Artificial simulated saliva,gastric and intestinal digestion and fermentation in vitro by human gut microbiota of intrapolysaccharide from Paecilomyces cicadae TJJ1213

In this study,the in vitro digestion and fermentation of two intra-polysaccharide fractions(IPS1 and IPS2)from Paecilomyces cicadae TJJ1213 were investigated.The constituent monosaccharides of IPS1 and IPS2 were not changed after simulated saliva,gastric and small intestinal digestion.However,they can be hydrolyzed and utilized by gut microbiota,and short-chain fatty acids(SCFAs)level were increased after IPS1 and IPS2 treatments.Furthermore,16 S r RNA sequencing analysis of fermentation samples were performed.Alpha-diversity,beta-diversity and taxonomic composition differences analysis revealed that IPS1 and IPS2 promoted the proliferation of beneficial bacteria and modulated the overall structure of gut microbiota.Taxonomic comparison analysis found that IPS1 increased the relative abundances of beneficial bacteria including Megamonas,Bifidobacterium and Lactobacillus,while IPS2 could increase the abundance of Bacteroides,Parabacteroides and Phascolarctobacterium.In addition,they can also decrease the levels of pathogenic bacteria containing Escherichia-Shigella,Klebsiella and Fusobacterium.These results indicated that IPS from Paecilomyces cicadae TJJ1213 could be used as potential candidates for new functional foods.

Three previously undescribed metabolites from Cordyceps cicadae JXCH-1,an entomopathogenic fungus

Three previously undescribed compounds,cordycicadione(1),cordycicadin F(2),and 7-hydroxybassiatin(3),were isolated from the cultures of Cordyceps cicadae JXCH1,an entomopathogenic fungus.Their structures and relative configurations were elucidated primarily by NMR spectroscopic analysis.The absolute configurations of 1 and 2 were determined by ECD calculations.Single-crystal X-ray diffraction method was adopted to determine the absolute configuration of 3.Compound 2 is a polycyclic polyketide with an unusual enol ether moiety and a spiro ring.The compounds obtained in this study were subjected to screening their inhibition against the proliferation of the human lung cancer cell line A549 and the production of nitric oxide in murine macrophages RAW264.7.

Separation of Insecticidal Ingredient of Paecilomyces cicadae (Miquel) Samson

In this study, the insecticidal active substance of the fruiting body of Paecilomyces cicadae （Miquel） Samson that can kill the larvae of Plutella xylostella （Lepidoptera： Plutellidae） was isolated and purified by combined separation with biology experiments for the first time. By means of bioactive guided isolation of 40% ethanol-H2O, dichloromethane, ethyl acetate, n-butyl ethanol, macroporous adsorption resin column, high performance liquid chromatography, and sephadex A-20 column, the insecticidal active compound was purified,and a white powder compound was obtained. The compound was active against the third instar larvae ofP. xylostella in 24, 48, and 72 h at the concentration of 10.0 mg mL-1 with a corrective mortality of （95.1 ± 1.8）, （97.7 ± 1.6）, and （99.1 ± 1.7）%, respectively. The compound was strongly sensitive to the larvae. It was stable at acid environment, and was not sensitive to pepsin, and other chracteristics were studied.

Protective Effect and Mechanism of Polysaccharides from Cordyceps cicadae on Acute Liver Injury Induced by D-GlaN in Mice

[Objectives]To observe the protective effect of Cordyceps cicadae polysaccharides on acute liver injury induced by D-galactosamine( D-GlaN) in mice,and to explore its mechanism. [Methods] Seventy-five male Kunming mice were randomly and evenly divided into 5 groups according to the digital table method: normal group( CK)( injected intraperitoneally with saline solution),model group( injected intraperitoneally with D-GlaN),low-dose C. cicadae polysaccharides group( administered with 0. 5 g/kg of C. cicadae polysaccharides solution by gavage),middle-dose C. cicadae polysaccharides group( administered with 1. 0 g/kg of C. cicadae polysaccharides solution by gavage) and high-dose C. cicadae polysaccharides group( administered with 2. 0 g/kg of C. cicadae polysaccharides solution by gavage). After 12 d of administration,the liver histopathological score,liver homogenate indexes( superoxide dismutase,SOD; malondialdehyde,MDA; glutathione peroxidase,GSH-Px; nitric oxide,NO) and serum markers( aspartate transaminase,AST; alanine transaminase,ALT; alkaline phosphatase,ALP; cholinesterase,CHE) of mice in each group were detected. The expression levels of nuclear factor-κB( NF-κB) and tumor necrosis factor-α( TNF-α) in liver tissues were detected by immunohistochemistry. [Results]The liver histopathological score and the MDA,NO,AST,ALT,ALP,NF-κB and TNF-α levels were significantly higher( P < 0. 05) and the SOD,GSH-Px and CHE levels were significantly lower( P <0. 05) in the model group compared with the normal group. Compared with those in the model group,the liver tissue histopathological scores in the low-,middle-and high-dose C. cicadae polysaccharides groups were all significantly reduced( P < 0. 05). With the increase of treatment dose,the liver tissue histopathological scores showed a significant decrease( P < 0. 05). Compared with the model group,the levels of MDA,NO,AST,ALT,ALP,NF-κB and TNF-α were significantly lower( P < 0. 05),and the levels of SOD,GSH-Px and CHE were significantly higher( P < 0. 05) in the low-,middle-and high-dose C. cicadae polysaccharides groups. With the increase of treatment dose,the levels of MDA,NO,AST,ALT,ALP,NF-κB and TNF-α declined significantly( P < 0. 05),while the levels of SOD,GSH-Px and CHE rose significantly( P < 0. 05). [Conclusions] C. cicadae polysaccharides have a significant protective effect on D-GlaN-induced acute liver injury in mice in a dose-dependent manner,and the mechanism may be related to the inhibition of NF-κB inflammatory signaling pathway.

Study on Optimization of Culture Conditions for Wild Isaria cicadae Miquel Liquid Strain

[Objectives] To study the effect of different culture conditions on the growth of wild I. cicadae Miq. liquid strain. [Methods]The I. cicadae Miq. strain was inoculated into the liquid culture medium with different carbon sources,nitrogen sources,micronutrients and p H,cultured in the constant temperature shaking incubator with rotating speed of 120 r/min at 19℃,and the mycelium pellet diameter,density and weight were compared between different treatments. [Results] The results showed that the optimum components of I. cicadae Miq.strain liquid included soluble starch,milk powder and vitamin B_(12),and the optimum p H was 5. 0-6. 0. [Conclusions] Soluble starch was the most suitable carbon source for the culture medium of I. cicadae Miq. liquid strain; milk powder was the most suitable nitrogen source for the culture medium of I. cicadae Miq. liquid strain; the most suitable p H was 5. 0-9. 0 for the mycelial growth of I. cicadae Miq.; the formula and mixture ratio of the optimum culture medium for the growth of liquid strain were determined.

Cordyceps cicadae extracts ameliorate renal malfunction in a remnant kidney model

Background and Objectives:Chronic kidney disease(CKD) is a growing public health problem with an urgent need for new pharmacological agents.Cordyceps cicadae is widely used in traditional Chinese medicine(TCM) and has potential renoprotective benefits.The current study aimed to determine any scientific evidence to support its clinical use.Methods:We analyzed the potential of two kinds of C.cicadae extract,total extract(TE) and acetic ether extract(AE),in treating kidney disease simulated by a subtotal nephrectomy(SNx) model.Sprague-Dawley rats were divided randomly into seven groups:sham-operated group,vehicle-treated SNx,Cozaar,2 g/(kg·d) TE SNx,1 g/(kg·d) TE SNx,92 mg/(kg·d) AE SNx,and 46 mg/(kg·d) AE SNx.Renal injury was monitored using urine and serum analyses,and hematoxylin and eosin(HE) and periodic acid-Schiff(PAS) stainings were used to analyze the level of fibrosis.The expression of type IV collagen(Col IV),fibronectin(FN),transforming growth factor-β1(TGF-β1),and connective tissue growth factor(CTGF) was detected by immunohistochemistry.Results:Renal injury,reflected in urine and serum analyses,and pathological changes induced by SNx were attenuated by TE and AE intervention.The depositions of Col IV and FN were also decreased by the treatments and were accompanied by reduced expression of TGF-β1 and CTGF.In some respects,2 g/(kg·d) of TE produced better effects than Cozaar.Conclusions:For the first time,we have shown that C.cicadae may inhibit renal fibrosis in vivo through the TGF-β1/CTGF pathway.Therefore,we conclude that the use of C.cicadae could provide a rational strategy for combating renal fibrosis.

Distinctive quality control method for solid-state fermented Isaria cicadae from strain Ic-17-7 and application in a rat model of type 2 diabetes

This work was aimed to establish a quality control method for evaluating the effects on glucose and lipids of the fruiting body of Isaria cicadae Miquel from strain Ic-17-7(Ic-17-7fb) using a rat model of type 2 diabetes(T2DM). Random amplified polymorphic DNA, sequence-characterized amplified region, and high-performance liquid chromatography(HPLC) were used for the quality control of Ic-17-7fb. The pharmacological effects on streptozocin(STZ)-induced high fat diet(HFD)-fed Albino Wistar rats were evaluated. The rats underwent the following treatments: control, metformin, Ic-17-7fb(0.166 and 0.5 g·kg;) or without treatment. The fasting blood glucose(FBG), blood glucose, total cholesterol(TC), triglyceride(TG), high-density lipoprotein(HDL-c), and low-density lipoprotein(LDL-c) were measured. Ic-17-7fb amplified a single specific band by S11-2-F3 and S11-2-R3 primers. An HPLC-based quality and quantity method was established for industrial application. The contents of adenosine and N;-(2-hydroxyethyl) adenosine(HEA) of the cultivated Ic-17-7fb were analyzed. All of the validation lots of cultured Ic-17-7fb passed the quantity control of the training set(0.90 mg·g;of adenosine and 0.89 mg·g;of HEA). After two weeks of administration, the average FBG was 4.89 ± 0.42(control), 26.10 ± 5.77(model), 23.63 ± 6.15(metformin), 17.96 ± 9.36(Ic-17-7fb for 0.166 g·kg;), and 19.69 ±8.71 mmol·L;(Ic-17-7fb for 0.5 g·kg;). The FBG of Ic-17-7fb(0.166 g·kg;) treatment significantly reduced by 31.19%, compared with the model after two weeks of administration(P < 0.01). Metformin, Ic-17-7fb(0.166 g·kg;), and Ic-17-7fb(0.5 g·kg;) reduced TC, TG, HDL-c, and LDL-c compared with the T2DM model treatment at the 6 th week of treatment(P < 0.05). This study established the first quality standard for Ic-17-7fb, which can be effectively applied in the treatment of T2DM. The reliable quality control method and pharmacological effect will broaden its application space.

Chemical Constituents from Mycelia and Spores of Fungus Cordyceps cicadae

Objective To isolate and identify the chemical constituents from mycelia and spores of the fungus Cordyceps cicadae,respectively.Methods The chemical constituents were isolated and purified by repeated silica gel,Sephadex LH-20,and reversed phase HPLC.The structures of these compounds were elucidated on the basis of extensive spectroscopic analysis,including 1 D and 2D NMR.Results Nine known sterols such as ergosterol（1）,ergosterol peroxide（2）,9,11-dehydroergosterol peroxide（3）,3β,5α,9α-trihydroxy-（22E,24R）-ergosta-7,22-dien-6-one（4）,3β,5α,9α,14α-tetrahydroxy-（22E,24R）-ergosta-7,22-dien-6-one（5）,5 α,6α-e poxy-（22 E,,24 R）-e rgosta-8（14）,22-diene-3α,7α-diol （6）, 3β,5α,6β-（22E,24R）-ergosta-7,22-dien-3,5,6-triol （7）, 3β,5α,6α-6- methoxyergosta-（22E,24R）-7,22-diene-3,5-diol （8）, 4-hydroxy-17R-methylincisterol （9）, together with a resorcinol derivative, 5-n-nonadecylresorcinol （10）, a cydo- desipeptide, beauvericin （11）, and a nucleoside, N^6-（2-hydroxyethyl）adenosine （12） were successively isolated from the cultivated C. cicadaemycelia and spores. Conclusion Compounds 3-10 are reported for the first time from the title sample, beauvericin exhibits significant cytotoxicity against human leukemia cell line and human lung cancer cell line.

Use of tree species by three species of Magicicada(Hemiptera:Cicadidae)in an Appalachian forest

Periodical cicadas(Magicicada spp.)are endemic to deciduous forests in the eastern United States.In successional forests,they must partition resources such as host trees to coexist.We measured tree size,emergence holes,oviposition scar bundles,and chorusing center abundances of Magicicada species on 12 common tree species in a deciduous forest to understand host-tree use.We predicted that the abundance of periodical cicadas and use of specific host-tree species would change depending on the Magicicada species and tree life stage.We considered the size of the tree(diameter at breast height)as a covariate to control for tree size and collected eggs for a greenhouse experiment to assess whether nymphs prefer to feed on Quercus rubra or Acer saccharum.More emergence holes were found below Quercus species than any other tree species.The abundance of periodical cicadas on host trees used for chorusing centers varied depending on the Magicicada species,but were most abundant on Quercus species.Oviposition scar bundles were also more frequent on Quercus.More nymphs were found on Quercus than Acer in the nymph preference study.Though periodical cicadas used Quercus hosts more than other tree species,their abundances on different host tree sizes and species differed significantly.Periodical cicada species may use specific host species and life stages as a way to partition resources and minimize competition among the Magicicada species during emergence years.

蝉花多糖超声波辅助提取工艺的优化

以去离子水为提取剂,采用单因素试验和正交试验考察了提取时间、提取温度和料液比对蝉花(Cordyceps cicadae)多糖提取率的影响。结果表明,蝉花多糖的最佳提取工艺条件为提取温度70℃、提取时间30 min、料液比1∶20(m/V,g∶mL),在这个条件下蝉花多糖提取率为2.37%。

GC法同时测定蝉花中三种脂肪酸含量

分析江苏茅山地区蝉花(Isaria cicadae Miquel)中脂肪酸的组成,建立了同时测定蝉花中十六酸甲酯、油酸甲酯和亚油酸甲酯3种脂肪酸含量的方法。采用气相色谱-质谱联用法分析蝉花中脂肪酸的组成,并用气相色谱法对其主要脂肪酸进行定量分析。结果表明,蝉花中主要有8种脂肪酸,十六酸甲酯、油酸甲酯、亚油酸甲酯分别在50~500μg/m L(r=0.999 9)、100~1 000μg/m L(r=0.999 5)、20~200μg/m L(r=0.999 3)线性关系良好,平均回收率分别为97.66%、98.13%、97.33%,RSD分别为1.02%、0.73%、1.03%(n=9)。

水溶性农药替代品种对茶园假眼小绿叶蝉的防治效果

使用国家茶叶产业体系新研发的水溶性农药替代品种茶蝉净,将其与凯恩、联苯菊酯、帕力特在恩施州的3个不同地区对假眼小绿叶蝉(Empoasca cicada Gothe)的防治效果进行了比较分析。结果表明,凯恩和茶蝉净具有更好的速效性,而帕力特在长效性上具有更大的优势,因此可以考虑通过联合用药来兼顾防治过程的速效和长效。针对茶园假眼小绿叶蝉的防治,以药剂的速效性、长效性及对天敌影响的大小综合评价,4种参试药剂的先后次序为帕力特、茶蝉净、凯恩、联苯菊酯。

Research and development of Cordyceps in Taiwan

Cordyceps is treasured entomopathogenic fungi that have been used as antitumor,immunomodulating,antioxidant,and pro-sexual agent.Cordyceps,also called DongChongXiaCao in Chinese,Yartsa Gunbu(Tibetan),means winter worm-summer grass.Natural Cordyceps sinensis with parasitic hosts is difficult to be collected and the recent findings on its potential pharmacological functions,resulted in skyrocketing prices.Therefore,finding a mass-production method or an alternative for C.sinensis products is a top-priority task.In this review,we describe current status of Cordyceps research and its recent developments in Taiwan.The content and pharmacological activities of four major industrial species of Cordyceps(C.sinensis,Cordyceps militaris,Cordyceps cicadae and Cordyceps sobolifera)used in Taiwan,were reviewed.Moreover,we highlighted the effect of using different methods of fermentation and production on the morphology and chemical content of Cordyceps sp.Finally,we summarized the bottle-necks and challenges facing Cordyceps research as well as we proposed future road map for Cordyceps industry in Taiwan.

HPLC法检测蝉拟青霉中N6-(2-羟乙基)-腺苷含量

为了建立蝉拟青霉(Paecilomyces cicadae)5704s中N6-(2-羟乙基)-腺苷(HEA)含量测定方法,以去离子水为溶剂,采用超声辅助水浴法提取HEA,基于HPLC法测定其含量。经优化确定色谱条件为Agilent ZORBAX Extend-C色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇∶5 mmol/L乙酸铵(含0.1%甲酸),梯度洗脱,检测波长为260 nm,流速为1.0 mL/min,柱温为25℃,进样量为10μL。结果显示,HEA浓度为0.25~100.00μg/mL时,峰面积和HEA含量之间线性关系良好,R^(2)=0.9999,仪器的检出限为0.25μg/mL,平均回收率为97.73%~108.95%,RSD为0.37%~1.29%,测得样品中HEA含量为0.78 mg/g。该方法简便、快捷、准确、稳定,适用于蝉拟青霉5704s菌丝体中HEA的测定。

A Possible Population-Driven Phase Transition in Cicada Chorus

We investigate the collective synchronization of cicada chirping. Using both experimental and phenomeno- logical numerical techniques, here we show that the onset of a periodic two-state acoustic synchronous behavior in cicada chorus depends on a critical size of population Nc=21, above which a typical chorus state appears periodically with a 30 second-silence state in between, and further clarify its possibility concerning a new class of phase transition, which is unusually driven by population. This work has relevance to acoustic synchronization and to general physics of phase transition.

Occurrence and Bioaccumulation of Micropollutants in Black Cicada

Seven organophosphoric acid triesters （OPEs） and eight polycyclic aromatic hydrocarbons （PAHs） in black cicadas were measured to determine the concentration levels and to investigate how the cicadas are affected by soil contamination. Adult cicadas, nymphal exoskeletons, and soils were sampled in Higashi Osaka, Japan. Four OPEs and six PAHs were detected in the adult cicadas. The total concentrations of OPEs and PAHs ranged from 27.2 ng/g to 824 ng/g and from 4.30 ng/g to 270 ng/g, respectively. Four OPEs and five PAHs were detected in the nymphal exoskeletons. The total concentrations of OPEs and PAHs ranged from 184 ng/g to 1830 ng/g and from 40.3 ng/g to 970 ng/g, respectively. In the soils, three OPEs and six PAHs were detected at the same concentration levels as those detected in the adult cicadas. Significant correlations were observed in the micropollutant concentrations between soils and adult cicadas, and between soils and nymphal exoskeletons. This trend indicates that black cicadas accumulate OPEs and PAHs from contaminated soils.

Cicada(Tibicen linnei) steers by force vectoring

To change flight direction, flying animals modulate aerodynamic force either relative to their bodies to generate torque about the center of mass, or relative to the flight path to produce centripetal force that curves the trajectory. In employing the latter, the direction of aerodynamic force remains fixed in the body flame and rotations of the body redirect the force. While both aforementioned techniques are essential for flight, it is critical to investigate how an animal balances the two to achieve aerial locomotion. Here, we measured wing and body kinematics of cicada （Tibicen linnei） in flee flight, including flight periods of both little and substantial body reorientations. It is found that cicadas employ a common force vectoring technique to execute all these flights. We show that the direction of the half-stroke averaged aerodynamic force relative to the body is independent of the body orientation, varying in a range of merely 20 deg. Despite directional limitation of the aerodynamic force, pitch and roll torque are generated by altering wing angle of attack and its mean position relative to the center of mass. This results in body rotations which redirect the wing force in the global flame and consequently change the flight traiectorv.

Summer in July

It is a summer afternoon.The clear blue sky is decorated[装饰]with floating clouds.Cicadas[蝉]are buzzing[嗡嗡叫]by the brook[溪流].

Gradient Wetting Transition from the Wenzel to Robust Cassie-Baxter States along Nanopillared Cicada Wing and Underlying Mechanism

Gradient wettability is important for some living organisms.Herein,the dynamic responses of water droplets impacting on the surfaces of four regions along the wing vein of cicada Cryptotympana atratafabricius are investigated.It is revealed that a gradient wetting behavior from hydrophilicity(the Wenzel state)to hydrophobicity and further to superhydrophobicity(the Cassie-Baxter state)appears from the foot to apex of the wing.Water droplets impacting on the hydrophilic region of the wing cannot rebound,whereas those impacting on the hydrophobic region can retract and completely rebound.The hydrophobic region exhibits robust water-repelling performance during the dynamic droplet impact.Moreover,a droplet sitting on the hydrophobic region can recover its spherical shape after squeezed to a water film as thin as 0.45 mm,and lossless droplet transportation can be achieved at the region.Based on the geometric parameters of the nanopillars at the hydrophilic and hydrophobic regions on the cicada wing,two wetting models are developed for elucidating the mechanism for the gradient wetting behavior.This work directs the design and fabrication of surfaces with gradient wetting behavior by mimicking the nanopillars on cicada wing surface.

Coding of Temporal Parameters of Sound Signals by Auditory Interneurons in the Cicada Cryptotympana atrata

Temporal information of the songs is generally species-specific and implies certain'meanings', which are very important in intra- and interspecific communicationsof animals. As to the mechanisms underlying song pattern recognition, it waspresumed that there were special time rate recognizers or temporally tuned neuronsin the auditory nervous system, but no evidence has been obtained so far. Someinvestigators suggested that there were 'rate band-pass' cells in the auditory neuronalcenters, which were found in the brain of frogs.