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Background: At present, some cancer patients experience hyperprogressive disease (HPD) after receiving immunotherapy. This study used the Response Evaluation Criteria in Solid Tumors 1.1 to evaluate the incidence of HPD in patients receiving immune checkpoint inhibitors (ICIs) for treating primary liver cancer (PLC) and to explore the risk factors for HPD., Methods: This retrospective, single-center study included patients with PLC who were treated with ICIs. The RECIST 1.1 was used to determine patients with HPD. Univariate and multivariate regression analyses were performed to explore the risk factors for HPD, and clinical variables with prognostic significance for HPD were included to establish a risk model., Results: Among 129 patients with PLC treated with ICIs, HPD occurred in 13 patients (10.1%). In the multivariate regression analysis, lymph node metastasis and lung metastasis were risk factors for HPD. The area under the curve of the risk model, established by including lymph node metastasis, lung metastasis, neutrophil-lymphocyte ratio, albumin, and performance status, was 0.801 (P<0.001). The progression-free survival of HPD patients was significantly worse than that of non-HPD patients (P<0.001)., Conclusions: In this study, 10.1% of patients with PLC had HPD. Compared with the non-HPD patients, lung metastasis and lymph node metastasis were independent risk factors of HPD.

Grafting is a technique that provides a substantial way to enhance nutrient utilization thereby improves plant growth and yield quality. Although it is commonly practised in horticultural crops, the impact of scion-rootstock interaction on nutrient distribution is still unclear. Here, 'Newhall' navel orange plants grafted on Trifoliate orange (T) as the original rootstock were inarched with trifoliate orange (N/Tt combination) or carrizo citrange (N/Tc combination) rootstock seedlings. The experimental plants were treated with isotope 10 B solution for 7 weeks to investigate the effect of different inarched rootstocks on B distribution and translocation by using a two-root system. From this study, the original rootstock played a more dominant role in B distribution to scion tissues than the inarching rootstock either in N/Tt or N/Tc combination. From inarched combinations, the carrizo citrange in the N/Tc combination had a higher ability to distribute B to new leaves, new twigs and old twigs than trifoliate orange in the N/Tt combination. However, the original rootstock of N/Tt distributed more B to scion tissues than N/Tc and the B concentration in old leaves and new leaves of N/Tt plants was significantly higher than that of N/Tc plants. These results suggest that scion B status is influenced by the B distribution of two inarching rootstocks in an inarching plant, as well as the affinity between the inarching rootstock and grafted plant. In addition, by either adding 10 B to the inarching rootstock or original rootstock, we could detect 10 B in the other rootstock root in both N/Tt and N/Tc combinations. The results further suggest that B can translocate from rootstock to leaves and then, re-translocate from scion to rootstock through the cycling of B transportation., (Copyright © 2021 Elsevier Masson SAS. All rights reserved.)

Marine natural products are abundant resources for antioxidants, but the antioxidant property of the soft corals-derived sinularin and dihydrosinularin were unknown. This study aimed to assess antioxidant potential and antiproliferation effects of above compounds on cancer cells, and to investigate the possible relationships between them. Results show that sinularin and dihydrosinularin promptly reacted with 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis (3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS), and hydroxyl ( • OH), demonstrating a general radical scavenger activity. Sinularin and dihydrosinularin also show an induction for Fe +3 -reduction and Fe +2 -chelating capacity which both strengthen their antioxidant activities. Importantly, sinularin shows higher antioxidant properties than dihydrosinularin. Moreover, 24 h ATP assays show that sinularin leads to higher antiproliferation of breast, lung, and liver cancer cells than dihydrosinularin. Therefore, the differential antioxidant properties of sinularin and dihydrosinularin may contribute to their differential anti-proliferation of different cancer cells.

Combined treatment is increasingly used to improve cancer therapy. Non-ionizing radiation ultraviolet-C (UVC) and sinularin, a coral Sinularia flexibilis -derived cembranolide, were separately reported to provide an antiproliferation function to some kinds of cancer cells. However, an antiproliferation function using the combined treatment of UVC/sinularin has not been investigated as yet. This study aimed to examine the combined antiproliferation function and explore the combination of UVC/sinularin in oral cancer cells compared to normal oral cells. Regarding cell viability, UVC/sinularin displays the synergistic and selective killing of two oral cancer cell lines, but remains non-effective for normal oral cell lines compared to treatments in terms of MTS and ATP assays. In tests using the flow cytometry, luminescence, and Western blotting methods, UVC/sinularin-treated oral cancer cells exhibited higher reactive oxygen species production, mitochondrial superoxide generation, mitochondrial membrane potential destruction, annexin V, pan-caspase, caspase 3/7, and cleaved-poly (ADP-ribose) polymerase expressions than that in normal oral cells. Accordingly, oxidative stress and apoptosis are highly induced in a combined UVC/sinularin treatment. Moreover, UVC/sinularin treatment provides higher G2/M arrest and γH2AX/8-hydroxyl-2'deoxyguanosine-detected DNA damages in oral cancer cells than in the separate treatments. A pretreatment can revert all of these changes of UVC/sinularin treatment with the antioxidant N -acetylcysteine. Taken together, UVC/sinularin acting upon oral cancer cells exhibits a synergistic and selective antiproliferation ability involving oxidative stress-dependent apoptosis and cellular DNA damage with low toxic side effects on normal oral cells.

Aims: F11R gene encodes junctional adhesion molecule-A protein (JAM-A), which is expressed in various types of cells and is involved in leukocyte extravasation during inflammation. Sjögren's syndrome (SS) is a chronic systemic inflammatory disease that involves lymphocytes invasion of exocrine glands. F11R has been studied in autoimmune diseases, but any association between F11R and SS has not yet been investigated. Therefore, experiments were undertaken to examine the relationships among F11R gene polymorphism, messenger RNA (mRNA) expression and SS patients., Methods: Three hundred and twenty-nine patients with SS, and 223 healthy controls were enrolled in their recruitment from the Kaohsiung Medical University Hospital. Genomic DNA was extracted from peripheral blood mononuclear cells and gene polymorphisms were genotyped by TaqMan real-time polymerase chain reaction (PCR). F11R mRNA expression was quantitated by quantitative real-time PCR with TaqMan Gene Expression Assay., Results: Our study showed the genotype -688A/C (rs6695707) was not found in relation to SS patients. The odds ratio of -436A/G (rs12567886) genotype was notably associated with less susceptibility of SS in human leukocyte antigen (HLA)-DR2 negative and HLA-DR3 negative individuals. F11R mRNA expression was lower in SS patients than in the cells of healthy controls., Conclusion: The result indicated that G allele of -436A/G genotype has the potential protective effect against SS disease condition. F11R mRNA was expressed significantly lower in SS patients., (© 2021 Asia Pacific League of Associations for Rheumatology and John Wiley & Sons Australia, Ltd.)

MRE11, the nuclease component of RAD50/MRE11/NBS1 DNA repair complex which is essential for repair of DNA double-strand-breaks in normal cells, has recently garnered attention as a critical factor in solid tumor development. Herein we report the crucial role of MRE11 in oral cancer progression in a nuclease-independent manner and delineate its key downstream effectors including CXCR4. MRE11 expression in oral cancer samples was positively associated with tumor size, cancer stage and lymph node metastasis, and was predictive of poorer patient survival and radiotherapy resistance. MRE11 promoted cell proliferation/migration/invasion in a nuclease-independent manner but enhanced radioresistance via a nuclease-dependent pathway. The nuclease independent promotion of EMT and metastasis was mediated by RUNX2, CXCR4, AKT, and FOXA2, while CXCR4 neutralizing antibody mitigated these effects in vitro and in vivo. Collectively, MRE11 may serve as a crucial prognostic factor and therapeutic target in oral cancer, displaying dual nuclease dependent and independent roles that permit separate targeting of tumor vulnerabilities in oral cancer treatment.

Single nucleotide polymorphisms (SNPs) in the promoter region of CD209 (cluster of differentiation 209) may influence expression levels, and higher expression of CD209 on immune cells correlate with severity of cartilage destruction in patients with rheumatoid arthritis (RA). Due to the lack of a comprehensive study, this study aimed to investigate the CD209 promoter variants and haplotypes in a Taiwanese population and the association with RA development. Deoxyribonucleic acid (DNA) of peripheral blood mononuclear cells from 126 RA patients and 124 healthy controls was purified, and the CD209 gene promoter was amplified by polymerase chain reaction and analyzed by Sanger sequencing. Results showed that a novel variant -96C>A polymorphism in CD209 promoter was identified in the Taiwanese population, and the frequency was significantly higher in RA patients than in controls (11.51% vs. 2.42%, P < .0001). The odds ratio (OR) for the development of RA was 5.88 (95% CI 2.35-14.74, P < .0001). Other known variants were also evaluated; for instance, -1180 T/T (rs7359874) was increased in RA patients, and the OR for the development of RA was 3.26, 95% CI 0.85-12.52, P = .07). Besides, the haplotype frequencies were calculated; -1180A-939C-871 T-336 T-139 T-96A and -1180 T-939 T-871C-336 T-139C-96A were increased in RA patients (P = .004 and 0.05, respectively). In summary, CD209-96A variant could be an important factor for the development of RA in the Taiwanese population., (© 2021 The Authors. Journal of Clinical Laboratory Analysis published by Wiley Periodicals LLC.)

Hepatocellular carcinoma (HCC) is a severe disease that accounts for 80% of liver cancers. Chemotherapy is the primary therapeutic strategy for patients who cannot be treated with surgery or who have late-stage HCC. C 2 -ceramide is an effective reagent that has been found to inhibit the growth of many cancer types. The metabolism of C 2 -ceramide plays a vital role in the regulation of cell death/cell survival. The phenoxyphenol compound 4-{2,3,5,6-tetrafluoro-4-[2,3,5,6-tetrafluoro-4-(4-hydroxyphenoxy)phenyl]phenoxy}phenol (diTFPP) was found to have a synergistic effect with C 2 -ceramide, resulting in considerable cell death in the HA22T HCC cell line. diTFPP/C 2 -ceramide cotreatment induced a two- to threefold increase in cell death compared to that with C2-ceramide alone and induced pyknosis. Annexin V/7-aminoactinomycin D (7AAD) double staining and Western blotting indicated that apoptosis was involved in diTFPP/C 2 -ceramide cotreatment-mediated cell death. We next analyzed transcriptome alterations in diTFPP/C 2 -ceramide-cotreated HA22T cells with next-generation sequencing (NGS). The data indicated that diTFPP treatment disrupted sphingolipid metabolism, inhibited cell cycle-associated gene expression, and induced autophagy and reactive oxygen species (ROS)-responsive changes in gene expression. Additionally, we assessed the activation of autophagy with acridine orange (AO) staining and observed alterations in the expression of the autophagic proteins LC3B-II and Beclin-1, which indicated autophagy activation after diTFPP/C 2 -ceramide cotreatment. Elevated levels of ROS were also reported in diTFPP/C 2 -ceramide-treated cells, and the expression of the ROS-associated proteins SOD1, SOD2, and catalase was upregulated after diTFPP/C 2 -ceramide treatment. This study revealed the potential regulatory mechanism of the novel compound diTFPP in sphingolipid metabolism by showing that it disrupts ceramide metabolism and apoptotic sphingolipid accumulation.

Many long noncoding RNAs (lncRNAs) have been annotated, but their functions remain unknown. The authors found a novel lnc-APUE (lncRNA accelerating proliferation by upregulating E2F1) that is upregulated in different cancer types, including hepatocellular carcinoma (HCC), and high lnc-APUE level is associated with short recurrence-free survival (RFS) of HCC patients. Gain- and loss-of-function analyses showed that lnc-APUE accelerated G1/S transition and tumor cell growth in vitro and allows hepatoma xenografts to grow faster in vivo. Mechanistically, lnc-APUE binds to miR-20b and relieves its repression on E2F1 expression, resulting in increased E2F1 level and accelerated G1/S phase transition and cell proliferation. Consistently, lnc-APUE level is positively associated with the expression of E2F1 and its downstream target genes in HCC tissues. Further investigations disclose that hepatocyte nuclear factor 4 alpha (HNF4 α ) binds to the lnc-APUE promoter, represses lnc-APUE transcription, then diminishes E2F1 expression and cell proliferation. HNF4 α expression is reduced in HCC tissues and low HNF4 α  level is correlated with high lnc-APUE expression. Collectively, a HNF4 α /lnc-APUE/miR-20b/E2F1 axis in which HNF4 α represses lnc-APUE expression and keeps E2F1 at a low level is identified. In tumor cells, HNF4 α downregulation leads to lnc-APUE upregulation, which prevents the inhibition of miR-20b on E2F1 expression and thereby promotes cell cycle progression and tumor growth., Competing Interests: The authors declare no conflict of interest., (© 2021 The Authors. Advanced Science published by Wiley‐VCH GmbH.)

Hepatocellular carcinoma (HCC) is a leading cause of death, resulting in over 700 thousand deaths annually worldwide. Chemotherapy is the primary therapeutic strategy for patients with late-stage HCC. Heteronemin is a marine natural product isolated from Hippospongia sp. that has been found to protect against carcinogenesis in cholangiocarcinoma, prostate cancer, and acute myeloid leukemia. In this study, heteronemin was found to inhibit the proliferation of the HCC cell lines HA22T and HA59T and induce apoptosis via the caspase pathway. Heteronemin treatment also induced the formation of reactive oxygen species (ROS), which are associated with heteronemin-induced cell death, and to trigger ROS removal by mitochondrial SOD2 rather than cytosolic SOD1. The mitogen-activated protein kinase (MAPK) signaling pathway was associated with ROS-induced cell death, and heteronemin downregulated the expression of ERK, a MAPK that is associated with cell proliferation. Inhibitors of JNK and p38, which are MAPKs associated with apoptosis, restored heteronemin-induced cell death. In addition, heteronemin treatment reduced the expression of GPX4, a protein that inhibits ferroptosis, which is a novel form of nonapoptotic programmed cell death. Ferroptosis inhibitor treatment also restored heteronemin-induced cell death. Thus, with appropriate structural modification, heteronemin can act as a potent therapeutic against HCC., Competing Interests: The authors have no conflicts of interest to declare., (Copyright © 2021 Wen-Tsan Chang et al.)

Sperm are specialized cells that require adenosine triphosphate (ATP) to support their function. Maintaining sperm energy homeostasis in vitro is vitally important to improve the efficacy of boar sperm preservation. Metformin can activate 5'-AMP-activated protein kinase (AMPK) to improve metabolic flexibility and maintain energy homeostasis. Thus, the aim of the present study was to investigate whether metformin can improve boar sperm quality through AMPK mediation of energy metabolism. Sperm motility parameters, membrane integrity, acrosome integrity, mitochondrial membrane potential (ΔΨ m ), ATP content, glucose uptake, and lactate efflux were analyzed. Localization and expression levels of AMPK and phospho-Thr 172 -AMPK (p-AMPK) were also detected by immunofluorescence and western blotting. We found that metformin treatment significantly increased sperm motility parameters, ΔΨ m , and ATP content during storage at 17 °C. Moreover, results showed that AMPK was localized at the acrosomal region, connecting piece, and midpiece of sperm and p-AMPK was distributed at the post-acrosomal region, connecting piece, and midpiece. When sperm were incubated with metformin for 4 h at 37 °C, sperm motility parameters, ΔΨ m , ATP content, p-AMPK, glucose uptake, and lactate efflux all significantly increased, whereas the addition of Compound C treatment, an inhibitor of AMPK, counteracted these positive effects. Together, our results suggest that metformin promotes AMPK activation, which contributes to the maintenance of energy hemostasis and mitochondrial activity, thereby maintaining boar sperm functionality and improving the efficacy of semen preservation.

Objective: To investigate the effects of exogenous NaHS on myelin basic protein (MBP) and learning and memory of hippocampal neurons in mice with spinocerebellar ataxia type 3 (SCA3) and its therapeutic significance. Methods: Twelve male normal mice were randomly selected as normal control group (NC Group), and 48 SCA3 mice were randomly selected as SCA3 model group (M Group), low dose group (NL Group, 10 μmol/kg), medium dose group (NM Group, 50μmol/kg) and high dose group (NH Group, 100 μmol/kg), 12 rats in each group. The drug treated groups were injected with NaHS intraperitoneally once a day for 4 weeks. The changes of learning and memory ability of SCA3 mice before and after the intervention of different doses of NaHS were determined by Morris water maze, the content of hydrogen sulfide (H 2 S) in hippocampus was measured by spectrophotometry, the expression of MBP was detected by immunohistochemistry, and the morphological changes of neuron myelin sheath were observed by electron microscope. Results: Compared with the control group, the learning and memory ability of SCA3 mice was decreased significantly (P＜0.05), and the content of H 2 S in hippocampus was decreased (P＜0.05). After different doses of exogenous NaHS treatment, the learning and memory ability was improved in different degrees (P＜0.05), and the contents of H 2 S and MBP in hippocampus of SCA3 mice were also improved in different degrees (P＜0.05). Conclusion: Exogenous NaHS may increase the contents of H 2 S and MBP in the hippocampus of SCA3 mice, which may have a protective effect on the neurons, and then improve the learning and memory ability of SCA3 mice, and provide a new idea for the treatment of SCA3.

Objectives: FOXA2 gene methylation links to the progression of cancers, but has not been documented in oral cancer. Herein, we explore the role of FOXA2 in the migration of oral cancer cells., Material and Methods: Methylation-specific PCR was applied for gene methylation. Wound healing and transwell experiments were tested for cell migration. FOXA2 expression in oral cancer tissues was addressed by immunohistochemistry, followed by statistical analysis of its association with clinical manifestations and patient survival., Results: FOXA2 bound to the promoter of CDH1 and enhanced the expression of its gene product E-cadherin, and decreased the cancer cell migration activity. High FOXA2 expression in oral cancer tissues was associated with high E-cadherin expression, decreased lymph node metastasis, and increased patient survival., Conclusion: FOXA2-E-cadherin link is involved in regulation of oral cancer cell metastasis and provides a new insight for the tumor suppressor activity of FOXA2 in oral cancer., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd. All rights reserved.)

Effects of elevated atmospheric CO 2 concentration and temperature on rice dry matter accumulation vary in planting regions and cropping systems. It remains unclear how dry matter productivity responds to factorial combination of elevated CO 2 and temperature in the double rice cropping system of China. Field experiments were conducted using open-top chambers (OTC) to simulate different scenarios of elevated CO 2 and/or temperature for three rotations of double rice in Jingzhou, Hubei Province. Liangyou 287 and Xiangfengyou 9 were used as rice cultivar for early rice and late rice, respectively. There were five treatments: UC, paddy field without OTC covering; CK, OTC with the similar temperature and CO 2 concentration to field environment; ET, OTC with 2 ℃ temperature elevation; EC, OTC with 60 μmol·mol -1 CO 2 elevation; ETEC, OTC with simu-ltaneous 2 ℃ temperature elevation and 60 μmol·mol -1 CO 2 elevation. We measured aboveground biomass, leaf area index (LAI) and net assimilation rate (NAR) of dry matter under different treatments. Our results showed that elevated CO 2 and/or temperature had no significant effects on NAR from transplanting to jointing, increased NAR from jointing to heading, but decreased NAR from heading to maturity (except for EC treatment in early rice). Elevated CO 2 and/or temperature promoted leaf area development at all growth stages, with ETEC showing the highest increase in LAI except at maturity. Warming and CO 2 enrichment jointly promoted dry matter accumulation at heading, with ETEC increasing aboveground biomass by 10.3%-39.8% and 23.6%-34.4% compared with CK in early rice and late rice, respectively. At maturity of early rice, elevated temperature partly offset the positive effects of elevated CO 2 on aboveground biomass, as shown by a reduction of 3.2%-14.1% under ETEC compared with EC. Contrarily at maturity of late rice, co-elevation of CO 2 and temperature further increased aboveground biomass, showing a synergistic interaction. Results from regression analysis showed that warming and CO 2 enrichment had positive effects on NAR at vegetative stages of double rice, while warming showed negative effects on NAR at reproductive stages. Considering the dissimilarities in growth characteristics, growing periods and ambient temperature, elevated CO 2 and temperature might increase dry matter production in the Chinese double rice cropping system.

Objective: The aim of this study was to explore the potential effect of miRNA-1297 on myocardial fibrosis (MF) and its underlying mechanism., Materials and Methods: MF model was established by cardiac perfusion of Angiotensin II (Ang-II) in mice. The primary myocardial fibroblasts were extracted from MF mice (Ang-II infusion group) and controls (sham group), respectively. The relative levels of miRNA-1297 and ULK1 in the in vivo and in vitro MF models were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Meanwhile, the protein expressions of fibrosis-related genes in MF mice and primary myocardial fibroblasts were determined by Western Blot. Subsequently, the Dual-Luciferase Reporter Gene Assay was applied to verify the downstream gene of miRNA-1297. In addition, a series of rescue experiments were conducted to elucidate the role of miRNA-1297/ULK1 in regulating MF., Results: Masson staining showed plenty of micro-vessels around myocardial tissues and significantly increased contents of intercellular collagen in Ang-II infusion group when compared with those in the sham group. Western blot results revealed that the protein expressions of Col1a1 and α-SMA were significantly upregulated in myocardial tissues of MF mice. QRT-PCR data illustrated that miRNA-1297 was remarkably downregulated in MF model. ULK1 was verified as the target gene of miRNA-1297, which was upregulated in the MF model. The overexpression of miRNA-1297 or the knockdown of ULK1 could downregulate the protein levels of Col1a1 and α-SMA in primary myocardial fibroblasts extracted from MF mice. Notably, ULK1 overexpression could reverse the regulatory effect of miRNA-1297 on MF., Conclusions: MiRNA-1297 suppresses myocardial fibrosis via down-regulating ULK1.

The hexosamine biosynthetic pathway (HBP), a branch of glucose metabolism, provides a substrate for glycosylation modification, which has a wide-ranging effect on cellular functions. Glutamine-fructose-6-phosphate transaminase 2 (GFPT2) has been reported to regulate the HBP as the first and rate-limiting enzyme. Given the inverse association between GFPT2 expression and survival of patients with serous ovarian cancer (SOC) observed in The Cancer Genome Atlas (TCGA) database, we attempted to investigate the role of GFPT2 and its related mechanisms in SOC. The results showed that GFPT2 was over-expressed in SOC tissues, and positive correlations with advanced stage (FIGO III/IV), suboptimal removal rate and poor survival were observed in 90 SOC patients. Cell migration and invasion were also inhibited in GFPT2 knockdown SKOV3 and HEY cells. The levels of O-linked β-N-acetylglucosamine (O-GlcNAc) and intranuclear β-catenin were evaluated and the observed increase in O-GlcNAcylation induced by GFPT2 may contribute to epithelial-mesenchymal transition (EMT). These data provide novel insights into the function of GFPT2 and O-GlcNAcylation in the EMT and thus the invasiveness SOC., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

Background: Rheumatoid arthritis (RA) is an autoimmune disease where both genetics and epigenetics are contributing factors. In order to discover genetic and epigenetic associations with RA and its phenotypes, we analysed RNA expression, DNA variations and DNA methylation of programmed cell death 1 (PDCD1) in a cohort of RA patients and healthy controls., Methods: RA patients (n = 206) and healthy controls (n = 234) were included for analysis of PDCD1 expression, PDCD1 polymorphisms and PDCD1 methylation. Differences in continuous variables between groups were compared by applying t tests. Associations between phenotypes and genotypes were evaluated with contingency tables. Sensitivity analyses were conducted to confirm the robustness of results, considering potential confounding factors and different treatment response definitions. Odds ratio (OR) and 95% confidence interval (95% CI) were calculated., Results: Higher expression of PDCD1 was found in RA compared to controls (P < 0.001), with similar PDCD1 polymorphisms in RA and controls. rs36084323 decreased inadequate response to conventional synthetic disease-modifying antirheumatic drugs (OR = 0.37, 95% CI = 0.19-0.72, P = 0.003), and rs41386349 increased rheumatoid factor seropositivity (OR = 11.89, 95% CI = 1.57-89.87, P = 0.003). Sensitivity analysis adjusting for further potential confounders and using different treatment response definition indicated similar results. Additionally, DNA methylation change at regulatory region of PDCD1 was detected in RA (P = 0.036)., Conclusion: Altogether, this was the first study to suggest genetic and epigenetic changes of PDCD1 in RA subsets and RA. Independent prospective cohorts are awaited to address the implications of these genetic and epigenetic changes in disease pathogenesis and phenotypes of RA., (© 2019 Stichting European Society for Clinical Investigation Journal Foundation.)

Using next-generation sequencing to decipher methylome and transcriptome and underlying molecular mechanisms contributing to rheumatoid arthritis (RA) for improving future therapies, we performed methyl-seq and RNA-seq on peripheral blood mononuclear cells (PBMCs) from RA subjects and normal donors. Principal component analysis and hierarchical clustering revealed distinct methylation signatures in RA with methylation aberrations noted across chromosomes. Methylation alterations varied with CpG features and genic characteristics. Typically, CpG islands and CpG shores were hypermethylated and displayed the greatest methylation variance. Promoters were hypermethylated and enhancers/gene bodies were hypomethylated, with methylation variance associated with expression variance. RA genetically associated genes preferentially displayed differential methylation and differential expression or interacted with differentially methylated and differentially expressed genes. These differentially methylated and differentially expressed genes were enriched with several signaling pathways and disease categories. 10 genes ( CD86, RAB20, XAF1 , FOLR3, LTBR , KCNH8 , DOK7, PDGFA, PITPNM2, CELSR1 ) with concomitantly differential methylation in enhancers/promoters/gene bodies and differential expression in B cells were validated. This integrated analysis of methylome and transcriptome identified novel epigenetic signatures associated with RA and highlighted the interaction between genetics and epigenetics in RA. These findings help our understanding of the pathogenesis of RA and advance epigenetic studies in regards to the disease.

Background: GADD45 genes are stress sensors in response to cellular stress response, activated signal pathways leading to the stimulation of inflammatory cytokines. This study is to examine the associations of GADD45a and GADD45b genes with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) patients., Methods: 230 patients of RA, 140 patients of SLE, and 191 healthy controls were enrolled. Genomic DNA was extracted from peripheral blood mononuclear cells and gene polymorphisms were genotyped by TaqMan assay. RNA expression was quantitated with real-time polymerase chain reaction., Results: The RNA expression of the GADD45b gene was significantly lower in RA patients than the control cases ( p = 0.03). The odds ratio of GADD45a genotype -589 CC (rs581000) was significantly low (OR = 0.36, 95% CI, 0.15-0.87) in DR4-negative RA patients. The odds ratio of GADD45b genotype -712CT (rs3795024) in DR4-negative RA patients was 0.41 (95% CI, 0.18-0.95). In clinical manifestation, the odds ratio of GADD45b -712CT genotype with anti-RNP antibody was 4.14 (95% CI, 1.10-15.63) in SLE patients. GADD45a genotype -589GG+GC was associated with rheumatoid factor (RF) in SLE patients., Conclusions: Genotypes GADD45a -589CC and GADD45b -712CT were shown to be less susceptible to RA and related to the disease state in SLE patients.

The ALPL gene is linked to hypophosphatasia, a rare genetic disease. Owing to the inverse relationships between ALPL expression and both the International Federation of Gynecology and Obstetrics (FIGO) stages and histological grades assigned to patients with serous ovarian cancer (SOC), this study was designed to explore the role and possible mechanisms of ALPL in cell motility of high grade SOC (HGSOC). The effects of ALPL overexpression on migration and invasion were detected in HGSOC cell lines SKOV3 and HEY. Gene ontology analysis for differential genes with ALPL overexpression identified several biological processes, including EMT, correlated with cell motility. Genes potentially implicated in EMT and associated with ALPL were screened using The Cancer Genome Atlas (TCGA) database. The WNT receptor Frizzled2 (FZD2) was identified and its role in HGSOC cell motility and survival was investigated. It was found that forced expression of ALPL could inhibit migration, invasion, and EMT in HGSOC cells. It also reduced the expression of FZD2 and its ligand WNT5A, accompanied by suppressed expression of their downstream target phosphorylated-STAT3 (pSTAT3). These effects were initiated via the FZD2 knockdown using siRNA and reversed by recombinant WNT5A protein. The relationship between FZD2 expression and poor HGSOC patient survival was also investigated. This data supports that ALPL might restrict the function of WNT5A-FZD2-STAT3 axis, a non-canonical WNT pathway for promoting EMT progression, which results in attenuated migration and invasion in HGSOC cells and improves survival in patients with SOC., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Non-small cell lung cancer (NSCLC) is a type of malignant cancer, and 85% of metastatic NSCLC patients have a poor prognosis. C₂-ceramide induces G2/M phase arrest and cytotoxicity in NSCLC cells. In this study, the autophagy-inducing effect of C₂-ceramide was demonstrated, and cotreatment with the autophagy inhibitor chloroquine (CQ) was investigated in NSCLC H460 and H1299 cells. The results suggested that C₂-ceramide exhibited dose-dependent anticancer effects in H460 and H1299 cells and autophagy induction. Zebrafish-based acridine orange staining confirmed the combined effects in vivo. Importantly, the combination of a sublethal dose of C₂-ceramide and CQ resulted in additive cytotoxicity and autophagy in both cell lines. Alterations of related signaling factors, including Src and SIRT1 inhibition and activation of the autophagic regulators LAMP2 and LC3-I/II, contributed to the autophagy-dependent apoptosis. We found that C₂-ceramide continuously initiated autophagy; however, CQ inhibited autophagosome maturation and degradation during autophagy progression. Accumulated and non-degraded autophagosomes increased NSCLC cell stress, eventually leading to cell death. This study sheds light on improvements to NSCLC chemotherapy to reduce the chemotherapy dose and NSCLC patient burden.

Bottle aging is the final stage before wines are drunk, and is considered as a maturation time when many chemical changes occur. To get a better understanding of the evolution of wines' flavor profile, the flavor compounds (phenolic and volatile compounds), dissolved oxygen (DO), and flavor characters (OAVs and chromatic parameters) of rosé and dry white wines bottled with different closures were determined after 18 months' bottle aging. The results showed the main phenolic change trends of rosé wines were decreasing while the trends of white wines were increasing, which could be the reason for their unique DO changing behaviors. Volatile compounds could be clustered into fluctuating, increasing, and decreasing groups using k-means algorithm. Most volatile compounds, especially some long-chain aliphatic acid esters (octanoates and decanoates), exhibited a lower decrease rate in rosé wines sealed with natural corks and white wines with screw caps. After 18 months of bottle aging, wines treated with natural corks and their alternatives could be distinguished into two groups based on flavor compounds via PLS-DA. As for flavor characters, the total intensity of aroma declined obviously compared with their initial counterparts. Rosé wines exhibit visual difference in color, whereas such a phenomenon was not observed in white wines.

The phenomenon of nuclear translocation of growth hormone receptor (GHR) in human, rat, and fish has been reported. To date, this phenomenon has not been described in a domestic animal (such as pig). In addition, the molecular mechanisms of GHR nuclear translocation have not been thoroughly elucidated. To this end, porcine hepatocytes were isolated and used as a cell model. We observed that porcine growth hormone (pGH) can induce porcine GHR's nuclear localization in porcine hepatocytes. Subsequently, the dynamics of pGH-induced pGHR's nuclear localization were analyzed and demonstrated that pGHR's nuclear localization occurs in a time-dependent manner. Next, we explored the mechanism of pGHR nuclear localization using different pGHR ligands, and we demonstrated that pGHR's nuclear translocation is GH(s)-dependent. We also observed that pGHR translocates into cell nuclei in a pGH dimerization-dependent fashion, whereas further experiments indicated that IMPα/β is involved in the nuclear translocation of the pGH-pGHR dimer. The pGH-pGHR dimer may form a pGH-GHR-JAK2 multiple complex in cell nuclei, which would suggest that similar to its function in the cell membrane, the nuclear-localized pGH-pGHR dimer might still have the ability to signal., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Withaferin A (WFA) is one of the most active steroidal lactones with reactive oxygen species (ROS) modulating effects against several types of cancer. ROS regulation involves selective killing. However, the anticancer and selective killing effects of WFA against oral cancer cells remain unclear. We evaluated whether the killing ability of WFA is selective, and we explored its mechanism against oral cancer cells. An MTS tetrazolium cell proliferation assay confirmed that WFA selectively killed two oral cancer cells (Ca9-22 and CAL 27) rather than normal oral cells (HGF-1). WFA also induced apoptosis of Ca9-22 cells, which was measured by flow cytometry for subG1 percentage, annexin V expression, and pan-caspase activity, as well as western blotting for caspases 1, 8, and 9 activations. Flow cytometry analysis shows that WFA-treated Ca9-22 oral cancer cells induced G2/M cell cycle arrest, ROS production, mitochondrial membrane depolarization, and phosphorylated histone H2A.X (γH2AX)-based DNA damage. Moreover, pretreating Ca9-22 cells with N -acetylcysteine (NAC) rescued WFA-induced selective killing, apoptosis, G2/M arrest, oxidative stress, and DNA damage. We conclude that WFA induced oxidative stress-mediated selective killing of oral cancer cells.

Type I epithelial ovarian cancer (EOC) is primarily resistant to platinum-based chemotherapies and needs novel therapeutics. Given the aberrant Wnt activation in type I EOC and the involvement of Dapper1 Antagonist of Catenin-1 (DACT1) in Wnt signalling, the role of DACT1 in tumourigenesis of type I EOC was evaluated. Firstly, all tested EOC cell lines and primary EOC tissues, especially type I EOC, were observed to have significantly lower DACT1 expression than normal controls. Next, 3AO cells, which arise from a patient with primary mucinous EOC and express low endogenous levels of DACT1, were transfected with a lentivirus carrying full-length DACT1 (3AO-DACT1), grew slower and formed smaller tumours in nude mice compared to 3AO-NC. Furthermore, 3AO-DACT1 had lower levels of key mediators of canonical Wnt signalling, Dvl2 and β-catenin, GSK-3β with phosphorylated Ser9, and the Wnt/β-catenin target genes, with significantly lower nuclear β-catenin levels. Additionally, 3AO-DACT which contained higher levels of lipidated LC3 (LC3-II) and Beclin1, but lower levels of p62/SQSTM1, were more sensitive to cis-platinum. And chloroquine partially rescued its cis-platinum resistance. We identified DACT1 as a negative regulator in type I EOC, protecting against malignant expansion by inhibiting canonical Wnt signalling and cis-platinum resistance by regulating autophagy.

We previously reported that the soft coral-derived bioactive substance, sinuleptolide, can inhibit the proliferation of oral cancer cells in association with oxidative stress. The functional role of oxidative stress in the cell-killing effect of sinuleptolide on oral cancer cells was not investigated as yet. To address this question, we introduced the reactive oxygen species (ROS) scavenger ( N -acetylcysteine [NAC]) in a pretreatment to evaluate the sinuleptolide-induced changes to cell viability, morphology, intracellular ROS, mitochondrial superoxide, apoptosis, and DNA damage of oral cancer cells (Ca9-22). After sinuleptolide treatment, antiproliferation, apoptosis-like morphology, ROS/mitochondrial superoxide generation, annexin V-based apoptosis, and γH2AX-based DNA damage were induced. All these changes were blocked by NAC pretreatment at 4 mM for 1 h. This showed that the cell-killing mechanism of oral cancer cells of sinuleptolide is ROS dependent., Competing Interests: Disclosure The authors report no conflicts of interest in this work.

Cervical cancer is the second most common female cancer worldwide. DNA methylation is one of a number of epigenetic regulation mechanisms leading to gene silencing in neoplastic cells. Aberrant methylation results in the silencing of tumor suppressor gene expression, and has been detected in a high percentage of human cancers. In the present study, the methylation status of three tumor suppressor genes, retinoic acid receptor β (RARβ), p16 and cadherin 1 (CDH1), and the inflammatory-associated cyclooxygenase-2 (COX-2) gene, was examined at distinct stages of cervical intraepithelial neoplasia (CIN). The results of the present study revealed that the COX-2 gene was unmethylated between CIN I and carcinoma specimens. The RARβ gene exhibited a minimal change in methylation frequency, whereas the CDH1 methylation level was increased <2-fold between CIN I and carcinoma. Notably, the methylation frequency of p16 was 13.2% in normal specimens; 18.2% in CIN I; 35.7% in CIN II; 31.6% in CIN III; and 15.4% in carcinoma. By contrast, the methylation frequency of p16 increased between CIN I and carcinoma in the absence of high-risk group papillomaviruses. The results of bisulfite sequencing indicated that the 10 CpG sites were all methylated in p16 gene methylation-positive individuals. The results of the present study demonstrate that the methylation frequency of p16 and CDH1 was progressively increased during the development of malignant stages in CIN, and may be an additional tool for current cytomorphology-based screening of cervical cell specimens.

Aim: The purpose of this study is to investigate the role of methylation in the histone deacetylases 6 (HDAC6) promoter and HDAC6 messenger RNA (mRNA) expression in the pathogenesis of systemic lupus erythematosus (SLE)., Method: Direct bisulfite-polymerase chain reaction (PCR) sequencing was performed to detect the HDAC6 promoter methylation in 33 patients with SLE and 35 healthy controls. The HDAC6 mRNA expression was measured in 93 SLE patients and 84 healthy controls by using the method of quantitative real-time PCR., Results: This study demonstrated that the methylation rates at HDAC6-680, -660 and -658 were significantly increased in the SLE patients compared with healthy controls (P = 0.041, 0.034 and 0.029, respectively). The SLE patients also had lower HDAC6 mRNA expression than the controls (P = 0.031). However, there was no significant difference in HDAC6 mRNA expression between patients with active and inactive SLE., Conclusion: The SLE patients had higher methylation in the HDAC6 promoter and lower HDAC6 mRNA expression than the controls. These changes may be related to the susceptibility of SLE. However, they are not associated with the disease activity of SLE., (© 2015 Asia Pacific League of Associations for Rheumatology and Wiley Publishing Asia Pty Ltd.)

Objective: Sinuleptolide, a soft corals-derived bioactive norditerpenoid, is a marine natural product with a potent anti-inflammatory effect. We evaluate the potential anti-oral cancer effects of sinuleptolide and investigate the possible mechanisms involved., Designs: Cell viability, cell cycle, apoptosis, reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and DNA damage analyses were performed., Results: In a cell viability assay, we found that sinuleptolide is dose-responsively antiproliferative against oral gingival cancer Ca9-22 cells but less harmful to normal human gingival fibroblast (HGF-1) cells (P<0.001). In cell cycle analysis, sinuleptolide induced subG1 accumulation at a higher dose and led to G2/M arrest of Ca9-22 cells (P<0.005). Apoptosis was significantly increased in sinuleptolide-treated Ca9-22 cells based on annexin V and poly(ADP-ribose) polymerase (PARP) expressions (P<0.05-0.0001). Based on flow cytometer analysis, sinuleptolide also induced the generation of ROS and decreased MMP in a dose-responsive manner (P<0.05-0.0001). DNA damage increased dose-responsively after sinuleptolide treatments (P < 0.001) based on comet and γH2AX assays., Conclusion: Sinuleptolide can induce an antiproliferation of oral cancer Ca9-22 cells involving apoptosis, oxidative stress and DNA damage, suggesting that sinuleptolide represents a potential chemotherapeutic drug for oral cancer treatment., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Aim: Although F11 receptor (F11R), also named junctional adhesion molecular A (JAM-A), participates in leukocyte migration, its role in autoimmune diseases has not been specifically disclosed. In this study, we examined the association of F11R expression with the development and clinical manifestations of rheumatoid arthritis (RA)., Method: RNA from peripheral blood mononuclear cells (PBMCs) and DNA from the peripheral blood in RA patients and a healthy control group were extracted. F11R messenger RNA (mRNA) expression was determined by quantitative real-time polymerase chain reaction. The F11R polymorphisms were determined by the TaqMan genotyping assay., Results: There was more F11R mRNA expression in the PBMCs of RA patients than those of the control group (P = 0.018). In F11R promoter -688 A > C, C carriers have lower titers of the anticyclic citrullinated peptide (anti-CCP) antibodies (P = 0.002) and fewer positive rates of Schirmer's tests (P = 0.009). The effect is independent of the existence of HLA-DR4. Different genotypes in F11R promoter -688 A > C and -436 A > G do not lead to changes of the gene expression in RA patients., Conclusion: RA patients have higher mRNA expression of F11R. In RA patients, F11R -688 C may be a protective factor for the development of anti-CCP antibodies and positive rates of Schirmer's tests., (© 2015 Asia Pacific League of Associations for Rheumatology and Wiley Publishing Asia Pty Ltd.)

Statement of Problem: The effects of different heat treatments on the internal oxidation and metal-ceramic bond in Pd-Ag alloys with different trace elements require further documentation., Purpose: The purpose of this in vitro study was to determine whether heat treatment affects the metal-ceramic bond strength of 2 Pd-Ag alloys containing different trace elements., Material and Methods: Thirteen cast specimens (25×3×0.5 mm) from each of 2 Pd-Ag alloy groups (W-1 and Argelite 61+3) were allocated to heat treatments before porcelain application: heating under reduced atmospheric pressure of 0.0014 MPa and 0.0026 MPa and heating under normal atmospheric pressure. Bond strengths were evaluated using a 3-point bending test according to ISO9693. Results were analyzed using 2-way ANOVA and Tukey HSD test (α=.05). Visual observation was used to determine the failure types of the fractured specimens. Scanning electron microscopy and energy dispersive spectroscopy were used to study morphologies, elemental compositions, and distributions in the specimens., Results: The W-1 group had a mean bond strength significantly higher than that of Argelite 61+3 (P<.001). Heating under reduced atmospheric pressures of 0.0014 MPa and 0.0026 MPa resulted in similar bond strengths (P=.331), and both pressures had significantly higher bond strengths than that of heating under normal atmospheric pressure (P=.002, P<.001). Heating under different air pressures resulted in Pd-Ag alloys that contained either Sn or In and Ga, with various degrees of internal oxidation and different quantities of metallic nodules., Conclusions: Heating under reduced atmospheric pressure effectively improved the bond strength of the ceramic-to-Pd-Ag alloys., (Copyright © 2015 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

Double-stranded breaks (DSBs) are cytotoxic DNA lesions caused by oxygen radicals, ionizing radiation, and radiomimetic chemicals. Increasing understanding of DNA damage signaling has provided an ever-expanding list of modulators reported to orchestrate DNA damage repair and ataxia telangiectasia mutated (ATM) is the master regulator and main transducer of the DSB response. Increasingly, it is being realized that DNA damage response is a synchronized and branched network that functionalizes different molecular cascades to activate special checkpoints, thus temporarily arresting progression of the cell cycle while damage is being assessed and processed. It is noteworthy that both nutrigenetics and nutrigenomics have revolutionized the field of molecular biology and rapidly accumulating experimental evidence has started to shed light on biological activities of a wide range of phytochemicals reported to modulate cell cycle, DNA repair, cell growth, differentiation and apoptosis as evidenced by cell-based studies. In this review, we have attempted to provide an overview of DNA damage signaling, how ATM signaling regulates tumor necrosis factors-related apoptosis inducing ligand (TRAIL)-induced intracellular network. We also illuminate on how resveratrol, epigallocatechin gallate, curcumin, jaceosidin, cucurbitacin, apigenin, genistein, and others trigger activation of ATM in different cancer cells as well as agents for ATM inactivation. Understanding the interplay of TRAIL-induced intracellular signaling and ATM modulation of downstream effectors is very important. This holds particularly for a reconceptualization of the apparently paradoxical roles and therapeutically targetable for enhancing the response to DNA damage-inducing therapy.

Group 9 transition metal complexes have been widely explored as therapeutic agents due to their unique geometry, their propensity to undergo ligand exchanges with biomolecules and their diverse steric and electronic properties. These metal complexes can offer distinct modes of action in living organisms compared to carbon-based molecules. In this study, we investigated the antimicrobial and anti-proliferative abilities of a series of cyclometallated iridium(III) complexes. The iridium(III) complex 1 inhibited the growth of S. aureus with MIC and MBC values of 3.60 and 7.19 μM, respectively, indicating its potent bactericidal activity. Moreover, complex 1 also exhibited cytotoxicity against a number of cancer cell lines, with particular potency against ovarian, cervical and melanoma cells. This cyclometallated iridium(III) complex is the first example of a substitutionally-inert, Group 9 organometallic compound utilized as a direct and selective inhibitor of S. aureus.

Purpose: To test the feasibility of a self-management intervention for lower urinary tract symptoms (LUTS) in adults with heart failure (HF) discharged from hospital., Design: Single blinded randomized controlled trial., Subjects and Setting: Thirty-one adults, aged 50 years and older, with an HF diagnosis and 1 or more LUTS were recruited during their hospitalization after passing a cognitive screen. Subjects received the intervention and completed postintervention measures in their own homes., Methods: During hospitalization, subjects were recruited, enrolled, and consented, and then completed baseline questionnaires and 24-hour pad test. After discharge, both groups received educational sessions on different topics by telephone in 4-weekly sessions. The specific aims were to determine: (1) subject recruitment and retention rates, (2) subjects' adherence to baseline and postintervention measures, and (3) subjects' and nurse interventionist's adherence to the protocol. The LUTS intervention effects on specific clinical outcomes were explored., Results: Potential subjects were recruited over 5 months at an enrollment ratio of 4.7:1. Approximately 68% completed the study. Average age was 66.3 ± 9.8 years (mean ± SD). The majority were female (54.8%) and white (51.6%). Most subjects had urinary incontinence (UI) (74.2%) and 77.4% rated their health as either fair or poor. The study was underpowered to determine statistical significance at P < .05 level. Thirty-three percent of the LUTS intervention group reported improved UI frequency postintervention, compared to 25% of the attention control group., Conclusion: Adults with HF experience LUTS, but little is known about how best to manage and treat it. This study showed that it is possible to recruit and retain adults who have HF and rate their health as fair or poor into a 4-week intervention study, although oversampling is needed due to attrition.

Background: Exercise is important for fitness and recovery of older adults after hospitalization for treatment of cardiovascular disease. Home-based, nurse-led exercise programs may be beneficial., Objective: The aim of this study was to test the effects of a low-intensity, home-based exercise protocol led by an advanced practice nurse on health-related quality of life (HRQOL), physical fitness, and left ventricular ejection fraction (LVEF) in older adults after hospital discharge with a cardiovascular disease diagnosis., Methods: The study was randomized and single blinded. Seventy-seven older adults (≥75 years old, mean = 80.68 years old) were included; 32 subjects in the intervention and 29 in the control group completed the study. The low-intensity, home-based exercise protocol is composed of 14-type joint exercises and walking for 12 weeks. The main outcome measures were assessments on the Medical Outcomes Study Short-Form-36, the Senior Fitness Test, and LVEF at baseline and 12 weeks after hospital discharge., Results: After 12 weeks, the intervention group showed significant improvements in HRQOL (physical functioning, role-physical, bodily pain, and vitality; p < .05) as well as on the Senior Fitness Test (chair stands, arm curls, Timed Up and Go, and 6-minute walk distance; p < .05); there was no significant improvement in LVEF (p = .56)., Conclusions: The low-intensity, home-based exercise led by an advanced practice nurse was effective in improving HRQOL and physical fitness. Adherence was high, and there were no adverse events related to exercise.

Background: The course of incident delirium and subsyndromal delirium (SSD) after cardiac surgery is not well studied., Objective: The aim of this study was to evaluate the course of incident delirium and SSD, their risk factors, and impact on patients' cognitive function after elective coronary artery bypass graft (CABG) surgery., Methods: Consecutive patients scheduled for an isolated CABG at a tertiary medical center in Taiwan were enrolled if they had no preoperative delirium symptoms. Delirium was assessed daily for 1 week after surgery using the Confusion Assessment Method. Subsyndromal delirium was defined as presenting with any core symptom below the diagnostic threshold for delirium. Cognitive function was assessed by the Mini-mental State Examination., Results: Of 38 participants, 7 had incident (first-time) delirium (18.4% incidence) and 13 had incident SSD (34.2% incidence). Whereas SSD usually lasted 1 day, delirium changed gradually to SSD to recovery and its symptomatology lasted longer. We identified 6 delirium risk factors: older age, more comorbidities, cardiac pulmonary bypass, blood transfusion, larger transfusion volume, and longer duration of intraoperative blood pressure less than 60 mm Hg. The frequencies of these risk factors for SSD were often intermediate between those of risk factors in groups with and without delirium. By hospital discharge, participants with delirium had the longest hospital stays and lowest cognitive scores, those with SSD had intermediate stays and scores, and those without delirium had the lowest stays and scores., Conclusion: Delirium and SSD after CABG are common. Greater number and severity of risk factors for delirium may predict increasingly poor outcomes, with the dose-response relationship between risk factors and outcomes for SSD intermediate between that for no symptoms and full delirium. Intervention trials are indicated, particularly for patients with a greater number and severity of predisposing and precipitating risk factors.

Cervical cancer screening guidelines do not comprehensively define what constitutes high risk. This study developed and validated simple risk-scoring schemes to improve Papanicolaou smear screening for women at high risk. Four cumulative risk score (CRS) schemes were derived respectively for the development of cervical intraepithelial neoplasia grade 1 (CIN1) and grade 2 or worse (CIN2+) using community-based case-control data (n = 1523). By calculating the area under the receiver operating characteristic (AU-ROC) curve, these schemes were validated in a Papanicolaou smear follow-up cohort (n = 967) and a hospital-based cytology screening population (n = 217). A high DNA load of high-risk human papillomavirus (HR-HPV) was the main predictor for CIN1 and CIN2+, although age, married status combined with the number of sexual partners, active and passive smoking and age at sexual debut also affected associated lesions. In the training set, only the HPV-testing-contained CIN2+ CRS scheme presented an excellent discrimination for identifying CIN2+ (AU-ROC = 0.866). Using a CRS cutoff value of 4 to identify CIN2+, the sensitivity and specificity of predicting CIN2+ for the 3- and 5-year follow-ups were 100% and 90.8%, and 83.3% and 90.4%, respectively, in the validation cohort. In the hospital-based validation population, the CRS scheme showed comparable discrimination for CIN2+ detection (sensitivity 88.2% and specificity 84.6%). Women with CRS ≥ 4 had a 5.4% and 9.1% of 3- and 5-year cumulative incidence, respectively, and a 40.5-fold hazard ratio of developing CIN2+. In conclusion, combined with HR-HPV testing and verified risk factors, a simple CRS scheme could effectively improve the implementation of CIN2+ screening., (© 2014 UICC.)

Data obtained through high-throughput technologies have gradually revealed that a unique stratified epithelial architecture of human skin along with the antioxidant-response pathways provided vital defensive mechanisms against UV radiation. However, it is noteworthy that skin is a major target for toxic insult by UV radiations that can alter its structure and function. Substantial fraction of information has been added into the existing pool of knowledge related to natural products mediated biological effects in UV exposed skin cells. Accumulating evidence has started to shed light on the potential of these bioactive ingredients as protective natural products in cosmetics against UV photodamage by exerting biological effects mainly through wide ranging intracellular signalling cascades of oxidative stress and modulation of miRNAs. In this review, we have summarized recently emerging scientific evidences addressing underlying mechanisms of UV induced oxidative stress and deregulation of signalling cascades and how natural products can be used tactfully to protect against UV induced harmful effects.

Radiotherapy effectively destroys cancer cells in many sites of the body, but several limitations remain. This study investigated alternative splicing, which is a common mechanism of increased diversity in mRNAs and proteins. The relationships of alternative splicing to DNA damage and radiation such as UV and ionizing radiation were analyzed. The DNA damage responses of many genes involved in alternative splicing were compared between non-radiation and radiation treatments. Drugs that affect radioresistence or radiosensitization by modulating the effects of alternative splicing and radiation were also reviewed.

The aim of this study was to apply a decision forest to analysis of the ultrasound characteristics and laboratory test indices of four types of primary glomerulopathy, and quantitative analysis of the four pathologic types using a combination of these two methods. The decision trees were derived from 41 clinical indices and 5 characteristic sonographic indices obtained for the left kidney. Fifty-six patients who had undergone ultrasound-guided renal biopsy were reviewed retrospectively, and on pathologic examination, the patients were diagnosed with primary glomerulopathy, which includes mesangial proliferative glomerulonephritis, membranous nephropathy, immunoglobulin A nephropathy and minimal change disease. In this study, eight characteristic indicators were correlated with pathologic type in the 56 cases of primary glomerulopathy. The order calculated by decision forests, from high to low, is proteinuria, length of kidney, serum creatinine, plasma albumin, area of kidney, total protein, thickness of renal parenchyma, 24-h urine protein. The glomerulopathy with the highest ++++ proteinuria is membranous nephropathy, which accounts for 39.2% (22/56) of the total sample; this was followed by minimal change disease, mesangial proliferative glomerulonephritis and immunoglobulin A nephropathy. On the basis of our analysis of 41 clinical indices, the key indices for quantitative analysis of primary glomerulonephritis are laboratory tests, and these include urine protein, serum creatinine, plasma albumin, total serum protein and 24-h urine protein. The three key sonographic features are measurement indices: renal length, renal area and renal parenchymal thickness. From the eight characteristic indicators, we observed that with respect to severity (from most severe to least severe), the four types of glomerulopathy are membranous nephropathy, minimal change disease, mesangial proliferative glomerulonephritis and immunoglobulin A nephropathy., (Copyright © 2014 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

Tobacco includes thousands of chemicals such as nicotine, which causes numerous diseases including oral cancer. We synthesized nicotinic acid based probes by chemical modification to identify the proteins expressed by the oral cancer cell line Ca9-22 that interact with the nicotinic functional group. Proteins belonging to human oral squamous cell carcinoma were pulled down by a probe carrier based on nicotinic acid, which was reacted with 3-aminopropyltriethoxysilane to compose nicotinic acid linked 3-aminopropyltriethoxysilane exposed on the SiO2 surface. Oral cancer cell lysates were incubated with the nicotinic acid chemical probes to identify the interactions between the nicotinic group and oral cancer cell line extracted proteins. The interactions between the chemical probes and proteins identified as their targets were confirmed by consulting chemicals databases. Interestingly, chaperone proteins (e.g., heat-shock proteins and endoplasmin) that were found to interact with nicotinic acid were identified as binding partners in ribosomal and nucleosome assembly complexes., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Focal hypermethylation in promoter regions of tumor suppressor genes against the background of global hypomethylation is a landmark of carcinogenesis. This study aimed to investigate the methylation status of retinoic acid receptor beta2 (RARβ2) and long interspersed nuclear elements (LINE-1) in different stages of esophageal squamous cell carcinoma (ESCC)., Method: The tumor and adjacent normal esophageal tissues from 125 male ESCC patients who underwent primary surgery were analyzed for the methylation status of RARβ2 promoter and LINE-1 through methylation-specific polymerase chain reaction and pyrosequencing., Results: RARβ2 hypermethylation was detected in 20% of the tumor samples, but not in the normal counterparts. The methylation frequency of LINE-1 was significantly lower in the tumor than in the normal parts (median: 67.7% vs. 80%, P < 0.0005). Ninety-eight patients (78.4%) had both RARβ2 hypermethylation and LINE-1 hypomethylation or either one. There was a trend toward higher risk of advanced T stage (P for trend = 0.05) or lymph node metastasis (P for trend = 0.02) when more adverse gene methylation profiles were present., Conclusion: Methylation status of RARβ2 and LINE-1 was related to the development and possibly the severity of ESCC., (© 2014 Wiley Periodicals, Inc.)

Cytokines are involved in the pathogenesis of autoimmune diseases. Oncostatin M receptor (OSMR) activates JAK/STAT and MAPK pathways leading to the stimulation of a variety of cytokines and inflammatory substances. Many pro-inflammatory cytokines are involved in the inflammatory process of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). In this study, we carried out experiments to examine the relationship of OSMR promoter polymorphisms with RA and SLE patients. 241 patients of RA, 143 patients of SLE and 203 healthy controls were enrolled in their recruitment from the Kaohsiung Medical University Hospital. Genomic DNA was extracted from peripheral blood mononuclear cells and gene polymorphism was genotyped by TaqMan real-time polymerase chain reaction. The OMSR promoter region -100 G/T (rs22922016) genotype was in no relation to the susceptibility of RA, but -100 T/T (rs22922016) genotype could prevent the patients with sicca syndrome and the existence of anti-Ro antibodies. In contrast, the -100 G/T+T/T (rs22922016) genotypes were significantly associated with an increased risk of SLE (odds ratio, OR=1.62, 95% confidence interval (CI), 1.01-2.62). 94.38% of SLE patients with arthritis were belonged to the -1687C/C (rs540558) genotype. The T allele of promoter region -100 T/T (rs22922016) has protective effect and could ameliorate the disease condition in RA patients, whereas the same T allele was a risk allele in the susceptibility of SLE. The disease severity of rheumatoid arthritis and systemic lupus erythematosus can be partially affected by the OSMR promoter polymorphisms.

In a previous study, we calculated the surface dynamics of noisy viscoelastic supported films by using an adiabatic approximation. An expression was derived for the time-dependent power spectral density (PSD), which was found to produce good agreement with experiment. In this study, we extend the treatment to viscoelastic free-standing films. Two sets of surface capillary normal modes, namely, the squeezing and bending modes, were found. The frequency dispersion relation of the former resembles that of supported films. The latter is distinctively different and diverges at long wavelengths. By incorporating the experimental conditions, we obtained satisfactory agreement between theory and experiment.

Background: Asthma is a common chronic inflammatory respiratory disease. Previous studies have suggested that the pathogenesis of asthma may be affected by epigenetic regulation. The purpose of this study is to characterize the effect of the methylation of each CpG site in the ADAM33 (a disintegrin and metalloproteinase 33) gene in adult asthma., Methods: A human CpG island microarray was used to examine 4 asthmatic cases and 4 healthy controls, and the results suggested that there might be differences in methylation within exon 9 of the ADAM33 gene. Therefore, we designed a case-control study with 50 asthmatic patients and 50 age- and sex-matched healthy controls to examine the relationship between the CpG methylation of the ADAM33 gene and asthma using bisulfite deoxyribonucleic acid modification and sequencing., Results: Bisulfite sequencing experiments showed that the 14 CpG sites in exon 9 of the ADAM33 gene were highly methylated (100%) in all individuals. The proportions of methylation of the 14 CpG sites in ADAM33 in the case group were not different from those of the control group. The methylation of exon 9 of this locus was not associated with age, sex, IgE levels, or lung function. This study found no association between the methylation of CpG sites in exon 9 of the ADAM33 gene and adult asthma., Conclusions: The 14 CpG sites were highly methylated in the case and control groups. Further investigation of exon 9 in ADAM33 in a larger population is needed to evaluate its role in asthma., (Copyright © 2012 S. Karger AG, Basel.)

Objectives: To investigate the role that monocytes and solute carrier family 11 member A1 (SLC11A1) gene polymorphisms play in the pathogenesis of reactive arthritis (ReA)., Methods: SLC11A1 274C/T and 823C/T polymorphisms were determined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). The phagocytic activity of monocytes was analysed by flow cytometry after they were co-cultured with Chlamydia trachomatis. The inclusions in the monocytes were demonstrated by immunohistochemistry staining. Bactericidal activity was determined by immunofluorescence staining with the recovered inclusions., Results: There was no significant difference in the phagocytic activity of monocytes between the ReA patients and healthy controls. The bactericidal activity of monocytes from the healthy controls was more efficient than that from the ReA patients. The patients with SLC11A1 823T tended to have a higher bactericidal activity of monocytes than those with SLC11A1 823 C/C. Moreover, the bactericidal activity of monocytes in the patients with SLC11A1 274T seemed to decrease in comparison with that in the patients with SLC11A1 274C/C., Conclusions: The bactericidal activity of monocytes in patients with ReA is lower than that in healthy controls. The SLC11A1 274C/T and 823C/T polymorphisms may be associated with the decreased bactericidal activity of the monocytes.

Ketoacidotic syndromes are frequently encountered in acute care medicine. This article focuses on ketosis and ketoacidotic syndromes associated with intoxications, alcohol abuse, starvation, and certain dietary supplements as well as inborn errors of metabolism. Although all of these various processes are characterized by the accumulation of ketone bodies and metabolic acidosis, there are differences in the mechanisms, clinical presentations, and principles of therapy for these heterogeneous disorders. Pathophysiologic mechanisms that account for these disorders are presented, as well as guidance regarding identification and management., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Digital image processing has been applied on various fields such as classification and qualitative analysis. In this work, a very simple quantitative approach was proposed for the first time. Based on the digital grayscale images of three-dimensional fluorescence spectra, several wavelet moment invariants were calculated, and used to establish the linear models for the quantitative analysis. This approach was applied to the quantitative analysis of Tryptophan, Tyrosine and Phenylalanine in mixture samples, and the correlation coefficients R(2) of the obtained linear models were more than 0.99, which were supported by the strict statistical parameters as well as leave-one-out and Jackknife cross-validations. Our study indicates that the selected wavelet moment invariants are immune from the noise and background signals, and the quantitative analysis can be performed accurately based on the overlapping peaks of compounds in mixture. This proposed approach provides a novel pathway for the analysis of three-dimensional spectra.

Objectives: To investigate the associations of DNA methylation levels and mRNA expressions of DNA cytosine-5-methyltransferase 1 (DNMT1) and methyl CpG-binding domain 2 (MBD2) with rheumatoid arthritis (RA)., Methods: The global methylation status of DNA was measured in 65 patients with RA and 64 healthy controls by the ELISA method. DNMT1 and MBD2 mRNA were also detected in 177 RA patients and 95 controls using the quantitative real-time polymerase chain reaction method., Results: The global methylation of DNA was significantly decreased in the RA patients compared to the controls (p=0.005, 95% CI=0.0835-0.4503). The patients with RA had higher expressions of DNMT1 and MBD2 mRNA than the controls (p<0.001, 95% CI=-0.0024 to -0.0053 and p<0.001, 95% CI=-0.0079 to -0.0167, respectively). We also found that the MBD2 mRNA level was not related to the disease activity of RA. However, the expression of DNMT1 mRNA tended to be associated with the disease activity of RA (p=0.08). The levels of DNA methylation and DNMT1 mRNA were significantly decreased in the patients with anti-CCP antibody compared with those without (p=0.005, 95% CI=-0.7333 to -0.1373 and p=0.003, 95% CI=-0.0071 to -0.0022, respectively). The differences in the methylation level and expressions of DNMT1 and MBD2 were not significant between the patients treated with and without anti-TNFα biological agents (Enbrel or Humira)., Conclusion: This study demonstrated that the RA patients have a significantly lower level of DNA methylation than the controls. Moreover, RA patients have higher expressions of DNMT1 and MBD2 mRNA. The anti-TNFα biological agents do not seem to affect DNA methylation and mRNA expressions of DNMT1 and MBD2 in RA patients., (Copyright © 2010 Elsevier B.V. All rights reserved.)