Subplenones A-J: Dimeric Xanthones with Antibacterial Activity from the Endophytic Fungus Subplenodomus sp. CPCC 401465.

Subplenones A-J (1-10), 10 new xanthone dimers, have been isolated and characterized from the endophytic fungus Subplenodomus sp. CPCC 401465, which resides within the Chinese medicinal plant Gentiana straminea. The isolation process was guided by antibacterial assays and molecular-networking-based analyses. The chemical structures of these compounds were elucidated through the interpretation of nuclear magnetic resonance (NMR) and high-resolution electrospray ionization mass spectrometry (HRESIMS) data. Furthermore, the relative configuration of the compounds was determined using NMR and single-crystal X-ray diffraction analyses, and the absolute configuration was established using electronic circular dichroism calculations. All of the isolated compounds exhibited significant inhibitory activity against Gram-positive bacteria. Notably, compounds 1, 5, and 7 displayed remarkable inhibitory activity against methicillin-resistant Staphylococcus aureus (MRSA) ATCC 700698, with a minimum inhibitory concentration (MIC) of 0.25 µg/mL, and against vancomycin-resistant Enterococcus faecium (VRE) ATCC 700221, with MIC values ranging from 0.5 to 1.0 µg/mL.

Metabolites from the plant endophytic fungus Penicillium sp. CPCC 401423 and their cytotoxic activity against MIA PaCa-2 cells.

Twenty-two metabolites were isolated from Penicillium sp. CPCC 401423 cultured on rice. The structures of all compounds were elucidated mainly by MS and NMR analysis as well as the necessary CD experimental evidence, of which penicillidione A (1), penicillidione B (2), (E)-4-[(4-acetoxy-3-methyl-2-butenyl)oxy]phenylacetic acid (3), (S)-2-hydroxy-2-{4-[(3-methyl-2-butenyl)oxy]phenyl} (4), (S)-4-(2,3-dihydroxy-3-methyl-butoxy)phenylacetic acid (5), (E)-4-[(3-carboxy-2-butenyl)oxy]benzoic acid (6), (Z)-4-[(4-hydroxy-3-methyl-2-butenyl)oxy]benzoic acid (7), open-cycled N-demethylmelearoride A (12), and penostatin M (16) were identified as new compounds. The cytotoxic activity against human pancreatic carcinoma cell line MIA PaCa-2a was detected. Among them, compounds 13-15 and 22 displayed significant cytotoxicity against MIA-PaCa-2 cells with IC50 values of 8.9, 36.5, 31.8, and 22.3 µM, respectively (positive control gemcitabine IC50 65.0 µM).

Mining and characterization of the PKS-NRPS hybrid for epicoccamide A: a mannosylated tetramate derivative from Epicoccum sp. CPCC 400996.

BACKGROUND: Genomic analysis indicated that the genomes of ascomycetes might carry dozens of biosynthetic gene clusters (BGCs), yet many clusters have remained enigmatic. The ascomycete genus Epicoccum, belonging to the family Didymellaceae, is ubiquitous that colonizes different types of substrates and is associated with phyllosphere or decaying vegetation. Species of this genus are prolific producers of bioactive substances. The epicoccamides, as biosynthetically distinct mannosylated tetramate, were first isolated in 2003 from Epicoccum sp. In this study, using a combination of genome mining, chemical identification, genetic deletion, and bioinformatic analysis, we identified the required BGC epi responsible for epicoccamide A biosynthesis in Epicoccum sp. CPCC 400996. RESULTS: The unconventional biosynthetic gene cluster epi was obtained from an endophyte Epicoccum sp. CPCC 400996 through AntiSMASH-based genome mining. The cluster epi includes six putative open reading frames (epiA-epiF) altogether, in which the epiA encodes a tetramate-forming polyketide synthase and nonribosomal peptide synthetases (PKS-NRPS hybrid). Sequence alignments and bioinformatic analysis to other metabolic pathways of fungal tetramates, we proposed that the gene cluster epi could be involved in generating epicoccamides. Genetic knockout of epiA completely abolished the biosynthesis of epicoccamide A (1), thereby establishing the correlation between the BGC epi and biosynthesis of epicoccamide A. Bioinformatic adenylation domain signature analysis of EpiA and other fungal PKS-NRPSs (NRPs) indicated that the EpiA is L-alanine incorporating tetramates megasynthase. Furthermore, based on the molecular structures of epicoccamide A and deduced gene functions of the cluster epi, a hypothetic metabolic pathway for biosynthesizing compound 1 was proposed. The corresponding tetramates releasing during epicoccamide A biosynthesis was catalyzed through Dieckmann-type cyclization, in which the reductive (R) domain residing in terminal module of EpiA accomplished the conversion. These results unveiled the underlying mechanism of epicoccamides biosynthesis and these findings might provide opportunities for derivatization of epicoccamides or generation of new chemical entities. CONCLUSION: Genome mining and genetic inactivation experiments unveiled a previously uncharacterized PKS - NRPS hybrid-based BGC epi responsible for the generation of epicoccamide A (1) in endophyte Epicoccum sp. CPCC 400996. In addition, based on the gene cluster data, a hypothetical biosynthetic pathway of epicoccamide A was proposed.

Modified Approach of Manufacturer's Power Curve Based on Improved Bins and K-Means++ Clustering.

The ideal wind turbine power curve provided by the manufacturer cannot monitor the practical performance of wind turbines accurately in the engineering stage; in this paper, a modified approach of the wind turbine power curve is proposed based on improved Bins and K-means++ clustering. By analyzing the wind speed-power data collected by the supervisory control and data acquisition system (SCADA), the relationship between wind speed and output is compared and elaborated on. On the basis of data preprocessing, an improved Bins method for equal frequency division of data is proposed, and the results are clustered through K-means++. Then, the wind turbine power curve correction is realized by data weighting and regression analysis. Finally, an example is given to show that the power curve of the same type of wind turbines, which, installed in different locations, are discrepant and different from the MPC, and the wind turbine power curve obtained by using this method can reflect the output characteristics of the wind turbine operating more effectively in a complex environment.

Norethindrone alters mating behaviors, ovary histology, hormone production and transcriptional expression of steroidogenic genes in zebrafish (Danio rerio).

The impact of progestins (i.e. synthetic forms of progesterone) on aquatic organisms has drawn increasing attention due to their widespread occurrence in the aquatic environments and potential effects on the endocrine system of fish. In this study, the effects of norethindrone (NET, a progestin) on the reproductive behavior, sex hormone production and transcriptional expressions were evaluated by exposing female zebrafish to NET at 0, 3.1, 36.2 and 398.6 ng L-1 for 60 days. Results showed that NET impaired the mating behaviors of female at 36.2 and 398.6 ng L-1 exhibited by males and increased the frequency of atretic follicular cells in the ovary exposed to NET at 398.6 ng L-1. As for sex hormones, plasma testosterone concentration in zebrafish increased, while estradiol concentration decreased. Up-regulation of genes (Npr, Mpra, Mprß, Fshß, Lß, Tshb, Nis and Dio2) was detected in the brain of fish exposed to NET at 398.6 ng L-1. The transcriptional levels of genes (Esr1, Vtg1, Ar, Cyp19a, Cyp11b and Ptgs2) were generally inhibited in the ovary of zebrafish by NET at 398.6 ng L-1. Moreover, the transcripts of genes (Vtg1, Esr1, Ar and Pgr) in the liver were reduced by NET at 36.2 and 398.6 ng L-1. Our findings suggest that NET can potentially diminish the of fish populations not only by damaging their reproductive organs, but also by altering their mating behavior through the changes in the expressions of genes responsible for the production of sex hormones.

The protective effects of polysaccharide extract from Xin-Ji-Er-Kang formula on Ang II-induced HUVECs injury, L-NAME-induced hypertension and cardiovascular remodeling in mice.

BACKGROUND: Xin-Ji-Er-Kang (XJEK) is a Chinese herbal formula, which has been reported to exert effective protection against cardiovascular diseases, including hypertension and myocarditis. METHODS: Cultured human umbilical vascular endothelial cells (HUVECs) were treated with angiotensin II (Ang II) and different concentrations of aqueous layer extracts (AqE). Subsequently nitric oxide (NO) and endothelial nitric oxide synthase (eNOS) expression levels were detected. In addition, fifty Kunming mice were randomized into control, Nω-nitro-L-arginine methyl ester (L-NAME), L-NAME+AqE, L-NAME+XJEK and L-NAME+fosinopril treatment groups. Following 8 weeks of treatment, the cardiac hemodynamic index was measured, relaxation of the aorta was examined and pathological changes were observed. Colorimetric analysis and enzyme linked immunosorbent assay (ELISA) were applied to determine the relevant indicators in plasma and cardiac tissues. RESULTS: The in vitro study results demonstrated that AqE could preserve endothelial function (NO, 21.05 ± 2.03 vs. 8.64 ± 0.59; eNOS, 1.08 ± 0.17 vs.0.73 ± 0.06). In addition, the in vivo results demonstrated that compared with the control group, treatment with AqE could enhance a high hemodynamic state (left ventricular systolic pressure, 116.76 ± 9.96 vs.114.5 ± 15.16), improve endothelial function (NO, 7.98 ± 9.64 vs. 1.66 ± 3.11; eNOS, 19.78 ± 3.18 vs.19.38 ± 3.85), suppress oxidative stress (OS) (superoxide dismutase, 178.17 ± 13.78 vs. 159.38 ± 18.86; malondialdehyde, 0.77 ± 0.13 vs.1.25 ± 0.36) and reverse cardiovascular remodeling. CONCLUSION: Polysaccharide from XJEK exerts protective effects against Ang II-induced injury in HUVECs and L-NAME-induced hypertension in mice and the underlying mechanism may be attributed to improving endothelial dysfunction, OS and the inflammation status in mice.

Proapoptotic and Growth-inhibitory Effects of Plumbagin on Human Gastric Cancer Cells Via Suppression of Signal Transducer and Activator of Transcription 3 and Protein Kinase B.

Context • Gastric cancer (GC) is the fourth most common cancer and the second leading cause of cancer-related deaths in the world. The current treatments include surgery and chemotherapy, either alone or in combination with radiotherapy, but the prognosis for patients with GC is usually poor. A safe and effective chemopreventive treatment for this malignant disease is urgently needed. Objective • The study intended to investigate the effects and underlying mechanisms of plumbagin, a quinonoid constituent that is derived from the roots of the medicinal plant Plumbago zeylanica, which exhibits potent anticancer properties against a number of cancers. Design • The in vitro study used the human GC cell line SGC-7901. Setting • All experiments were conducted at the Hubei University of Chinese Medicine and Tongji Medical College, Huazhong University of Science and Technology (Wuhan, China). Intervention • SGC-7901 cells were cultured in 30-mm dishes and treated with plumbagin at concentrations of 0, 5, 10, to 20 µmol/L. The cells were incubated with 10 µmol/L plumbagin for different amounts of time (0, 2, 4, 8, 12, and 24 h) in contact with the cancer cells. Outcome Measures • The cell viability was examined using a cell counting kit-8 viability assay, and the cell proliferation rate was determined using a 5-ethynyl-2'-deoxyuridine incorporation assay. The cell cycle distribution was assessed by flow cytometry using propidium iodide staining, and Western blotting was used to assess the expression of BAX, BCL-2, and caspase-3 and to identify any downregulation in the activation of transcription 3 (STAT3), protein kinase B (Akt), and extracellular signal-regulated kinase (ERK1/2). Results • The plumbagin concentrations of 5-20 mmol/L reduced the viability of the GC cells in a dependent manner. Plumbagin suppressed the expression of BAX, BCL-2, pro-caspase-3, and cleaved-caspase-3. It also restrained the expression and phosphorylation of STAT3 and decreased the phosphorylation of Akt1 but did not change the total protein or phosphorylation levels of ERK1/2. Conclusions • Plumbagin inhibits cell apoptosis in human GC cells, and that effect may be related with its ability to suppress phosphorylation of STAT3 and Akt. Given those 2 effects, plumbagin may be a promising agent in the treatment of gastric cancer.

Protective effects of Xinji'erkang on myocardial infarction induced cardiac injury in mice.

BACKGROUND: Myocardial infarction (MI) is a major risk factor responsible for morbidity and mortality. Xinji'erkang (XJEK) has been clinically used as an effective medication in the treatment of coronary heart disease and myocarditis. The purpose of this study was to investigate the cardioprotective effect of Xinji'erkang on MI mice. METHODS: Forty male mice were randomly assigned into four groups as follows (n = 10): sham, model, MI with administration of XJEK and fosinopril for four weeks. At the end of studies, hemodynamic parameters and electrocardiography (ECG) were recorded. Heart and body mass were measured and heart weight/body weight (HW/BW) ratio was calculated as index of hypertrophy. The hypertrophy of heart and aorta was examined using the hematoxylin and eosin (HE) staining, and the collagen deposition was evaluated using Van Gieson (VG) staining. Serum nitric oxide level (NO), superoxide dismutase (SOD) activity and malondialdehyde (MDA) concentration were assayed by colorimetric analysis. The expressions of endothelial NO synthetase (eNOS) expression in serum and cardiac tissues were determined using ELISA assay and immunohistochemistry. Angiotensin II (Ang II) in serum and cardiac tissues was measured using ELISA assay. Besides, tumor necrosis factor-α (TNF-α), interleukin1ß (IL-1ß) and interleukin10 (IL-10) were observed in cardiac tissues with ELISA assay as well. RESULTS: The administration of XJEK significantly improved cardiac dysfunction and abnormal ECG with reduced HW/BW ratio and ameliorated cardiomyocyte hypertrophy and collagen deposition compared to MI, which was partly due to the decreased SOD and increased MDA in serum. Moreover, XJEK treatment also improved endothelial dysfunction (ED) with not only enhanced eNOS activities in serum and cardiac tissues and elevated NO levels in serum, but also decreased Ang II content in serum and cardiac tissues. Finally, protein expressions of pro-inflammation cytokines, TNF-α and IL-1ß in the cardiac tissues with XJEK treatment were significantly decreased compared to model. On the contrary, IL-10, an anti-inflammatory cytokine concentrated in cardiac tissues was significantly enhanced compared to model. CONCLUSION: Xinji'erkang exerts cardioprotective effect on myocardial infarction in mice, which may be due to the improvement of endothelial dysfunction and the reduction of oxidative stress and inflammation response.

Short-Circuit Fault Detection and Classification Using Empirical Wavelet Transform and Local Energy for Electric Transmission Line.

In order to improve the classification accuracy of recognizing short-circuit faults in electric transmission lines, a novel detection and diagnosis method based on empirical wavelet transform (EWT) and local energy (LE) is proposed. First, EWT is used to deal with the original short-circuit fault signals from photoelectric voltage transformers, before the amplitude modulated-frequency modulated (AM-FM) mode with a compactly supported Fourier spectrum is extracted. Subsequently, the fault occurrence time is detected according to the modulus maxima of intrinsic mode function (IMF2) from three-phase voltage signals processed by EWT. After this process, the feature vectors are constructed by calculating the LE of the fundamental frequency based on the three-phase voltage signals of one period after the fault occurred. Finally, the classifier based on support vector machine (SVM) which was constructed with the LE feature vectors is used to classify 10 types of short-circuit fault signals. Compared with complementary ensemble empirical mode decomposition with adaptive noise (CEEMDAN) and improved CEEMDAN methods, the new method using EWT has a better ability to present the frequency in time. The difference in the characteristics of the energy distribution in the time domain between different types of short-circuit faults can be presented by the feature vectors of LE. Together, simulation and real signals experiment demonstrate the validity and effectiveness of the new approach.

Mechanical Fault Diagnosis of High Voltage Circuit Breakers Based on Variational Mode Decomposition and Multi-Layer Classifier.

Mechanical fault diagnosis of high-voltage circuit breakers (HVCBs) based on vibration signal analysis is one of the most significant issues in improving the reliability and reducing the outage cost for power systems. The limitation of training samples and types of machine faults in HVCBs causes the existing mechanical fault diagnostic methods to recognize new types of machine faults easily without training samples as either a normal condition or a wrong fault type. A new mechanical fault diagnosis method for HVCBs based on variational mode decomposition (VMD) and multi-layer classifier (MLC) is proposed to improve the accuracy of fault diagnosis. First, HVCB vibration signals during operation are measured using an acceleration sensor. Second, a VMD algorithm is used to decompose the vibration signals into several intrinsic mode functions (IMFs). The IMF matrix is divided into submatrices to compute the local singular values (LSV). The maximum singular values of each submatrix are selected as the feature vectors for fault diagnosis. Finally, a MLC composed of two one-class support vector machines (OCSVMs) and a support vector machine (SVM) is constructed to identify the fault type. Two layers of independent OCSVM are adopted to distinguish normal or fault conditions with known or unknown fault types, respectively. On this basis, SVM recognizes the specific fault type. Real diagnostic experiments are conducted with a real SF6 HVCB with normal and fault states. Three different faults (i.e., jam fault of the iron core, looseness of the base screw, and poor lubrication of the connecting lever) are simulated in a field experiment on a real HVCB to test the feasibility of the proposed method. Results show that the classification accuracy of the new method is superior to other traditional methods.

Targeting NF-κΒ and TNF-α Activation by Electroacupuncture to Suppress Collagen-induced Rheumatoid Arthritis in Model Rats.

BACKGROUND: Rheumatoid arthritis is an autoimmune disease that can cause chronic inflammation of the joints and other areas of the body. Acupuncture is an emerging alternative therapy for rheumatoid arthritis; however, the molecular mechanism underlying the beneficial effect of acupuncture has not been elucidated. OBJECTIVE: The study aimed to determine the effects of electroacupuncture (EA) on the expression of nuclear factor kappa Β (NF-κΒ) and tumor necrosis factor α (TNF-α) in model rats and to elucidate its anti-inflammatory mechanism in rheumatoid arthritis. DESIGN: The research team conducted a randomized animal study evaluating the efficacies of acupuncture on rheumatoid arthritis. Setting • All processes for the study were conducted at the Hubei University of Chinese Medicine and the Tongji Medical College of the Huazhong University of Science and Technology. Intervention • Twenty rats were randomly selected from 80 male Wistar rats and designated as the normal control (NC) group. The remaining 60 rats were prepared as rheumatoid arthritis models by administering intradermal injections of bovine, type 2 collagen emulsified in incomplete Freund's adjuvant for 3 wk. The 60 rats were randomly divided into 3 groups: (1) a rheumatoid arthritis model (model group); (2) a real-acupuncture-plus model (RA group); and (3) a sham-acupuncture-plus model (SA group), with 20 rats randomly assigned to each group. Zusanli (ST-36), Xuanzhong (GB-39), and Shenshu (BL-23) were applied for treatment of the RA group, whereas sham acupoints were selected for the SA group. OUTCOME MEASURES: The study used ultrastructural observations, an arthritis index, the expression levels of TNF-α and NF-κΒ (p65) in synovial cells, and the content of serum inflammatory cytokines to measure results. RESULTS: The arthritis index, the expression levels of NF-κΒ (p65) and TNF-α in synovial tissues, and the contents of interleukin (IL) 1-ß (IL-1ß), IL-6, and IL-8 increased for the 3 model groups compared with measurements for the NC group (P < .01). Those parameters were lower for the RA group than for the model group (P < .05). However, no statistically significant difference was observed between the model and SA groups. CONCLUSIONS: Acupuncture mediates the anti-inflammatory NF-kB pathway to reduce disease severity in collagen-induced arthritis.

A bibliometric analysis of chemotherapy and pain in pediatric patients over the last decade.

Background: Chemotherapy is an important treatment for children with cancer, and chemotherapy-induced pain is an important role in affecting patients' quality of life. In our study, bibliometric analysis was used to identify current research hotspots and future research trends of chemotherapy and pain in children over the last decade. Our findings can provide a reference for the research in the field of chemotherapy and pain in children. Method: Publications of chemotherapy and pain in children were collected from the Web of Science Core Collection database. CiteSpace was used to analyze publication characteristics from 2013 to 2022. Results: We identified 1,130 eligible publications in the field of chemotherapy and pain in children, with an increasing trend of publications over the last decade. In the field of chemotherapy and pain in children, the United States had the most publication with 346, followed by China with 135. The author with the most published papers was Pamela S Hinds (n = 8) from the United States. The journals that published the most papers were the Journal of pediatric hematology oncology (n = 44) and Medicine (n = 44). The Journal of Clinical Oncology was cited the most frequency (n = 422). St. Jude Children's Research Hospital had the most publication (n = 23). The specific keywords related to the field of chemotherapy and pain in children were "children", "chemotherapy", "management", "childhood cancer", "randomized controlled trial" and "efficacy". Emerging research focuses predominantly on symptomatic and supportive interventions for chemotherapy and pain in children. Conclusion: Attention to chemotherapy and pain in children with cancer was insufficient. This bibliometric analysis showed the upward trend of chemotherapy and pain in children over the last decade. More studies are needed to improve the quality of life in children with chemotherapy-induced pain. This study may provide useful information to guide future research on chemotherapy and pain in children.

Late-stage cascade of oxidation reactions during the biosynthesis of oxalicine B in Penicillium oxalicum.

Oxalicine B (1) is an α-pyrone meroterpenoid with a unique bispirocyclic ring system derived from Penicillium oxalicum. The biosynthetic pathway of 15-deoxyoxalicine B (4) was preliminarily reported in Penicillium canescens, however, the genetic base and biochemical characterization of tailoring reactions for oxalicine B (1) has remained enigmatic. In this study, we characterized three oxygenases from the metabolic pathway of oxalicine B (1), including a cytochrome P450 hydroxylase OxaL, a hydroxylating Fe(II)/α-KG-dependent dioxygenase OxaK, and a multifunctional cytochrome P450 OxaB. Intriguingly, OxaK can catalyze various multicyclic intermediates or shunt products of oxalicines with impressive substrate promiscuity. OxaB was further proven via biochemical assays to have the ability to convert 15-hydroxdecaturin A (3) to 1 with a spiro-lactone core skeleton through oxidative rearrangement. We also solved the mystery of OxaL that controls C-15 hydroxylation. Chemical investigation of the wild-type strain and deletants enabled us to identify 10 metabolites including three new compounds, and the isolated compounds displayed potent anti-influenza A virus bioactivities exhibiting IC50 values in the range of 4.0-19.9 µmol/L. Our studies have allowed us to propose a late-stage biosynthetic pathway for oxalicine B (1) and create downstream derivatizations of oxalicines by employing enzymatic strategies.

Verbalide A~F: new phthalide derivatives from the endophytic fungus Preussia sp. CPCC 400972.

There are six new phthalide derivatives Verbalide A ~ F (1-6) together with another known derivative (7) isolated from the endophytic fungus Preussia sp. CPCC 400972. Their structures were established by comprehensive spectroscopic analyses, including NMR and HRESIMS. In addition, compounds 1-7 exhibited excellent inhibitory effect against influenza A virus.

[Warm needling reduces oxidative damage and inflammation of cartilage in knee osteoarthritis rats].

OBJECTIVE: To observe the effect of warm needling on the expression of oxidative stress related factors and pro-inflammatory factors in cartilage of mono sodium iodoacetate (MIA) induced knee osteoarthritis (KOA) model rats, so as to explore its mechanisms underlying improvement of KOA. METHODS: Sixty male Sprague Dawley rats were randomly divided into 5 groups: control, model, acupuncture, moxibustion,warm needling, with 12 rats in each group. Rats of the acupuncture, moxibustion,warm needling groups received manual acupuncture or moxibustion or both stimulation of "Zusanli" (ST36) for 15 minutes, once a day for 21 days beginning from the third day after modeling. The foot volume was measured by drainage method, and the plantar mechanical contraction reflex threshold (mechanical pain threshold, MPT) measured by using an electronic pain meter. After 21 days of treatment, the histopathological changes of knee joint were observed by HE staining, and Mankin score was calculated to evaluate the degree of cartilage destruction. The malondialdehyde (MDA) level was measured by colorimetry, and immunofluorescence staining was used to observe the expression of NOX2, SOD2 or IL-1ß. RESULTS: Compared with the control group, the knee joint swelling volume from the 3rd day after modeling, Mankin score, MDA level, and the number of NOX2 and IL-1ß positive cells were significantly increased (P<0.01, P<0.05), while the MPT from the 3rd day after modeling, and the number of SOD2 positive cells were considerably decreased (P<0.01) in the model group. After the interventions, the increased levels of the knee joint swelling volume from the 12th day after modeling, and the Mankin score, MDA level, NOX2 and IL-1ß positive cells, and the levels of decreased MPT from the 9th day after modeling and SOD2 positive cell number were reversed in the acupuncture, moxibustion,warm needling groups (P<0.05, P<0.01), and the effects of warm needling were significantly superior to those of simple manual acupuncture and simple moxibustion in down-regulating knee joint volume, Mainkin score, MDA le-vel, and NOX2 and IL-1ß positive cells, and in up-regulating MPT from the 12th day after modeling, and the number of SOD2 positive cells (P<0.05). No significant differences were found between the acupuncture and moxibustion groups in the levels of all the indexes mentioned above (P>0.05). HE staining showed rough and damaged articular surface, with subchondral neovascularization and moderate connective tissue hyperplasia, and abundant lymphocyte and monocyte infiltration in the model group, which was milder in the acupuncture, moxibustion groups particularly in the warm needling group after 21 days' interventions. CONCLUSION: Warm needling can relieve knee joint pain, swelling and inflammatory damage in KOA rats, which may be associated with its function in inhibiting oxidative stress and inflammation in the cartilage of KOA. The therapeutic effect of warm needling is better than that of manual acupuncture and moxibustion alone.

Imaging Study on Acupuncture Inhibiting Inflammation and Bone Destruction in Knee Osteoarthritis Induced by Monosodium Iodoacetate in Rat Model.

OBJECTIVE: We aim to explore whether acupuncture inhibits inflammation and bone destruction in rat model monosodium iodoacetate (MIA)-induced knee osteoarthritis (KOA) by 18F-fluorodeoxyglucose (18F-FDG) small-animal positron emission tomography (PET) and micro-computed tomography (CT) imaging. METHODS: KOA was induced in rats by intra-articular injection MIA (2 mg/50 µL) through the right knee of the rats. Forty male Sprague Dawley rats weighing 280 to 340 g (12 weeks old) were randomly divided into four groups including Control group, KOA group, KOA plus manual acupuncture group (KOA+MA), KOA plus sham acupuncture group (KOA+SA). The acupuncture treatment lasted for three weeks (one-day rest after six days of treatment). Paw withdrawal threshold test and open-field test were used to assess mechanical allodynia and locomotor activity respectively for once a week. Hematoxylin and eosin (H&E) staining was used to assess the damage of the cartilage, synovium and infrapatellar fat pad (IFP). 18F-FDG PET was performed to quantify joint inflammation. The influence on the subchondral bone in these rats was confirmed by micro-CT. RESULTS: Mechanical hyperalgesia, joint inflammation, and obvious bone destruction were observed in the KOA group. H&E staining of the knee joint found that manual acupuncture played a protective effect in cartilage, synovium and IFP destruction. However, compared with KOA group, the results in sham acupuncture had no significant difference. After manual acupuncture treatment in KOA rats, inflammation was significantly suppressed shown by 18F-FDG PET imaging. Micro-CT analysis of the knee joint revealed that manual acupuncture protected bone by inhibiting osteophyte development and subchondral bone remodeling. CONCLUSION: The results of 18F-FDG PET and micro-CT showed that manual acupuncture inhibited inflammation and bone destruction, which provides reliable evidence for the effectiveness of acupuncture in hindering development of KOA, and provides reliable evidence for clinical application of acupuncture.

Discovery and Activation of the Cryptic Cluster from Aspergillus sp. CPCC 400735 for Asperphenalenone Biosynthesis.

Postgenomic analysis manifested that filamentous fungi contain numerous natural product biosynthetic gene clusters in their genome, yet most clusters remain cryptic or down-regulated. Herein, we report the successful manipulation of strain Aspergillus sp. CPCC 400735 that enables its genetic engineering via targeted overexpression of pathway-specific transcriptional regulator AspE. The down-regulated metabolic pathway encoded by the biosynthetic gene cluster asp was successfully up-activated. Analyses of mutant Ai-OE::aspE extracts led to isolation and characterization of 13 asperphenalenone derivatives, of which 11 of them are new compounds. All of the asperphenalenones exhibited conspicuous anti-influenza A virus effects with IC50 values of 0.45-2.22 µM. Additionally, their identification provided insight into biosynthesis of asperphenalenones and might benefit studies of downstream combinatorial biosynthesis. Our study further demonstrates the effective application of targeted overexpressing pathway-specific activator and novel metabolite discovery in microorganisms. These will accelerate the exploitation of the untapped resources and biosynthetic capability in filamentous fungi.

Cytochalasins from coastal saline soil-derived fungus Aspergillus flavipes RD-13 and their cytotoxicities.

Chemical investigation of coastal saline soil-derived fungus Aspergillus flavipes RD-13 led to the isolation of two new seco-cytochalasins (1) and (2) along with nine known analogs. Their structures were elucidated by comprehensive spectral analysis, and the absolute configurations of these two new ones were determined through Rh2(OCOCF3)4-induced CD experiment and chemical interconversions. Moreover, the absolute configuration of a known compound named cytochalasins Z18 (3) was also determined for the first time. Structurally, compounds 1, 2 and 3 were the open ring derivatives of compounds 5, 8, and 4, respectively. All compounds were evaluated for their cytotoxic activities on A549, H1299 and H520 cells and 4 exhibited the strongest inhibitory activities towards the above cell lines with IC50 values of 0.15, 0.23 and 0.43 µg/mL, respectively. Preliminary structure-activity relationship analysis suggested the importance of macrocyclic ring in cytochalasins to confer cytotoxicity.

A Combination of Genome Mining with an OSMAC Approach Facilitates the Discovery of and Contributions to the Biosynthesis of Melleolides from the Basidiomycete Armillaria tabescens.

Genome mining revealed that the genomes of basidiomycetes may include a considerable number of biosynthetic gene clusters (BGCs), yet numerous clusters remain unidentified. Herein, we report a combination of genome mining with an OSMAC (one strain, many compounds) approach to characterize the spectrum of melleolides produced by Armillaria tabescens CPCC 401429. Using F1 fermentation medium, the metabolic pathway of the gene cluster mel was successfully upregulated. From the extracts of the wild-type strain, two new melleolides (1 and 2), along with five new orsellinic acid-derived lactams (10-14), were isolated, and their structures were elucidated by LC-HR-ESIMS/MS and 2D-NMR. Several melleolides exhibited moderate anti-carcinoma (A549, NCI-H520, and H1299) effects with IC50 values of 4.0-48.8 µM. RNA-sequencing based transcriptomic profiling broadened our knowledge of the genetic background, regulation, and mechanisms of melleolide biosynthesis. These results may promote downstream metabolic engineering studies of melleolides. Our study demonstrates the approach is effective for discovering new secondary metabolites from Armillaria sp. and will facilitate the mining of the unexploited biosynthetic potential in other basidiomycetes.

Acupuncture for chronic sciatica: protocol for a multicenter randomised controlled trial.

BACKGROUND: Chronic Sciatica is a disabling condition causing considerable medical, social and financial implications. Currently, there is no recognised long-term effective treatment to alleviate sciatica. Acupuncture has been widely used for treating chronic pains with persistent analgesic effects. We aim to evaluate the efficacy and safety of acupuncture for chronic sciatica with follow-up in 52 weeks. METHODS AND ANALYSIS: This is a multicenter randomised sham-controlled trial. A total of 216 patients with chronic sciatica will be enrolled and randomly assigned to the acupuncture or sham acupuncture group. There will be 10 treatment sessions applied in 4 weeks with frequency decreased over time. Patients will complete follow-ups during 52 weeks. The primary outcomes are changes in leg pain intensity and disability from baseline to week 4. Secondary outcomes include back pain intensity, frequency and bothersomeness, quality of life, and global perceived effect. Adverse events will be recorded in detail. ETHICS AND DISSEMINATION: Ethical approval of this trial was granted from the ethics committee of Beijing University of Chinese Medicine and all study centres (No. 2020BZYLL0803). Written informed consent will be obtained from enrolled patients. Trial results will be disseminated in peer-reviewed publications. TRIAL REGISTRATION NUMBER: ChiCTR2100044585 (Chinese Clinical Trial Registry, http://www.chictr.org.cn, registered on 24 March 2021); preresults.