Isoform specific roles of Broad‐Complex in larval development in Leptinotarsa decemlineata

Broad‐Complex (BrC) is a downstream target of both 20‐hydroxyecdysone and juvenile hormone signalling. BrC regulates morphogenetic changes between nymphal instars in hemimetabolans, whereas it controls pupal commitment, pupal morphogenesis and inhibits adult differentiation in holometabolans. Among five BrC cDNAs (Z1–Z4 and Z6) identified in the Colorado potato beetle, we found in this work that Z1, Z2 and Z6 were mainly expressed at the last (fourth) instar and prepupal stages, whereas the levels of Z3 and Z4 increased during the penultimate (third) instar stage, peaked at the last instar larval phase and gradually decreased at the prepupal and pupal periods. When knocking down all BrC isoforms by RNA interference (RNAi) at the penultimate instar stage, around 20% of the resultant larvae remained as moribund beetles. These moribund BrC RNAi larvae were completely or partially wrapped in old cuticle. Likewise, a portion of larvae treated for a single double‐stranded RNA of Z3, Z4 or Z6 displayed a degree of similar aberrancies, increasing in the order of isoforms Z6 < Z3 < Z4. When silencing all BrC isoforms at the last instar period, most of the RNAi larvae did not normally pupate or emerge as adults. Separately silencing each of the five zinc finger domains revealed that approximately 70% of the Z1 RNAi larvae remained as prepupae, around 60% of the Z6 RNAi specimens formed aberrant prepupae or pupae and about 60% of the Z2 RNAi beetles became deformed pupae. After removal of the old exuviae, these deformed larvae in which either Z1, Z2 or Z6 was depleted possessed adult prothorax and mesothorax, developing antenna, mouthparts and wing discs. Moreover, less than 50% of the resultant pupae finally emerged as adults when either of Z1, Z2 or Z6 was knocked down. Therefore, our findings reveal, for the first time, that the two roles of BrC in insect groups (ie directing morphogenetic changes during juvenile development and regulating larval–pupal–adult metamorphosis) are played by different BrC isoforms in Leptinotarsa decemlineata.     

Amplification and quantification of cold‐associated microRNAs in the Colorado potato beetle (Leptinotarsa decemlineata) agricultural pest

The Colorado potato beetle [Leptinotarsa decemlineata (Say)] is an important insect pest that can inflict considerable damage to potato plants. This insect can survive extended periods of cold exposure, and yet the molecular switches underlying this phenomenon have not been fully elucidated. A better characterization of this process would highlight novel vulnerabilities associated with L. decemlineata that could serve as targets for the management of this devastating pest. Using high‐throughput sequencing, the current work reveals a cold‐associated signature group of microRNAs (miRNAs) in control (15 °C) and ?5 °C‐exposed L. decemlineata. The results show 42 differentially expressed miRNAs following cold exposure including miR‐9a‐3p, miR‐210‐3p, miR‐276‐5p and miR‐277‐3p. Functional analysis of predicted targets associated with these cold‐responsive miRNAs notably linked these changes with vital metabolic and cellular processes. Overall, this study highlights the miRNAs probably responsible for facilitating cold adaptation in L. decemlineata and implicates miRNAs as a key molecular target to consider in the development of novel pest management strategies against these insects.     

Ovariectomy induces oxidative stress and impairs bone antioxidant system in adult rats

BACKGROUND: There is increasing evidence suggesting the role of free radicals in bone resorption and bone loss. Ovariectomized rats have been used as the animal model for the study of osteoporosis. Oxidative stress due to reactive oxygen species (ROS) can cause oxidative damage to cells. Cells have a number of defense mechanisms to protect themselves from the toxicity of ROS. Even though, there are studies portraying the role of free radicals in bone loss, the defense mechanism adapted by bone in ovariectomized animals remains obscure. We investigated the impact of ovariectomy (OVX) on the bone antioxidant system in rats. METHODS: Sixty days after bilateral OVX, the rats were killed and the femora were removed, the tissue homogenates were made and used for the estimation of superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione S transferase (GST), lipid peroxidation (LPO) and hydrogen peroxide (H(2)O(2)). RESULTS: The levels of LPO and H(2)O(2) were increased and enzymatic antioxidants like SOD, GPx, GST were decreased in ovariectomized animals when compared to sham-operated control rats. The calculated correlation coefficient demonstrated strong correlation (0.905) between the production of H(2)O(2) and LPO in the femur bone. A strong inverse (-0.945) correlation was observed between H(2)O(2) production and SOD activity. CONCLUSIONS: OVX induces oxidative stress in the femur bone of rats.     

The Keap1–Nrf2–ARE Pathway As a Potential Preventive and Therapeutic Target: An Update

The Keap1–Nrf2–ARE ((Kelch‐like ECH‐Associating protein 1) nuclear factor erythroid 2 related factor 2‐antioxidant response element) pathway is one of the most important defense mechanisms against oxidative and/or electrophilic stresses, and it is closely associated with inflammatory diseases, including cancer, neurodegenerative diseases, cardiovascular diseases, and aging. In recent years, progress has been made in strategies aimed at modulating the Keap1–Nrf2–ARE pathway. The Nrf2 activator DMF (Dimethylfumarates) has been approved by the FDA as a new first‐line oral drug to treat patients with relapsing forms of multiple sclerosis, while a phase 3 study of another promising candidate, CDDO‐Me, was terminated for safety reasons. Directly inhibiting Keap1–Nrf2 protein–protein interactions as a novel Nrf2‐modulating strategy has many advantages over using electrophilic Nrf2 activators. The development of Keap1–Nrf2 protein–protein interaction inhibitors has become a topic of intense research, and potent inhibitors of this target have been identified. In addition, inhibiting Nrf2 activity has attracted an increasing amount of attention because it may provide an alternative cancer therapy. This review summarizes the molecular mechanisms and biological functions of the Keap1–Nrf2–ARE system. The main focus of this review is on recent progress in studies of agents that target the Keap1–Nrf2–ARE pathway and the therapeutic applications of such agents.     

Surface modification of titanium with hydroxyapatite–heparin–BMP‐2 enhances the efficacy of bone formation and osseointegration in vitro and in vivo

Surface‐modified titanium (Ti) samples with hydroxyapatite (HAp) and heparin (Hep)–bone morphogenetic protein‐2 (BMP‐2) complex (Ti/HAp/Hep/BMP‐2) were prepared, and their efficacies on the enhancements of bone formation and osseointegration in vitro and in vivo were examined as compared to Ti/HAp and Ti/Hep/BMP‐2. The modified surfaces were characterized by X‐ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM) and contact angle goniometry. In vitro studies revealed that MG‐63 human osteosarcoma cell lines grown on Ti/HAp/Hep/BMP‐2 increased the amounts of alkaline phosphatase (ALP) activity, calcium deposition and the levels of OCN mRNA gene expression as compared to those grown on Ti/HAp, Ti/Hep/BMP‐2 or pristine Ti. Moreover, Ti/HAp/Hep/BMP‐2 exhibited higher bone volume (BV), bone volume/tissue volume (BV/TV), removal torque value and bone–implant contact (BIC) than Ti/HAp, Ti/Hep/BMP‐2 or pristine Ti in vivo. Histological evaluations showed that many desirable features of bone remodelling existed at the interface between Ti/HAp/Hep/BMP‐2 and the host bone. Consequently, Ti/HAp/Hep/BMP‐2 may have potential for clinical use as dental or orthopaedic implants. Copyright © 2014 John Wiley & Sons, Ltd.     

SARS-CoV-2疫苗接种成人血清SARS-CoV-2IgM抗体、IgG抗体和中和抗体水平分析

目的 评估成人接种严重急性呼吸综合征冠状病毒2(SARS-CoV-2)疫苗后SARS-CoV-2 IgM、IgG和中和抗体的形成情况.方法 选取接受SARS-CoV-2疫苗接种的成人80名(疫苗接种组),检测疫苗接种完成后7 d内和第14天的血清SARS-CoV-2 IgM抗体、IgG抗体及中和抗体水平.以未接种SAR...     

Nuclear factor erythroid‐derived 2–related factor 2 activates glutathione S‐transferase expression in the midgut of Spodoptera litura (Lepidoptera: Noctuidae) in response to phytochemicals and insecticides

Detoxication enzymes play an important role in insect resistance to xenobiotics such as insecticides and phytochemicals. We studied the pathway for activating the expression of glutathione S‐transferases (GSTs) in response to selected xenobiotics. An assay of the promoter activity of GST epsilon 1 (Slgste1) of Spodoptera litura led to the discovery of a cis‐regulating element. An antioxidant response element was activated in response to indole‐3‐carbinol (I3C) and chlorpyrifos (CPF) and was able to bind with the xenobiotic sensor protein nuclear factor erythroid‐derived 2–related factor 2 (SlNrf2). SlNrf2 and Slgste1 were responsive to reactive oxygen species induced by I3C and CPF in a S. litura cell line, as well as in S. litura midguts. SlNrf2 RNA interference (RNAi) reduced the message RNA levels of Slgste1 and the peroxidase activity of GSTs in response to I3C, xanthotoxin, CPF and deltamethrin. SlNrf2 RNAi and inhibitor treatment of GST activity decreased the viability of I3C‐treated cells. These results indicate that SlNrf2 activates the expression of GSTs in response to oxidative stresses caused by exposure to xenobiotics.     

Reliability and validity of the Algase Wandering Scale – version 2 for Japanese people with dementia

In Japan, where older people already make up more than 23% of the population and the proportion is still growing, the burden on those caring for people with dementia is an increasing problem. This burden is magnified by wandering behavior, a peripheral symptom. Thus, there is a need for an objective measure of wandering behavior to determine what constitutes effective care. In this study, we translated the Algase Wandering Scale – Version 2 into Japanese, and examined its reliability and validity. Ambulatory residents with dementia were selected from two nursing homes and two wards specializing in dementia care in hospitals in Japan. Nurses and care workers taking care of these residents answered questionnaires regarding the residents. From the results, the Algase Wandering Scale – Version 2, Japanese version, was examined for inter‐rater reliability, stability, internal consistency, and concurrent validity. The results of the analysis in the present study demonstrated that the Algase Wandering Scale – Version 2, Japanese version, has reliability and validity, and that it can measure the presence or absence of wandering and its severity. Surveys of residents with various wandering patterns in many facilities and verification of construct validity are warranted in the future.     

Agar–gelatin hybrid sponge‐induced three‐dimensional in vitro ‘liver‐like’ HepG2 spheroids for the evaluation of drug cytotoxicity

Agar–gelatin hybrid sponges were used as scaffolds to induce the formation of three‐dimensional (3D) spheroids of HepG2 cells. Agar and gelatin in 2:1 ratio were used to make films and sponges. The cell adhesive properties of the films were evaluated by the attachment kinetics. The growth kinetics of HepG2 cells was studied using MTT assay and morphology of the 3D spheroids was observed through inverted optical microscopy. The liver cell‐specific functions of the 3D spheroids were evaluated in terms of albumin secretion and urea synthesis. Paracetamol was used as a model drug to investigate the use of these 3D spheroids in the preliminary cytotoxicity evaluation of drugs. The results showed that the agar–gelatin hybrid sponges induced the formation of 3D HepG2 spheroids with significant liver‐specific functions. These spheroids exhibited higher amounts of albumin and urea synthesis than the control monolayer culture. These 3D spheroids were found to be more sensitive to the drug (TCIC₅₀ value of 4.6 mM ) than the control monolayer (TCIC₅₀ value of 6.2 mM ). The study shows that agar–gelatin‐induced HepG2 3D spheroids can be used for the preliminary evaluation of the toxicity of drugs and chemicals. Copyright © 2009 John Wiley & Sons, Ltd.     

伴t(1;19)(q23;p13)和E2A-PBX1成人急性T淋巴细胞白血病一例报告附文献复习

目的 总结1例伴有t(1;19)和E2A-PBX1融合基因阳性的成人T细胞急性淋巴细胞白血病(ALL)患者的诊疗体会.方法 对1例成人T细胞ALL患者进行染色体核型分析,流式细胞术检测免疫表型,同时进行融合基因多重RT-PCR扩增.结果 患者染色体核型为47,XY,9p+,15p+,17q-,der(19),t(1;19)(q23;p13)[5]/46,XY[15].E2A-PBX1融合基因阳性表达.给予hyperCVAD(环磷酰胺+长春新碱+阿霉素+地塞米松)方案治疗后患者获血液学完全缓解,染色体核型复查为46,XY[10],E2A-PBX1融合基因检测阴性.结论 E2A-PBX1+ t(1;19)也可以发生于T细胞ALL,E2a-PBX1白血病细胞在不同的癌基因协同信号或微环境下转化方向可变.     

伴t(1;19)(q23;p13)和E2A-PBX1成人急性T淋巴细胞白血病一例报告附文献复习

目的 总结1例伴有t(1;19)和E2A-PBX1融合基因阳性的成人T细胞急性淋巴细胞白血病(ALL)患者的诊疗体会.方法 对1例成人T细胞ALL患者进行染色体核型分析,流式细胞术检测免疫表型,同时进行融合基因多重RT-PCR扩增.结果 患者染色体核型为47,XY,9p+,15p+,17q-,der(19),t(1;19)(q23;p13)[5]/46,XY[15].E2A-PBX1融合基因阳性表达.给予hyperCVAD(环磷酰胺+长春新碱+阿霉素+地塞米松)方案治疗后患者获血液学完全缓解,染色体核型复查为46,XY[10],E2A-PBX1融合基因检测阴性.结论 E2A-PBX1+ t(1;19)也可以发生于T细胞ALL,E2a-PBX1白血病细胞在不同的癌基因协同信号或微环境下转化方向可变.     

Trends in the incidence of use of noninsulin glucose‐lowering drugs between 2006 and 2013 in France

This study aimed at describing trends in the incidence of use of noninsulin glucose‐lowering drugs (NIGLDs) between 2006 and 2013 in France. Repeated cross‐sectional studies on NIGLD new users were performed annually from 2006 to 2013 within the Echantillon Généraliste des Bénéficiaires (EGB) database, a 1/97th representative sample of the population covered by the French healthcare insurance system. NIGLD included metformin, sulfonylureas, α‐glucosidase inhibitors, thiazolidinediones, dipeptidylpeptidase‐4 (DPP‐4) inhibitors, glinides and glucagon‐like peptide‐1 analogues. New users were defined as patients with no delivery of any NIGLD (first‐line new users) or no delivery of a NIGLD of the same class (add‐on/switch new users) in the preceding year. Incidence rates of use and corresponding 95% confidence intervals (95% CI) were estimated per 1000 persons. Among the 507 043 persons included in the EGB in 2006, 2036 were identified as NIGLD first‐line new users and 2192 as add‐on/switch new users, which corresponded to an incidence of use of 4.0‰ (95%CI 3.8–4.2) and 4.3‰ (4.1–4.5), respectively. First‐line incidence increased to 5.3‰ (5.1–5.5) in 2010 and then decreased to 4.2‰ (4.0–4.4) in 2013; add‐on/switch incidence increased to 8.0‰ (7.8–8.2) in 2010 and then decreased to 5.3‰ (5.1–5.5) in 2013. This reduction was mainly related to DPP‐4 inhibitors, whose use as add‐on/switch NIGLDs was roughly halved between 2010 and 2013. Concomitantly, the use of sulfonylureas and glinides increased. In conclusion, after reaching a peak in 2010, the incidence of use of NIGLDs has markedly decreased in France. Since then, prescribers seem to have reverted to older and well‐known therapies.     

Effects of variation in the human α2A‐ and α2C‐adrenoceptor genes on cognitive tasks and pain perception

Background: The mechanisms underlying interindividual variability in pain perception and cognitive responses are undefined but highly heritable. α_2C‐ and α_2A‐adrenergic receptors regulate noradrenergic activity and are important mediators of pain perception and analgesia. We hypothesized that common genetic variants in these genes, particularly the ADRA2C 322–325 deletion variant, affect pain perception or cognitive responses. Methods: We studied 73 healthy subjects (37 Caucasians and 36 African–Americans) aged 25.4 ± 4.6 years. Pain response to a cold pressor test was measured using a 10 cm visual analog scale and again on the next day, after three infusions of the selective α₂‐agonist dexmedetomidine. Standardized cognitive tests were administered at baseline and after each infusion. The contribution of ADRA2C deletion genotype, dexmedetomidine concentration, and other covariates to pain perception and cognitive responses was determined using multiple linear regression models. Secondary analysis examined the effects of ADRA2A and other ADRA2C variants on pain perception. Results: ADRA2C Del homozygotes had higher pain scores in response to cold at baseline (6.3 ± 1.8 cm) and after dexmedetomidine (5.6 ± 2.2 cm) than insertion allele carriers (4.6 ± 2.1 cm [baseline] and 3.8 ± 1.9 cm [after dexmedetomidine]; adjusted P‐values = 0.019 and 0.004, respectively). Cognitive responses were unrelated to ADRA2C Ins/Del genotype. None of the other ADRA2A and ADRA2C variants was significantly related to cold pain sensitivity before dexmedetomidine; after dexmedetomidine, ADRA2A rs1800038 was marginally associated (P = 0.03). Conclusion: The common ADRA2C del322–325 variant affected pain perception before and after dexmedetomidine but did not affect other cognitive responses, suggesting that it contributes to interindividual variability in pain perception.