Mechanistic insight into the impact of interaction between goethite and humic acid on the photooxidation and photoreduction of bifenthrin.

Numerous application of pyrethroid insecticides has led to their accumulation in the environment, threatening ecological environment and human health. Its fate in the presence of iron-bearing minerals and natural organic matter under light irradiation is still unknown. We found that goethite (Gt) and humic acid (HA) could improve the photodegradation of bifenthrin (BF) in proper concentration under light irradiation. The interaction between Gt and HA may further enhance BF degradation. On one hand, the adsorption of HA on Gt may decrease the photocatalytic activity of HA through decreasing HA content in solution and sequestering the functional groups related with the production of reactive species. On the other hand, HA could improve the photocatalytic activity of Gt through extending light absorption, lowing of bandgap energy, hindering the recombination of photo-generated charges, and promoting the oxidation and reduction reaction on Gt surface. The increased oxygen vacancies on Gt surface along with the reduction of trivalent iron and the nucleophilic attack of hole to surface hydroxyl group contributed to the increasing photocatalytic activity of Gt. Electron paramagnetic resonance and quenching studies demonstrated that both oxidation species, such as hydroxyl radical (•OH) and singlet oxygen (1O2), and reducing species, such as hydrogen atoms (H•) and superoxide anion radical (O2•-), contributed to BF degradation in UV-Gt-HA system. Mass spectrometry, ion chromatography, and toxicity assessment indicated that less toxic C23H22ClF3O3 (OH-BF), C9H10ClF3O (TFP), C14H14O2 (OH-MBP), C14H12O2 (MBP acid), C14H12O3 (OH-MBP acid), and chloride ions were the main degradation products. The production of OH-BF, MPB, and TFP acid through oxidation and the production of MPB and TFP via reduction were the two primary pathways of BF degradation.

Mechanism of interaction between ascorbic acid and soil iron-containing minerals for peroxydisulfate activation and organophosphorus flame retardant degradation.

Soil constituents may play an important role in peroxydisulfate (PDS)-based oxidation of organic contaminants in soil. Iron-containing minerals (Fe-minerals) have been found to promote PDS activation for organics degradation. Our study found that ascorbic acid (H2A) could enhance PDS activation by soil Fe-minerals for triphenyl phosphate (TPHP) degradation. Determination and characterization analyses of Fe fractions showed that H2A could induce the reductive dissolution of solid Fe-minerals and the increasing of oxygen vacancies/hydroxyl groups content on Fe-minerals surface. The increasing of divalent Fe (Fe(II)) accelerated PDS activation to generate reactive oxygen species (ROS). Electron paramagnetic resonance (EPR) and quenching studies showed that sulfate radicals (SO4•-) and hydroxyl radicals (HO•) contributed significantly to TPHP degradation. The composition and content of Fe-minerals and soil organic matter (SOM) markedly influenced ROS transformations. Surface-bond and structural Fe played the main role in the production of Fe(II) in reaction system. The high-concentration SOM could result in ROS consumption and degradation inhibition. Density functional theory (DFT) studies revealed that H2A is preferentially adsorbed at α-Fe2O3(012) surface through Fe-O-C bridges rather than hydrogen bonds. After absorption, H atoms on H2A may further be migrated to adjacent O atoms on the α-Fe2O3(012) surface. With the transformation of H atoms to the α-Fe2O3(012) surface, the Fe-O-C bridge is broken and one electron is transferred from the O to Fe atom, inducing the reduction of trivalent Fe (Fe(III)) atom. MS/MS2 analysis, HPLC analysis, and toxicity assessment demonstrated that TPHP was transformed to less toxic 4-hydroxyphenyl diphenyl phosphate (OH-TPHP), diphenyl hydrogen phosphate (DPHP), and phenyl phosphate (PHP) through phenol-cleavage and hydroxylation processes, and even be mineralized in reaction system.

Screening for genes, miRNAs and transcription factors of adipogenic differentiation and dedifferentiation of mesenchymal stem cells.

BACKGROUND: The purpose of present study was to reveal the molecular mechanisms responsible for both adipogenic differentiation and dedifferentiation of mesenchymal stem cells (MSCs). METHODS: Microarray data GSE36923 were obtained from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) between adipogenically differentiated cells vs undifferentiated bone marrow-derived MSCs, adipogenically differentiated cells vs dedifferentiated cells samples at day 7 and adipogenically differentiated cells vs dedifferentiated cells samples at day 35 were screened, and overlapped DEGs across the three groups were analyzed. The underlying functions of the upregulated and downregulated DEGs were investigated by Gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analysis. The protein-protein interaction network was constructed, and hub genes were obtained subsequently. Hub genes were verified with GSE113253 dataset, and then miRNA-gene network and TF-gene network were constructed. RESULTS: A total of 284 upregulated DEGs and 376 downregulated DEGs overlapped across the three groups. PPAR signaling pathway, AMPK signaling pathway, insulin signaling pathway, carbon metabolism, pyruvate metabolism, fatty acid metabolism, regulation of lipolysis in adipocytes, biosynthesis of amino acids, citrate cycle (TCA cycle) and 2-Oxocarboxylic acid metabolism were the top 10 pathways involving in the upregulated DEGs, and graft-versus-host disease, allograft rejection, viral myocarditis, cell adhesion molecules, phagosome, type I diabetes mellitus, antigen processing and presentation, autoimmune thyroid disease, intestinal immune network for IgA production and rheumatoid arthritis were the top 10 pathways in downregulated DEGs. After validation, the 8 hub genes were IL6, PPARG, CCL2, FASN, CEBPA, ADIPOQ, FABP4 and LIPE. Ten key miRNAs were hsa-mir-27a-3p, hsa-mir-182-5p, hsa-mir-7-5p, hsa-mir-16-5p, hsa-mir-1-3p, hsa-mir-155-5p, hsa-mir-21-3p, hsa-mir-34a-5p, hsa-mir-27a-5p and hsa-mir-30c-5p, and 10 key TFs were TFDP1, GTF2A2, ZNF584, NRF1, ZNF512, NFRKB, CEBPG, KLF16, GLIS2 and MXD4. CONCLUSION: Our study constructed miRNA-gene network and TF-gene network involved in both adipogenic differentiation and dedifferentiation of MSCs, contributing to enhancing the efficiency of MSCs transplantation in soft tissue defect repair and developing more potent remedies for adipogenesis-related skeletal disorders.

Anxiety disorders and adhesive capsulitis: a bidirectional Mendelian randomization study.

Background: Previous epidemiological investigations and related research efforts consistently have outlined an observable association between anxiety disorders and adhesive capsulitis (AC). However, the intricate nature of the causal connection between these entities has yet to be fully clarified. Therefore, this investigative study aims to thoroughly examine and delineate the causal interrelationship between anxiety disorders and AC using a bidirectional, two-sample Mendelian randomization (MR) approach. Methods: To pursue this inquiry, datasets related to anxiety disorders and AC were meticulously obtained from a publicly accessible genomewide association study. Instrumental variables, in the form of single nucleotide polymorphisms, were subsequently identified, undergoing a rigorous screening process that included intensity adjustment and the amelioration of linkage disequilibrium. The primary analytical tool for scrutinizing causal ramifications was the inverse variance weighting (IVW) methodology, complemented by supplementary analytical techniques such as weighted median, MR-Egger, simple mode, and weighted mode. Additionally, evaluations of heterogeneity and pleiotropy were meticulously conducted. Heterogeneity was assessed using Cochran's Q-test in conjunction with the IVW and MR-Egger methods, while pleiotropy was appraised through the MR-Egger intercept and MR-PRESSO analysis methods. A leave-one-out analysis was undertaken to enhance the reliability of our findings. Finally, AC was utilized to infer reverse causality concerning the risk of anxiety disorders. Results: The random effects IVW analysis results yielded statistical significance (P = 9.362 × 10-6), demonstrating a causal link between anxiety disorders and elevated susceptibility to AC, reflected in an odds ratio of 1.267 (95% confidence interval: 1.141-1.407). Conversely, the inverse MR analysis predominantly produced null findings. Furthermore, sensitivity analyses underscored the robustness of our conclusions. Conclusion: In summary, our meticulously conducted study unequivocally supports the presence of a causal connection between anxiety disorders and an increased propensity for AC. Unfortunately, the reverse MR analysis failed to provide compelling evidence indicative of a reciprocal genetic causative relationship between AC and anxiety disorders.

Surface-enhanced Raman spectroscopy detection of organic molecules and in situ monitoring of organic reactions by ion-induced silver nanoparticle clusters.

Surface-enhanced Raman spectroscopy (SERS) finds wide applications in the field of organic molecule detection. However, reliable SERS detection of organic molecules and in situ monitoring of organic reactions under natural conditions by metal colloids are still challenging due to the formation of unstable nanoparticle clusters in solution and the low solubility of the organic molecules. Here, we approach the problems by introducing calcium ions to aggregate silver nanoparticles to form stable hot spots and acetone to promote uniform distribution of organic molecules on the nanoparticle surface. Significantly, our method exhibits stable SERS detection of up to 6 types of organic molecules in liquid. With acetone signals as an internal standard, we are able to determine molecule concentrations as well as monitor 3 kinds of organic reactions in situ. Our method shows potential for biomedical analysis, environmental analysis, and organic catalysis research.

KIF2C: a novel link between Wnt/ß-catenin and mTORC1 signaling in the pathogenesis of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is the most common primary liver malignancy and is the fourth-leading cause of cancer-related deaths worldwide. HCC is refractory to many standard cancer treatments and the prognosis is often poor, highlighting a pressing need to identify biomarkers of aggressiveness and potential targets for future treatments. Kinesin family member 2C (KIF2C) is reported to be highly expressed in several human tumors. Nevertheless, the molecular mechanisms underlying the role of KIF2C in tumor development and progression have not been investigated. In this study, we found that KIF2C expression was significantly upregulated in HCC, and that KIF2C up-regulation was associated with a poor prognosis. Utilizing both gain and loss of function assays, we showed that KIF2C promoted HCC cell proliferation, migration, invasion, and metastasis both in vitro and in vivo. Mechanistically, we identified TBC1D7 as a binding partner of KIF2C, and this interaction disrupts the formation of the TSC complex, resulting in the enhancement of mammalian target of rapamycin complex1 (mTORC1) signal transduction. Additionally, we found that KIF2C is a direct target of the Wnt/ß-catenin pathway, and acts as a key factor in mediating the crosstalk between Wnt/ß-catenin and mTORC1 signaling. Thus, the results of our study establish a link between Wnt/ß-catenin and mTORC1 signaling, which highlights the potential of KIF2C as a therapeutic target for the treatment of HCC.

Super-enhancer-driven AJUBA is activated by TCF4 and involved in epithelial-mesenchymal transition in the progression of Hepatocellular Carcinoma.

Background and Aims: Aberrant transcriptional programs are highly regulated processes that play important roles in the development and progression of hepatocellular carcinoma (HCC). Emerging evidence suggests that super-enhancers (SEs) often drive critical oncogene expression. However, SE-associated genes in HCC pathogenesis are still poorly understood. Methods: We performed integrative ChIP-seq and Hi-C analyses of HCC cells and identified ajuba LIM protein (AJUBA) as a SE-associated gene. We evaluated AJUBA expression in HCC using immunohistochemistry, immunoblotting, and qRT-PCR. ChIP and luciferase reporter assays were performed to demonstrate that transcription factor 4 (TCF4) bound to AJUBA-associated SEs. We then assessed the role of AJUBA in HCC using both in vitro and in vivo assays. Epithelial-mesenchymal transition (EMT) was examined using immunofluorescence and immunoblotting assays. Furthermore, we used immunoprecipitation and BiFC assays to explore the underlying mechanisms. Results: We identified AJUBA as a SE-associated oncogene in HCC regulated by TCF4. High AJUBA expression was related to an aggressive phenotype and unfavorable outcome in HCC patients. AJUBA knockdown significantly reduced cell migration and invasion capacities both in vitro and in vivo. Furthermore, AJUBA overexpression in HCC recruited tumor necrosis factor associated factor 6 (TRAF6), enhancing the phosphorylation of Akt and increasing Akt activity toward GSK-3ß, thus promoting EMT. Conclusions: Our results provide functional and mechanistic links between the SE-associated gene AJUBA and tumor EMT in aggressive HCC.

Correction to: KIFC1 is activated by TCF-4 and promotes hepatocellular carcinoma pathogenesis by regulating HMGA1 transcriptional activity.

In the original publication of this article [1], the indicated stages (I-II III-IV) were missing in the clinical stage part at both Table 1 and Table 2.

Correlation of iris collagen and in-vivo anterior segment structures in patients in different stages of chronic primary angle-closure in both eyes.

Purpose: To compare the density of iris collagen and anterior segment parameters in eyes with chronic primary angle closure glaucoma (CPACG) and their fellow eyes with confirmed or suspected primary angle closure (PAC/PACS). Methods: Nineteen patients with CPACG in one eye and PAC/PACS in the fellow eye requiring trabeculectomy in the CPACG eye and iridectomy in the fellow eye were recruited. Anterior segment optical coherence tomography (AS-OCT) measurements were conducted under light and dark conditions. Iris specimens, obtained by iridectomy/trabeculectomy, were analyzed by sirius red polarization for quantifying type I/III collagen density. AS-OCT parameters and type I/III collagen densities were compared between the two eyes. Results: The iris curvatures were flatter in CPACG eyes in light and dark conditions (P < 0.05). The iris areas in light condition and iris thicknesses in dark condition were smaller in CPACG eyes (P < 0.05). The density of collagen type I in CPACG eyes was lower (P = 0.048). The light-to-dark changes in CPACG eyes and PAC/PACS eyes, respectively, were -0.679 ± 0.701 and -1.627 ± 0.802 mm for pupil diameters, and 0.069 ± 0.113 and 0.258 ± 0.157 mm2 for iris areas, which differed significantly (P < 0.001). With the decrease of type I collagen, anterior chamber width increased and iris areas decreased in dark condition. Conclusion: Dynamic changes in pupil diameter and iris areas differed significantly between CPACG eyes and their fellow PAC/PACS eyes. Decreased type I collagen density in iris tissue was associated with decreased iris area and increased anterior chamber width, which may contribute to disease progression.

KIFC1 is activated by TCF-4 and promotes hepatocellular carcinoma pathogenesis by regulating HMGA1 transcriptional activity.

BACKGROUND: Kinesins play important roles in the development and progression of many human cancers. The functions and underlying mechanisms of kinesin family member C1 (KIFC1), a member of the kinesin-14 family, in the pathogenesis of hepatocellular carcinoma (HCC) have not been fully elucidated. METHODS: In this study, 168 HCC samples were first analyzed to examine the association between KIFC1 expression and patient clinicopathological features and prognosis. The role of KIFC1 in HCC cell proliferation and metastasis was investigated both in vivo and in vitro. The upstream regulation and downstream targets of KIFC1 were studied by qRT-PCR, western blotting, coimmunoprecipitation, chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays. RESULTS: KIFC1 was highly expressed in HCC tissues and positively associated with advanced stages and poor prognosis. KIFC1 knockdown suppressed HCC cell proliferation and invasion both in vitro and in vivo. Furthermore, KIFC1 knockdown decreased invadopodia formation and reduced epithelial-mesenchymal transition (EMT). HMGA1, an architectural transcriptional factor, was identified to interact with KIFC1. HMGA1 could bind to the promoters of Stat3, MMP2 and EMT-related genes and promote gene transcription. KIFC1 enhanced HMGA1 transcriptional activity and facilitated HCC proliferation and invasion. Moreover, KIFC1 was activated by TCF-4, and KIFC1 inhibition enhanced HCC cell sensitivity to paclitaxel. CONCLUSIONS: Our findings suggest that KIFC1, activated by TCF-4, functions as an oncogene and promotes HCC pathogenesis through regulating HMGA1 transcriptional activity and that KIFC1 is a potential therapeutic target to enhance the paclitaxel sensitivity of HCC.

The Relationship Between Binocular Visual Field Loss and Various Stages of Monocular Visual Field Damage in Glaucoma Patients.

PURPOSE: To investigate the effects of monocular visual field loss severity on binocular visual field (BVF) loss in primary angle-closure glaucoma, primary open-angle glaucoma, and normal tension glaucoma patients. MATERIALS AND METHODS: In this observational cross-sectional study, 250 glaucoma patients and 31 healthy participants were assigned to groups according to the stage of monocular visual field loss in both eyes; normal, early, moderate, or severe. BVF assessments were determined via integrated visual field and Esterman binocular visual evaluations. Monocular and BVF parameters were compared within and among groups. RESULTS: In patients with one eye at normal or early stage and the other at severe stage, the average integrated mean deviations (MDs) were [mean (SD)], -1.67 (1.39), and -3.27 (2.05) dB, respectively, and the average Esterman scores were >95% [99.17% (1.89%), 96.08% (3.99%), respectively]. Where both eyes had progressed to moderate or severe damage (moderate/moderate, moderate/severe, or severe/severe), the average integrated MDs were worse than -6 dB, and the mean Esterman scores in the moderate/moderate and the moderate/severe damage groups were still >90% [94.20% (5.96%), 94.32% (4.95%), respectively], but it dropped rapidly from >90% to 68.44% (26.27%) when both eyes were at severe stage. CONCLUSIONS: The BVF can remain relatively intact provided one eye is at the normal or early stage. Significant BVF defects measured by integrated MD were evident when both eyes had progressed to moderate or severe stage, and significant Esterman BVF defects were only detected when both eyes had advanced to a severe stage.

Immunosuppressive Effect of Geniposide on Mitogen-Activated Protein Kinase Signalling Pathway and Their Cross-Talk in Fibroblast-Like Synoviocytes of Adjuvant Arthritis Rats.

Geniposide (GE), an iridoid glycoside compound derived from Gardenia jasminoides Ellis fruit, is known to have anti-inflammatory and immunoregulatory activities. The aim of this study was to investigate the protective mechanism of GE in the regulation of the mitogen-activated protein kinase (MAPK) signalling pathway and the cross-talk among the MAPK signalling pathway in fibroblast-like synoviocytes (FLS) of adjuvant arthritis (AA) rats. AA was induced by injecting with Freund's complete adjuvant. Male SD rats and FLS were subjected to treatment with GE (30, 60 and 120 mg/kg) in vivo from day 14 to 21 after immunization and GE (25, 50 and 100 µg/mL) in vitro, respectively. The proliferation of FLS was assessed by MTT. IL-4, IL-17, IFN-γ, and TGF-ß1 were determined by ELISA. Key proteins in the MAPK signalling pathway were detected by Western blot. GE significantly reduced the proliferation of FLS, along with decreased IFN-γ and IL-17 and increased IL-4 and TGF-ß1. In addition, GE decreased the expression of p-JNK, p-ERK1/2 and p-p38 in FLS of AA rats. Furthermore, disrupting one MAPK pathway inhibited the activation of other MAPK pathways, suggesting cross-talk among MAPK signalling. In vivo study, it was also observed that GE attenuated histopathologic changes in the synovial tissue of AA rats. Collectively, the mechanisms by which GE exerts anti-inflammatory and immunoregulatory effects may be related to the synergistic effect of JNK, ERK1/2 and p38. Targeting MAPK signalling may be a new therapeutic strategy in inflammatory/autoimmune diseases.

Dynamic changes of anterior segment in patients with different stages of primary angle-closure in both eyes and normal subjects.

PURPOSE: To compare changes in anterior segment parameters under light and dark (light-to-dark) conditions among eyes with chronic primary angle-closure glaucoma (CPACG), fellow eyes with confirmed or suspect primary angle-closure (PAC or PACS), and age-matched healthy eyes. METHODS: Consecutive patients with CPACG in one eye and PAC/PACS in the fellow eye, as well as age-matched healthy subjects were recruited. Anterior segment optical coherence tomography measurements were conducted under light and dark conditions, and anterior chamber, lens, and iris parameters compared. Demographic and biometric factors associated with light-to-dark change in iris area were analyzed by linear regression. RESULTS: Fifty-seven patients (mean age 59.6±8.9 years) and 30 normal subjects matched for age (60.6±9.3 years) and sex ratio were recruited. In regards to differences under light-to-dark conditions, angle opening distance at 500 µm (AOD500µm) and iris area during light-to-dark transition were smaller in CPACG eyes than fellow PACS/PAC eyes and normal eyes (P<0.017). Pupil diameter change was largest in normal eyes, and larger in PACS/PAC eyes than CPACG eyes (P<0.017). There was an average reduction of 0.145 mm2 in iris area for each millimeter of pupil diameter increase in CPACG eyes, 0.161 mm2 in fellow PAC/PACS eyes, and 0.165 mm2 in normal eyes. Larger iris curvature in the dark and diagnosis of PACG were significantly associated with less light-to-dark iris area changes. CONCLUSIONS: Dynamic changes in iris parameters with light-to-dark transition differed significantly among CPACG eyes, fellow PAC/PACS eyes, and normal eyes. Greater iris curvature under dark conditions was correlated with reduced light-to-dark change in iris area and pupil diameter, which may contribute to disease progression.

Willingness to Use Mobile Health in Glaucoma Patients.

INTRODUCTION: Development of mobile health (m-health) in China carries tremendous potential, especially for glaucoma, one of the major chronic ophthalmic diseases afflicting millions of people. However, little research has been undertaken to investigate the willingness of glaucoma patients to use m-health and the factors influencing their decisions. SUBJECTS AND METHODS: This was a cross-sectional study. A self-administered or face-to-face interview survey was performed on 1,487 patients with glaucoma at the outpatient glaucoma service, Zhongshan Ophthalmic Center, Sun Yat-sen University. Questionnaires captured patients' demographic data, WeChat access, and willingness to receive m-health. The data were analyzed by single factor chi-square test. Multiple logistic regression revealed the motivators and barriers to accept m-health adoption. RESULTS: One thousand ninety-seven valid questionnaires were obtained. Seven hundred twenty-five respondents (66.1%) were willing to participate in m-health programs. 65.4% were younger than 60 years old. 40.9% had travel time from home to hospital of >3 h. 63.6% had more than four follow-up visits for glaucoma. 86.5% experienced trouble events during clinic visits. The overall WeChat usage rate was 61.7%. Age, travel time, number of visits, trouble events in clinic, and WeChat access in patients with glaucoma were related to the willingness to use m-health (p < 0.05). CONCLUSIONS: Most patients with glaucoma were willing to participate in m-health programs, which are essential to increasing and improving access to care.

The guanine nucleotide exchange factor Net1 facilitates the specification of dorsal cell fates in zebrafish embryos by promoting maternal ß-catenin activation.

Wnt/ß-catenin signaling is essential for the initiation of dorsal-ventral patterning during vertebrate embryogenesis. Maternal ß-catenin accumulates in dorsal marginal nuclei during cleavage stages, but its critical target genes essential for dorsalization are silent until mid-blastula transition (MBT). Here, we find that zebrafish net1, a guanine nucleotide exchange factor, is specifically expressed in dorsal marginal blastomeres after MBT, and acts as a zygotic factor to promote the specification of dorsal cell fates. Loss- and gain-of-function experiments show that the GEF activity of Net1 is required for the activation of Wnt/ß-catenin signaling in zebrafish embryos and mammalian cells. Net1 dissociates and activates PAK1 dimers, and PAK1 kinase activation causes phosphorylation of S675 of ß-catenin after MBT, which ultimately leads to the transcription of downstream target genes. In summary, our results reveal that Net1-regulated ß-catenin activation plays a crucial role in the dorsal axis formation during zebrafish development.

A Small Molecule That Protects the Integrity of the Electron Transfer Chain Blocks the Mitochondrial Apoptotic Pathway.

In response to apoptotic stimuli, mitochondria in mammalian cells release cytochrome c and other apoptogenic proteins, leading to the subsequent activation of caspases and apoptotic cell death. This process is promoted by the pro-apoptotic members of the Bcl-2 family of proteins, such as Bim and Bax, which, respectively, initiate and execute cytochrome c release from the mitochondria. Here we report the discovery of a small molecule that efficiently blocks Bim-induced apoptosis after Bax is activated on the mitochondria. The cellular target of this small molecule was identified to be the succinate dehydrogenase subunit B (SDHB) protein of complex II of the mitochondrial electron transfer chain (ETC). The molecule protects the integrity of the ETC and allows treated cells to continue to proliferate after apoptosis induction. Moreover, this molecule blocked dopaminergic neuron death and reversed Parkinson-like behavior in a rat model of Parkinson's disease.

Determination of geniposide in adjuvant arthritis rat plasma by ultra-high performance liquid chromatography tandem mass spectrometry method and its application to oral bioavailability and plasma protein binding ability studies.

A specific, sensitive and high throughput ultra-high performance liquid chromatography-electrospray ionization tandem mass spectrometric method (UHPLC-ESI-MS/MS) was established and validated to assay geniposide (GE), a promising anti-inflammatory drug, in adjuvant arthritis rat plasma: application to pharmacokinetic and oral bioavailability studies and plasma protein binding ability. Plasma samples were processed by de-proteinised with ice-cold methanol and separated on an ACQUITY UPLC™ HSS C18 column (100 mm × 2.1mm i.d., 1.8 µm particle size) at a gradient flow rate of 0.2 mL/min using acetonitrile-0.1% formic acid in water as mobile phase, and the total run time was 9 min. Mass detection was performed in selected reaction monitoring (SRM) mode with negative electro-spray ionization includes the addition of paeoniflorin (Pae) as an internal standard (IS). The mass transition ion-pair was followed as m/z 387.4 → 122.4 for GE and m/z 479.4 → 449.0 for IS. The calibration curves were linear over the concentration range of 2-50,000 ng/mL with lower limit of quantification of 2 ng/mL. The intra-day and inter-day precisions (RSD, %) of the assay were less than 8.4%, and the accuracy was within ± 6.4% in terms of relative error (RE). Extraction recovery, matrix effect and stability were satisfactory in adjuvant arthritis rat plasma. The UHPLC-ESI-MS/MS method was successfully applied to a pharmacokinetic study of GE after oral administration of depurated GE at 33, 66, 132 mg/kg and intravenous injection at 33, 66, 132 mg/kg in adjuvant arthritis (AA) rats. In addition, it was found that GE has rapid absorption and elimination, low absolute bioavailability, high plasma protein binding ability in AA rats after oral administration within the tested dosage range. It suggested that GE showed slow distribution into the intra- and extracellular space, and the binding rate was not proportionally dependent on plasma concentration of GE when the concentration of GE was below 5.0 µg/mL.

Anti-inflammatory effects and pharmacokinetics study of geniposide on rats with adjuvant arthritis.

The aim of this study was to explore the anti-inflammatory effects of Geniposide (GE), an iridoid glycoside compound extracted from Gardenia jasminoides Ellis (GJ) fruit in adjuvant-induced arthritis (AA) rats and its pharmacokinetic (PK) basis. AA was induced by injecting with Freund's complete adjuvant (FCA). Male SD rats were subjected to treatment with GE (30, 60 and 120mg/kg) from day 17 to 24 after immunization. Fibroblast-like synoviocyte (FLS) proliferation was assessed by MTT. Interleukin (IL)-1, IL-6, TNF-α and IL-10 were determined using double-sandwich enzyme-linked immunosorbent assay (ELISA). Expression of p38 mitogen-activated protein kinases (p38MAPKs) related proteins in FLS was detected by Western blotting. PK profiles were simultaneously detected by ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) in AA rat plasma after oral administration of GE on day 17 after immunization. As a result, GE promoted the recovery of arthritis and inhibited the colonic inflammation damage in AA rats by decreasing the expression level of TNF-α, IL-1 and IL-6, increasing the production of IL-10 and inhibiting the expression of phospho-p38 (p-p38) related proteins in FLS. PK parameters (AUC, Cmax and t1/2) tended to be associated with dosage-related decreasing of efficacy index.

[Evaluation of the influence of humidity and temperature on the drug stability by initial average rate experiment].

OBJECTIVE: To evaluate the influence of temperature and humidity on the drug stability by initial average rate experiment, and to obtained the kinetic parameters. METHODS: The effect of concentration error, drug degradation extent, humidity and temperature numbers, humidity and temperature range, and average humidity and temperature on the accuracy and precision of kinetic parameters in the initial average rate experiment was explored. The stability of vitamin C, as a solid state model, was investigated by an initial average rate experiment. RESULTS: Under the same experimental conditions, the kinetic parameters obtained from this proposed method were comparable to those from classical isothermal experiment at constant humidity. The estimates were more accurate and precise by controlling the extent of drug degradation, changing humidity and temperature range, or by setting the average temperature closer to room temperature. CONCLUSION: Compared with isothermal experiments at constant humidity, our proposed method saves time, labor, and materials.