioinformatics tools. RESULTS: A novel missense mutation, c.G711T (p.L237F), was identified in the low-density lipoprotein receptor-related protein 6 (LRP6) gene in all affected individuals. Bioinformatics analysis predicted the mutation to be deleterious, with the mutant LRP6 protein displaying a tertiary structural change that might disturb the Wnt/ß-catenin signalling pathway. CONCLUSIONS: The identification of the mutation in the LRP6 gene and autosomal dominant inheritance with TA in the generations is consistent with the mutation being responsible for TA in the family, and furthers the association of LRP6 with nonsyndromic TA.

Delivery Mechanism of the Pharmaceutical Complex of Genistein-Adenine Based on Spectroscopic and Molecular Modelling at Atomic Scale.

Genistein (GS) exhibits various biological activities, but its clinical application is limited because of the low bioavailability. In this study, a GS-adenine pharmaceutical complex was prepared through solvent evaporation to improve the bioavailability of GS, and a molecular model of a two-component supramolecular pharmacological transport mechanism was established. The structure of GS-adenine was characterized, in addition, interaction patterns between GS and adenine were investigated using density functional theory. The results showed that the solubility of GS-adenine was five times higher than that of GS, and the cumulative release rate of GS-adenine was 86 %. The results of fluorescence spectroscopy and molecular dynamic simulations showed that GS-adenine bound to the Sudlow's site I of HSA mainly through hydrophobic interactions. This study provides a useful reference for synthesizing pharmaceutical complexes to improve solubility and for exploring the mechanism of multiple pharmaceutical components in vivo.

A novel mutation of MSX1 inherited from maternal mosaicism causes a severely affected child with nonsyndromic oligodontia.

Mutations of MSX1 have been associated with nonsyndromic hypodontia. To seek the causal gene mutation sites in a family with nonsyndromic oligodontia, whole-exome sequencing (WES) was performed to seek the causative locus of the family. The candidate mutation was further identified by Sanger sequencing afterward. Two mutations of MSX1 were found both in the proband and her mother. One novel heterozygous missense mutation (c.C667G, p.R223G) of MSX1 inherited from the asymptomatic mother with mosaic mutation was located in the highly conserved fragment of exon 2. The other was a synonymous mutation (c.C348T, p.G116G) in exon 1, which had been reported. The novel maternal heterozygous missense mutation (c.C667G, p.R223G) was likely to be the major reason for nonsyndromic oligodontia in the family. This is the first mosaic variant that has been recorded of the MSX1 gene. Our study expands the phenotype-genotype correlation associated with MSX1 variants. Our study also suggests that the determination of the mosaicism is essential for precise genetic counseling if a disease appears to arise de novo.

Folium Sennae and emodin reverse airway smooth muscle contraction.

The objective of this project was to find a bronchodilatory compound from herbs and clarify the mechanism. We found that the ethanol extract of Folium Sennae (EEFS) can relax airway smooth muscle (ASM). EEFS inhibited ASM contraction, induced by acetylcholine, in mouse tracheal rings and lung slices. High-performance liquid chromatography assay showed that EEFS contained emodin. Emodin had a similar reversal action. Acetylcholine-evoked contraction was also partially reduced by nifedipine (a selective inhibitor of L-type voltage-dependent Ca2+ channels, LVDCCs), YM-58483 (a selective inhibitor of store-operated Ca2+ entry, SOCE), as well as Y-27632 (an inhibitor of Rho-associated protein kinase). In addition, LVDCC- and SOCE-mediated currents and cytosolic Ca2+ elevations were inhibited by emodin. Emodin reversed acetylcholine-caused increases in phosphorylation of myosin phosphatase target subunit 1. Furthermore, emodin, in vivo, inhibited acetylcholine-induced respiratory system resistance in mice. These results indicate that EEFS-induced relaxation results from emodin inhibiting LVDCC, SOCE, and Ca2+ sensitization. These findings suggest that Folium Sennae and emodin may be new sources of bronchodilators.

Paracetamol inhibits Ca2+ permeant ion channels and Ca2+ sensitization resulting in relaxation of precontracted airway smooth muscle.

The purpose of this study was to screen a bronchodilator from old drugs and elucidate the underlying mechanism. Paracetamol (acetaminophen) is a widely used analgesic and antipyretic drug. It has been reported that it inhibits the generation of prostaglandin and histamine, which play roles in asthma. These findings led us to explore whether paracetamol could be a potential bronchodilator. Paracetamol inhibited high K+- and acetylcholine (ACH)-induced precontraction of mouse tracheal and bronchial smooth muscles. Moreover, the ACH-induced contraction was partially inhibited by nifedipine (selective blocker of LVDCCs), YM-58483 (selective inhibitor of store-operated Ca2+ entry (SOCE), canonical transient receptor potential 3 (TRPC3) and TRPC5 channels) and Y-27632 (selective blocker of ROCK, a linker of the Ca2+ sensitization pathway). In single airway smooth muscle cells, paracetamol blocked the currents sensitive to nifedipine and YM-58483, and inhibited intracellular Ca2+ increases. In addition, paracetamol inhibited ACH-induced phosphorylation of myosin phosphatase target subunit 1 (MYPT1, another linker of the Ca2+ sensitization pathway). Finally, in vivo paracetamol inhibited ACH-induced increases of mouse respirator system resistance. Collectively, we conclude that paracetamol inhibits ASM contraction through blocking LVDCCs, SOCE and/or TRPC3 and/or TRPC5 channels, and Ca2+ sensitization. These results suggest that paracetamol might be a new bronchodilator.

[Occurrences and dietary exposure analysis for acaricides in the 6th Chinese total diet study].

OBJECTIVE: To study the residue levels of acaricides in Chinese dietary samples and dietary intakes of Chinese residents. METHODS: A total of 288 mixed dietary samples from 24 provinces in the 6 th Chinese total diet study were analyzed for residue levels of acaricides by liquid chromatography-high resolution mass spectrometry. Additionally, based on the dietary consumption of local residents, the dietary intake of acaricides was estimated. RESULTS: Among the tested dietary samples, the detection rates of pyridaben, spirodiclofen and propargite were 11. 8%(34/288), 8. 7%(25/288), and 6. 3%(18/288), respectively. They were mainly detected in vegetable and fruit samples. The average residue level of pyridaben in the vegetable samples was higher than that in other dietary samples with the content of 6. 26 µg/kg. The average residue level of spirodiclofen in the fruits samples was higher than that in other dietary samples with the content of 3. 92 µg/kg. The average residue level of propargite in the vegetable samples was higher than that in other dietary samples with the content of 0. 90 µg/kg. According to the dietary exposure analysis, the average dietary exposure levels of pyridaben, spirodiclofen and propargite in the general population of China were 48. 31, 1. 62 and 2. 25 ng/(kg·d), respectively. According to the dietary contribution rate, the three acaricides were mainly from vegetable samples. CONCLUSION: Although acaricides were detected in varying degrees in the Chinese dietary samples, the general population's health risk caused by the dietary intake in China is at a low level.

A novel PAX9 mutation found in a Chinese patient with hypodontia via whole exome sequencing.

OBJECTIVE: To investigate a novel gene mutation in a Chinese patient with non-syndromic hypodontia. SUBJECTS AND METHODS: Mutation analysis was carried out by whole exome sequencing. Bioinformatics tools were used for the biophysical predictions of the mutative protein. Luciferase reporter assay was performed to analyse the effects of mutation on protein function. PAX9 and BMP4 gene expression from mutant cells was detected by qRT-PCR. RESULTS: A novel heterozygous mutation (c.G1057A) was detected in the patient but was not found in the controls. The novel missense mutation led to a Val111Met substitution in the paired box domain which was completely conserved evolutionarily, as analysed by dbNSFP. The mutation was predicted to be disease-causing and harmful using MutationTaster and CADD, respectively. Protean of Lasergene showed that this mutation may lead to ß-region shortening in the mutant protein compared to the wild type. Luciferase reporter assay indicated that the mutated protein reduced the transactivation activity of PAX9. This mutation led to increased levels of PAX9 transcript and reduced levels of BMP4 transcript, likely due to compensatory activation and lower transactivation activity of mutant PAX9. CONCLUSION: This novel mutation (c.G1057A) in PAX9 caused hypodontia by altering PAX9 gene function and downregulating BMP4 gene expression.

[Ethacrynic acid inhibits airway smooth muscle contraction in mice].

The aim of this study was to investigate the inhibitory effect and the underlying mechanism of ethacrynic acid (EA) on the contraction in mice. BL-420S force measuring system was used to measure the tension of mouse tracheal rings. The whole cell patch clamp technique was utilized to record the channel currents of airway smooth muscle (ASM) cells. The calcium imaging system was used to determine the intracellular Ca2+ concentration ([Ca2+]i) in ASM cells. The results showed that EA significantly inhibited the high K+ (80 mmol/L) and acetylcholine (ACh, 100 µmol/L)-induced contraction of mouse tracheal rings in a dose-dependent manner. The maximal relaxation percentages were (97.02 ± 1.56)% and (85.21 ± 0.03)%, and the median effective concentrations were (40.28 ± 2.20) µmol/L and (56.22 ± 7.62) µmol/L, respectively. EA decreased the K+ and ACh-induced elevation of [Ca2+]i from 0.40 ± 0.04 to 0.16 ± 0.01 and from 0.50 ± 0.01 to 0.39 ± 0.01, respectively. In addition, EA inhibited L-type voltage-dependent calcium channel (LVDCC) and store-operated calcium channel (SOCC) currents in ASM cells, and Ca2+ influx. Moreover, EA decreased the resistance of the respiratory system (Rrs) in vivo in mice. These results indicated that EA inhibits LVDCC and SOCC, which results in termination of Ca2+ influx and decreases of [Ca2+]i, leading to relaxation of ASM. Taken together, EA might be a potential bronchodilator.

Generation and Role of Oscillatory Contractions in Mouse Airway Smooth Muscle.

BACKGROUND/AIMS: Tetraethylammonium chloride (TEA) induces oscillatory contractions in mouse airway smooth muscle (ASM); however, the generation and maintenance of oscillatory contractions and their role in ASM are unclear. METHODS: In this study, oscillations of ASM contraction and intracellular Ca2+ were measured using force measuring and Ca2+ imaging technique, respectively. TEA, nifedipine, niflumic acid, acetylcholine chloride, lithium chloride, KB-R7943, ouabain, 2-Aminoethoxydiphenyl borate, thapsigargin, tetrodotoxin, and ryanodine were used to assess the mechanism of oscillatory contractions. RESULTS: TEA induced depolarization, resulting in activation of L-type voltage-dependent Ca2+ channels (LVDCCs) and voltage-dependent Na+ (VNa) channels. The former mediated Ca2+ influx to trigger a contraction and the latter mediated Na+ entry to enhance the contraction via activating LVDCCs. Meanwhile, increased Ca2+-activated Cl- channels, inducing depolarization that resulted in contraction through LVDCCs. In addition, the contraction was enhanced by intracellular Ca2+ release from Ca2+ stores mediated by inositol (1,4,5)-trisphosphate receptors (IP3Rs). These pathways together produce the contractile phase of the oscillatory contractions. Furthermore, the increased Ca2+ activated the Na+-Ca2+ exchanger (NCX), which transferred Ca2+ out of and Na+ into the cells. The former induced relaxation and the latter activated Na+/K+-ATPase that induced hypopolarization to inactivate LVDCCs causing further relaxation. This can also explain the relaxant phase of the oscillatory contractions. Moreover, the depolarization induced by VNa channels and NCX might be greater than the hypopolarization caused by Na+/K+-ATPase alone, inducing LVDCC activation and resulting in further contraction. CONCLUSIONS: These data indicate that the TEA-induced oscillatory contractions were cooperatively produced by LVDCCs, VNa channels, Ca2+-activated Cl- channels, NCX, Na+/K+ ATPase, IP3Rs-mediated Ca2+ release, and extracellular Ca2+.

Female Reproductive Performance in the Mouse: Effect of Oral Melatonin.

Although melatonin has some of the broadest ranges of actions on the physiology of vertebrates, especially on their reproductive processes, the mechanism by which melatonin regulates animal reproduction is still incompletely understood. This study was designed to determine the effect of oral melatonin on the reproductive performance of female mice. Female ICR mice (7 weeks old) were given melatonin-containing water (3, 30 and 300 µg/mL; melatonin) or water only (control) until 10 weeks of age. Then, some of the mice were successfully mated (confirmed by vaginal plugs), and the number of live births and their weights were recorded. Some mice were used for a histological analysis of the number of follicles in the ovaries. Others were used for oocyte collection after superovulation, and in vitro fertilization (IVF) was performed. The mRNA expression of the apopotosis-related genes (BAX, BCL2) in the IVF embryos were analyzed. After melatonin administration, the mice showed similar serum melatonin levels to that of the control. The number of antral follicles per mm² unit area in the 30 µg/mL melatonin-treated group (14.60) was significantly higher than that of the control (7.78), which was lower than that of the 3 µg/mL melatonin-treated group (12.29). The litter size was significantly higher in the 3 µg/mL melatonin-treated group (15.5) than in the control (14.3). After IVF, the hatched blastocyst formation rate in the 30 µg/mL melatonin-treated group (85.70%) was significantly higher than that of the control (72.10%), and it was the same for the BCL2/BAX expression ratio. Although oral melatonin did not appear to have an effect on the serum melatonin rhythm in the mouse, melatonin did increase litter size at the 3 µg/mL dose level, and improved the developmental competency of IVF embryos at the 30 µg/mL level.