DsFoxO knockout affects development and fecundity of Drosophila suzukii.

Forkhead box O (FoxO), a key transcription factor in many species, participates in numerous physiological and pathological processes of organisms through a variety of signaling pathways. In the present study, we established DsFoxO knockout (DsFoxO-KO) strain using CRISPR/Cas9, and the influence on development and fecundity of mutant strain were evaluated. To clarify the corresponding mechanism, a transcriptome analysis was conducted subsequently. The results showed that the survival rates of the DsFoxO-KO strain in larval, pupal, and adult stages were all significantly lower than those of control. The duration of the pupal stage was similar between the two strains; however, durations of egg, larva, adult preoviposition period (APOP), and total APOP (TPOP) in the DsFoxO-KO strain were all significantly longer compared to those of the control strain. The fecundity of the DsFoxO-KO strain was 20.31 eggs/female, which was significantly lower than that of the control strain (430.47 eggs/female). With the transcriptome analysis, 612 differentially expressed genes (DEGs) were identified. Following COG and GO analyses, we found that most of the DEGs were associated with the metabolic process. According to the KEGG database, the mTOR signaling, MAPK signaling, Wnt signaling, and Toll and Imd signaling pathways; insect hormone biosynthesis; autophagy; and apoptosis were altered in the DsFoxO-KO strain. These results demonstrated that knockout of DsFoxO in D. suzukii significantly influenced its development and fecundity, while transcriptome analysis provided insights to explore the corresponding molecular mechanism. These findings highlighted the critical role of FoxO in D. suzukii and might contribute to the development of novel management strategies for these flies in the future.

Controlled SPION-Exosomes Loaded with Quercetin Preserves Pancreatic Beta Cell Survival and Function in Type 2 Diabetes Mellitus.

Introduction: Quercetin has an ideal therapeutic effect on islet function improvement in type 2 diabetes mellitus (T2DM). However, the therapeutic benefit of quercetin is hindered by its poor bioavailability and limited concentration in pancreatic islets. In this study, superparamagnetic iron oxide nanoparticle (SPION)-modified exosomes were prepared to load quercetin, hoping to endow quercetin with enhanced water solubility and active targeting capacity with the help of magnetic force (MF). Methods: Transferrin-modified SPIONs (Tf-SPIONs) were synthesized by exploiting N-hydroxysuccinimidyl (NHS) conjugation chemistry, and quercetin-loaded exosomes (Qu-exosomes) were acquired by electroporation. Tf-SPION-modified quercetin-loaded exosomes (Qu-exosome-SPIONs) were generated by the self-assembly of transferrin (Tf) and the transferrin receptor (TfR). The solubility of quercetin was determined by high-performance liquid chromatography (HPLC) analysis. The pancreatic islet targeting capacity and insulin secretagogue and antiapoptotic activities of Qu-exosome-SPIONs/MF were evaluated both in vitro and in vivo. Results: The Qu-exosome-SPIONs were well constructed and harvested by magnetic separation with a uniform size and shape in a diameter of approximately 86.2 nm. The water solubility of quercetin increased 1.97-fold when loaded into the SPION-modified exosomes. The application of SPIONs/MF endowed the Qu-exosomes with favorable targeting capacity. In vitro studies showed that Qu-exosome-SPIONs/MF more effectively inhibited or attenuated ß cell apoptosis and promoted insulin secretion in response to elevated glucose (GLC) compared with quercetin or Qu-exosome-SPIONs. In vivo studies demonstrated that Qu-exosome-SPIONs/MF displayed an ideal pancreatic islet targeting capacity, thereby leading to the restoration of islet function. Conclusion: The Qu-exosome-SPIONs/MF nano-delivery system significantly enhanced the quercetin concentration in pancreatic islets and thereby improved pancreatic islet protection.

Gut bacterium promotes host fitness in special ecological niche by affecting sugar metabolism in Drosophila suzukii.

As an important fruit pest of global significance, Drosophila suzukii occupies a special ecological niche, with the characteristics of high sugar and low protein contents. This niche differs from those occupied by other fruit-damaging Drosophila species. Gut bacteria substantially impact the physiology and ecology of insects. However, the contribution of gut microbes to the fitness of D. suzukii in their special ecological niche remains unclear. In this study, the effect of Klebsiella oxytoca on the development of D. suzukii was examined at physiological and molecular levels. The results showed that, after the removal of gut microbiota, the survival rate and longevity of axenic D. suzukii decreased significantly. Reintroduction of K. oxytoca to the midgut of D. suzukii advanced the development level of D. suzukii. The differentially expressed genes and metabolites between axenic and K. oxytoca-reintroduced D. suzukii were enriched in the pathways of carbohydrate metabolism. This advancement was achieved through an increased glycolysis rate and the regulation of the transcript level of key genes in the glycolysis/gluconeogenesis pathway. Klebsiella oxytoca is likely to play an important role in increasing host fitness in their high-sugar ecological niche by stimulating the glycolysis/gluconeogenesis pathway. As a protein source, bacteria can also provide direct nutrition for D. suzukii, which depends on the quantity or biomass of K. oxytoca. This result may provide a new target for controlling D. suzukii by inhibiting sugar metabolism through eliminating the effect of K. oxytoca and thus disrupting the balance of gut microbial communities.

Neuroprotective Effects of Deuterium-Depleted Water (DDW) Against H2O2-Induced Oxidative Stress in Differentiated PC12 Cells Through the PI3K/Akt Signaling Pathway.

Oxidative stress plays an important role in the pathogenesis of neurodegenerative diseases, including Alzheimer's disease and Parkinson's disease. Induction of endogenous antioxidants to act against oxidative stress-mediated neuronal damage seems to be a reasonable strategy for delaying the progression of such diseases. In this study, we investigated the neuroprotective effect of deuterium-depleted water (DDW) against H2O2-induced oxidative stress in differentiated PC12 cells and the possible signaling pathways involved. The differentiated PC12 cell line was pretreated with DDW containing different concentrations (50-100 ppm) of deuterium and then treated with H2O2 to induce oxidative stress and neurotoxicity. We assessed cell survival, reactive oxygen species (ROS) generation, TUNEL assay, catalase (CAT), copper and zinc-containing superoxide dismutase (CuZn-SOD) and superoxide dismutase (SOD) activity and performed Western blot analysis to investigate the neuroprotective effect of DDW. The results indicated that DDW could attenuate H2O2-induced apoptosis, reduce ROS formation, and increase CAT, CuZn-SOD and SOD activity in H2O2-treated PC12 cells. Western blot analysis revealed that DDW treatment significantly increased the expression of p-Akt, Bcl-2 and GSK-3ß. However, the protective effect of DDW on cell survival and the DDW-mediated increases in p-Akt, Bcl-2 and GSK-3ß were abolished by pretreatment with the phosphatidylinositol-3-kinase (PI3K) inhibitor LY294002. In summary, DDW may protect differentiated PC12 cells against H2O2-induced oxidative stress through the PI3K/Akt signaling pathway.

Oviposition Suitability of Drosophila Suzukii (Diptera: Drosophilidae) for Nectarine Varieties and Its Correlation with the Physiological Indexes.

The nectarine is an important fruit, which is attacked by Drosophila suzukii in Europe and the United States but there are no reports of it attacking nectarines in China. Here, we determined the oviposition preference of D. suzukii six on intact and sliced nectarine varieties in China and how physical and physiological indexes of the fruit correlate with these preferences. D. suzukii were allowed to oviposit on two early-, two middle- and two late-maturing varieties of nectarine-Shuguang and Chunguang, Fengguang and Zhong you 4, Zhong you 7 and Zhong you 8, respectively and the number of larvae also followed the order. The firmness, soluble solids content and the nutritional components of the amino acid, protein, soluble sugar and pectin contents of each variety were measured. D. suzukii preferred the early Shuguang variety, followed by the early Chunguang variety and then the middle Zhong you 4 and Fengguang varieties. Taken together, results show that D. suzukii shows preferences for earlier rather than later varieties of nectarines in China and that these preferences are related to the fruit's physical and physiological traits. Results suggest that mixed cultivation of early-, middle- and late-maturing nectarine varieties should be avoided in order to prevent fly dispersal and infestation by D. suzukii.

Evidence for the Participation of Acid-Sensing Ion Channels (ASICs) in the Antinociceptive Effect of Curcumin in a Formalin-Induced Orofacial Inflammatory Model.

Curcumin, a major bioactive component of turmeric, has diverse therapeutic effects such as anti-inflammatory, antioxidant, anticancer, and antinociceptive activities. The acid-sensing ion channels (ASICs), which can be activated by acute drops in the extracellular pH, play an important role in nociception. However, very little is known about the interaction between ASICs and curcumin in nociception of inflammation. In our study, we investigated whether the antinociceptive effects of curcumin are mediated via ASICs using an orofacial nociceptive model and in vitro western blotting, immunofluorescence, whole-cell patch-clamp recordings in the trigeminal system. Intraperitoneally administered curcumin at a dose of 50 mg/kg can reduce hyperalgesia in both the phases of a formalin-induced orofacial nociceptive model. Curcumin reduced the amplitude of ASICs currents in a dose-dependent manner in trigeminal ganglion (TG) neurons, and curcumin also reduced the protein quantity but did not change the distribution of ASICs in TG. Thus, our results indicate that curcumin can reduce formalin-induced ASICs activation and thus inhibit ASICs-mediated inflammatory pain hypersensitivity.

Lung morphometry changes in prevention of airway remodeling by protocatechuic aldehyde in asthmatic mice.

Airway remodeling can lead to irreversible airflow obstruction and persistent airway hyper-responsiveness, which is the pathological basis of refractory asthma. To investigate the preventive effect of protocatechuic aldehyde on airway remodeling in asthmatic mice by lung morphometry methods. BALB/c mice were used to establish model of airway remodeling by ovalbumin (OVA) inhalation. Bronchoalveolar lavage fluid (BALF) were collected for eosinophils (EOS) count and detection of interleukin 4 (IL-4), interleukin-13 (IL-13) and interferon (IFN-γ) content. The left lung pathological sections were performed HE, AB-PAS and Masson staining. The epithelial lamina thickness of the left main bronchus (Re), the smooth muscle layer thickness (Rm), the number of goblet cells and goblet cell area percentage (%Ac) and gas side of the road and vascular collagen deposition (%Aco, %Avc) situation were measured. Protocatechuic aldehyde gavage made the reduction of BALF EOS count. IL-4 and IL-13 levels also decreased, while the IFN-γ level increased. The left main bronchus Re, Rm, goblet cell count, Ac% and Aco% and Avc% reduced. Protocatechuic aldehyde can significantly control airway inflammation and prevent airway remodeling.

Lack of genetic associations between PPAR-γ gene rs1801282 polymorphism and Alzheimer's disease in general population: a meta-analysis.

Published studies have evaluated the association between PPAR-γ rs1801282 polymorphism and Alzheimer's disease (AD) susceptibility. However, a definitive conclusion remains elusive. The aim of this study was to derive a more precise estimation of this association. We searched PubMed, Embase, Alzgene database, Chinese National Knowledge Infrastructure (CNKI), China Biological Medicine Database and Wanfang Databases for related studies. Twelve case-control studies with a total of 4874 cases and 5439 controls were finally identified to be eligible studies in this meta-analysis. The association was assessed by summarizing the odds ratios (ORs) with the corresponding 95% confidence intervals (CIs). Overall, there was no significant association between PPAR-γ rs1801282 polymorphism and Alzheimer's disease risk in all genetic models (the allele model G vs. C: OR=1.09, 95% CI 0.99-1.19, p=0.07; the homozygous model GG vs. CC: OR=1.04, 95% CI 0.75-1.44, p=0.80; heterozygote model GC vs. CC: OR=1.11, 95% CI 1,00-1.22, p=0.05; the dominant model GG+GC vs. CC: OR=1.10, 95% CI 1.00-1.22, p=0.05; the recessive model GG vs. GC+CC: OR=1.02, 95% CI 0.74-1.41, p=0.90). In subgroup analysis by ethnicity, no significant difference was found in both Asians and Caucasians. In summary, the present meta-analysis suggests that the PPAR-γ rs1801282 polymorphism may not be associated with genetic susceptibility of AD in general population.

Smooth muscle progenitor cells involved in the development of airway remodeling in a murine model of asthma.

BACKGROUND: The mechanisms regulating airway remodeling changes remain poorly understood. Recently, a smooth muscle progenitor cell was identified in the peripheral circulatory system that plays an important role in the reconstruction of injured blood vessels. However, to the best of our knowledge, there is no report in the medical literature regarding the role of smooth muscle progenitor cells (SPCs) in asthma. OBJECTIVE: The aim of this study was to investigate the relationship between SPCs and the development of airway remodelling in a murine model of asthma. METHODS: Chronic asthma with airway remodeling was generated by sensitizing and stimulating BALB/c mice with atomized ovalbumin (OVA). Bronchoalveolar lavage fluid (BALF) was collected for eosinophils (EOS) counting and histological analysis. The Ficoll method was used to isolate mononuclear cells from peripheral blood. Smooth muscle myosin heavy chain (SM-MHC) and highly glycosylated type I transmembrane protein (CD34⁺) were selected as two markers to detect the expression of SPCs by Flow Cytometry. RESULTS: Long-term inhalation of OVA produced thickening of the epithelial and smooth muscle layer, goblet cell hyperplasia, collagen deposition around smooth muscle, luminal exudates and inflammatory cell infiltration. The number of SPCs in the asthma group was significantly higher than in the control group. CONCLUSION: Long-term inhalation of OVA results in airway remodeling and the smooth progenitor muscle cell are involved in the development of airway remodeling.

Cyclin D1 (CCND1) G870A polymorphisms and cervical cancer susceptibility: a meta-analysis based on ten case-control studies.

Many studies have evaluated the association between cyclin D1 (CCND1) G870A polymorphism and cervical cancer susceptibility. However, these studies showed inconsistent results. The aim of this study was to derive a more precise estimation of this association. We searched PubMed and Embase for related studies that had been published in English, and ten case-control studies with a total of 2,864 cases and 3,898 controls were finally identified to be eligible studies in the meta-analysis. The association was assessed by summarizing the odds ratios (ORs) with the corresponding 95 % confidence intervals (CIs). Overall, there was no significant association between cyclin D1 (CCND1) G870A polymorphism and cervical cancer risk (for the allele model A vs. G: OR = 1.02, 95 % CI 0.88-1.19, p = 0.76; for the co-dominant model AA vs. GG: OR = 1.03, 95 % CI 0.75-1.41, p = 0.85; for the dominant model AA + GA vs. GG: OR = 1.00, 95 % CI 0.78-1.28, p = 0.99; for the recessive comparison AA vs. GA + GG: OR = 1.06, 95 % CI 0.85-1.32, p = 0.62). In subgroup analysis by ethnicity, no significant difference was found in both Asians and Caucasians. In summary, the present meta-analysis provides evidence that genotypes for the cyclin D1 (CCND1) G870A polymorphism may be not associated with genetic susceptibility of cervical cancer.

Deuterium-depleted water (DDW) inhibits the proliferation and migration of nasopharyngeal carcinoma cells in vitro.

Recent studies have demonstrated that natural water that has 65% of the deuterium concentration depleted, can exhibit anti-tumor properties. However, the anti-tumor effects of DDW on various nasopharyngeal carcinoma (NPC) cells have not previously been reported. In the present study, NPC cell lines and normal preosteoblast MC3T3-E1 cells were grown in RPMI1640 media containing different deuterium concentrations (50-150 ppm). The effects of DDW on the proliferation and migration of NPC and MC3T3-E1 cells were investigated using the MTT, plate colony formation, and Transwell assays, as well as Boyden chamber arrays, flow cytometry (FCM), western blot and immunofluorescence. We found that DDW was an effective inhibitor of NPC cell proliferation, plated colony formation, migration and invasion. In contrast, the growth of normal preosteoblast MC3T3-E1 cells was promoted when they were cultured in the presence of DDW. Cell cycle analysis revealed that DDW caused cell cycle arrest in the G1/S transition, reduced the number of cells in the S phase and significantly increased the population of cells in the G1 phase in NPC cells. Western blot analysis revealed that treatment with DDW significantly increased the expression of NADPH:quinone oxidoreductase-1 (NQO1), while immunofluorescence assay analysis revealed that treatment with DDW decreased the expression of PCNA and matrix metalloproteinase 9 (MMP9) in NPC cells. These results demonstrated that DDW is a novel, non-toxic adjuvant therapeutic agent that suppresses NPC cell proliferation, migration, and invasion by inducing the expression of NQO1 and causing cell cycle arrest, as well as decreasing PCNA and MMP9 expression.

[Effects of polysaccharide of radix ranunculi ternati on immunomodulation and anti-oxidation].

OBJECTIVE: To study effects of polysaccharide of Radix Ranunculi Ternati (PRT) on immunological function and anti-oxidation activity of mouse. METHOD: Cell proliferations of splenocyte, thymocyte and peritoneal macrophage were measured by MTT colorimetry. The phagocytic function of peritoneal macrophage was measured by neutral red colorimetric method. The disoxidation power of PRT was measured by Prussian blue method. The clearing effect of PRT on hydroxyl radical was measured by salicylic acid capture method. The clearing effect of PRT on superoxide anion free radical was measured by pyrogallol auto oxidation method. RESULT: PRT among 25-400 mg x L(-1) could enhance thymocytes and spleen lymphocyte proliferation and macrophage phagocytosis. PRT(200 mg x L(-1)) has the strongest macrophage proliferation. PRT in different concentration has shown some disoxidation effects. PRT in 8 g x L(-1) has nearly the same ability of clearing x OH by Vit C with the same concentration. The clearance rate of PRT on O2*- is 95.39%. CONCLUSION: PRT can enhance the cell proliferation capability of thymocytes, spleen lymphocytes and peritoneal macrophages. PRT can enhance macrophage phagocytosis in a dose-response relationship. PRT has saome disoxidation power and strong ability of clearing x OH and O2*-.

[Clinical observation of teacher tablets in treatment of pharyngitis].

OBJECTIVE: To evaluate the efficacy of teacher tablets in the treatment of pharyngitis. METHOD: One hundred and thirty six patients with acute pharyngitis or chronic pharyngitis in attack were randamly divided into two groups: treated group (n=68), the patients were given teacher tablets for 7 days, control group (n=68), the patients were given Qinlian capsule for 7 days. Before and after the experimental medicine-taking test, general condition, clinical symptoms and features of examinations on laryngo-pharynx, throat swab bacterial culture were measured. RESULT: After 7 day medicine-taking experiment, teacher tablets can improve clinical symptoms (at an efficacy rate of more than 60%) and features (at an efficacy rate of more than 80%) of laryngopharynx, in treated group, the inhibition ratios of alpha streptococcus, neisseria and staphylococcus aureus are more than 50%. There are no significant difference between treated group and control groups in those detected index. CONCLUSION: Teacher tablets is effective for pharyagitis.

In vitro arsenic trioxide induces apoptosis in T cells of asthmatic patients by a Bcl-2 related mechanism.

This study examined the effects of arsenic trioxide on apoptosis and interleukin-4 release in T cells of asthmatic patients in vitro and investigated the role of Bcl-2 in the active mechanism. T cells were isolated from asthmatic patients (n = 21) and healthy controls (n = 20), and then treated with arsenic trioxide and dexamethasone. Cell apoptosis was measured using fluorescence microscopy, flow cytometry and a cytochrome c ELISA kit. Interleukin-4 levels in the serum and in supernatants from T cells were quantified by ELISA. Flow cytometric analysis and immunofluorescence studies were performed to determine Bcl-2 expression. T cells of the asthmatic patients (i. e. without treatment) exhibited decelerated spontaneous apoptosis after 24 h incubation in vitro when compared to T cells of the healthy controls. With dexamethasone treatment, an increase in apoptosis of T cells was not significantly different between both groups, irrespective of the method used. Arsenic trioxide treatment, however, significantly increased the apoptosis of T cells of the asthmatic group and showed a slight effect on the control group. In asthmatic patients, elevated levels of interleukin-4 and up-regulated Bcl-2 expression were detected. Moreover, in vitro, T cells of asthmatic patients spontaneously released more interleukin-4 and exhibited more Bcl-2 expression than T cells from the control group. Arsenic trioxide treatment significantly decreased interleukin-4 release and down-regulated Bcl-2 expression in asthmatic patients, while it only slightly affected healthy controls. Dexamethasone treatment decreased interleukin-4 release in both groups examined. It did not significantly influence Bcl-2 expression. These results suggest that arsenic trioxide induces T cell apoptosis and decreases interleukin-4 release in T cells of asthmatic patients in vitro and that down-regulation of Bcl-2 expression may be an important mechanism.

[Effects of arsenic trioxide on apoptosis of peripheral T-lymphocytes from asthmatic patients and normal subjects in vitro].

OBJECTIVE: To study the effects of arsenic trioxide on apoptosis of peripheral T-lymphocytes from asthmatic patients and normal subjects in vitro. METHODS: The T-lymphocytes were isolated from the blood of 21 asthmatic patients and 20 healthy controls and treated with arsenic trioxide and dexamethasone. Cell apoptosis was observed by fluorescence microscope and measured with flow cytometry and Cytochrome C ELISA kit. RESULTS: The T-lymphocytes from the asthmatic patients, when compared to those from of the healthy control, exhibited decelerated spontaneous apoptosis after a 24-hour incubation in vitro. Dexamethasone treatment significantly increased the percentage of apoptotic T-lymphocytes from both asthmatic patients and normal subjects in comparable magnitude. Arsenic trioxide treatment, in contrast, significantly increased the percentage of apoptotic T-lymphocytes from asthmatic patients, but slightly affected the cells from the control group. CONCLUSIONS: Spontaneous apoptosis of T-lymphocytes can be decelerated in asthmatic patients, whose T-lymphocytes are more sensitive to arsenic trioxide-induced apoptosis than those of normal subjects, but the T-lymphocytes from normal subjects and asthmatic patients are equally sensitive to dexamethasone.