FAM3D inhibits gluconeogenesis in high glucose environment via DUSP1/ZFP36/SIK1 axis.

Hyperglycemia is the most important factor leading to the complications of type 2 diabetes mellitus (T2DM). The primary condition for the treatment of T2DM is to change the glucose and lipid metabolism disorders in the liver and other insulin-sensitive tissues. The current study aims to unearth the potential molecular mechanism of inhibiting liver gluconeogenesis to provide a new theoretical basis for the treatment of T2DM. High glucose (HG) induction of HepG2 cells followed by treatment with sequence-similar family 3 member D (FAM3D). Dual specificity phosphatases 1 (DUSP1), zinc finger protein 36 (ZFP36), salt-induced kinase 1 (SIK1), p-SIK1, posphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) gene and protein expression level were detected by quantitative real-time polymerase chain reaction and western blot. The PEPCK and G6Pase activities were detected by enzyme linked immunosorbent assay. Glucose production assay to determine glucose content. The RNA binding protein immunoprecipitation assay was used to detect the binding of ZFP36 to SIK1. FAM3D facilitated the expression of DUSP1 but suppressed the expression of gluconeogenesis-related factors in an HG environment. The expression of ZFP36 was up-regulated in an HG environment. ZFP36 could reverse the inhibition of gluconeogenesis caused by FAM3D. HG-induced upregulation of ZFP36 was downregulated by overexpression of DUSP1. ZFP36 bound to SIK1, and downregulation of ZFP36 promoted SIK1 expression and inhibits gluconeogenesis. Our study demonstrated FAM3D inhibited gluconeogenesis through the DUSP1/ZFP36/SIK1 axis in an HG environment, which provided a new theoretical basis for exploring the pathogenesis and treatment strategy of T2DM.

Manchester-Oxford Foot Questionnaire Chinese version (MOXFQ-Ch): a validity and cross-cultural adaptation.

PURPOSE: To perform the Manchester-Oxford Foot Questionnaire Chinese Version (MOXFQ-Ch) through a cross-cultural adaptation and validation of the original questionnaire. MATERIAL AND METHODS: Three hundred and sixty nine patients (241 women/128 men, 48.75 ± 8.17 years old) with a diagnosis of hallux valgus foot (237 both feet; 74 right foot; 58 left foot) participated in this observational study. A translation and cross-cultural adaptation of the Manchester-Oxford Foot Questionnaire to Chinese was developed. Each participant completed twice the Manchester-Oxford Foot Questionnaire. In addition, the psychometric characteristics of Manchester-Oxford Foot Questionnaire Chinese Version were analyzed for internal consistency, construct validity and criterion validity (EuroQoL-5D; Short-Form 12v2; Foot Functional Index were used). RESULTS: The internal consistency of the different sub-scales and total value of the Manchester-Oxford Foot Questionnaire Chinese Version ranged between 0.976 (walking/standing) and 0.991 (pain). Moreover, in the item response analysis, the results ranged between 0.973 (walking/standing) and 0.998 (pain). The standard error of the measurement scores ranged between 1.071 (social interaction) and 1.864 (Manchester-Oxford Foot Questionnaire Chinese Version total value). The values of Minimal Detectible Change 90 were 4342. In addition, values of the root mean square error of approximation and Goodness-of-fit index were: 0.076 and 0.916 respectively. For criterion validity, were used the questionnaires: Short-Form 12v2, Foot Function Index (Chinese Version), and EuroQol-5D. Correlations with the three factors show a "r" value ranged between 0.104 (Factor 1 - SF12v2-Sub-scale) to 0.819 (FFI-Ch). CONCLUSIONS: The Manchester-Oxford Foot Questionnaire has been translated and culturally adapted from the original version into Chinese. Manchester-Oxford Foot Questionnaire Chinese Version has shown excellent internal consistency, external validity ranges from moderate to excellent (depending on the questionnaire used) and, demonstrated a structure of one factor psychometrically supported. Consequently, the Manchester-Oxford Foot Questionnaire Chinese Version could be introduced into Chinese-speaking clinical and research settings to assess and monitor patients with hallux valgus Implications for rehabilitation Manchester-Oxford Foot Questionnaire has been cross-cultural adapted from the original version to Chinese. Manchester-Oxford Foot Questionnaire Chinese Version have been reported satisfactory psychometric properties and consistent results. Chinese speaking clinician and researcher, could use the Manchester-Oxford Foot Questionnaire Chinese Version to assess and follow up patients with food and ankle disease.

Chinese cross-cultural adaptation and validation of the Foot Function Index as tool to measure patients with foot and ankle functional limitations.

PURPOSE: To perform a cross-cultural adaptation and validation of the Foot Function Index (FFI) questionnaire to develop the Chinese version. MATERIALS AND METHODS: Three hundred and six patients with foot and ankle neuromusculoskeletal diseases participated in this observational study. Construct validity, internal consistency and criterion validity were calculated for the FFI Chinese version after the translation and transcultural adaptation process. RESULTS: Internal consistency ranged from 0.996 to 0.998. Test-retest analysis ranged from 0.985 to 0.994; minimal detectable change 90: 2.270; standard error of measurement: 0.973. Load distribution of the three factors had an eigenvalue greater than 1. Chi-square value was 9738.14 (p < 0.001). Correlations with the three factors were significant between Factor 1 and the other two: r = -0.634 (Factor 2) and r = -0.191 (Factor 1). Foot Function Index (Taiwan Version), Short-Form 12 (Version 2) and EuroQol-5D were used for criterion validity. Factors 1 and 2 showed significant correlation with 15/16 and 14/16 scales and subscales, respectively. CONCLUSIONS: Foot Function Index Chinese version psychometric characteristics were good to excellent. Chinese researchers and clinicians may use this tool for foot and ankle assessment and monitoring. Implications for rehabilitation A cross-cultural adaptation of the FFI has been done from original version to Chinese. Consistent results and satisfactory psychometric properties of the Foot Function Index Chinese version have been reported. For Chinese speaking researcher and clinician FFI-Ch could be used as a tool to assess patients with foot disease.

Cryptoporic acid S, a new drimane-type sesquiterpene ether of isocitric acid from the fruiting bodies of Cryptoporus volvatus.

A new drimane-type sesquiterpene with an isocitric acid moiety, cryptoporic acid S (1), together with six known compounds, cryptoporic acid D (2), ß-sitosterol (3), ß-daucosterol (4), stigmast-4-en-3-one (5), ergosterol (6), and (22E,24R)-ergosta-7,22-diene-3ß,5α,6ß-triol (7), was isolated from the fruiting bodies of Cryptoporus volvatus. The structures of these compounds were established on the basis of UV, IR, MS, 1D and 2D NMR analysis. In the meanwhile, compounds 1 and 2 were evaluated for antioxidant activity using the methods of 2,2-diphenyl-1-picrylhydrazyl free radical scavenging activity (DPPH-RSA) and ferric reducing antioxidant power (FRAP) assay, and they exhibited moderate antioxidant activities.

Foot and ankle ability measure to measure functional limitations in patients with foot and ankle disorders: a Chinese cross-cultural adaptation and validation.

PURPOSE: To perform a cultural adaptation and validation study (internal and external) of the FAAM questionnaire to create the Chinese version of the questionnaire (FAAM-Ch). MATERIALS AND METHODS: Two independent professional native translators performed a translation from English to Chinese and reverse translation. Psychometric properties analysis: Internal consistency of measure was analysed through the Cronbach's α coefficients. After extraction by maximum likelihood (EML), the structure factor and construct validity was analysed; to extract a factor, it was necessary to complete the following three requirements: ≥10% of variance, Eigenvalue >1.0 and scree plot inflection point. Standard error measurement (SEM) and minimal detectable change 90 (MDC90) were calculated. FFI-Taiwan version, SF12v2, and EuroQol5D were used for criterion validity analysis. RESULTS: The internal consistency (Cronbach's α) for specific FAAM-Ch subscales was 0.879 (ADL) and 0.901 (Sport); test-retest analysis (interclass correlation) item ranging between 0.758 and 0.970 (ADL: 0758-0946; Sport: 0.911-0.970). Measures error: 3.449% (MDC90) and 1.478% (SEM). Chi-squared value =15228.74 and gl 406) (p < 0.001) and the Kaiser-Meyer-Oklin values (0.919). The correlation level with the FFI is strong, with SF12v2 is between poor and strong and with EuroQoL5d is between moderate and strong Conclusions: FAAM-Chinese version has satisfactory "transversal" psychometric properties, facilitating the inclusion of FAAM-Chinese into research and clinical practice. Implications for Rehabilitation Cross-cultural adaptation of the FAAM-Ch has been performed from the original version. The psychometric properties of the FAAM-Ch indicate satisfactory and consistent results (particularly in the internal consistence, reliability and criterion validity) with the original version. FAAM-Ch can be used by Chinese speaking clinicians and researches.

An unusual sesquiterpene coumarin from the seeds of Ferula sinkiangensis.

A sesquiterpene coumarin, sinkiangenorin E, consisting of a novel bicyclo[4.3.1]decane-type sesquiterpene system, was isolated from the seeds of Ferula sinkiangensis. The structure of sinkiangenorin E including the relative stereochemistry and the absolute configuration was elucidated on the basis of spectroscopic data. The new compound showed cytotoxic activity against AGS cells (IC50, 12.7 µM) and inhibiting effect against influenza A H1N1 (IC50, 4.0 µM), which provided important clues for the study on the bioactivities of this type of sesquiterpene coumarins.

[A new bisabolane-type sesquiterpenoid from Coreopsis tinctoria].

To study the chemical constituents of the inflorescences of Coreopsis tinctoria from Xinjiang, isolation and purification of constituents were carried out by column chromatography on macroporous resin (D101) , MCI gel, MDS gel, silica gel, Sephadex LH-20 and semi-preparative HPLC. The structures of the compounds were identified by physicchemical properties and spectral data analysis. Fourteen compounds were isolated and identified as coretinterpenoid A (1), coretinphenol (2), quercetin (3), quercetin-3-O-ß-glucopyranoside (4), luteolin (5), taxifolin (6), 7, 3', 5'-trihydroxyflavanone (7), isookanin (8), isookanin-7-O-ß-D-glucopyranoside (9), 5, 7, 3', 5'-tetrahydroxyflavanone-7-O-ß-D-glucopyranoside (10), butein (11), okanin (12), sulfuretin (13), and linocinnamarin (14). Compound 1 was a new isabolane-type sesquiterpenoid and compounds 4, 10 and 13 were isolated from this plant for the first time.

[Chemical constituents of lateral roots of Aconitum carmichaelii Debx].

In order to find the cardiotonic constituents of lateral roots of Aconitum carmichaelii Debx., the investigation was carried out. Silica gel column chromatography, Sephadex LH-20, medium-pressure MCI and reverse phase ODS column chromatography were used to separate the 90% EtOH extract of the lateral roots of Aconitum carmichaelii Debx. The structures of the isolated compounds have been identified by chemical properties and spectroscopic analyses. Ten compounds were isolated and their structures were elucidated as benzoic acid-5-hydroxy-2-benzoyl-amino methyl ester (1), honokiol (2), pinoresinol (3), salicylic acid (4), p-hydroxy-cinnamic acid (5), songorine (6), karakoline (7), mesaconitine (8), hypaconitine (9) and 14-benzoylhypaconitine (10), separetely. Compound 1 is a new compound and its structure has been established by NMR, HR-ESI-MS, UV, IR and X-Ray. Compound 2-5 are isolated from the lateral roots of Aconitum carmichaelii Debx. for the first time.

Telomere 3' overhang-specific DNA oligonucleotides induce autophagy in malignant glioma cells.

Telomere 3' overhang-specific DNA oligonucleotides (T-oligos) induce cell death in cancer cells, presumably by mimicking telomere loop disruption. Therefore, T-oligos are considered an exciting new therapeutic strategy. The purpose of this study was to elucidate how T-oligos exert antitumor effects on human malignant glioma cells in vitro and in vivo. We demonstrated that T-oligos inhibited the proliferation of malignant glioma cells through induction of nonapoptotic cell death and mitochondria hyperpolarization, whereas normal astrocytes were resistant to T-oligos. Tumor cells treated with T-oligos developed features compatible with autophagy, with development of autophagic vacuoles and conversion of an autophagy-related protein, microtubule-associated protein 1 light chain 3 from type I (cytoplasmic form) to type II (membrane form of autophagic vacuoles). A reverse-phase protein microarray analysis and Western blotting revealed that treatment with T-oligos inhibited the mammalian target of the rapamycin (mTOR) and the signal transducer and activator of transcription 3 (STAT3). Moreover, pretreatment with T-oligos significantly prolonged the survival time of mice inoculated intracranially with malignant glioma cells compared with that of untreated mice and those treated with control oligonucleotides (P=0.0065 and P=0.043, respectively). These results indicate that T-oligos stimulate the induction of nonapoptotic autophagic also known as type II programmed cell death and are thus promising in the treatment of malignant glioma.

Signaling pathway requirements for induction of senescence by telomere homolog oligonucleotides.

Cellular senescence is a major defense against cancer. In human fibroblasts, suppressing both the p53 and pRb pathways is necessary to bypass replicative senescence as well as senescence induced by ectopic expression of a dominant negative form of the telomere repeat binding factor 2, TRF2(DN). We recently reported that exposure to oligonucleotides homologous to the telomere 3' overhang (T-oligos) activates both the p53 and pRb pathways and leads to senescence in primary human fibroblasts. To further characterize T-oligo-induced senescence, we compared established isogenic fibroblast cell lines lacking functional p53 and/or pRb pathways to the normal parental line. Here, we report that, as in physiologic senescence, inactivation of both the p53 and pRb pathways is necessary to suppress T-oligo-induced senescence. Moreover, T-oligo rapidly induces senescence in a malignant fibroblast-derived cell line, demonstrating the potential of using T-oligo as a novel anticancer therapeutic. Our data support the hypothesis that exposure of the TTAGGG tandem repeat telomere 3' overhang sequence is the event that initiates signaling through DNA damage response pathways after experimental telomere disruption, serial passage, or acute genomic damage of normal cells.

Fas and c-kit are involved in the control of hair follicle melanocyte apoptosis and migration in chemotherapy-induced hair loss.

Chemotherapy alters the structure and function of hair follicle melanocytes. Molecular mechanisms controlling melanocyte responses during chemotherapy-induced hair loss, however, remain largely unknown. Using immunohistology and multicolor confocal microscopy, we show here that cyclophosphamide administration to C57BL/6 mice alters the activity and fate of hair follicle melanocytes. After 24-48 h, hair bulb melanocytes expressing Fas undergo apoptosis. The number of apoptotic follicular melanocytes is significantly reduced (p<0.01) in cyclophosphamide-treated Fas knockout mice compared to wild-type controls, suggesting that Fas signaling contributes to chemotherapy-induced melanocyte death. After 3-5 d, surviving hair bulb melanocytes express c-kit receptor, proliferate, and appear to migrate up the outer root sheath. Tyrosinase-positive and melanogenically active cells then appear in the epidermis. By Western blotting and immunohistochemistry, expression levels of the c-kit ligand, stem cell factor, in skin and epidermis are strongly increased after cyclophosphamide treatment. Cyclophosphamide-induced migration of the hair follicle melanocytes into epidermis is completely abrogated by administration of c-kit neutralizing antibody. These data suggest that chemotherapy induces a complex response in the hair follicle melanocytes, which includes apoptosis, proliferation, and migration. Pharmacologic manipulation of Fas and c-kit signaling pathways might be useful for the correction of skin hyperpigmentation as a side-effect of chemotherapy.

Induction of a p95/Nbs1-mediated S phase checkpoint by telomere 3' overhang specific DNA.

Telomere shortening induces a nonproliferative senescent phenotype, believed to reduce cancer risk, and telomeres are involved in a poorly understood manner in responses to DNA damage. Although telomere disruption induces p53 and triggers apoptosis or cell cycle arrest, the features of the disrupted telomere that trigger this response and the precise mechanism involved are poorly understood. Using human cells, we show that DNA oligonucleotides homologous to the telomere 3' overhang sequence specifically induce and activate p53 and activate an S phase checkpoint by modifying the Nijmegen breakage syndrome protein, known to mediate the S phase checkpoint after DNA damage. These responses are mediated, at least in part, by the ATM kinase and are not attributable to disruption of cellular telomeres. Based on these and earlier data, we propose that these oligonucleotides mimic a physiological signal, exposure of the telomere 3' overhang due to opening of the normal telomere loop structure, and hence evoke these protective antiproliferative responses in the absence of DNA damage or telomere disruption.

Evidence that exposure of the telomere 3' overhang sequence induces senescence.

Normal human cells cease proliferation after a finite number of population doublings, a phenomenon termed replicative senescence. This process, first convincingly described by Hayflick and Moorhead [Hayflick, L. & Moorhead, P. S. (1961) Exp. Cell Res. 25, 595-621] for cultured human fibroblasts 40 years ago, is suggested to be a fundamental defense against cancer. Several events have been demonstrated to induce the senescent phenotype including telomere shortening, DNA damage, oxidative stress, and oncogenic stimulation. The molecular mechanisms underlying senescence are poorly understood. Here we report that a 1-week exposure to oligonucleotide homologous to the telomere 3'-overhang sequence TTAGGG (T-oligo) similarly specifically induces a senescent phenotype in cultured human fibroblasts, mimicking serial passage or ectopic expression of a dominant negative form of the telomeric repeat binding factor, TRF2(DN). We propose that exposure of the 3' overhang due to telomere loop disruption may occur with critical telomere shortening or extensive acute DNA damage and that the exposed TTAGGG tandem repeat sequence then triggers DNA-damage responses. We further demonstrate that these responses can be induced by treatment with oligonucleotides homologous to the overhang in the absence of telomere disruption, a phenomenon of potential therapeutic importance.

Modulation of base excision repair alters cellular sensitivity to UVA1 but not to UVB1.

Oxidative DNA damage has been implicated in some of the biological properties of UVA but so far not in the acute photosensitivity or cellular sensitivity. In contrast to pyrimidine dimers, oxidative DNA damage is predominantly processed by base excision repair (BER). In order to further clarify the role of oxidative DNA damage and its repair in the acute cellular response to UV light, we studied UVA1 and UVB sensitivities in three different cell model systems with modified BER. 8-Oxoguanine-DNA-glycosylase 1-/- (OGG1-/-) mouse embryonal fibroblasts and human fibroblasts in which BER was inhibited by incubation with methoxyamine were hypersensitive to UVA1, in particular to low doses. This hypersensitivity could be partially corrected by reexpression of OGG1 in OGG1-/- cells. The Chinese hamster ovary (CHO) cells with upregulated AP-endonuclease 1 exhibited reduced UVA1 sensitivity. UVB sensitivity was not altered in any of the cell models. These results indicate that DNA damage, in particular oxidative DNA damage, contributes to cellular UVA1 sensitivity and underline a pivotal role of its repair in the cellular responses to UVA1.