Chemical components in cultivated Cordyceps sinensis and their effects on fibrosis / 中草药·英文版

OBJECTIVE@#Cultivated Cordyceps sinensis powder has been used as clinical drug and healthy food to nourish the lung and kidney, which solves the problem of serious shortage of wild C. sinensis. This study aims to explore the chemical components and compared their anti-fibrotic effects in cultivated C. sinensis.@*METHODS@#Nucleosides, sterols and polysaccharides were separated and purified from cultivated C. sinensis, and analyzed by high performance liquid chromatography, gas chromatography-mass spectrometry and chemical chromogenic methods, respectively. In high glucose-induced rat mesangial cell models, fibronectin and type 1 collagen were used as evaluation indicators.@*RESULTS@#There were 10 kinds of nucleosides and one sterol in cultivated C. sinensis. The contents of nucleosides, sterols and polysaccharides in the cultivated C. sinensis were close to 2%, 0.55% and 4.4%, respectively. Furthermore, nucleoside, sterol and polysaccharide components exhibited varying degrees of anti-fibrotic activity. The nucleoside components and sterol components inhibited the expression of extracellular matrix more effectively in the three main components.@*CONCLUSION@#Cultivated C. sinensis remains the similar compounds with the wild C. sinensis, and nucleosides and sterols may be the main active substances that contribute to its anti-fibrotic effects. The project of this study may provide valuable information on further optimization of more effective remedies with few side effects based on cultivated C. sinensis.

Differential transcription of mating-type genes during sexual reproduction of natural Cordyceps sinensis / 中国中药杂志

Natural Cordyceps sinensis as an insect-fungal complex, which is developed after Ophiocordyceps sinensis infects a larva of Hepialidae family. Seventeen genotypes of O. sinensis have been identified in natural C. sinensis. This paper summarized the literature reports and GenBank database regarding occurrence and transcription of the mating-type genes of MAT1-1 and MAT1-2 idiomorphs in natural C. sinensis, in Hirsutella sinensis(GC-biased Genotype #1 of O. sinensis), to infer the mating pattern of O. sinensis in the lifecycle of natural C. sinensis. The mating-type genes and transcripts of MAT1-1 and MAT1-2 idiomorphs were identified in the metagenomes and metatranscriptomes of natural C. sinensis. However, their fungal sources are unclear because of co-colonization of several genotypes of O. sinensis and multiple fungal species in natural C. sinensis. The mating-type genes of MAT1-1 and MAT1-2 idiomorphs were differentially present in 237 H. sinensis strains, constituting the genetic control of the O. sinensis reproduction. Transcriptional control of the O. sinensis reproduction includes: differential transcription or silencing of the mating-type genes of MAT1-1 and MAT1-2 idiomorphs, and the MAT1-2-1 transcript with unspliced intron I that contains 3 stop codons. Research on the H. sinensis transcriptome demonstrated differential and complementary transcriptions of the mating-type genes of MAT1-1 and MAT1-2 idiomorphs in Strains L0106 and 1229, which may become mating partners to accomplish physiological heterothallism. The differential occurrence and transcription of the mating-type genes in H. sinensis are inconsistent with the self-fertilization hypothesis under homothallism or pseudohomothallism, but instead indicate the need of mating partners of the same H. sinensis species, either monoecious or dioecious, for physiological heterothallism, or heterospecific species for hybridization. Multiple GC-and AT-biased genotypes of O. sinensis were identified in the stroma, stromal fertile portion(densely covered with numerous ascocarps) and ascospores of natural C. sinensis. It needs to be further explored if the genome-independent O. sinensis genotypes could become mating partners to accomplish sexual reproduction. S. hepiali Strain FENG experienced differential transcription of the mating-type genes with a pattern complementary to that of H. sinensis Strain L0106. Additional evidence is needed to explore a hybridization possibility between S. hepiali and H. sinensis, whether they are able to break the interspecific reproductive isolation. Genotypes #13～14 of O. sinensis feature large DNA segment reciprocal substitutions and genetic material recombination between 2 heterospecific parental fungi, H. sinensis and an AB067719-type fungus, indicating a possibility of hybridization or parasexuality. Our analysis provides important information at the genetic and transcriptional levels regarding the mating-type gene expression and reproduction physiology of O. sinensis in the sexual life of natural C. sinensis and offers crucial reproductive physiology evidence, to assist in the design of the artificial cultivation of C. sinensis to supplement the increasing scarcity of natural resource.

Cordycepin, a metabolite of Cordyceps militaris, inhibits xenograft tumor growth of tongue squamous cell carcinoma in nude mice / 南方医科大学学报

OBJECTIVE@#To evaluate the inhibitory effect of cordycepin on oral cancer xenograft in nude mice and explore the underlying mechanisms.@*METHODS@#Sixteen BALB/c mice bearing subcutaneous human tongue squamous cell carcinoma (TSCC) TCA-8113 cell xenografts were randomized into model group and cordycepin treatment group for daily treatment with saline and cordycepin for 4 weeks. After the treatment, the tumor xenografts were dissected and weighed to assess the tumor inhibition rate. Histological changes in the heart, spleen, liver, kidney, and lung of the mice were evaluated with HE staining, and tumor cell apoptosis was examined using TUNEL staining; The expressions of Bax, Bcl-2, GRP78, CHOP, and caspase-12 in the xenografts were detected using RT-qPCR and Western blotting.@*RESULTS@#Cordycepin treatment resulted in a tumor inhibition rate of 56.09% in the nude mouse models, induced obvious changes in tumor cell morphology and significantly enhanced apoptotic death of the tumor cells without causing pathological changes in the vital organs. Cordycepin treatment also significantly reduced Bcl-2 expression (P < 0.05) and increased Bax, GRP78, CHOP, and caspase-12 expressions at both the RNA and protein levels in the tumor tissues.@*CONCLUSION@#Cordycepin treatment can induce apoptotic death of TCA-8113 cell xenografts in nude mice via the endogenous mitochondrial pathway and endoplasmic reticulum stress pathways.

Protective Mechanism of Cordyceps sinensis Treatment on Acute Kidney Injury-Induced Acute Lung Injury through AMPK/mTOR Signaling Pathway / 中国结合医学杂志

OBJECTIVE@#To investigate protective effect of Cordyceps sinensis (CS) through autophagy-associated adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathway in acute kidney injury (AKI)-induced acute lung injury (ALI).@*METHODS@#Forty-eight male Sprague-Dawley rats were divided into 4 groups according to a random number table, including the normal saline (NS)-treated sham group (sham group), NS-treated ischemia reperfusion injury (IRI) group (IRI group), and low- (5 g/kg·d) and high-dose (10 g/kg·d) CS-treated IRI groups (CS1 and CS2 groups), 12 rats in each group. Nephrectomy of the right kidney was performed on the IRI rat model that was subjected to 60 min of left renal pedicle occlusion followed by 12, 24, 48, and 72 h of reperfusion. The wet-to-dry (W/D) ratio of lung, levels of serum creatinine (Scr), blood urea nitrogen (BUN), inflammatory cytokines such as interleukin- β and tumor necrosis factor- α, and biomarkers of oxidative stress such as superoxide dismutase, malonaldehyde (MDA) and myeloperoxidase (MPO), were assayed. Histological examinations were conducted to determine damage of tissues in the kidney and lung. The protein expressions of light chain 3 II/light chain 3 I (LC3-II/LC3-I), uncoordinated-51-like kinase 1 (ULK1), P62, AMPK and mTOR were measured by Western blot and immunohistochemistry, respectively.@*RESULTS@#The renal IRI induced pulmonary injury following AKI, resulting in significant increases in W/D ratio of lung, and the levels of Scr, BUN, inflammatory cytokines, MDA and MPO (P<0.01); all of these were reduced in the CS groups (P<0.05 or P<0.01). Compared with the IRI groups, the expression levels of P62 and mTOR were significantly lower (P<0.05 or P<0.01), while those of LC3-II/LC3-I, ULK1, and AMPK were significantly higher in the CS2 group (P<0.05 or P<0.01).@*CONCLUSION@#CS had a potential in treating lung injury following renal IRI through activation of the autophagy-related AMPK/mTOR signaling pathway in AKI-induced ALI.

Structural characteristics and antitumor activity of polysaccharides from Cordyceps species / 药学学报

Polysaccharides is one of the main bioactive components of Cordyceps species, because of the potential clinical value with stronger anti-tumor, such as anti-neuroblastoma, anti-melanoma, anti-lung cancer, anti-colon cancer and so on, its have received widespread attention in biomedical field and increasing research in last decades. According to structural elucidation, this review gives a systematic literature overview on antitumor mechanism of Cordyceps species-derived polysaccharides from three aspects, including inhibition of tumor cell growth, enhancement of immunomodulatory activity and reduction of tumor metastasis. Finally, it also puts forward some scientific problems for follow up research.

Cordyceps sinensis extract protects against the ovariectomy-induced bone loss via the action on osteoclasts / 药学实践杂志

Objective To explore the effects of Cordyceps sinensis extract (CSE) on osteoporosis and RANKL-mediated osteoclastogenesis. Methods Bone marrow-derived macrophages (BMMs) was isolated from the bone marrow of C57BL/6 mice. CSE was added in osteoclast differentiation. Osteoclasts were stained by tartrate-resistant acid phosphatase (TRAP). The nearly mature osteoclasts were planted on hydroxyapatite plates and the area of bone lacunae was observed by microscope. The F-actin belt was stained by DAPI and phylloeptide and the number of nuclei was observed by confocal microscopy. The expressions of DC-STAMP, ATP6V0D2, TRAP, CTSK, and NFATC1 were detected by q-PCR. The protein expression of the MAPK pathway was detected by Western Blot. The in vivo experiments were carried out by administering CSE to the ovariectomized mice daily through gavage. After 6 weeks of intervention, mouse femurs were taken for morphological analysis. Peripheral blood was taken for ELISA. Results CSE represses osteoclastogenesis, bone resorption, F-actin belts formation, osteoclast specific gene expressions and MAPK signaling pathways in vitro. In vivo study indicated that CSE prevents OVX-induced osteoporosis and preserves bone volume by repressing osteoclast activity and function. It also increases the serum ALP, BGP content, and reduces TRAP content. Conclusion CSE can attenuate osteoclast formation and OVX-induced osteoporosis, suggesting potential clinical therapeutic effects for osteoporosis.

Arsenic Content in Cordyceps： A Review / 中国实验方剂学杂志

Cordyceps， a unique precious Chinese herbal medicine in China， has the effects of tonifying the kidney and lungs， stopping bleeding， and resolving phlegm. It has definite clinical effects on diabetes， respiratory diseases （such as chronic obstructive pulmonary disease）， chronic kidney diseases， slow arrhythmia， and hyperlipidemia. We searched China National Knowledge Infrastructure （CNKI）， Wanfang Data， VIP， SpringerLink （Biomedical Sciences）， Embase， Proquest （Life Sciences）， PubMed， Cochrane Library， and SinoMed for the research on the arsenic content in Cordyceps. According to statistical analysis， arsenic in Cordyceps mainly exists in the non-toxic or low-toxic form. The arsenic-containing compounds in Cordyceps may be an arsenic-sugar compound with anticancer effect. The inorganic arsenic content in Cordyceps is lower than the arsenic standard stipulated by the State Food and Drug Administration. It remains uncertain whether different measurement methods can affect the results. The total arsenic content varies in Cordyceps from different producing areas， and the general rule of total arsenic content in Cordyceps produced in different provinces， districts， and counties cannot be obtained from the results of the existing studies. The arsenic content in Cordyceps is mainly concentrated in the insect part， which is significantly different from that in stroma， and no arsenic is detected in the mycelia. The soil arsenic content in the growing areas is generally high， which may be a main reason for the high arsenic content in Cordyceps. As a valuable medicinal material for tonifying and nourishing， Cordyceps still plays an irreplaceable role in disease prevention， health care， chronic disease management， and rehabilitation.

Systematic screening and structural characterization of dipeptides using offline 2D LC-LTQ-Orbitrap MS:A case study of Cordyceps sinensis / 药物分析学报

Cordyceps sinensis(C.sinensis)is a widely used and highly valuable traditional Chinese medicine.Several dipeptides have been detected in C.sinensis,but current scientific knowledge of its chemical makeup remains limited.In this study,an improved approach that integrates offline two-dimensional liquid chromatography(2D LC)separation,precursor ion list,library screening,and diagnostic ion filtering was established to systematically screen and characterize dipeptides in C.sinensis.Offline 2D LC integrating hydrophilic interaction LC and reverse phase separations was established to eliminate interference and identify the target dipeptides.A library containing the potential 400 dipeptides was created,and a precursor ion list with all theoretical precursor ions was adopted to trigger the MS/MS scan with high sensitivity.To identify dipeptides,the type and connection sequence of amino acids were determined according to the product ions.Ile and Leu residues were differentiated for the first time according to the characteristic ion at m/z 69.07.Ultimately,170 dipeptides were identified or tentatively characterized from C.sinensis,and most are reported for the first time in this species herein.In addition,the identified dipeptides were also applied for discrimination among the three Cordyceps species,and 11 markers were identified.The obtained results provide a deeper understanding of the chemical basis of C.sinensis.

Mechanism of Cordyceps in Treating Bronchial Asthma and Chronic Renal Failure with Concept of “Same Treatment for Different Diseases” Based on Network Pharmacology and Molecular Docking Technology / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Cordyceps in treating bronchial asthma and chronic renal failure with the concept of "same treatment for different diseases" in traditional Chinese medicine （TCM） by network pharmacology and molecular docking technology. MethodThe active components and potential targets of Cordyceps were collected from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and SwissTargetPrediction. The disease targets were obtained from Therapeutic Target Database （TTD）， DrugBank， GeneCards and other databases. The common targets were obtained from the intersection of potential targets and disease targets. The protein-protein interaction （PPI） network was constructed by STRING11.5， and the ''component-target-diseas'' network of Cordyceps was established by Cytoscape 3.9.0. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses were carried out by Metascape， and molecular docking was performed by Autodock 4.2. ResultSixty common targets of disease and drug were screened out. The core targets mainly involved protein kinase B1 （Akt1）， non-receptor tyrosine kinase， sarcoma virus protein （SRC）， TP53， matrix metalloproteinase-9 （MMP-9）， and prostaglandin endoperoxide synthase 2 （PTGS2）. The potential targets were mainly enriched in the signaling pathways of renin-angiotensin system （RAS）， RAP1， phosphoinositide 3 kinase/protein kinase B （PI3K/Akt）， mitogen-activated protein kinase （MAPK）， etc. ConclusionThe active components of Cordyceps inhibited inflammatory response and reduced fibrosis and cell apoptosis in a multi-target and multi-pathway manner. The findings of this study preliminarily revealed the potential targets and modern biological mechanism of Cordyceps in treating bronchial asthma and chronic renal failure with the concept of ''same treatment for different diseases''， and provided references for in-depth experimental verification and clinical application.

Cordyceps sinensis extracts attenuates HBx-induced mesangial cell proliferation and extracellular matrix synthesis by suppression of the PI3K/Akt pathways / 中国临床药理学与治疗学

AlM: To investigate the effect of C. sinensis extracts on HBx (Hepatitis B virus X protein) induced cell proliferation and extracellular matrix (ECM) accumulation and the underlying mechanism in cultured human mesangial cells (HMCs). METHODS: Human mesangial cells were stable transfected with pCMV-HBx to establish an HBx over-expression model, and a control group was transfected with an empty vector. Cell proliferation was determined by MTT assay and DNA synthesis assay. Western blotting was used to measure the expression of PI3K/Akt signaling pathway-related proteins and extracellular matrix. RESULTS: HBx transfection induced cell proliferation, matrix accumulation. HBx-transfected mesangial cells had increased activity of the PI3K/Akt pathways, and treatment with C. sinensis suppressed this effect. CONCLUSlON: C.sinensis attenuates the HBx-induced human mesangial cell proliferation and matrix production in human mesangial cells via inhibition of the PI3K/Akt pathways.

Identification of Cordyceps cicadae and Tolypocladium dujiaolongae based on ITS sequences and chemical pattern recognition method / 中国中药杂志

Based on ITS sequences, the molecular identification of Cordyceps cicadae and Tolypocladium dujiaolongae was carried out, and high-performance liquid chromatography(HPLC) fingerprint combined with chemical pattern recognition method was established to differentiate C. cicadae from its adulterant T. dujiaolongae. The genomic DNA from 10 batches of C. cicadae and five batches of T. dujiaolongae was extracted, and ITS sequences were amplified by PCR and sequenced. The stable differential sites of these two species were compared and the phylogenetic tree was constructed via MEGA 7.0. HPLC was used to establish the fingerprints of C. cicadae and T. dujiaolongae, and similarity evaluation, cluster analysis(CA), principal component analysis(PCA), and partial least squares discriminant analysis(PLS-DA) were applied to investigate the chemical pattern recognition. The result showed that the sources of these two species were different, and there were 115 stable differential sites in ITS sequences of C. cicadae and T. dujiao-longae. The phylogenetic tree could distinguish them effectively. HPLC fingerprints of 18 batches of C. cicadae and 5 batches of T. dujiaolongae were established. The results of CA, PCA, and PLS-DA were consistent, which could distinguish them well, indicating that there were great differences in chemical components between C. cicadae and T. dujiaolongae. The results of PLS-DA showed that six components such as uridine, guanosine, adenosine, and N~6-(2-hydroxyethyl) adenosine were the main differential markers of the two species. ITS sequences and HPLC fingerprint combined with the chemical pattern recognition method can serve as the identification and differentiation methods for C. cicadae and T. dujiaolongae.

Development of an eco-friendly and fast HPLC method for quantitative analysis of four nucleosides in Cordyceps and related products / 中国天然药物

An eco-friendly and fast HPLC method was developed for the determination of adenosine, inosine, guanosine and uridine in Cordyceps and related products (fermented mycelia of Hirsutella sinensis andPaecilomyces hepiali). The sample was ultrasonically extracted using 0.5% phosphoric acid solutions for 2.5 min. Sample separation was performed on a Poroshell SB-Aq column (50 mm × 4.6 mm, 2.7 μm) using eco-friendly mobile phase consisting of formic acid and ammonium formate aqueous solution at a flow rate of 1.0 mL·min

Establishment and Application of Identification Method for Fishy Odor of Cordyceps Based on HS-SPME/GC-QQQ-MS/MS / 中国实验方剂学杂志

Objective:To establish and apply a new practical analytical method for identifying the fishy odor of Cordyceps based on headspace-solid phase microextraction-gas chromatography-triple quadrupole mass spectrometry （HS-SPME/GC-QQQ-MS/MS） technique. Method:The InertCap Pure-WAX capillary column （0.25 mm×30 m， 0.25 μm） was used for chromatographic separation. The injection port temperature was set at 250 ℃. The injection mode was split injection with a ratio of 5∶1. High purity helium was used as the carrier gas and control mode was set to constant pressure. The column flow rate was 1.43 mL∙min^-1， the linear velocity was 43.3 cm∙s^-1， and the purge flow rate was 3.0 mL∙min^-1. The chromatographic column temperature program as follows：maintained the initial temperature at 50 ℃ for 5 min， and increased the temperature at a rate of 10 ℃∙min^-1 to 250 ℃， held for 10 min. The column equilibrium time was 2.0 min. The ion source of mass spectrographic analysis was electron ionization with ion source temperature of 200 ℃， and the monitoring mode was set to multiple reaction monitoring. Result:Seven batches of Cordyceps samples were collected， including 3 batches from Sichuan， 3 batches from Qinghai and 1 batch from Tibet. There were six batches of counterfeits， including 3 batches from Sichuan， 2 batches from Guizhou and 1 batch in Xinjiang. A total of 81 volatile compounds were screened out in Cordyceps， which could be divided into 13 types （esters， ketones， aldehydes and others） according to the compound structure， indicating that the fishy odor of Cordyceps was a complex odor. There was no significant difference in the types of volatile compounds of Cordyceps from different regions， which suggested that these volatile compounds in Cordyceps produced in Tibet （Naqu）， Qinghai （Yushu and Guoluo） and Sichuan （Litang， Rangtang and Seda） were relatively consistent. However， the contents of some volatile compounds in Cordyceps produced in different regions were quite different， and 16 volatile compounds with significant difference were screened out， including 1-methoxy-2-propyl acetate， <italic>γ</italic>-octalactone， hexyl acetate and others， those compounds maybe could been used as the quality markers for identification of regions of Cordyceps. There was a large difference in volatile compounds between Cordyceps and its counterfeits， and 34 volatile compounds were screened out， including ethyl acetate， acetophenone， 2-ethyl-1-hexanol and others， those compounds maybe could been used as the quality markers for authenticity identification of Cordyceps. Conclusion:In summary， the established method for identifying the fishy odor of Cordyceps in this paper has the characteristics of high sensitivity， accuracy and simplicity， which can provide reference for the analysis of volatile compounds in other Chinese herbal medicines.

Fermented Cordyceps Powder Combined with ACEI/ARB in Treatment of Diabetic Kidney Disease： A Systematic Review / 中国实验方剂学杂志

［Abstract］Objective:To evaluate the efficacy and safety of fermented cordyceps powder combined with angiotensin-converting enzyme inhibitors （ACEI）/angiotensin Ⅱ receptor blocker （ARB） in the treatment of diabetic kidney disease （DKD）. Method:The randomized controlled trials （RCTs） concerning the treatment of DKD with fermented cordyceps powder plus ACEI/ARB were retrieved from Pubmed， Embase， Cochrane library， China National Knowledge Infrastructure （CNKI）， Chinese BioMedical Literature Database on disc （CBMdisc）， Wanfang Data Knowledge Service Platform， and Chongqing Weipu Database for Chinese Technical Periodicals （VIP）. The quality of the included articles was evaluated by the Cochrane Collaboration's tool， followed by data analysis using RevMan 5.3. Result:A total of 48 RCTs were included， involving 4 562 cases. As revealed by Meta-analysis， the effective rate of fermented cordyceps powder combined with ACEI/ARB was higher than that of ACEI/ARB ［risk ratio （RR）=1.20， 95% confidence interval （CI） （1.15，1.24）， <italic>P</italic><0.000 01］. Moreover， such combination effectively reduced urinary albumin excretion rate ［standardized mean difference （SMD）=-2.61，95%CI （-3.17，-2.05），<italic>P</italic><0.000 01］，24-h proteinuria［SMD=-1.75，95%CI （-2.15，-1.35），<italic>P</italic><0.000 01］， serum creatinine（Scr）［mean difference （MD）=-14.57，95%CI （-17.94，-11.21），<italic>P</italic><0.000 01］， blood urea nitrogen（BUN）［MD=-1.05，95%CI （-1.29，-0.81），<italic>P</italic><0.000 01］， cystatin C （Cys-C） ［MD=-0.52，95%CI （-0.68，-0.36），<italic>P</italic><0.000 01］， fasting blood glucose（FBG）［MD=-0.59，95%CI （-0.93，-0.25），<italic>P</italic>=0.000 6］， hemoglobin A1c（HbA1c）［MD=-0.50，95%CI（-0.75，-0.24），<italic>P</italic>=0.000 1］， tumor necrosis factor-<italic>α</italic>（TNF）-<italic>α </italic>［SMD=-1.68，95%CI （-2.21，-1.15），<italic>P</italic><0.000 01］， C-reactive protein（CRP） ［SMD=-1.35，95%CI （-1.77，-0.93），<italic>P</italic><0.000 01］， and interleukin-6 （IL-6） ［SMD=-1.52，95%CI （-1.98，-1.07），<italic>P</italic><0.000 01］. There was no significant difference in the incidence of adverse events between the two groups ［RR=0.77，95%CI （0.49，1.21），<italic>P</italic>=0.25］. Conclusion:Fermented cordyceps powder combined with ACEI/ARB is more effective than ACEI/ARB in the treatment of DKD， which is worthy of clinical promotion and use. More multi-center RCTs with a large sample size are needed for verification.

Identification of geographical origins of Cordyceps based on data of amino acids with self-organizing map neural network / 中国中药杂志

In this study, data of amino acids of Cordyceps samples from Qinghai and Tibet was analyzed with self-organizing map neural network. A model of XY-Fused network was established with the content of 8 major amino acids and total amino acids for the identification of geographical origins of Cordyceps from Qinghai and Tibet. It had the prediction accuracy of 83.3% for the test set. In addition, data mining indicated that methionine was a special kind of amino acid in Cordyceps which could serve as a marker to identify its geographical origins. On this basis, the content ratio of methionine to total amino acids was proposed to be a quantifiable indicator to distinguish Cordyceps from Qinghai and Tibet.

Comparison of endogenetic microbial community diversity between wild Cordyceps sinensis,artificial C. sinensis and habitat soil / 中国中药杂志

To obtain the difference of the fungal and bacterial community diversity between wild Cordyceps sinensis, artificial C. sinensis and their habitat soil, Illmina Hiseq high-throughput sequencing technology was applied. The results show that Proteobacteria was the dominant bacterial phylum in C. sinensis, Actinobacteria was the dominant bacterial phylum in soil microhabitat, Ophiocordyceps sinensis was the predominant dominant fungus of C. sinensis. The α diversity analysis showed that the fungal diversity of stroma was lower than other parts, and the fungal diversity of wild C. sinensis was lower than that of artificial C. sinensis. The β diversity analysis showed that the fungal and bacterial community diversity of soil microhabitat samples was significantly different from that of C. sinensis. The fungal community diversity was less different between wild and artificial C. sinensis, especially in sclerotia. LEfSe analysis showed a lot of species diversity between wild and artificial C. sinensis. Those different species between wild C. sinensis, artificial C. sinensis and their habitat soil provide ideas for further research on breed and components of C. sinensis.

Construction and optimization of cordycepin-producing Saccharomyces cerevisiae / 生物工程学报

Cordycepin is the key active component of medicinal fungus Cordyceps militaris, and it shows multiple functional activities such as anti-tumor and anti-virus. Cordycepin was conventionally produced by liquid fermentation of C. militaris, but the long production cycle and the low productivity constrained its development and application. In this study, two key genes for cordycepin biosynthesis (ScCNS1 and ScCNS2) were introduced into Saccharomyces cerevisiae S288C, producing 67.32 mg/L cordycepin at 240 h. Analysis of gene expression profiles indicated that ZWF1, PRS4, ADE4, ScCNS1 and ScCNS2 which encode enzymes involved in pentose phosphate pathway, purine metabolism and cordycepin biosynthesis pathway, were significantly up-regulated in the late phage of fermentation. Optimization of fermentation medium determined that 50 g/L initial glucose followed by feeding, supplemented with 5 mmol/L Cu²⁺ and 1.0 g/L adenine were the best condition. Fed-batch fermentation using the engineered yeast in a 5 L stirred fermenter produced 137.27 mg/L cordycepin at 144 h, with a productivity up to 0.95 mg/(L·h) reached, which was 240% higher than that of the control.

Synergistic Effect of Acoustic and Vacuum Drying to Antioxidant Attributes of Cordyceps militaris

Acoustic drying allows the utilization of lower temperatures than conventional methodology in the drying process.Vacuum drying is one of the most energy demanding processes. Water evaporation also takes place at lower temperatures under vacuumand hence the product processing temperature can be significantly lower, offering higher product quality. Cordyceps militarisis a well-known entamophagus fungus with wonderful health benefits such as adaptogenic, aphrodisiac, anti-oxidant, anti-aging, neuroprotective, nootropic, immunomodulatory, anti-cancer and hepatoprotective role by its phytochemical constituents. This study focused on the synergistic effects of acoustic and vacuum drying on antioxidant properties of Cordyceps militaris. We noticed that acoustic drying at power 800 W in frequency 40kHz combined with vacuum drying at pressure -0.8 bar were suitable for dehydration of this valuable material. From this approach, a combination of acoustic and vacuum drying created a synergistic effect consuming less energy than single drying method because it can be performed at low temperature while maintaining the product quality and wholesomeness. Moisture content is partly removed by acoustic drying and further dehydration in a vacuum dryer to reduce moisture to a stable level

Development, in vitro biocompatibility, and antitumor efficacy of acetic acid-modified Cordyceps sinensis polysaccharide nanoparticle drug delivery system

Docetaxel-loaded acetic acid conjugated Cordyceps sinensis polysaccharide (DTX-AA-CSP) nanoparticles were prepared through dialysis and their release rates in vitro, particle sizes, zeta potentials, drug loading capacities, and encapsulation efficiencies were characterized for the synthesis of AA-modified CSPs from traditional Chinese medicine Cordyceps sinensis (Berk.) Sacc. Then, the AA-modified CSPs were characterized by 1H-NMR and FT-IR. Furthermore, the biocompatibility of the delivery carrier (AA-CSP nanoparticles) was assessed on human umbilical vein endothelial cells. In vitro antitumor activity studies on DTX-AA-CSP nanoparticles were conducted on the human liver (HepG2) and colon cancer cells (SW480). The DTX-AA-CSP nanoparticles were spherical and had an average size of 98.91±0.29 nm and zeta potential within the −19.75±1.13 mV. The encapsulation efficiency and loading capacity were 80.95%±0.43% and 8.09%±0.04%, respectively. In vitro, DTX from the DTX-AA-CSP nanoparticles exhibited a sustained release, and the anticancer activities of DTX-AA-CSP nanoparticles against SW480 and HepG2 were significantly higher than those of marketed docetaxel injection (Taxotere®) in nearly all the tested concentrations. The AA-CSP nanoparticles showed good biocompatibility. This study provided a promising biocompatible delivery system for carrying antitumor drugs for cancer therapy

Advances in biosynthesis of cordycepin from Cordyceps militaris / 生物工程学报

Cordycepin as the main active ingredient of Cordyceps militaris, a traditional medicinal fungus in China, has many physiological functions such as anti-cancer, anti-tumor and anti-virus activity. The most potential route for effective cordycepin production has been considered as liquid fermentation of C. militaris though with low productivity at present. Thus, it is urgent to apply both process engineering strategy and metabolic engineering strategy to enhance the productivity of cordycepin. In this review, the effects of medium components (i.e. the carbon/nitrogen source, precursor substances and metal ions) and operation factors (i.e. pH, dissolved oxygen and light) on cordycepin biosynthesis in liquid fermentation system are summarized. Besides, separation of cordycepin, the gene cluster involved and predicted biosynthesis pathways of cordycepin are also discussed, providing possible solutions of finally realizing efficient production of cordycepin.