Aging related obesity and type 2 diabetes mellitus suppress neuromuscular communication and aggravate skeletal muscle dysfunction in rhesus monkeys.

Age-related functional deterioration in skeletal muscle raises the risk for falls, disability, and mortality in the elderly, particularly in obese people or those with type 2 diabetes mellitus (T2D). However, the response of the skeletal muscle to transitioning from obesity to diabetes remains poorly defined, despite that obesity is classified as a stage of pre-diabetes. We screened and selected spontaneously obese and diabetic rhesus monkeys and examined altered protein expression in skeletal muscle of healthy aging (CON), obesity aging (OB), and type 2 diabetes mellitus aging (T2D) rhesus monkeys using Tandem Mass Tags (TMT)-based quantitative proteomic analysis. In total, we identified 142 differentially expressed proteins. Muscle-nerve communication proteins were firstly suppressed at obese-stage. With the disintegration of skeletal muscle, mitochondrial complex I and other energy homeostasis relate proteins were significantly disordered at T2D stage. Indicating that aging related obesity suppressed muscle-nerve communication and contribute to T2D related functional deterioration of skeletal muscles in elderly rhesus monkeys. Some alterations of muscular functional regulator are detected in both obesity and T2D samples, suggesting some T2D related skeletal muscular hypofunctions are occurring at obesity or pre-obesity stage. Muscle-nerve communication proteins and muscular function related proteins could be potential therapy target or early diagnose marker of for skeletal muscular hypofunctions in aging obesity populations.

Two undescribed benzopyran derivatives from Ranunculus sceleratus L. with their in vitro anti-inflammatory activity in lipopolysaccharide-activated RAW 264.7 cells.

Previous studies on Ranunculus sceleratus L. have shown the existence of coumarins and their anti-inflammatory effect. Phytochemical work was conducted to investigate the bioactive compounds, leading to the isolation of two undescribed benzopyran derivatives, namely ranunsceleroside A (1) and B (3), together with two known coumarins (2, 4) from the whole plant of R. sceleratus L. All compounds were structurally identified by extensive spectroscopic analysis and then investigated for their inhibitory effect on nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and interleukin-6 (IL-6) production induced by lipopolysaccharide (LPS) in RAW 264.7 murine macrophages, repectively. As a result, compound 1-4 presented inhibitory effects on the production of NO, TNF-α, IL-1ß, and IL-6 in a concentration-dependent manner, which provides a potential chemical basis for the traditional use of R. sceleratus L. as an anti-inflammatory plant.

Antihyperuricemic and Renal Protective Effect of Cordyceps chanhua (Ascomycetes) Fruiting Bodies in Acute Hyperuricemia and Chronic Gout Rodent Models.

Cordyceps chanhua has been widely used in traditional Chinese medicine. The uric acid-lowering effect of artificially cultivated fruiting bodies of C. chanhua (FBCC) was studied using the acute hyperuricemia (AH) and chronic gout (CG) animal models. The AH mice and CG rats were randomly divided into 6 groups: the negative control group, model group, positive control group, low-dose group, medium-dose group, and high-dose group of FBCC, respectively. Serum uric acid, creatinine, urea nitrogen, and liver xanthine oxidase (XOD) activity were detected. Renal tubulointerstitial injury and urate crystals in CG rats were evaluated. The results showed that the uric acid content in AH mice with the high-dose FBCC group decreased statistically (P < 0.05). In the CG rats, the serum uric acid level in all FBCC groups and the serum creatinine value in the high-dose group exhibited a significant decrease (P < 0.05); the scores of renal tubulointerstitial damage and urate deposit were reduced in the high-dose group of FBCC. FBCC can reduce uric acid and improve renal function, demonstrating it as a beneficial supplement for uric acid-lowering and gout-relieving drugs.

In vitro and in vivo development of interspecies Asian elephant embryos reconstructed with pig enucleated oocytes.

Interspecies somatic cell nuclear transfer (iSCNT) has an immense potential to rescue endangered animals and extinct species like mammoths. In this study, we successfully established an Asian elephant's fibroblast cell lines from ear tissues, performed iSCNT with porcine oocytes and evaluated the in vitro and in vivo development of reconstructed embryos. A total of 7780 elephant-pig iSCNT embryos were successfully reconstructed and showed in vitro development with cleavage rate, 4-cell, 8-cell and blastocyst rate of 73.01, 30.48, 5.64, and 4.73%, respectively. The total number of elephant-pig blastocyte cells and diameter of hatched blastocyte was 38.67 and 252.75 µm, respectively. Next, we designed species-specific markers targeting EDNRB, AGRP and TYR genes to verify the genome of reconstructed embryos with donor nucleus/species. The results indicated that 53.2, 60.8, and 60.8% of reconstructed embryos (n = 235) contained elephant genome at 1-cell, 2-cell and 4-cell stages, respectively. However, the percentages decreased to 32.3 and 32.7% at 8-cell and blastocyst stages, respectively. Furthermore, we also evaluated the in vivo development of elephant-pig iSCNT cloned embryos and transferred 2260 reconstructed embryos into two surrogate gilts that successfully became pregnant and a total of 11 (1 and 10) fetuses were surgically recovered after 17 and 19 days of gestation, respectively. The crown-rump length and width of elephant-pig cloned fetuses were smaller than the control group. Unfortunately, none of these fetuses contained elephant genomes, which suggested that elephant embryos failed to develop in vivo. In conclusion, we successfully obtained elephant-pig reconstructed embryos for the first time and these embryos are able to develop to blastocyst, but the in vivo developmental failure needs further investigated.

Concomitant viral and bacterial pneumonia among patients in ICU with mechanical respiratory support.

INTRODUCTION: The use of mechanical ventilators in the intensive care unit (ICU) is often associated with higher risk of respiratory tract infections, including ventilator-associated pneumonia (VAP). Concomitant bacterial-viral infection was reported to worsen patient's clinical condition. This study evaluated the rate of concomitant bacterial-viral infections in patients with VAP and analyzed their clinical outcomes. METHODOLOGY: In this retrospective observational study 107 patients diagnosed with VAP and admitted in ICU with mechanical ventilator support between April 2018 and May 2019 in the Department of Respiratory Medicine, Dachang Hospital, Shanghai, China were included. 27 most commonly involved lower respiratory tract infection (LRTI) pathogens (bacteria and virus) and seven genetic markers of antibiotic resistance were detected and analyzed using Biofire® FilmArray® Pneumonia Panel plus (bioMérieux SA, Paris, France). RESULTS: Of the 107 patients, 45 (42.1%) patients had bacterial infection alone (bacterial group), 26 (24.3%) had virus infection alone (viral group) and 24 (22.4%) patients had concomitant bacterial-viral infection (mixed group). Sixty-nine (64.5%) and 50 (46.7%) patient samples were positive for bacterial (bacterial and mixed groups) and viral (viral and mixed groups) detection, respectively. Streptococcus pneumonia (11.2%) and Influenza A (17, 15.9%), were the predominantly identified bacterial and viral species. The blaCTX-M (21.5%) was the predominant resistance gene detected. Twenty-four (22.4%) patients were positive for concomitant bacterial-viral infection; Staphylococcus aureus and Influenza A were the most common bacterial-viral combination identified. CONCLUSIONS: Concomitant bacterial-viral infection was higher compared to previously published studies and the increased duration of mechanical ventilation was associated with increased disease severity.

Chemical Constituent Analysis of Ranunculus sceleratus L. Using Ultra-High-Performance Liquid Chromatography Coupled with Quadrupole-Orbitrap High-Resolution Mass Spectrometry.

Ranunculus sceleratus L.(RS) has shown various pharmacological effects in traditional Chinese medicine. In our previous study, the positive therapeutic effect on α-naphthylisothiocyanate induced intrahepatic cholestasis in rats was obtained using TianJiu treatment with fresh RS. However, the chemical profile of RS has not been clearly clarified, which impedes the research progress on the therapeutic effect of RS. Herein, an ultra-high performance liquid chromatography coupled with quadrupole Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) method was developed to rapidly separate and identify multiple constituents in the 80% methanol extract of RS. A total of sixty-nine compounds (19 flavonoids, 22 organic acids, 6 coumarins, 4 lignans, 14 nitrogenous compounds, and 4 anthraquinones) were successfully characterized. A total of 12 of these compounds were unambiguously identified by standard samples. Their mass spectrometric fragmentation pathways were investigated. It is worth noting that flavonoids and lignans were identified for the first time in RS. In this study, we successfully provide the first comprehensive report on identifying major chemical constituents in RS by UHPLC-Q-Orbitrap HRMS. The obtained results enrich the RS chemical profile, paving the way for further phytochemical study, quality control, and pharmacological investigation of RS.

Inonotsuoxide B suppresses hepatic stellate cell activation and proliferation via the PI3K/AKT and ERK1/2 pathway.

Hepatic stellate cells (HSCs) serve a pivotal role in the formation and degradation of the extracellular matrix during liver fibrosis. Inonotsuoxide B is a tetracyclic triterpenoid that can be extracted from Inonotus obliquus and has been previously reported to inhibit the growth of liver and gastric cancer cells. However, its effect on liver fibrosis remain poorly understood. Therefore, in the present study, the potential antiproliferative effects of inonotsuoxide B on HSCs was investigated. Initially, cells were divided into the following five groups: Control; platelet-derived growth factor (PDGF)-BB (10 ng/ml); and PDGF-BB + inonotsuoxide B (5, 10 and 20 µg/ml) groups. Inonotsuoxide B treatment (5, 10 and 20 µg/ml) was revealed to reverse PDGF-BB-induced HSC proliferation. Furthermore, the protein expression of α-smooth-muscle actin (α-SMA) and type I collagen was significantly decreased in the inonotsuoxide B (10 and 20 µg/ml) groups compared with the PDGF-BB group. Inonotsuoxide B (5, 10 and 20 µg/ml) was also revealed to suppress PDGF-BB-induced α-SMA mRNA expression and activation of the PI3K/AKT and ERK signaling pathways in HSCs. These findings suggest that inonotsuoxide B suppresses the proliferation and activation of HSCs by inhibiting the PI3K/AKT and ERK1/2 signaling pathways.

Clinical Randomized Controlled Study of Acupuncture Treatment on Children with Autism Spectrum Disorder (ASD): A Systematic Review and Meta-Analysis.

This study aimed to summarize the effectiveness and safety of acupuncture in the treatment of autism spectrum disorder (ASD) through literature analysis and evaluation. All studies were retrieved from various databases as follows: English databases, such as PubMed, Cochrane Library, Ovid, and Web of Science, and Chinese databases, such as China National Knowledge Infrastructure (CNKI), WanFang Data (WF), and Technology Periodical Database (VIP). The Cochrane Collaboration's Bias Risk Assessment Scale was used to assess the studies' risk of bias. The effects of acupuncture treatment for ASD were determined using the following indicators: childhood autism rating scale (CARS), autism behavior check list (ABC), Reynell developmental language scale (RDLS), and functional independence measure of children (WeeFIM). The risk map of bias of these studies' quality and the meta-analysis results of the indicators was prepared with RevMan 5.2 software. Finally, 16 studies were included, five of which were in English and 11 were in Chinese. The 16 studies included 1332 patients. The CARS results for subgroup analysis were as follows: acupuncture subgroup (MD = -2.65, 95% CI (-3.22, -2.07)) and acupuncture plus massage subgroup (MD = -10.35, 95% CI (-11.34, -9.36)). The ABC results were as follows: (MD = -6.70, 95% CI (-9.10, -4.29)). The analysis results of sensory, relating, language, body and object use, and social/self-help in the subitems of ABC were as follows: sensory (MD = -2.67, 95% CI (-2.90, -2.44)), relating (MD = -3.28, 95% CI (-3.55, -3.02)), language (MD = -2.45, 95% CI (-2.73, -2.16)), body and object use (MD = -1.19, 95% CI (-1.38, -1.00)), and social/self-help (MD = -2.09, 95% CI (-2.30, -1.89)). For the analysis results of comprehension and expression ages in the subitems of RDLS, the comprehension age results were as follows: (MD = 0.08, 95% CI (-0.06, 0.22), P = 0.27). Those of expression age were as follows: (MD = 0.15, 95% CI (0.04, 0.26), P=0.009). The WeeFIM results were as follows: (MD = 3.70, 95% CI (2.38, 5.02)). This study suggested that acupuncture could effectively treat ASD. However, acupuncture methods and prescriptions at this stage remain heterogeneous, and acupuncture treatment operations require standardization. Studies using rigorous and standard research designs are needed to draw stronger conclusions about the advantages of using acupuncture to treat children and adolescents with ASD.

Influence of lacing bars on the buckling capacity of four-legged latticed columns considering geometric imperfections.

The buckling behavior of latticed columns had been widely investigated based on the theory of Euler, Engesser and Timoshenko shear beam. Although these methods had been formulated and proved to be accurate in case of special assumptions, the influences of lacing bars on the buckling behavior of latticed columns were unclear. This paper modeled a general four-legged latticed column to study the influence of the cross-section characteristics of lacing bars along with their imperfections on the buckling capacity of latticed columns. Three loading conditions and four geometric imperfect models were built to testify the performance of lacing bars. To calculate the buckling load of latticed columns with imperfections accurately, advanced nonlinear analytical procedures using Newton-Raphson incremental-iterative method (ANAP-NR) and Risk arc-length incremental-iterative method (ANAP-Risk) were developed, and then validated by FE software ABAQUS. The current data in the paper show the maximum variation on the critical buckling load of latticed columns, caused by the cross-section area, the bending moment of inertia outer lacing plane, and the imperfections of lacing bars, could reach 68%, 30%, and 25%. The analytical results indicate the great importance of lacing bars on the buckling capacity of latticed columns.

Inonotsuoxide B regulates M1 to M2 macrophage polarization through sirtuin-1/endoplasmic reticulum stress axis.

We explored the effect of tetracyclic triterpenoid inonotsuoxide B (IB) extracts of Inonotus obliquus on M1 to M2 macrophage polarization and its possible underlying mechanism. Lipopolysaccharide (LPS)-activated M1 macrophages exert pro-inflammatory effects and release inflammatory cytokines including interleukin (IL)-1ß and tumor necrosis factor (TNF)-α. The model and various groups were treated with different IB concentrations (2.5, 5, and 10 µg/mL) to observe changes in the M1 and M2 phenotypes, gene expression of NAD-dependent deacetylase sirtuin-1 (Sirt1), and endoplasmic reticulum stress (ERS). SIRT1-siRNA and thapsigargin (TG), an ERS agonist, were used to examine the relationship between SIRT1/ERS and the effect of IB on M1 to M2 RAW264.7 macrophage phenotypic changes. We found that IB had no effect on RAW264.7 cell proliferation at 10 µg/mL. Increasing concentrations of IB (2.5, 5, and 10 µg/mL) decreased the number of phenotypic M1 macrophages and, consequently, decreased the release of the inflammatory cytokines, IL-1ß and TNF-α. Furthermore, IB treatment increased the level of phenotypic M2 macrophages, which increased the release of anti-inflammatory cytokines such as arginase (Arg)-1 and found in inflammatory zone 1 (FIZZ1) in a dose-dependent manner. Further, we found that IB increased the expression of SIRT1 and inhibited that of ERS. Inhibition of Sirt1 expression by siRNA significantly increased that of ERS marker genes and IL1ß. Excessive ERS levels inhibited the IB-induced transformation of phenotypic M1 macrophage to the M2 macrophage phenotype. Therefore, IB, an extract of I. obliquus, may regulate macrophage polarization through the SIRT1/ERS signaling pathway.

TianJiu therapy for α-naphthyl isothiocyanate-induced intrahepatic cholestasis in rats treated with fresh Ranunculus sceleratus L.

ETHNOPHARMACOLOGICAL RELEVANCE: TianJiu (TJ) therapy, one type of cold moxibustion, applies to specific acupuncture points with herbal patches of hot nature, providing a constant irritant to the skin until the presence of hyperemia and blistering. Traditional and clinical reports suggest that TJ is an effective therapy for the treatment of jaundice with fresh Ranunculus sceleratus L. (RS), in which protoanemonin is one of the main irritant constituents. However, the therapeutic effect of TJ treatment with fresh RS against intrahepatic cholestasis has not been studied in animal experiments. AIM OF THE STUDY: Present study was undertaken to investigate the effect of TJ treatment with fresh RS against intrahepatic cholestasis in rats and provide an experimental basis for the underlying mechanism of TJ therapy. MATERIALS AND METHODS: Male intrahepatic cholestatic Sprague-Dawley rats induced by 2% α-naphthylisothiocyanate (ANIT, 80 mg/kg B.W.) were treated by TJ therapy with fresh RS. The levels of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), direct bilirubin (DBIL), total bilirubin (TBIL), total bile acid (TBA), hepatic malondialdehyde (MDA) and nitric monoxide (NO), as well as hepatic body ratio, bile flow and hepatic histopathological assay were measured and evaluated to investigate the therapeutic effect of TJ treatment with fresh RS. Phytochemical analysis of fresh and dried RS was performed by gas chromatography-mass spectrometer (GC-MS). RESULTS: After TJ treatment with fresh RS, the abnormally elevated levels of serum AST, ALT, ALP, DBIL, TBIL and TBA, as well as hepatic MDA and NO at 108 h were reduced significantly (versus model group, P < 0.01). The hepatic body ratio, bile flow and hepatic pathological change of cholestatic rats at 108 h in TJ group were restored when compared with those of model group. Thirty-one compounds including lactones, flavonoids and phenolic acids were identified and determined by GC-MS analysis. The content of protoanemonin in fresh RS (9.49%) was about 25-fold higher than that in dried RS (0.38%). CONCLUSIONS: TJ treatment with fresh RS exhibited good therapeutic effect on ANIT-induced intrahepatic cholestasis in rats, which may be due to the attenuated oxidative stress in the liver tissue. It is rational for the ancients to choose fresh RS as TJ herbal patches because of its abundant protoanemonin with the character of irritant. The qualitative and quantitative results of GC-MS analysis provided the chemical basis of TJ therapy with fresh RS, which can be regarded as a simple and efficient method for the treatment of cholestasis hepatitis.

Achyranthis bidentatae radix enhanced articular distribution and anti-inflammatory effect of berberine in Sanmiao Wan using an acute gouty arthritis rat model.

ETHNOPHARMACOLOGICAL RELEVANCE: Sanmiao Wan (SMW) has been a basic prescription employed for the treatment for gout in the clinic since Yuan dynasty. Achyranthis bidentatae radix (ABR) is designed as a lower-guiding drug in SMW to augment the articular accumulation of active ingredients and improve the anti-inflammatory effect. AIM OF THE STUDY: Present study was undertaken to investigate the dose-response relationship of berberine in SMW between the articular concentration and anti-inflammatory effect in the knee joint under the lower-guiding of ABR. MATERIALS AND METHODS: Rats were divided into control group, model group and SMW without or with low, medium and high doses of ABR groups. Rat model of acute gouty arthritis (AGA) was established by intra-articular injection of 0.2 mL monosodium urate crystal (20 mg/mL) inside knee joint cavity on day 2 during drug treatment slots. Knee joint swelling, synovial hyperplasia and inflammatory cell infiltration were investigated for anti-inflammatory study. The concentrations of berberine in rat plasma and tissues were determined by UPLC-MS/MS method. The effect of ABR on the expression levels of P-glycoprotein (P-gp) and MDR1 mRNA in the synovial tissues of knee joints in AGA rats was examined by Western blot and RT-qPCR assay, respectively. RESULTS: The distribution of berberine increased by 6.53%, 44.31% and 212.96% in the knee joint and 474.93%, 631.01% and 1063.3% in the ankle for SMW with low, medium and high doses of ABR groups, compared with SMW without ABR group. Similarly, the plasma level of berberine increased by 19.81%, 143.4% and 681.13%. On the contrary, the distribution of berberine evidently decreased 3.23, 10.61 and 46.21-fold in heart and 3.68, 6.74 and 24.78-fold in lung. SMW with different doses of ABR groups exhibited better efficiency than SMW without ABR group on ameliorating knee joint swelling, inhibiting synovial hyperplasia and alleviating inflammatory cell infiltration of AGA rats. The treatment with ABR could down-regulate the MDR1 mRNA and P-gp expressions of synovial tissues of knee joints in AGA rats. CONCLUSIONS: The enhanced articular distribution of berberine in SMW was attributed to the lower-guiding effect of ABR, which could evidently increase the plasma concentration of berberine, improve the supply of blood of inflamed joint, reduce the distribution of berberine in heart and lung and significantly inhibit the MDR1 mRNA and P-gp expression of synovial tissues of knee joints in AGA rats. The dose-response relationship of berberine between the enhanced articular concentration and improved anti-inflammatory effect in the knee joint under the lower-guiding of ABR was observed for the first time.

Salutaxel, a Conjugate of Docetaxel and a Muramyl Dipeptide (MDP) Analogue, Acts as Multifunctional Prodrug That Inhibits Tumor Growth and Metastasis.

Salutaxel (3) is a conjugate of docetaxel (7) and a muramyl dipeptide (MDP) analogue. Docetaxel (7) has been recognized as a highly active chemotherapeutic agent against various cancers. MDP and its analogues are powerful potentiators of the antitumor actions of various tumor-necrotizing agents. This article documents the discovery of compound 3 and presents pharmacological proof of its biological function in tumor-bearing mice. Drug candidate 3 was superior to compound 7 in its ability to prevent tumor growth and metastasis. Compound 3 suppressed myeloid-derived suppressor cell (MDSC) accumulation in the spleens of tumor-bearing mice and decreased various serum inflammatory cytokines levels. Furthermore, compound 3 antagonized the nucleotide-binding oligomerization domain-like receptor 1 (NOD1) signaling pathway both in vitro and in vivo.

Anti-angiogenic activity and antitumor efficacy of amphiphilic twin drug from ursolic acid and low molecular weight heparin.

Heparin, a potential blood anti-coagulant, is also known for its binding ability to several angiogenic factors through electrostatic interactions due to its polyanionic character. However, the clinical application of heparin for cancer treatment is limited by several drawbacks, such as unsatisfactory therapeutic effects and severe anticoagulant activity that could induce hemorrhaging. Herein, low molecular weight heparin (LMWH) was conjugated to ursolic acid (UA), which is also an angiogenesis inhibitor, by binding the amine group of aminoethyl-UA (UA-NH2) with the carboxylic groups of LMWH. The resulting LMWH-UA conjugate as an amphiphilic twin drug showed reduced anticoagulant activity and could also self-assemble into nanomicelles with a mean particle size ranging from 200-250 nm. An in vitro endothelial tubular formation assay and an in vivo Matrigel plug assay were performed to verify the anti-angiogenic potential of LMWH-UA. Meanwhile, the in vivo antitumor effect of LMWH-UA was also evaluated using a B16F10 mouse melanoma model. LMWH-UA nanomicelles were shown to inhibit angiogenesis both in vitro and in vivo. In addition, the i.v. administration of LMWH-UA to the B16F10 tumor-bearing mice resulted in a significant inhibition of tumor growth as compared to the free drug solutions. These findings demonstrate the therapeutic potential of LMWH-UA as a new therapeutic remedy for cancer therapy.

Analysis of radical scavenging active components in the fermented mycelia of Ophiocordyceps formosana.

The rates of 1,1-diphenyl-2-picryhydrazyl (DPPH)-free radical scavenging and hydroxyl radical eliminating were employed as indexes to qualitatively and quantitatively analyse the radical scavenging activity of the extracts with different solvents from fermented mycelia of Hirsutella huangshanensis RCEF0868, the anamorph of Ophiocordyceps formosana (FMOF). The results showed that both the aqueous extract and the methanol extract had significant radical scavenging activity. The half maximal inhibitory concentrations (IC50) of the aqueous extract on DPPH-free radical and hydroxyl radical are 0.85 and 1.37 mg/mL, respectively, while the IC50 of methanol extract is 1.23 and 2.91 mg/mL, respectively. The combined liquid chromatography-diode array detector-high resolution mass spectrometer(HRMS) analysis and activity determination revealed that the molecular formulas of three radical scavengers in aqueous extract were C6H14O6(1), C14H17N5O8(2) and C20H33N5O9(3). In addition to the compound 1, the methanol extract had another three radical scavengers as C30H22O11(4), C30H22O10(5) and C30H18O10(6). All the six kinds of components exhibited the DPPH radical scavenging activity, and compounds 1, 4 and 5 also showed the hydroxyl radical eliminating activity at the same time. By querying the database of natural products and reference to the relevant reports, compound 3 may be a new entity. The others were respectively cordycepic acid, succinoadenosine, 4α-oxyrugulosin, rugulosin and skyrin and, among them, succinoadenosine and 4α-oxyrugulosin were found in entomogenous fungi for the first time.

Identification and determination of myriocin in Isaria cicadae and its allies by LTQ-Orbitrap-HRMS.

A hybrid linear ion trap-quadrupole-Orbitrap high-resolution mass spectrometry (LTQ-Orbitrap-HRMS) was used to qualitatively and quantitatively analyse the myriocin in Isaria cicadae and its allies. The samples were prepared with 95% methanol for 30 min by ultrasonic-assisted extraction. The target compound was purified by ODS solid-phase extraction (SPE) column. The enriched samples were identified by mass spectrometry. The results showed that the contents of myriocin in both wild and artificial Isaria cicadae were below the detection limit, while a strain of Ophiocordyceps longissima and Cordyceps cicadae Shing (Dujiaolong), both closely related to the Isaria cicadae, and its asexual mycelia are rich in myriocin. It suggests that it may be wrong to consider C. cicadae as I. cicadae's teleomorph in Genbank or Mycobank in many published reports based on chemical classification, and the species rich in myriocin is probably not Isaria cicadae.

Determination and comparative analysis of 13 nucleosides and nucleobases in natural fruiting body of Ophiocordyceps sinensis and its substitutes.

Nucleosides and nucleobases are one of the most important indicators of quality control. A sensitive and reliable high performance liquid chromatography-ultraviolet method was applied to analyse 13 nucleosides and nucleobases simultaneously in 15 batches of nine Ophiocordyceps species and its allies in China. Principal component analysis (PCA) and cluster analysis were conducted by SPSS 22.0 software (IBM Corp., Armonk, NY, USA). The 15 samples of Cordyceps were differentiated successfully based on their nucleoside and nucleobase content. Total nucleosides content in mycelium was significantly higher than that in the natural fruiting bodies of Ophiocordyceps sinensis (NFOS). Five nucleosides or nucleobases - adenine (A), guanosine (Gu), uracil (U), uridine (Ur) and guanine (G) - were the major components contributed to the total variance according to PCA. The profiles of the 13 tested nucleosides and nucleobases (including adenosine, cytidine, guanosine, inosine, thymidine, uridine, cordycepin, adenine, cytosine, guanine, thymine, uracil and hypoxanthine) can discriminate different samples and can be candidate indicators applied for the quality control of Ophiocordyceps and its allies.

Orcinol glucoside produces antidepressant effects by blocking the behavioural and neuronal deficits caused by chronic stress.

This study focused on the antidepressant potential of orcinol glucoside (OG) and its possible mechanisms of action. We established a depressed rat model using 3 consecutive weeks of chronic unpredictable mild stress (CUMS). The antidepressant-like effect of OG was revealed using the sucrose preference test, the open field test, the forced swimming test (FST), and the tail suspension test (TST). The activity of the hypothalamic-pituitary-adrenal (HPA) axis was evaluated by detecting the serum corticosterone (CORT) concentrations and mRNA expression of corticotrophin-releasing hormone (CRH) in the hypothalamus. The protein expression levels of brain-derived neurotrophic factor (BDNF) and total phosphorylated-ERK1/2 were detected by western blot. The results showed that OG treatment (1.5, 3, or 6mg/kg) alleviated the depression-like behaviour of rats under CUMS, as indicated by the increased sucrose preference and the decreased immobility in both the FST and TST, although the rearing frequency in the open field test increased only in the group that received the lowest dose (1.5mg/kg OG). Rats that received OG treatment exhibited reduced serum CORT levels and CRH mRNA expression in the hypothalamus, suggesting that the hyperactivity of the HPA axis in CUMS rats was reversed by OG treatment. Moreover, OG treatment upregulated the protein levels of BDNF and phosphorylated-ERK1/2 in the hippocampus, even above control levels. Our findings suggest that OG improved depressive behaviour in CUMS rats by downregulating HPA axis hyperactivity and increasing BDNF expression and ERK1/2 phosphorylation in the hippocampus.