Biosynthesis and Assembly Logic of Fungal Hybrid Terpenoid Natural Products.

In recent decades, fungi have emerged as significant sources of diverse hybrid terpenoid natural products, and their biosynthetic pathways are increasingly unveiled. This review mainly focuses on elucidating the various strategies underlying the biosynthesis and assembly logic of these compounds. These pathways combine terpenoid moieties with diverse building blocks including polyketides, nonribosomal peptides, amino acids, p-hydroxybenzoic acid, saccharides, and adenine, resulting in the formation of plenty of hybrid terpenoid natural products via C-O, C-C, or C-N bond linkages. Subsequent tailoring steps, such as oxidation, cyclization, and rearrangement, further enhance the biological diversity and structural complexity of these hybrid terpenoid natural products. Understanding these biosynthetic mechanisms holds promise for the discovery of novel hybrid terpenoid natural products from fungi, which will promote the development of potential drug candidates in the future.

Biosynthesis of Cosmosporasides Reveals the Assembly Line for Fungal Hybrid Terpenoid Saccharides.

Fungal hybrid terpenoid saccharides constitute a new and growing family of natural products with significant biomedical and agricultural activities. One representative family is the cosmosporasides, which feature oxidized terpenoid units and saccharide moieties; however, the assembly line of these building blocks has been elusive. Herein, a cos cluster from Fusarium orthoceras was discovered for the synthesis of cosmosporaside C (1) by genome mining. A UbiA family intramembrane prenyltransferase (UbiA-type PT), a multifunctional cytochrome P450, an α,ß-hydrolase, an acetyltransferase, a dimethylallyl transferase (DMAT-type PT) and a glycosyltransferase function cooperatively in the assembly of the scaffold of 1 using primary central metabolites. The absolute configuration at C4, C6 and C7 of 1 was also established. Our work clarifies the unexpected functions of UbiA-type and DMAT-type PTs and provides an example for understanding the synthetic logic of hybrid terpenoid saccharides in fungi.

The value of CT radiomics features to predict visceral pleural invasion in ≤3 cm peripheral type early non-small cell lung cancer.

OBJECTIVE: To investigate predictive value of CT-based radiomics features on visceral pleural invasion (VPI) in ≤3.0 cm peripheral type early non-small cell lung cancer (NSCLC). METHODS: A total of 221 NSCLC cases were collected. Among them, 115 are VPI-positive and 106 are VPI-negative. Using a stratified random sampling method, 70% cases were assigned to training dataset (n = 155) and 30% cases (n = 66) were assigned to validation dataset. First, CT findings, imaging features, clinical data and pathological findings were retrospectively analyzed, the size, location and density characteristics of nodules and lymph node status, the relationship between lesions and pleura (RAP) were assessed, and their mean CT value and the shortest distance between lesions and pleura (DLP) were measured. Next, the minimum redundancy-maximum relevance (mRMR) and least absolute shrinkage and selection operator (LASSO) features were extracted from the imaging features. Then, CT imaging prediction model, texture feature prediction model and joint prediction model were built using multifactorial logistic regression analysis method, and the area under the ROC curve (AUC) was applied to evaluate model performance in predicting VPI. RESULTS: Mean diameter, density, fractal relationship with pleura, and presence of lymph node metastasis were all independent predictors of VPI. When applying to the validation dataset, the CT imaging model, texture feature model, and joint prediction model yielded AUC = 0.882, 0.824 and 0.894, respectively, indicating that AUC of the joint prediction model was the highest (p < 0.05). CONCLUSION: The study demonstrates that the joint prediction model containing CT morphological features and texture features enables to predict the presence of VPI in early NSCLC preoperatively at the highest level.

Heterologous and Engineered Biosynthesis of Nematocidal Polyketide-Nonribosomal Peptide Hybrid Macrolactone from Extreme Thermophilic Fungi.

Fungal polyketide-nonribosomal peptide (PK-NRP) hybrid macrolactones are a growing family of natural products with biomedical and agricultural activities. One of the most important families is the thermolides, which are produced by extreme thermophilic fungi and exhibit strong nematocidal activity. We show here that the genes ThmABCE from Talaromyces thermophilus NRRL 2155 are critical for thermolide synthesis. Two separate single-module hrPKS (ThmA) and NRPS (ThmB) enzymes collaborate to synthesize the core macrolactone backbone (6 or 7), and the NRPS ThmB-CT domain catalyzes the key macrocyclization step in PK-NRP intermediate release via ester bond formation, representing a novel function of fungal NRPS C domains. We also show that heterologous and engineered expression of the Thm genes in the type strains of Aspergillus nidulans and Escherichia coli not only dramatically enhances the yields of thermolides but also affords different esterified analogues, such as butyryl- (thermolides J and K, 15 and 16), hexanoyl-, and octanyl- derivatives or mixed thermolides. Thermolides L and M (18 and 19), discovered via genome mining-based combinatorial biosynthesis, represent the first l-phenylalanine-based thermolides. Our work shows a unique biosynthetic mechanism of PK-NRP hybrid macrolactones from extremophiles, which led to the discovery of novel compounds and furthers our biosynthetic knowledge.

Population genetic dynamics of Himalayan-Hengduan tree peonies, Paeonia subsect. Delavayanae.

According to the present taxonomical treatment, Paeonia subsect. Delavayanae consists of only two species (P. delavayi and P. ludlowii) endemic to the Himalayan-Hengduan Mountains. Although P. ludlowii can be distinguished from P. delavayi on the basis of a series of morphological characters, the species delimitation remains controversial because the more widespread one, P. delavayi, exhibits considerable morphological diversity. Both chloroplast DNA markers and nuclear microsatellites or simple sequence repeats (nSSR) are used herein to reveal genetic diversity and relationships of the two taxa included in this subsection, and ecological niche modeling (ENM) is employed to get insights into their paleodistribution. Our results show that genetic boundaries between the two currently recognized species are unclear, probably due to recent divergence. Paeonia ludlowii is budding from P. delavayi, probably by genetic isolation but also by shifting its niche to the harsher upland Tibetan conditions. Paeonia delavayi itself would be, however, under active speciation, showing significant genetic differentiation and morphological diversity. Whereas P. ludlowii would have endured the Pleistocene glacial periods by in situ persistence in local, small refugia, a 'dual' model seems to apply for P. delavayi (in situ persistence and retreat to refugia). The rarity of P. ludlowii and high evolutionary potential of P. delavayi imply high priority for in situ conservation of both taxa. The Himalayan-Hengduan Mountains are an ideal arena for differentiation within subsect. Delavayanae of Paeonia, by means of expansions/contractions/displacements, vertical migrations, and local survival/extinctions in response to the Neogene climate fluctuations and geological changes.

Oxidative damage and antioxidative indicators in 48 h germinated rice embryos during the vitrification-cryopreservation procedure.

KEY MESSAGES: Cu/Zn SOD and other genes may be critical indicators of a stress response to reactive oxygen species (ROS) accumulation in 48 h germinated rice embryos subjected to vitrification cryopreservation. In the current study, reactive oxygen species (ROS) accumulation was investigated in 48 h germinated rice embryos during the vitrification-cryopreservation process. We found that vitrification-cryopreservation significantly affected ROS levels, especially superoxide anion levels, in 48 h germinated rice embryos. Malonaldehyde content in the apical meristems of germinated embryos was significantly positively correlated with the rate of superoxide anion generation and the highest levels of malonaldehyde content were reached after vitrification treatment. Cell viability in 48 h germinated embryos was significantly negatively correlated with the rate of superoxide anion generation, malonaldehyde content, and electrolyte leakage. Spatial and temporal patterns in ROS accumulation in these embryos existed during the vitrification procedure. Among the vitrification-cryopreservation treatments we assessed, the preculture treatment was found to stimulate superoxide anion generation and to activate the response system in the apical meristems of germinated embryos. Loading treatments motivated the catalase and ascorbate peroxidase activities. During the vitrification-dehydration treatment, oxidative stress reached the highest levels causing an antioxidative response. This response involved antioxidant enzymes promoting detoxification of ROS. Based on a comprehensive correlation analysis involving ROS accumulation, cell viability, the activities of antioxidant enzymes, and gene expression profiles, Cu/Zn SOD, CAT1, APX7, GR2, GR3, MDHAR1, and DHAR1 may be critical indicators of oxidative stress affected by the vitrification-cryopreservation treatments. The investigation of these antioxidative responses in 48 h germinated rice embryos may, therefore, provide useful information with respect to plant vitrification-cryopreservation.

CRYOPRESERVATION OF IN VITRO-GROWN SHOOT TIPS OF CHINESE MEDICINAL PLANT Atractylodes macrocephala KOIDZ. USING A DROPLET-VITRIFICATION METHOD.

BACKGROUND: Atractylodes macrocephala Koidz. is an important medicinal species from China that has been used for thousands of years for its special pharmacological antioxidant, hepatoprotective, anti-inflammatory, anti-allergic, antithrombotic, antiviral, and anticarcinogenic activities. OBJECTIVE: The aim of this research was to develop an efficient droplet-vitrification protocol for A. macrocephala shoot tips which could be used as a strategy for long-term conservation within gene banks. MATERIALS AND METHODS: The duration of preculture, loading, and PVS2 steps, as well as the recovery medium formulation, were optimized to achieve high levels of survival and regrowth for A. macrocephala shoot tips after liquid nitrogen exposure. RESULTS: Survival and regrowth levels after cryopreservation in the cultivar 'Baizhu' were as high as 76% and 62%, respectively. Thermal analysis using differential scanning calorimetry suggested that the PVS2 treatment plays a critical role for successful cryopreservation. CONCLUSION: The droplet-vitrification method established in this study could be used to cryopreserve A. macrocephala.

The meaning of life according to patients with advanced lung cancer: a qualitative study.

PURPOSE: This qualitative study explores the meaning of life and end-of-life coping strategies among patients in China with advanced lung cancer. METHODS: We conducted in-depth interviews with 21 hospitalized patients with advanced lung cancer and analysed the data using the 7-step Colaizzi method. RESULTS: The analysis revealed themes in patients' experiences and feelings about living with a terminal illness. These include: 1) The core of the meaning of life is "self-iteration," which includes self-recognition and cherishing life; 2) The existence form of the meaning of life is "yu-wei," including self-reliance and altruism; 3) The meaning of life is embodied in three levels: the past, present, and future. The past includes gratitude, guilt and remorse, and avoidance; the present includes using the support system, positive response, independence, and integrity; the future includes accompanying relatives, preparing for death, living a high quality of life, and worrying. CONCLUSION: Meaning of life is a multidimensional and diverse concept among patients with advanced lung cancer. Medical care providers and family members can provide targeted professional guidance and psychological support according to patients' characteristics to help them discover their meaning of life, improve their quality of life, and achieve a positive end-of-life perspective.

Berberine bridge enzyme-like oxidase-catalysed double bond isomerization acts as the pathway switch in cytochalasin synthesis.

Cytochalasans (CYTs), as well as their polycyclic (pcCYTs) and polymerized (meCYTs) derivatives, constitute one of the largest families of fungal polyketide-nonribosomal peptide (PK-NRP) hybrid natural products. However, the mechanism of chemical conversion from mono-CYTs (moCYTs) to both pcCYTs and meCYTs remains unknown. Here, we show the first successful example of the reconstitution of the CYT core backbone as well as the whole pathway in a heterologous host. Importantly, we also describe the berberine bridge enzyme (BBE)-like oxidase AspoA, which uses Glu538 as a general acid biocatalyst to catalyse an unusual protonation-driven double bond isomerization reaction and acts as a switch to alter the native (for moCYTs) and nonenzymatic (for pcCYTs and meCYTs) pathways to synthesize aspochalasin family compounds. Our results present an unprecedented function of BBE-like enzymes and highly suggest that the isolated pcCYTs and meCYTs are most likely artificially derived products.

[Effects of mitophagy on aerobic exercise intervention for depression in rats].

Objective: This experiment was designed to observe the effects of aerobic exercise on depressive behavior in rats induced by chronic unpredictable mild stress (CUMS), and to explore the possible mechanism by detecting the proteins related to mitochondrial autophagy. Methods: SD rats were randomly divided into three groups: blank control group (C, n=12), depression model group (D, n=12) and post-depression exercise group (D+E, n=12). Group D and D+E were modeled with CUMS for 28 days, and group D+E underwent aerobic exercise intervention for 4 weeks after model establishment. Then the behavior of rats was evaluated. The concentrations of whole brain dopamine and norepinephrine were determined by ELISA kits. The morphology and structure of mitochondria in the frontal lobe were observed with the transmission electron microscope (TEM). Mitochondrial autophagy lysosomes were localized by immunofluorescence colocalization. The expressions of LC3 and P62 proteins in the frontal lobe were measured with Western Blotting. The relative content of mitochondrial DNA was detected using Real-time PCR. Results: ①Compared with group C, the sucrose preference ratio in group D was decreased significantly(P＜0.01); Compared with group D, the sucrose preference ratio in group D+E was increased significantly (P＜0.01). ②In the open field experiment, compared with group C, group D had a significant decrease in activity, average speed and total distance (P＜0.05); Compared with group D, the average rate of activity in group D+E was significantly higher (P＜0.05). ③ELISA results showed that the levels of whole brain dopamine and norepinephrine were significantly lower in group D rats than those in group C (P＜0.05). ④Under transmission electron microscopy, compared with group C, group D had different degrees of mitochondrial swelling, decreased crest density, and intermembrane space dilation.; Compared with group D, a significant increase in mitochondrial autophagosomes and autophagic lysosomes was observed in neurons in group D+E. Increased co-localization of mitochondria with lysosomes in the D+E group could be observed under fluorescence microscopy. ⑤Compared with group C, the expression of P62 was increased significantly(P＜0.05), and LC3II/LC3I ratio was decreased significantly (P＜0.05) in group D; Compared with group D, LC3II/LC3I ratio was significantly higher in group D+E than that in group D (P＜0.05). ⑥Compared with group C, the relative number of mitochondrial DNA in the frontal lobe of group D was increased significantly (P＜0.05). Conclusion: Aerobic exercise has a significant improvement effect on depression induced by CUMS in rats, and its mechanism may be related to the upregulation of the level of linear autophagy.

Nuclear and chloroplast SSR markers in Paeonia delavayi (Paeoniaceae) and cross-species amplification in P. ludlowii.

PREMISE OF THE STUDY: Microsatellite primers were developed for Paeonia delavayi and P. ludlowii (Paeoniaceae) to study their population genetics and phytogeography. METHODS AND RESULTS: Nine polymorphic nuclear microsatellite loci were isolated from an enriched library of P. delavayi and primers were designed. The number of alleles per locus ranged from two to 16; the observed and expected heterozygosities ranged from 0.014 to 0.687 and 0.042 to 0.875, respectively. Six polymorphic chloroplast microsatellite loci were identified in P. delavayi and primers were provided. The number of alleles per locus ranged from two to six and the polymorphic information content ranged from 0.08 to 0.716. Both nuclear and chloroplast primers were successfully applicable to P. ludlowii. CONCLUSIONS: The markers developed here will facilitate analyses of genetic diversity, population genetic structure, phytogeographical patterns, and conservation for P. delavayi and P. ludlowii.

[Research progress on substitutes for autogenous soft tissue grafts in mucogingival surgery].

Mucogingival surgery is a general term for periodontal surgeries that correct aberrant periodontal soft tissues. Conventional mucogingival surgeries with pedicle flap or autologous soft tissue graft for treatment of gingival recession and insufficient keratinized tissues are always related to disadvantages such as need for a second surgery site, limited supplies, and complaints for postoperative discomfort. In this regard, research and application of soft tissue substitutes have gained increasing attention. Various kinds of soft tissue substitutes, including acellular dermal matrix and xenogeneic collagen matrix, have been developed and applied to clinical treatment. This review aims to summarize advances in research of the characteristics and clinical effectiveness of several soft tissue substitutes and provide references for clinical application.

A new eudesmane sesquiterpene lactone from Curcuma wenyujin.

The aim of the study was to investigate the sesquiterpene constituents from the rhizomes of Curcuma wenyujin Y. H. Chen et C. Ling. The isolation and purification of the constituents from the 50% EtOH extracts of the rhizomes were performed with repeated column chromatography over sillica gel and macroporous resin. Eight sesquiterpenes were obtained and identified as wenyujinlactone A (1), neolitamone A (2), zedoarondiol (3), isozedoarondiol (4), aerugidiol (5), curcumol (6), curdione (7) and (1R, 10R)-epoxy-(-)-1, 10-dihydrocurdine (8) by means of spectral analysis. Among them, compound 1 was found to be a new eudesmane sesquiterpene lactone, whilst compounds 2-5 were obtained from this plant for the first time.

[Analysis of allergen variation for 3292 patients with allergic rhinitis].

OBJECTIVE: To investigate the variation of common allergens in patients with allergic rhinitis in recent 4 years in Tianjin First Center Hospital. METHODS: The medical records of skin prick test on 3292 patients with allergic rhinitis between 2009 and 2012 were restrospectively analyzed. The changing trend of various allergens in 4 years and distribution differences were compared. The differences of the top 5 allergens in under age group, adult group and different gender group were further analyzed by SPSS 19.0 software. RESULTS: The positive rate of dermatophagoides farinae was increasing year by year, from 45.1% in 2009 to 66.3% in 2012, and the positive rate of dermatophagoides pteronyssinus increased from 42.0% in 2009 to 58.6% in 2012, the difference was statistically significant (χ(2) value was 68.70, 41.55, all P < 0.01). The positive rate of dermatophagoides farinae and dermatophagoides pteronyssinus in adult group and male group showed significant upward trend year by year (χ(2) value was 75.85, 69.93, 274.25, 42.62, all P < 0.01), but not in adult group and female group. The positive rate of quinoa, mugwort and humulus scandens decreased year by year between 2009 and 2011(χ(2) value was 22.08, 11.64, 203.19, all P < 0.01), but increased again in 2012(χ(2) value was 21.55, 29.38, 12.40, all P < 0.01). CONCLUSIONS: There is a tendency of change of allergens in patients with AR. This phenominon may be helpful for doctors to choose the type of skin prick liquid.

[Role of vasculogenic mimicry and endothelium-dependent vessel in metastasis of laryngeal cancer].

OBJECTIVE: To investigate the contribution of vasculogenic mimicry (VM) and endothelium-dependent vessel (EDV) to invasion and metastasis of laryngeal squamous cell carcinoma (LSCC). METHODS: A total of 203 cases with LSCC was reviewed and followed up. VM and EDV in LSCC tissues were assessed by double staining with anti-CD31 immunohistochemistry and periodic acid-schiff. Kruskal-Wallis test and one-way ANOVA were used to analyze the relationship between VM, EDV and clinical pathology parameters of LSCC. Kaplan-Meier analysis was used to evaluate overall survival (OS) of patients with LSCC. RESULTS: VM related to pTNM stage, lymph node metastasis and pathology grade of LSCC, while EDV related to primary sites, pTNM stage, T stage and distant metastasis of LSCC. Univariate analysis showed VM (P = 0.014), pTNM stage (P = 0.009), T stage (P = 0.013), nodal status (P = 0.013), histopathology grade (P = 0.038), tumor size (P = 0.028), and radiotherapy (P < 0.0001) related to OS. VM (P = 0.011), primary sites (P = 0.049), tumor size (P = 0.001) and radiotherapy (P < 0.0001) related to disease free survival. Multivariate analysis indicated that VM was an adverse predictor for both OS and disease free survival. CONCLUSIONS: Both VM and EDV existed in LSCC. VM contributed to progression of LSCC through promoting lymph node metastasis. VM is an independent predictor for the prognosis of LSCC.

Vasculogenic mimicry is a key prognostic factor for laryngeal squamous cell carcinoma: a new pattern of blood supply.

BACKGROUND: Recurrence and local lymph node metastasis affected the prognosis of patients with laryngeal cancer. The aim of this study was to analyze clinical and pathological significance of vasculogenic mimicry (VM) in laryngeal squamous cell carcinoma (LSCC) and evaluate its contribution to prognosis. METHODS: Data of 168 cases of LSCC were reviewed retrospectively to reveal clinical pathology and prognostic significance of VM. CD31 and periodic acid-Schiff double staining was used to identify VM. RESULTS: VM in LSCC contributed to lymph node metastasis (P = 0.003) and clinical progression. VM correlated to histopathology grade (P = 0.001) of LSCC. VM was an adverse prognostic factor for both disease-specific survival (P = 0.039) and metastasis-free survival (P = 0.042) by univariate survival analyses. And it was an independent prognostic factor for only disease-specific survival (P = 0.003) by multivariate survival analyses. CONCLUSIONS: VM existed in LSCC. LSCC with VM has more potential to invasion and metastasis.

[Clinical research of vestibular autorotation test for patients with vertebrobasilar insufficiency].

OBJECTIVE: To explore the diagnostic values of vestibular autorotation test (VAT) for patients with vertebrobasilar insufficiency (VBI). METHODS: VAT and videonystagmography ( VNG) were performed on 73 patients with VBI and 48 patients with peripheral vestibular lesions (contrast group). Parameters analyzed included Gain, phase and asymmetry of VAT, as well as the canal paresis (CP) of caloric test and results of optokinetic-pursuit tests in VNG. Positive result of the test could be defined if anyone of the parameters was abnormal. RESULTS: For VAT test, Gain was enhanced in VBI group and was reduced in contrast group. In VBI group and contrast group, Gain enhanced showed in 47 (64.4%) cases and 5 (10.4%) cases, respectively (chi2 = 31.19, P < 0.01). Simultaneity, Gain reduced in 11 cases (15.5%) and 22 cases (45.8%), respectively (chi2 = 13.82, P < 0.01). But there was no statistics significant for results of the parameters of phase, asymmetry and integration between two groups. For VNG test, results with optokinetic-pursuit tests were more abnormal in VBI group than that in contrast group, which showed central lesions characteristics. Forty-four cases (60.3%) in VBI group and 10 cases (20.8%) in control group showed central lesions results with optokinetic-pursuit tests and visual fixation test (chi2 = 15.89, P < 0.01). Unilateral or bilateral CP showed in 33 cases (68.6%) in control group and 51 cases (69.9%) in VBI group with caloric test. CONCLUSIONS: Gain of VAT is mostly enhanced in VBI group, and Gain as a main characteristic is reduced in patients with peripheral vestibular lesions. The Gain parameter is availability for assessing characteristics of vestibular lesions. Phase and asymmetry can be used to assess the vestibular function but can not indicate the characteristics of vestibular lesions.