Effects of beauvericin on the blood cells of Bombyx mori.

In this study, silkworms were treated by injection of the bioactive depsipeptide beauvericin (BEA) to explore its effect on the cellular immunity of larvae of the silkworm Bombyx mori. The results showed that: The LC50 of BEA for silkworms on the 3rd day of the 4th instar was 362.36 µM. The total count of circulating hemocytes in the silkworms decreased at 12 h after injection with 350 µM BEA, and reached the minimum value at 72 h post-treatment; at 48 h post-treatment, a large number of nodules formed by the aggregation of blood cells of the silkworms were observed under the light microscope. The survival rate of hemocytes in the larvae treated with BEA was significantly reduced in a dose-dependent manner in vivo and in vitro. The encapsulation of Q-Sepharose Fast Flow (QFF) gel particles by hemocytes in the treatment group was significantly higher than that in the control group at 1.5 h and 3 h post-treatment (P < 0.05). Moreover, the melanization ratio of QFF gel particles kept increasing with treatment time. The melanization rate at 24 h after treatment was significantly higher than that at other times (P < 0.05), reaching 55.33 %. Under the scanning electron microscope, BEA-treated larvae showed protrusions on the surface of their blood cells in vivo. Under the transmission electron microscope, it was observed that silkworm hemocytes were vacuolated. This study demonstrated that BEA had an effect on the blood cells of silkworms, and has thrown some light on the inhibitory effect and mechanism of BEA on insect cellular immunity.

Epiberberine regulates lipid synthesis through SHP (NR0B2) to improve non-alcoholic steatohepatitis.

Epiberberine (EPI), extracted from Rhizome Coptidis, has been shown to attenuate hyperlipidemia in vivo. Herein we have studied the mechanism by which EPI is active against non-alcoholic steatohepatitis (NASH) using, mice fed on a methionine- and choline-deficient (MCD) diet and HepG2 cells exposed to free fatty acids (FFA). We show that small heterodimer partner (SHP) protein is key in the regulation of lipid synthesis. In HepG2 cells and in the livers of MCD-fed mice, EPI elevated SHP levels, and this was accompanied by a reduction in sterol regulatory element-binding protein-1c (SREBP-1c) and FASN. Therefore, EPI reduced triglyceride (TG) accumulation in steatotic hepatocytes, even in HepG2 cells treated with siRNA-SHP, and also improved microbiota. Thus, EPI suppresses hepatic TG synthesis and ameliorates liver steatosis by upregulating SHP and inhibiting the SREBP1/FASN pathway, and improves gut microbiome.

Berberine inhibits gastric cancer development and progression by regulating the JAK2/STAT3 pathway and downregulating IL-6.

AIM: Gastric cancer is a prevalent malignant tumor that seriously affects human health. Berberine (BBR), an alkaloid from Chinese herbal medicines, inhibits the proliferation of various cancers. We evaluated the effects and related mechanisms of BBR on gastric cancer. MAIN METHODS: The MTT assay, flow cytometry, scratch assays, transwell experiments and xenograft nude mice models were used to investigate the antineoplastic effects of BBR. RNA-Seq, qRT-PCR, WB and ELISA were used to investigate the underlying mechanisms of BBR on gastric cancer metastasis. KEY FINDINGS: BBR treatment inhibited the proliferation of MKN-45 and HGC-27 cells, induced their apoptosis, G0/G1 cell arrest, and suppressed the migration as well as invasion of GC cells in vitro. Moreover, BBR inhibited in vivo tumor growth in MKN-45 xenograft mice. RNA-seq showed that interactions between cytokines and their receptors was one of the greatest enrichment modulated pathways and IL-6 was a key target. IL-6 knockdown significantly inhibited the activities of MKN-45 cells. Mechanistically, these findings imply that BBR inhibits GC cell proliferation by modulating the signaling pathways related to IL-6/JAK2/STAT3. SIGNIFICANCE: This study provides a theoretical basis for the use of BBR in gastric cancer prevention.

Running exercise protects spinophilin-immunoreactive puncta and neurons in the medial prefrontal cortex of APP/PS1 transgenic mice.

The medial prefrontal cortex (mPFC) is thought to be closely associated with emotional processes, decision making, and memory. Previous studies have identified the prefrontal cortex as one of the most vulnerable brain regions in Alzheimer's disease (AD). Running exercise has widely been recognized as a simple and effective method of physical activity that enhances brain function and slows the progression of AD. However, the effect of exercise on the mPFC of AD is unclear. To address these issues, we investigated the effects of 4 months of exercise on the numbers of spinophilin-immunoreactive puncta and neurons in the mPFC of 12-month-old APPswe/PSEN1dE9 (APP/PS1) transgenic AD model mice using stereological methods. The spatial learning and memory abilities of mice were tested using the Morris water maze. Four months of running exercise delayed declines in spatial learning and memory abilities. The stereological results showed significantly lower numbers of spinophilin-immunoreactive puncta and neurons in the mPFC of APP/PS1 mice than in the wild-type control group. The numbers of spinophilin-immunoreactive puncta and neurons in the mPFC of running APP/PS1 mice were significantly greater than those in the APP/PS1 control mice. In addition, running-induced improvements in spatial learning and memory were significantly associated with running-induced increases in spinophilin-immunoreactive puncta and neurons numbers in the mPFC. Running exercise could delay the loss of spinophilin-immunoreactive puncta and neurons in the mPFC of APP/PS1 mice. This finding might provide an important structural basis for exercise-induced improvements in the spatial learning and memory abilities of individuals with AD.

Integrating network pharmacology deciphers the action mechanism of Zuojin capsule in suppressing colorectal cancer.

BACKGROUND AND PURPOSE: Zuojin capsule (ZJC), a classical prescription, is outstanding in improving the conditions of patients with gastrointestinal diseases and colorectal cancer (CRC). Although ZJC has multi-ingredient and multi-target characteristics, its pharmacological effect on colorectal cancer and the underlying mechanism remain unclear. METHOD: Here, the activity of ZJC against CRC was evaluated by the experiments with CRC cells and HCT-116 xenografted mice. The key genes of CRC were obtained from the cancer genome atlas (TCGA). The genes potentially targeted by ZJC were collected from traditional Chinese medicine systems pharmacology (TCMSP) database. The underlying pathways related to selected targets were analyzed through gene ontology (GO) and pathway enrichment analyses. Western blot (WB), cellular thermal shift assay (CETSA), molecular docking and quantitative real-time PCR (QRT-PCR) were carried out to confirm the validity of the targets. RESULTS: In vitro and in vivo results indicated that ZJC may inhibit CRC cells and tumor growth. The network pharmacological analysis indicated that 22 compounds, 51 targets and 20 pathways were involved in the compound-target-pathway network. Our results confirmed that ZJC inhibited cycle progression, migration and induced apoptosis by targeting candidate genes (CDKN1A, Bcl2, E2F1, PRKCB, MYC, CDK2, and MMP9). We found that ZJC could directly change the protein level by regulating the protein stability and transcriptional activity of the target. CONCLUSIONS: In summary, combined network pharmacology and biological experiments proved that the main ingredients of ZJC such as quercetin, (R)-Canadine, palmatine, rutaecarpine, evodiamine, beta-sitosterol and berberine can target CDKN1A, Bcl2, E2F1, PRKCB, MYC, CDK2 and MMP9 to combat colorectal cancer. The results of this study provide a basic theory for the clinical trials of Zuojin Capsules against colorectal cancer.

Alterations of Gut Microbiome in Tibetan Patients With Coronary Heart Disease.

Coronary heart disease (CHD) is closely related to gut microbiota, which may be significantly affected by ethnicity and the environment. Knowledge regarding the gut microbiome of Tibetan CHD patients living in the Qinghai-Tibet Plateau is very limited. In this study, we characterized the physiological parameters and gut microbiota from 23 healthy Tibetans (HT), 18 CHD patients, and 12 patients with non-stenosis coronary heart disease (NCHD). We analyzed the alterations of the gut microbiome in CHD patients and investigated the relationship between these alterations and the pathological indicators. We found no changes in trimethylamine N-oxide, however, a significant increase in lipopolysaccharides and white blood cells, and a decrease in high-density lipoprotein were observed in the blood of CHD patients, compared to that in the HT group. The gut microbiota of the NCHD group had a significantly higher Shannon index than that of the HT group. Adonis analysis showed that both microbial compositions and functions of the three groups were significantly separated. The Dialister genus was significantly lower and Blautia, Desulfovibrio, and Succinivibrio were significantly higher in abundance in CHD patients compared with the HT group, and the changes were significantly correlated with physiological indexes, such as increased lipopolysaccharides. Moreover, enrichment of genes decreased in four pathways of amino acid metabolism, such as arginine biosynthesis and histidine metabolism, although two lipid metabolism pathways, including fatty acid degradation and arachidonic acid metabolism, increased in the CHD group. Additionally, occupation and a family history of CHD were shown to be risk factors and affected the gut microbiota in Tibetans. Our study will provide insights into the understanding of CHD, leading to better diagnosis and treatment of Tibetan patients.

Palmatine induces G2/M phase arrest and mitochondrial-associated pathway apoptosis in colon cancer cells by targeting AURKA.

Studies have shown that palmatine (PAL) has anti-cancer effects. However, the activity and potential mechanisms of PAL against colorectal cancer remain elusive. The results showed that PAL significantly inhibited the proliferation of colon cancer cells in vitro and in vivo without significant effect on non-tumorigenic colon cells. Target prediction and clinical sample database analysis suggested that PAL may contribute to colon cancer cells phase arrest and apoptosis by targeting aurora kinase A (AURKA). Inhibition and overexpression of AURKA proved that PAL induces G2/M phase arrest and apoptosis in colon cancer cells by targeting AURKA. Moreover, PAL promoted intracellular Reactive oxygen species (ROS) production and decreased mitochondrial membrane potential (ΔΨm). PAL reduced the levels of AURKA, Bcl-xl and Bcl2 proteins, and promoted the expression of pro-apoptotic proteins P53, P73, Caspase3 and Caspase9, as well as the increase of cytochrome c (cyt. c) in cell lysates in vitro and in vivo. Together, our study confirmed that PAL induced G2/M phase arrest and mitochondrial-associated pathway apoptosis in colon cancer cells by targeting AURKA. PAL may provide a novel solution for the treatment of colon cancer by serving as a new AURKA inhibitor.

Fluoxetine Promotes Hippocampal Oligodendrocyte Maturation and Delays Learning and Memory Decline in APP/PS1 Mice.

Oligodendrogenesis dysfunction impairs memory consolidation in adult mice, and an oligodendrocyte abnormality is an important change occurring in Alzheimer's disease (AD). While fluoxetine (FLX) is known to delay memory decline in AD models, its effects on hippocampal oligodendrogenesis are unclear. Here, we subjected 8-month-old male amyloid precursor protein (APP)/presenilin 1 (PS1) mice to the FLX intervention for 2 months. Their exploratory behaviors and general activities in a novel environment, spatial learning and memory and working and reference memory were assessed using the open-field test, Morris water maze, and Y maze. Furthermore, changes in hippocampal oligodendrogenesis were investigated using stereology, immunohistochemistry, immunofluorescence staining, and Western blotting techniques. FLX delayed declines in the spatial learning and memory, as well as the working and reference memory of APP/PS1 mice. In addition, APP/PS1 mice exhibited immature hippocampal oligodendrogenesis, and FLX increased the numbers of 2'3'cyclic nucleotide 3'-phosphodiesterase (CNPase)+ and newborn CNPase+ oligodendrocytes in the hippocampi of APP/PS1 mice. Moreover, FLX increased the density of SRY-related HMG-box 10 protein (SOX10)+ cells and reduced the percentage of oligodendrocyte lineage cells displaying the senescence phenotype (CDKN2A/p16INK4a) in the hippocampus of APP/PS1 mice. Moreover, FLX had no effect on the serotonin (5-HT) 1A receptor (5-HT1AR) content or number of 5-HT1AR+ oligodendrocytes, but it reduced the content and activity of glycogen synthase kinase 3ß (GSK3ß) in the hippocampus of APP/PS1 transgenic mice. Taken together, FLX delays the senescence of oligodendrocyte lineage cells and promotes oligodendrocyte maturation in the hippocampus of APP/PS1 mice. FLX may regulate GSK3ß through a mechanism other than 5-HT1AR and then inhibit the negative effect of GSK3ß on oligodendrocyte maturation in the hippocampus of an AD mouse model.

Fluoxetine delays the cognitive function decline and synaptic changes in a transgenic mouse model of early Alzheimer's disease.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder associated with cognitive decline. Previous studies have reported that the syndrome of AD begins with subtle alterations in hippocampal synapses prior to frank neuronal degeneration. It has recently been reported that fluoxetine (FLX) shows positive effects on AD patients who have depression and anxiety. However, it is unclear whether FLX can affect the pathogenesis of AD mice in the early stage of AD. To address this question, 8-month-old male APP/PS1 double-transgenic AD mice were administered a 10-week course of FLX (10 mg/kg/day) injections. Then, spatial learning and memory were evaluated using a Morris water maze test. Immunohistological staining and an unbiased stereological method were used to estimate the total number of dendritic spine synapses in the hippocampus. We found that FLX significantly shortened the mean escape latencies of the 10-month-old mice; reduced the elevated levels of soluble Aß40, Aß42, and amyloid plaques in the hippocampus; and prevented the decrease in dendritic spine synapses and in postsynaptic protein PSD-95 density in the dentate gyrus, CA1/2 and CA3 regions of the hippocampus. Our results indicate that reversing synaptic impairment might be considered a promising therapeutic approach for alleviating the cognitive deficits associated with early AD. Moreover, our results suggest that FLX may be a safe and effective drug for delaying the progress of AD, which might provide a starting point for further research into new preventative measures and treatments for AD.

Biogenic selenium and its hepatoprotective activity.

Elemental selenium nanoparticles (SeNPs) have multiple biological activities. In this study, we investigated the protective effects of biogenic SeNPs (BioSeNPs) on CCl4-induced liver damage in mice. The results showed that: (i) when compared to sodium selenite (SS), BioSeNPs has a similar tissue distribution after intragastrical administration to mice; (ii) BioSeNPs and SS showed comparable efficacy in increasing the activities of glutathione peroxidase and thioredoxin reductase in liver cell lines, mice blood and liver; (iii) pretreatment with BioSeNPs inhibiting the elevation of activities of various enzymes significantly which included aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, lactate dehydrogenase and liver lipid peroxide (p < 0.05 or p < 0.01) in CCl4-treated mice; (iv) activities of antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase) were significantly increased (p < 0.05 or p < 0.01) after a pretreatment with BioSeNPs in CCl4-treated mice; (v) histopathological damages in the liver from CCl4-treated mice were ameliorated by a pretreatment with BioSeNPs. In conclusion, these results have shown that BioSeNPs is able to protect the liver from CCl4-induced hepatic damage via increasing the antioxidant capacity and inhibiting oxidative damage. BioSeNPs may have the potential to be used as a trace element food supplement inducing antioxidant bioactivities.

Sulfasalazine inhibits inflammation and fibrogenesis in pancreas via NF-κB signaling pathway in rats with oxidative stress-induced pancreatic injury.

BACKGROUND: Pathogenesis and effective therapeutics of chronic pancreatic inflammation and fibrosis remain uncertain. PURPOSE: To investigate the effects of sulfasalazine (SF) on pancreatic inflammation and fibrogenesis. METHODS: Chronic pancreatic injury in rats was induced by diethyldithiocarbamate (DDC) and interfered by SF through intraperitoneal injection. The rats were divided into five groups: group N, normal control group, rats were treated with dilated water only; group DS1, rats received SF (10 mg/kg) 2 hours before DDC treatment; group DS2, rats were treated with DDC and then SF (100 mg/kg, twice a week); group DS3, rats were treated with DDC, then SF (100 mg/kg, thrice a week); and group DDC, rats were treated with DDC only. Pancreatic inflammation and fibrosis were determined by hematoxylin and eosin staining and Sirius red staining. The genes and proteins related to NF-κB pathway and fibrogenesis including NF-κB/p65, TNF-α, ICAM-1, α-SMA, and Con 1 were detected by immunohistochemical staining, reverse transcription polymerase chain reaction, and Western blotting. RESULTS: Rats in the DDC and DS1 groups showed the highest histological scores after DDC treatment, but the scores of DS2 and DS3 groups decreased significantly when compared with the DDC group. Sirius red staining showed collagen formation clearly in DDC and DS1 rats rather than in DS2 and DS3 rats. NF-κB/p65, ICAM-1, and α-SMA were strongly expressed in DDC and DS1 rats, while DS2 and DS3 rats showed mild to moderate expression by immunohistochemistry. Reverse transcription polymerase chain reaction showed increased levels of NF-κB/p65, ICAM-1, TNF-α, α-SMA, and Con 1 mRNA in DDC and DS1 rats in comparison to normal controls. The mRNA levels of these molecules in DS2 and DS3 rats were significantly lower than those in DS1 and DDC rats. Western blotting demonstrated that the NF-κB/p65, ICAM-1, and α-SMA expressions in pancreatic tissues of the rats of the DDC group were more clear than those of the normal control, DS2, and DS3 rats. CONCLUSION: SF inhibits pancreatic inflammation and fibrogenesis via NF-κB signaling pathway.

Identification of Candidate Genes for Seed Glucosinolate Content Using Association Mapping in Brassica napus L.

Rapeseed contains glucosinolates, a toxic group of sulfur-containing glucosides, which play critical roles in defense against herbivores and microbes. However, the presence of glucosinolates in rapeseed reduces the value of the meal as feed for livestock. We performed association mapping of seed glucosinolate (GS) content using the 60K Brassica Infinium single nucleotide polymorphism (SNP) array in 520 oilseed rape accessions. A total of 11 peak SNPs significantly associated with GS content were detected in growing seasons of 2013 and 2014 and were located on B. napus chromosomes A08, A09, C03, and C09, respectively. Two associated regions of GS content covered by these markers were further verified, and three B. napus homologous genes involved in the biosynthesis and accumulation of GS were identified. These genes were multigene family members and were distributed on different chromosomes. Moreover, two genes (BnGRT2 and BnMYB28) associated with GS content were validated by the qRT-PCR analysis of their expression profiles. The further identification and functionalization of these genes will provide useful insight into the mechanism underlying GS biosynthesis and allocation in B. napus, and the associated SNPs markers could be helpful for molecular maker-assisted breeding for low seed GS in B. napus.

Reduction of selenite to red elemental selenium by Rhodopseudomonas palustris strain N.

The trace metal selenium is in demand for health supplements to human and animal nutrition. We studied the reduction of selenite (SeO3⁻²) to red elemental selenium by Rhodopseudomonas palustris strain N. This strain was cultured in a medium containing SeO3⁻² and the particles obtained from cultures were analyzed using transmission electron microscopy (TEM), energy dispersive microanalysis (EDX) and X ray diffraction analysis (XRD). Our results showed the strain N could reduce SeO3⁻² to red elemental selenium. The diameters of particles were 80-200 nm. The bacteria exhibited significant tolerance to SeO3⁻² up to 8.0 m mol/L concentration with an EC50 value of 2.4 m mol/L. After 9 d of cultivation, the presence of SeO3²â» up to 1.0 m mol/L resulted in 99.9% reduction of selenite, whereas 82.0% (p<0.05), 31.7% (p<0.05) and 2.4% (p<0.05) reduction of SeO3⁻² was observed at 2.0, 4.0 and 8.0 m mol/L SeO3²â» concentrations, respectively. This study indicated that red elemental selenium was synthesized by green technology using Rhodopseudomonas palustris strain N. This strain also indicated a high tolerance to SeO3⁻². The finding of this work will contribute to the application of selenium to human health.

The effect of Beauveria brongniartii and its secondary metabolites on the detoxification enzymes of the pine caterpillar, Dendrolimus tabulaeformis.

The mortality of pine caterpillar, Dendrolimus tabulaeformis Tsai et Liu (Lepidoptera: Lasiocampidae), larvae treated with Beauveria brongniartii (Saccardo) Petch (Hypocreales: Clavicipitaceae) conidia and cell-free culture supernatants enriched for the secondary metabolites of the fungus was investigated. In addition, the effects of the treatments on the activities of two insect-related defense response proteins, glutathione S-transferase (GST) and esterase (EST), were measured over time. Bioassays were performed using a range of fungal spore (6 × 105 through 6 × 107 spores/mL) and supernatant extract concentrations (5.5-550 µg/mL). The results showed that the mortalities of D. tabulaeformis larvae were closely related to the concentration of the conidia and the metabolites of B. brongniartii. The differences among the treatments all reached a significant level. The activities of the two detoxifying enzymes, GST and EST, in the larvae increased simultaneously post-treatment. After infection with the conidial suspensions, the highest GST activity appeared at 3 days, and the activities of the caterpillars infected with 6 × 106 spores/mL and 6 × 107 spores/mL were significantly higher than in the control. Using α-naphthyl, the highest activity of EST also appeared at 3 days, and the differences for the three different concentrations were significant. A similar trend of change in the EST activity was observed using ß-naphthyl. After treatment with the secondary metabolite solution, the highest GST activity appeared at 6 hr, and significant differences were found both for the different durations (2, 4, 6, 12, 24, and 48 hr) and in the three concentration groups. When using α-naphthyl, the EST activity peak appeared at 24 hr, and the differences were significant among the durations of 2, 4, 6, 12, 24, and 48 hr. The effect of the concentration of the secondary metabolite solution notably induced the EST activity in the insects, and a similar result was obtained using ß-naphthyl. The data suggest that B. brongniartii produces secondary metabolites that disable the immune mechanisms of D. tabulaeformis, allowing the fungus to overcome and then kill its host. It was concluded that both the conidial suspensions and the metabolites of B. brongniartii were toxic to D. tabulaeformis larvae.

Cellular apoptosis of hemocytes from Dendrolimus tabulaeformis Tsai et Liu larvae induced with the secondary metabolites of Beauveria brongniartii (Sacc.) Petch.

To investigate the effect of the secondary metabolites of entomopathogenic fungus on the hemocyte immunity of host insect, the secondary metabolite complex (SMC) of Beauveriabrongniartii was used in three concentrations (5.5, 55, and 550 µg/mL), and the 4(th) instar larvae of the pine caterpillar Dendrolimustabulaeformis were employed as host insects. The larvae were inoculated with the SMC solutions by injection in bioassays. Apoptosis of the larval hemocytes was observed using fluorescence microscopy (FM), transmission electron microscopy (TEM), and flow cytometry (FCM). The FM results showed that in the treated groups, larval hemocytes exhibited symptoms of early apoptosis at 6 h post-treatment by radiating a non-uniform kelly fluorescence and exhibited symptoms of late apoptosis at 12 h post-treatment by radiating a non-uniform orange fluorescence. Under TEM, the following ultra-structural changes associated with apoptosis of the larval hemocytes were observed in the treated groups: the nuclei were hypertrophied, slight folds were on the nuclear envelope, the chromatin became concentrated, the mitochondrial cristae disappeared or were disorderly, most cells developed blebs, and fibrillar aggregation appeared and accumulated in the cytoplasm. Apoptosis of the larval hemocytes was detected by FCM at 6 h post-treatment; the percentage of early apoptotic cells in the SMC 5.5, 55, and 550 µg/mL treatment groups were 11.93%, 13.10%, and 18.42%, respectively. Late apoptosis first occurred at 12 h post-treatment; the highest rate of apoptosis was 36.54 ± 4.37% at 24 h post-treatment in the SMC 55 µg/mL treatment group. In general, the cellular apoptosis rate was positively correlated with the SMC concentration and the time post-treatment. These results indicate that secondary metabolites of B. brongniartii are able to attack the hemocytes of D. tabulaeformis larvae and induce cellular apoptosis, thereby providing new evidence that secondary metabolites of mycopathogens can act on host immune systems.

[Use of entomopathogenic fungi to degrade wax secreted by Ceroplastes japonicus].

OBJECTIVE: We used entomopathogenic fungi to degrade insect wax. METHODS: We used four fungal strains, Lecanicilliurn lecanii V3. 4504, V3. 4505, Beauveria bassiana FDB01, and Metarhizium anisopliae TSL06. Wax coverings of female adults of Ceroplastes japonicus Green (Insecta: Hemiptera: Coccoidea) were used as the sole carbon source in the mineral medium. RESULTS: All of the 4 strains could grow, reproduce, produce enzymes, and degrade wax. During a 7-day culture, the highest lipase activities of the 4 strains, V3. 4504, V3. 4505, FDB01, and TSL06 were 0.128 +/- 0.017, 0.056 +/- 0.002, 0.124 +/- 0.011, and 0.149 +/- 0.005 U/mL, respectively. The dehydrogenases activities of the 4 strains were 0.075 +/- 0.003, 0.074 +/- 0.003, 0.061 +/- 0.04, and 0. 066 +/- 0. 002 U/mL respectively. The degradation rates of wax by the 4 strains were 18.20 +/- 0.019, 11.00 +/- 0.011, 15.4 +/- 0.017, and 23.10 +/- 0.031%, respectively. CONCLUSION: The 4 strains could depredate wax of C. japonicus.

Observer-based reliable stabilization of uncertain linear systems subject to actuator faults, saturation, and bounded system disturbances.

A matrix inequality approach is proposed to reliably stabilize a class of uncertain linear systems subject to actuator faults, saturation, and bounded system disturbances. The system states are assumed immeasurable, and a classical observer is incorporated for observation to enable state-based feedback control. Both the stability and stabilization of the closed-loop system are discussed and the closed-loop domain of attraction is estimated by an ellipsoidal invariant set. The resultant stabilization conditions in the form of matrix inequalities enable simultaneous optimization of both the observer gain and the feedback controller gain, which is realized by converting the non-convex optimization problem to an unconstrained nonlinear programming problem. The effectiveness of proposed design techniques is demonstrated through a linearized model of F-18 HARV around an operating point.

[Effect of multi-generation culture on virulence of Lecanicilliurn lecanii in different media].

OBJECTIVE: The strain No. V3.4504 of Lecanicilliurn lecanii (Zimmermann), an entomopathogenic fungus, was studied on the effect of successive multi-generation culture in seven different media on its colony growth characteristics, extracellular enzyme activities and the virulence against scale insects. METHODS: The strain No. V3.4504 of L. lecanii was original isolated from a natural infected scale insect. The two species of scale insects used were Rhodococcus sariuoni Borchsenius and Ceroplastes japonicus Green. Seven media were used and fungus colony characteristics, growth rate and sporulation, extracellular protease and chitinase activity, and infective effect against the two species of scale insects were conducted. RESULTS: The fungus cultured on PDA medium for successive nine generations showed the most fast in colony growth, the minimal in sporulation, straight decline of extracellular protease and chitinase activity with generation increasing, and the minimal mortality of the scale insects. There was no significant effect to promote virulence of the fungus by increasing peptone into medium. On the media D, E and F, that with the body materials of the two scale insects, although the fungus appeared lower in the colony growth rate, its sporulation was higher upward 8.83 x 10(6) - 9.13 x 10(6) spores/cm2, extracellular protease and chitinase activities averagely reached 2.16 - 2.13 U/g and 1.01 - 1.03 U/g respectively, and the mortalities of the two scale insects were 55% - 58% and 39% - 42% respectively. Cultured three generations in vitro of the two scale insects, the fungus exhibited the highest activities in its protease and chitinase that were 3.08 - 2.92 U/g and 1.45 - 1.42 U/g respectively and the best infection effect against the two scale insects with mortalities of 71.30% and 58.89% respectively. A linear correlation was found between extracellular protease and chitinase activities of the fungus and the mortalities of the scale insects. CONCLUSION: Cultured on PDA medium successive multiple generations made retrogradation of the strain No. V3.4504 of of L. lecanii. It was significant effect on keeping the vigor and higher virulence of the fungus adding the body materials of the scale insects into the medium. The vitro by using live scale insects as medium materials was the best way for the rejuvenation of the entomopathogenic fungus and promoting its virulence.

Downregulation of tazarotene induced gene-2 (TIG2) in skin squamous cell carcinoma.

Tazarotene, an RARbeta/gamma-selective synthetic retinoid, was found to induce three genes, tazarotene-induced genes (TIG) 1, 2 and 3. TIG2 was abundantly expressed in non-lesional psoriatic skin, but at lower levels in psoriatic lesions. The protein precursor encoded by TIG2, called prochemerin, can be converted into bioactive chemerin. Chemerin can then bind to chemerin receptor, resulting in chemoattracting dendritic cells and macrophages, and serving as a bridge between innate and adaptive immunity. Our objective was to investigate the role of TIG2 in skin squamous cell carcinoma (SCC). The levels of TIG2-protein and transcript in normal skin tissues, uninvolved skin and SCC lesions were determined by immunohistochemistry and in situ hybridization. TIG2-protein and transcript were detected in all layers of normal epidermis and uninvolved skin adjacent to SCC lesions. In addition, TIG2 was also expressed in the corneum, granular layers and the upper and middle layers of stratum spinosum of the marginal part of SCC lesions. On the other hand, TIG2-protein and transcript were barely detectable around the keratin pearls of SCC 1-2, and were not detectable at all in SCC 3-4.

[Isolation and identification of toxins inhibiting Dentrolimus tabulaeformis from an antagonistic strain of Beauveria].

OBJECTIVE: We used submerged fermentation to cultivate a strain of the entomopathogenic fungus Beauveria tenella isolated from the infected larvae of Dentrolimus tabulaeformis in Pinus tabulaeformis forest in Chengde of Hebei Province in China. METHODS: We used ethyl acetate to extract antagonistic components from the fermentation broth and used silica gel column chromatography and GC/MS to separate and identify the components. RESULTS: Six compounds were obtained by silica gel column chromatography. The sixth compound had higher activity to kill the larvae of Dentrolimus tabulaeformis with a corrected mortality rate of 80%. Seventeen compounds were separated and identified by GC/MS in the 6th group, of which 3compounds were more than 10%, 2-Piperidinone (14.02%), 2-coumaranone (47.10%), and Pyrrolo[1,2-a]Pyrazine-1,4-dione, hexahydro (21.05%). CONCLUSION: 2-Piperidinone and 2-coumaranone had insecticidal activity (corrected mortality rate reached 83.32% and 91.61% respectively) and were the most important toxic substances to control pests.