Cadmium disrupts spermatogenic cell cycle via piRNA-DQ717867/p53 pathway.

Cadmium (Cd) is a harmful environmental pollutant that disrupts public health, including respiratory, digestive, and reproductive systems. In this study, male rats were exposed to CdCl2 at a dose of 3 mg/kg by oral for 28 days to investigate the impact on spermatogenesis. Testis tissue samples were collected after sacrifice, and piRNA expression levels were measured using piRNA microarray and qPCR. PiRNAs, specialized molecules involved in spermatogenesis, were examined. CdCl2 exposure led to disrupted piRNA expression, particularly in piRNA-DQ759395 in rats. This piRNA was found to have a binding site with p53, and a similar piRNA-DQ717867 was discovered in mice. In GC-2spd cells, CdCl2 exposure increased piRNA-DQ717867 expression, which resulted in cell cycle arrest and abnormal expression of cell cycle-related proteins. The activation of p53-related pathways and disruptions in cell cycle regulation were also observed. Antagomir-717867 transfections and PFT-a pretreatment in GC-2spd cells supported the involvement of piRNA-DQ717867 in regulating cell cycle-related proteins. This study suggests that Cd exposure induces abnormal expression of piRNA-DQ759395 in rat testis and that piRNA-DQ717867 may regulate p53, causing cell cycle abnormalities in GC-2spd cells. These findings help understand the mechanisms of male reproductive toxicity caused by Cd exposure and emphasize the role of piRNAs in cell cycle regulation and male reproductive health.

Prime editing in mice with an engineered pegRNA.

CRISPR editing involves double-strand breaks in DNA with attending insertions/deletions (indels) that may result in embryonic lethality in mice. The prime editing (PE) platform uses a prime editing guide RNA (pegRNA) and a Cas9 nickase fused to a modified reverse transcriptase to precisely introduce nucleotide substitutions or small indels without the unintended editing associated with DNA double-strand breaks. Recently, engineered pegRNAs (epegRNAs), with a 3'-extension that shields the primer-binding site of the pegRNA from nucleolytic attack, demonstrated superior activity over conventional pegRNAs in cultured cells. Here, we show the inability of three-component CRISPR or conventional PE to incorporate a nonsynonymous substitution in the Capn2 gene, expected to disrupt a phosphorylation site (S50A) in CAPN2. In contrast, an epegRNA with the same protospacer correctly installed the desired edit in two founder mice, as evidenced by robust genotyping assays for the detection of subtle nucleotide substitutions. Long-read sequencing demonstrated sequence fidelity around the edited site as well as top-ranked distal off-target sites. Western blotting and histological analysis of lipopolysaccharide-treated lung tissue revealed a decrease in phosphorylation of CAPN2 and notable alleviation of inflammation, respectively. These results demonstrate the first successful use of an epegRNA for germline transmission in an animal model and provide a solution to targeting essential developmental genes that otherwise may be challenging to edit.

The theoretical design of manganese catalysts with a Si-N-Si-C-Si-C six-membered ring core-based bowl-shaped quadridentate ligand for the hydrogenation of CO/CN bonds.

Herein, a new series of bowl-shaped quadridentate ligands with a Si-N-Si-C-Si-C six-membered ring core and their manganese catalysts were designed using the density functional theory (DFT) method for the hydrogenation of unsaturated CX (XN, O) bonds. The frameworks of these ligands named by LYG (LYG = P(R1)2CH2Si(CH2)(CH3)NSi(CH3)(CH2Si(CH3)CH2P(R3)2)CH2P(R2)2) have a Si-N-Si-C-Si-C six-membered ring core at the bottom of the bowl structure and each Si atom links with one phosphorus arm (-CH2PR2). The Mn catalyst Mn(CO)-LYG was constructed to catalyze the hydrogenation of CO/CN bonds. The calculated results indicate that due to the bowl-shaped structure of LYG quadridentate ligands, these Mn catalysts could be advantageous not only in the tuneup of catalytic activity and stereoselectivity by modifying three phosphorus arms but also in the homogeneous catalyst immobilization by linking with the Si-N-Si-C-Si-C six-membered ring core using different supports. This work might provide theoretical insights to design new framework transition-metal catalysts for the hydrogenation of CX bonds.

Calpain Promotes LPS-induced Lung Endothelial Barrier Dysfunction via Cleavage of Talin.

Acute lung injury (ALI) is characterized by lung vascular endothelial cell (EC) barrier compromise resulting in increased endothelial permeability and pulmonary edema. The infection of gram-negative bacteria that produce toxins like LPS is one of the major causes of ALI. LPS activates Toll-like receptor 4, leading to cytoskeleton reorganization, resulting in lung endothelial barrier disruption and pulmonary edema in ALI. However, the signaling pathways that lead to the cytoskeleton reorganization and lung microvascular EC barrier disruption remain largely unexplored. Here we show that LPS induces calpain activation and talin cleavage into head and rod domains and that inhibition of calpain attenuates talin cleavage, RhoA activation, and pulmonary EC barrier disruption in LPS-treated human lung microvascular ECs in vitro and lung EC barrier disruption and pulmonary edema induced by LPS in ALI in vivo. Moreover, overexpression of calpain causes talin cleavage and RhoA activation, myosin light chain (MLC) phosphorylation, and increases in actin stress fiber formation. Furthermore, knockdown of talin attenuates LPS-induced RhoA activation and MLC phosphorylation and increased stress fiber formation and mitigates LPS-induced lung microvascular endothelial barrier disruption. Additionally, overexpression of talin head and rod domains increases RhoA activation, MLC phosphorylation, and stress fiber formation and enhances lung endothelial barrier disruption. Finally, overexpression of cleavage-resistant talin mutant reduces LPS-induced increases in MLC phosphorylation in human lung microvascular ECs and attenuates LPS-induced lung microvascular endothelial barrier disruption. These results provide the first evidence that calpain mediates LPS-induced lung microvascular endothelial barrier disruption in ALI via cleavage of talin.

Purine synthesis suppression reduces the development and progression of pulmonary hypertension in rodent models.

AIMS: Proliferation of vascular smooth muscle cells (VSMCs) is a hallmark of pulmonary hypertension (PH). Proliferative cells utilize purine bases from the de novo purine synthesis (DNPS) pathways for nucleotide synthesis; however, it is unclear whether DNPS plays a critical role in VSMC proliferation during development of PH. The last two steps of DNPS are catalysed by the enzyme 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/inosine monophosphate cyclohydrolase (ATIC). This study investigated whether ATIC-driven DNPS affects the proliferation of pulmonary artery smooth muscle cells (PASMCs) and the development of PH. METHODS AND RESULTS: Metabolites of DNPS in proliferative PASMCs were measured by liquid chromatography-tandem mass spectrometry. ATIC expression was assessed in platelet-derived growth factor-treated PASMCs and in the lungs of PH rodents and patients with pulmonary arterial hypertension. Mice with global and VSMC-specific knockout of Atic were utilized to investigate the role of ATIC in both hypoxia- and lung interleukin-6/hypoxia-induced murine PH. ATIC-mediated DNPS at the mRNA, protein, and enzymatic activity levels were increased in platelet-derived growth factor-treated PASMCs or PASMCs from PH rodents and patients with pulmonary arterial hypertension. In cultured PASMCs, ATIC knockdown decreased DNPS and nucleic acid DNA/RNA synthesis, and reduced cell proliferation. Global or VSMC-specific knockout of Atic attenuated vascular remodelling and inhibited the development and progression of both hypoxia- and lung IL-6/hypoxia-induced PH in mice. CONCLUSION: Targeting ATIC-mediated DNPS compromises the availability of purine nucleotides for incorporation into DNA/RNA, reducing PASMC proliferation and pulmonary vascular remodelling and ameliorating the development and progression of PH.

Cyclic diguanylate analogues: Facile synthesis, STING binding mode and anti-tumor immunity delivered by cytidinyl/cationic lipid.

Herein 2-cyanoethoxy-N,N,N',N'-tetraisopropyl-phosphorodiamidite(10, PIII, 3.5 eq.) could synergistically react with 3',5'-dihydroxyl groups in a dinucleotide(PV) at the cyclization step for the synthesis of cyclic dinucleotides (CDNs) (c-di-GMP, cGAMP etc.) and their phosphorothioated analogues. A dynamic PIII-PV coordination mechanism has been proposed for the cyclization procedure which is confirmed by the variant 31P NMR data and molecular simulation. Among the mono-phosphorothioated CDNs, two stereoisomers showed different capacity for STING activation and the reason was predicted by molecular modeling. While compound 12b1 showed most potent ability to elicit cytokines (IFNß, IL-6, Cxcl9 and Cxcl10) induction compared to another stereoisomer. Also, 12b1 significantly inhibited the tumor growth in the EO771 model with both 0.1 µg (i.t.) and 2 µg (i.v.) administration through the aid of a Mix delivery system developed by our group, and achieved a 31% long-term survival rate of tumor-bearing mice. 12b1/Mix significantly improved the percentage of CD8+ or CD4+ effector memory T (Tem, CD44highCD62Llow) cells and CD8+ central memory T (Tcm, CD44highCD62Lhigh) cells in the blood of EO771 mice, inducing the immune memory against EO771 tumor cells. Relatively lower dose regimens of 12b1(0.1 µg)/Mix displayed better tumor suppression by more potent STING pathway activation and higher levels of cytokines induction in the tumor.

Constructing Carbon Nanotube Hybrid Fiber Electrodes with Unique Hierarchical Microcrack Structure for High-Voltage, Ultrahigh-Rate, and Ultralong-Life Flexible Aqueous Zinc Batteries.

Flexible aqueous zinc batteries are promising candidates as safe power sources for fast-growing portable and wearable electronics. However, the low working voltage, poor rate capability, and cycling stability have greatly restricted their development and applications. Here, a new family of flexible bimetallic phosphide/carbon nanotube hybrid fiber electrodes with unique macroscopic microcrack structure and microscopic porous nanoflower structure is reported. The hierarchical microcrack structure not only facilitates the penetration of electrolyte for effective exposure of active sites, but also can serve as buffers to relieve the stress concentrations of the fiber electrode under deformations, enabling impressive electrochemical performance and mechanical flexibility. Particularly, the fabricated flexible aqueous zinc batteries demonstrate high working voltage plateau and specific capacity (≈1.7 V, 258.9 mAh g-1 at 2 A g-1 ), ultrahigh rate capability (135.8 mAh g-1 at 50 A g-1 , fully charged in only 9.8 s) and impressive power density of 79 000 W kg-1 . Moreover, the flexible batteries show ultralong cycling life with 74.6% capacity retention after 20 000 cycles. The fiber batteries are also highly flexible and can be easily knitted into soft electronic textiles to power a smartphone, which are particularly promising for the next-generation of flexible and wearable electronics.

Non-pharmaceutical treatments to relieve pain or reduce opioid analgesic intake and improve quality of life after total hip replacement: a meta analysis.

To reduce pain after total hip replacement (THR), researchers are interested in drug-free interventions. However, there is still a lack of consensus on their prevention efficacy. We performed a meta-analysis to evaluate the use of nonpharmaceutical interventions for postoperative pain management after THR. We searched the Cochrane Library, MEDLINE, EMBASE, Web of Science, PEDRO, and ClinicalTrials.gov databases for articles published between and 1991 and 2020. The main outcome measures were postoperative pain, opioid consumption, and quality of life (QoL). In total, 1,942 patients were studied. We found moderate evidence indicating postoperative pain relief measured by the Western Ontario and McMaster Universities Arthritis Index Scale, with mean differences (MDs) of -0.28 (95% confidence interval [CI], -0.49 to -0.07; P=0.01; I2 =0%) within three months, -0.19 (95% CI, -0.40 to 0.02; P=0.07; I2 =0%) between 3-6 months, and -0.13 (95% CI, -0.35 to 0.08; P=0.21; I2 =0%) between 6-12 months. Additionally, we found that acupuncture therapy could reduce opioid analgesic consumption (MD, -0.98; 95% CI, -1.18 to -0.79; fentanyl [mg/h]; P<0.01; I2 =72.2%) and significantly improve pain relief with an MD of 0.90 (95% CI, 0.47 to 1.33; P<0.01; I2 =0%) measured using the visual analog scale. Electrotherapy slightly improved perceived pain with an MD of 0.22 (95% CI, -0.27 to 0.70; P=0.37; I2 =0%). Moreover, moderate evidence has shown that preoperative exercises improve QoL. This meta-analysis suggested that continuous passive motion did not improve pain or QoL. Postoperative exercise was associated with pain relief and improved QoL. Acupuncture therapy after THR has been shown to reduce opioid analgesic consumption.

A theoretical study on the hydrogenation of CO2 to methanol catalyzed by ruthenium pincer complexes.

Herein, a density functional theory (DFT) study was performed to investigate thoroughly the cascade reaction mechanism for the hydrogenation of carbon dioxide to methanol catalyzed by ruthenium pincer complex [RuH2(Me2PCH2SiMe2)2NH(CO)]. Three catalytic stages involving the hydrogenation of carbon dioxide (stage I), formic acid (stage II) and formaldehyde (stage III) were studied. The calculated results show that the dominant H2 activation strategy in the hydrogenation of CO2 to methanol may not be the methanol-assisted H2 activation, but the formate-assisted H2 activation. In this cascade reaction, all energy spans of stage I, II and III are 20.2 kcal mol-1 of the formate-assisted H2 activation. This implies that it could occur under mild conditions. Meanwhile, the catalyst is proposed to be efficient for the transfer hydrogenation using isopropanol as the hydrogen resource, and the ruthenium pincer complexes [RuH2(Me2PCH2SiMe2)2NH(CO)], [RuH2(Ph2PCH2SiMe2)2NH(CO)] and [RuH2(Me2PCH2SiMe2)2NH(CO)] exhibit similar catalytic activities for the hydrogenation of CO2 to methanol.

Facet-engineering of NH2-UiO-66 with enhanced photocatalytic hydrogen production performance.

Sluggish charge transfer is the major problem which restricts the development of metal-organic framework (MOF)-based photocatalysts. Recently, facet-engineering has been proven to be an effective method for solving this issue. However, due to difficulties in regulating the exposed facets of MOFs, there are few reports about the facet-engineering of MOF-based photocatalysts. Here, we firstly report facet-engineering for promoting the photocatalytic activity of NH2-UiO-66 crystals. In this study, by regulating the influence of kinetics and thermodynamics, cubic, tetra-decahedral, and octahedral forms of NH2-UiO-66 are synthesized. The photocatalytic hydrogen evolution rate of tetra-decahedral NH2-UiO-66 with co-exposed (100) and (111) crystal facets reaches 64.06 µmol g-1 h-1, which is approximately 2 and 1.5 times greater than that of the cubic and octahedral forms of NH2-UiO-66, respectively. The density functional theory (DFT) calculation and ultrafast spectroscopy results indicate that a slight staggering exists in the band structure of (100) and (111) facets, causing the facets junction to appear. The facet junction promotes the charge separation efficiency and prolongs the lifetime of the charge carriers, thereby giving tetra-decahedral NH2-UiO-66 optimal photocatalytic performance. This study demonstrates the feasibility and potential of facet-engineering for photocatalytic applications of MOFs.

A metal-organic framework-derived Zn1-xCdxS/CdS heterojunction for efficient visible light-driven photocatalytic hydrogen production.

ZCS-C (ZnCdS/CdS) QDs were synthesized via low-temperature vulcanization using zeolitic imidazolate framework-8 (ZIF-8) nanoparticles as a precursor, cation exchange, and heterojunction construction. Without any precious metal as a cocatalyst, the photocatalytic hydrogen production rate of ZCS-C-3 QDs reached 2.7 mmol g-1 h-1 under visible light irradiation. The optimized sample exhibited an outstanding chemical stability and recyclability, which is superior to most of the reported Zn1-xCdxS-based photocatalysts.

g-C3N4/ZnCdS heterojunction for efficient visible light-driven photocatalytic hydrogen production.

To suppress the aggregation behavior caused by the high surface energy of quantum dots (QDs), ZnCdS QDs were grown in situ on a g-C3N4 support. During the growth process, the QDs tightly adhered to the support surface. The ZnCdS QDs were prepared by low-temperature sulfurization and cation exchange with a zeolitic imidazolate framework precursor under mild conditions. The heterojunction of g-C3N4/ZnCdS-2 (CN/ZCS-2, with a g-C3N4 to ZIF-8 ratio of 2.0) not only showed excellent optical absorption performance, abundant reactive sites, and a close contact interface but also effectively separated the photogenerated electrons and holes, which greatly improved its photocatalytic hydrogen production performance. Under visible light irradiation (wavelength > 420 nm) without a noble metal cocatalyst, the hydrogen evolution rate of the CN/ZCS-2 heterojunction reached 1467.23 µmol g-1 h-1, and the durability and chemical stability were extraordinarily high.

DW14006 as a direct AMPKα1 activator improves pathology of AD model mice by regulating microglial phagocytosis and neuroinflammation.

Alzheimer's disease (AD) is a progressively neurodegenerative disease with typical hallmarks of amyloid ß (Aß) plaque accumulation, neurofibrillary tangle (NFT) formation and neuronal death extension. In AD brain, activated microglia phagocytose Aß and neuronal debris, but also aggravate inflammation stress by releasing inflammatory factors and cytotoxins. Improving microglia on Aß catabolism and neuroinflammatory intervention is thus believed to be a promising therapeutic strategy for AD. AMP-activated protein kinase (AMPK) is highly expressed in microglia with AMPKα1 being tightly implicated in neuroinflammatory events. Since indirect AMPKα1 activators may cause side effects with undesired intracellular AMP/ATP ratio, we focused on direct AMPKα1 activator study by exploring its potential function in ameliorating AD-like pathology of AD model mice. Here, we reported that direct AMPKα1 activator DW14006 (2-(3-(7-chloro-6-(2'-hydroxy-[1,1'-biphenyl]-4-yl)-2-oxo-1,2-dihydroquinolin-3-yl)phenyl)acetic acid) effectively improved learning and memory impairments of APP/PS1 mice, and the underlying mechanisms have been intensively investigated. DW14006 reduced amyloid plaque deposition by promoting microglial o-Aß42 phagocytosis and ameliorated innate immune response by polarizing microglia to an anti-inflammatory phenotype. It selectively enhanced microglial phagocytosis of o-Aß42 by upgrading scavenger receptor CD36 through AMPKα1/PPARγ/CD36 signaling and suppressed inflammation by AMPKα1/IκB/NFκB signaling. Together, our work has detailed the crosstalk between AMPKα1 and microglia in AD model mice, and highlighted the potential of DW14006 in the treatment of AD.

Vascular Metabolic Mechanisms of Pulmonary Hypertension.

Pulmonary hypertension (PH) is a severe and progressive disease characterized by increased pulmonary vascular resistance leading to right heart failure and death. In PH, the cellular metabolisms including those of the three major nutrients (carbohydrate, lipid and protein) are aberrant in pulmonary vascular cells. Glucose uptake, glycolysis, insulin resistance, sphingolipid S1P, PGE2, TXA2, leukotrienes and glutaminolysis are upregulated, and phospholipid-prostacyclin and L-arginine-nitric oxide pathway are compromised in lung vascular cells. Fatty acid metabolism is disordered in lung endothelial cells and smooth muscle cells. These molecular mechanisms are integrated to promote PH-specific abnormal vascular cell proliferation and vascular remodeling. This review summarizes the recent advances in the metabolic reprogramming of glucose, fatty acid, and amino acid metabolism in pulmonary vascular remodeling in PH and the mechanisms for how these alterations affect vascular cell fate and impact the course of PH.

A polystyrene-degrading Acinetobacter bacterium isolated from the larvae of Tribolium castaneum.

Polystyrene (PS) has been widely used in various fields, whereas this thermoplastic material is generally considered to be resistant to biodegradation. Tribolium castaneum (Coleoptera: Tenebrionidae), a common pest of stored agricultural products, is a powerful model organism for general insect research. In this study, the larvae of T. castaneum were observed chewing and eating extruded polystyrene foam (XPS). Investigation of the gut microbiome of plastic- and bran-fed T. castaneum larvae showed that Acinetobacter sp. was strongly associated with PS ingestion. Additionally, one bacterial strain capable of PS degradation, was successfully isolated from the gut of these larvae and identified as Acinetobacter sp. AnTc-1 by its 16S rDNA sequence. Gel penetration chromatography (GPC), 1H nuclear magnetic resonance (1H NMR) spectroscopy, thermo gravimetric analysis (TGA) and scanning electron microscope (SEM) were employed to characterize the PS degradation. After incubation with AnTc-1 for 60 days, the mass weight (12.14%) and molecular weight (13%/25%, weight-average molecular weight (Mw)/number-average molecular weight (Mn)) of PS powder were significantly reduced. The results indicated that the isolated strain of Acinetobacter sp. AnTc-1 has PS-degrading capacity. The isolated strain may play a role in the larval gut for biodegradation of PS and has potential to be applied for petroleum-based plastic degradation study and development of remediation approaches.

Vincamine as a GPR40 agonist improves glucose homeostasis in type 2 diabetic mice.

Vincamine, a monoterpenoid indole alkaloid extracted from the Madagascar periwinkle, is clinically used for the treatment of cardio-cerebrovascular diseases, while also treated as a dietary supplement with nootropic function. Given the neuronal protection of vincamine and the potency of ß-cell amelioration in treating type 2 diabetes mellitus (T2DM), we investigated the potential of vincamine in protecting ß-cells and ameliorating glucose homeostasis in vitro and in vivo. Interestingly, we found that vincamine could protect INS-832/13 cells function by regulating G-protein-coupled receptor 40 (GPR40)/cAMP/Ca2+/IRS2/PI3K/Akt signaling pathway, while increasing glucose-stimulated insulin secretion (GSIS) by modulating GPR40/cAMP/Ca2+/CaMKII pathway, which reveals a novel mechanism underlying GPR40-mediated cell protection and GSIS in INS-832/13 cells. Moreover, administration of vincamine effectively ameliorated glucose homeostasis in either HFD/STZ or db/db type 2 diabetic mice. To our knowledge, our current work might be the first report on vincamine targeting GPR40 and its potential in the treatment of T2DM.

TSPA as a novel ATF6α translocation inducer efficiently ameliorates insulin sensitivity restoration and glucose homeostasis in db/db mice.

Activating transcription factor 6α (ATF6α) as a transducer in unfolded protein response (UPR), plays an important role in liver glucose metabolism and insulin resistance. Thus, targeting ATF6α activation has been proposed to be a potential strategy for anti-T2DM drug discovery. Here, we determined that small molecule 2-[5-[1-(4-chlorophenoxy)ethyl]-4-phenyl-4H-1,2,4-triazol-3-yl]sulfanyl-N-(1,5-dimethyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazol-4-yl)acetamide (TSPA) functioned as an ATF6α translocation inducer effectively promoting ATF6α translocation into nucleus and ameliorating glucose homeostasis on db/db mice. TSPA promoted ATF6α translocation into nucleus without incresing C/EBP-homologous protein (CHOP) expression. TSPA restored the tunicamycin (TM)-stimulated insulin receptor (IR) desensitization through ATF6α activation, inhibited gluconeogenesis and efficiently improved glucose homeostasis on db/db mice. Furthermore, TSPA protected insulin pathway involving p38/X-box binding protein 1s (Xbp1s)/ER chaperones signaling pathway. Our current study has determined that ATF6α was a promising therapeutic target and also highlighted the potential of TSPA in the treatment of type 2 diabetes mellitus (T2DM).

HS218 as an FXR antagonist suppresses gluconeogenesis by inhibiting FXR binding to PGC-1α promoter.

INTRODUCTION: Farnesoid X receptor (FXR) as a member of nuclear receptor is tightly associated with glucose metabolism. Accumulated evidence has addressed the potential of FXR antagonist in the treatment of type 2 diabetes mellitus (T2DM), although the related mechanisms remain unclear. Here, we determined a specific FXR antagonist HS218 (N-benzyl-N-(3-(tert-butyl)-4-hydroxyphenyl)-2,4-dichlorobenzamide), which exhibited high activities in suppressing gluconeogenesis and ameliorating glucose homeostasis in db/db and HFD/STZ-induced T2DM mice. We would like to investigate the mechanisms underlying FXR antagonism in the regulation of gluconeogenesis by using HS218 as a probe. METHODS: HS218 was evaluated by glucose output assay. Binding affinity of HS218 to the ligand binding domain of FXR (FXR-LBD) was detected by Surface Plasmon Resonance (SPR) technology-based Biacore and fluorescence quenching assays. Mammalian one-hybrid and transactivation assays were carried out to detect the antagonistic effect of HS218 on FXR. Real-time PCR assay was performed to measure the expressions of FXR-target and gluconeogenic genes. Anti-diabetic efficiencies of HS218 were determined in db/db and HFD/STZ-induced T2DM mice. Assays by promoter 5'-deletion analysis and Chromatin immunoprecipitation (ChIP) were performed to detect the binding of FXR to peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) promoter. Western blot assay was used to determine the protein level in either cells or the liver tissues of mice. RESULTS: We determined that HS218 as a new FXR specific antagonist could FXR-dependently suppress gluconeogenesis in mouse primary hepatocytes, and effectively improve glucose homeostasis in db/db and HFD/STZ-induced T2DM mice. HS218 decreased gluconeogenesis by inhibiting the FXR-induced increase in the promoter activity of the key gluconeogenic gene PGC-1α, leading to the repression of PGC-1α and its target gene peroxisome proliferator-activated receptor α (PPARα). CONCLUSIONS: To our knowledge, our work might be the first report on the mechanism underlying FXR antagonist in the regulation of gluconeogenesis, and all results have also highlighted the potential of HS218 in the treatment of T2DM.

An Optimization Study on Listening Experiments to Improve the Comparability of Annoyance Ratings of Noise Samples from Different Experimental Sample Sets.

Annoyance ratings obtained from listening experiments are widely used in studies on health effect of environmental noise. In listening experiments, participants usually give the annoyance rating of each noise sample according to its relative annoyance degree among all samples in the experimental sample set if there are no reference sound samples, which leads to poor comparability between experimental results obtained from different experimental sample sets. To solve this problem, this study proposed to add several pink noise samples with certain loudness levels into experimental sample sets as reference sound samples. On this basis, the standard curve between logarithmic mean annoyance and loudness level of pink noise was used to calibrate the experimental results and the calibration procedures were described in detail. Furthermore, as a case study, six different types of noise sample sets were selected to conduct listening experiments using this method to examine the applicability of it. Results showed that the differences in the annoyance ratings of each identical noise sample from different experimental sample sets were markedly decreased after calibration. The determination coefficient (R²) of linear fitting functions between psychoacoustic annoyance (PA) and mean annoyance (MA) of noise samples from different experimental sample sets increased obviously after calibration. The case study indicated that the method above is applicable to calibrating annoyance ratings obtained from different types of noise sample sets. After calibration, the comparability of annoyance ratings of noise samples from different experimental sample sets can be distinctly improved.

Discovery of cycloalkyl-fused N-thiazol-2-yl-benzamides as tissue non-specific glucokinase activators: Design, synthesis, and biological evaluation.

Glucokinase (GK) activators are being developed for the treatment of type 2 diabetes mellitus (T2DM). However, existing GK activators have risks of hypoglycemia caused by over-activation of GK in islet cells and dyslipidemia caused by over-activation of intrahepatic GK. In the effort to mitigate risks of hypoglycemia and dyslipidemia while maintaining the promising efficacy of GK activator, we investigated a series of cycloalkyl-fused N-thiazol-2-yl-benzamides as tissue non-specific partial GK activators, which led to the identification of compound 72 that showed a good balance between in vitro potency and enzyme kinetic parameters, and protected ß-cells from streptozotocin-induced apoptosis. Chronic treatment of compound 72 demonstrated its potent activity in regulation of glucose homeostasis and low risk of dyslipidemia with diabetic db/db mice in oral glucose tolerance test (OGTT). Moreover, acute treatment of compound 72 did not induce hypoglycemia in C57BL/6J mice even at 200 mg/kg via oral administration.