Spatio-temporal variation and environmental drivers of chlorophyll a concentration and diatom community in four small urban lakes of Kunming, China.

Chlorophyll a (Chla) and diatom community structure are two indicators of lake water quality. In this study, we investigated the environmental parameters, chlorophyll a, and diatom community of four small urban lakes in Kunming (Beitan, Beihu, Nanhu and Longtan lakes in the campus of Yunnan Normal University) between March 2017 and December 2019. The results showed that the concentrations of total nitrogen (TN), total phosphorus (TP), and Chla in the four lakes showed significant seasonal fluctuation. The Chla concentration in Nanhu Lake, which had the highest nutrient level among the four lakes, was significantly higher than that in the other three lakes and largely affected by TN. In comparison, water temperature significantly contributed to the increases in Chla concentration in the other three lakes. Water temperature and TN were significantly correlated with Chla across the four lakes. Diatom assemblages in Beitan, Nanhu, and Longtan lakes were dominated by planktonic diatoms, and benthic diatoms were dominant in the shallowest lake Beihu, suggesting that water depth significantly affected the proportion of planktonic diatoms and dominant taxa. Water depth, TN, TP, transparency, and water temperature affected the spatio-temporal changes of diatom community structure, with water temperature as the major factor in causing the seasonal variation in diatom community, and TN and TP as the major drivers for community variation among lakes within the same season.

Effectiveness of oestrogen pretreatment in patients with expected poor ovarian response (POSEIDON groups 3 and 4) undergoing GnRH antagonist protocol: study protocol for a randomised controlled trial.

INTRODUCTION: Women characterised by diminished ovarian reserve are considered to have poor ovarian response (POR) according to Patient-Oriented Strategies Encompassing IndividualizeD Oocyte Number (POSEIDON) criteria. Patients in this population often have a poor prognosis for treatment with assisted reproductive technology. In previous studies, oestrogen pretreatment before ovarian stimulation has been shown to have a beneficial effect. However, recent studies presented conflicting conclusions. This study aims to evaluate the effectiveness of oestrogen pretreatment in patients with expected POR (POSEIDON groups 3 and 4) undergoing gonadotrophin releasing hormone antagonist (GnRH-ant) protocol. METHODS AND ANALYSIS: A prospective superiority randomised parallel controlled trial will be conducted at a tertiary university-affiliated hospital. A total of 316 patients will be randomly divided into two groups at a ratio of 1:1. In the intervention group, oral oestrogen pretreatment will be administered from day 7 after ovulation until day 2 of the next menstrual cycle. Afterwards, a flexible GnRH-ant protocol will be initiated. The control group will receive no additional intervention beyond routine ovarian stimulation. The primary outcome is the number of oocytes retrieved. Secondary outcomes include the total number of retrieved metaphase II oocytes, average daily dose of gonadotropin, total gonadotropin dose and duration of ovarian stimulation, cycle cancellation rate, top quality embryos rate, blastocyst formation rate, embryo implantation rate, clinical pregnancy rate, early miscarriage rate and endometrial thickness on trigger day. All data will be analysed according to the intention-to-treat and per-protocol principles. ETHICS AND DISSEMINATION: The ethical approval has been confirmed by the reproductive ethics committee of the affiliated hospital of Shandong University of Traditional Chinese Medicine (SDUTCM/2022.9.20). In addition, written informed consent will be obtained from all the participants before the study. The results will be disseminated via publications. TRIAL REGISTRATION NUMBER: ChiCTR2200064812.

The role of Erzhi Tiangui formula in expected poor ovarian responders undergoing in vitro fertilization-embryo transfer: A multicenter, randomized, double-blind, placebo-controlled trial.

INTRODUCTION: Advanced age is one of the primary risk factors for infertility. Poor ovarian response (POR) to exogenous gonadotropin is a prominent characteristic of advanced-age women undergoing in vitro fertilization and embryo transfer (IVF-ET), which results in fewer retrieved oocytes and poor pregnancy outcomes. Traditional Chinese medicine (TCM) has been shown to improve female fertility. Erzhi Tiangui (EZTG) formula, in the form of granules with 10 herbal ingredients, demonstrated potential benefits in improving oocyte and embryo quality and ovarian reserve. Thus, this study aims to evaluate the efficacy and safety of EZTG formula. METHOD: The study is a multicenter, double-blind, placebo-controlled, randomized controlled trial (RCT), which will be conducted at 10 reproductive centers of tertiary hospitals. This study will enroll 480 women with expected POR of advanced age (≥35 years old) who fulfill the 2011 Bologna criteria. Participants will be assigned to either the EZTG group or the placebo group at random in an equal ratio. Each individual will receive conventional IVF-ET with EZTG granules or placebo as a complementary treatment. The primary outcome is the number of oocytes retrieved. Adverse events and safety assessments will be also conducted. DISCUSSION: This study aims to provide robust evidence of the efficacy and safety of EZTG formula as a complementary treatment for advanced-age women with expected POR undergoing IVF-ET.

[Progress of researches on mechanisms of acupuncture therapy for lowering blood pressure].

Acupuncture treatment can regulate blood pressure (BP) through multiple levels and ways. In the present paper, we reviewed the progress of researches on the underlying mechanisms of acupuncture in lowering BP from 1) regulation of activities of the neuroendocrine, 2) improvement of metabolic abnormality, and 3) alternation of gene expression in the heart and BP-regulation-related centers of the brain. The neuroendocrine mechanism mainly involves the inhibition of neuroinflammatory reaction in some higher brain regions, reduction of neuronal apoptosis, and suppression of the sympathetic cardiovascular regulatory functional areas of the brain stem, regulation of neurotransmitters and autonomic balance, activation of brain areas related to BP regulation, and promotion of functional connection between brain networks. The improvement of metabolic abnormality mainly refers to amelioration of imbalance of intestinal flora and target metabolites related to hypertension. The alteration of gene expression mainly manifests as up- and down-regulation of expression of genes related to oxidative stress, inflammation and vascular endothelial function in the myocardium, hypothalamus, rostral ventrolateral medulla. We reviewed the new research progress on the mechanism of acupuncture for hypertension, in order to provide evidence and research ideas for the treatment of related cardiovascular diseases by using acupuncture therapy in the future.

Lycium barbarum polysaccharide alleviates oxygen glucose deprivation-induced PC-12 cells damage by up-regulating miR-24.

Objective: This research was aimed to detect the functions of Lycium barbarum polysaccharides (LBPs) on oxygen and glucose deprivation (OGD) injury and potential mechanisms at PC-12 cells. Methods: CCK-8, flow cytometry and reactive oxygen species (ROS) assays were used to detect OGD, LBPs and miR-24 effects on cell viability, apoptosis, and oxidative stress. MiR-24 was transfected and texted by transfection and qRT-PCR. Moreover, the related-protein levels of apoptosis, autophagy and pathways were tested by Western blotting. Results: LBPs significantly enhanced cell viability , inhibited cell apoptosis, autophagy and ROS level in OGD injury. In addition, miR-24 expression was declined in OGD-treated cells, while it was elevated when added LBPs. The preventive effects of LBPs on PC-12 cell damage induced by OGD were reversed by down-regulating miR-24. Furthermore, miR-24 inhibitor declined LBPs-induced change in Wnt/ß-catenin and JAK1/STAT3 pathways in OGD-injuried cells. Conclusions: LBPs exhibited preventive effects via up-regulating miR-122 and activating Wnt/ß-catenin and JAK1/STAT3 pathways in OGD-induced PC-12 cells.

Extraction Optimization, Structural Characterization, and Antioxidant Activities of Polysaccharides from Cassia Seed (Cassia obtusifolia).

In order to explore Cassia seed polysaccharides (CSPs) as natural antioxidants for application in the functional-food industry, microwave-assisted extraction (MAE) was optimized for the extraction of CSPs by using a response surface methodology. Furthermore, the chemical structures and antioxidant activities of CSPs extracted by MAE and hot water extraction were investigated and compared. The maximum extraction yield of CSPs extracted by MAE (8.02 ± 0.19%) was obtained at the optimized extraction parameters as follows: microwave power (415 W), extraction time (7.0 min), and ratio of water to raw material (51 mL/g). Additionally, the contents of the uronic acids, molecular weight, ratio of constituent monosaccharides, intrinsic viscosities, and degrees of esterification of CSPs were significantly affected by the MAE method. Moreover, CSPs exhibited remarkable 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) ABTS, 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl DPPH, nitric oxide, and hydroxyl radical scavenging activities as well as reducing power. The high antioxidant activities observed in CSPs extracted by MAE could be partially attributed to its low molecular weights and high content of unmethylated galacturonic acid. Results indicate that the MAE method could be an efficient technique for the extraction of CSPs with high antioxidant activity, and CSPs could be further explored as functional food ingredients.

Extraction Optimization, Physicochemical Characteristics, and Antioxidant Activities of Polysaccharides from Kiwifruit (Actinidia chinensis Planch.).

In order to evaluate effects of extraction techniques on the physicochemical characteristics and antioxidant activities of kiwifruit polysaccharides (KPS), and further explore KPS as functional food ingredients, both microwave-assisted extraction (MAE) and ultrasonic-assisted extraction (UAE) were optimized for the extraction of KPS. Furthermore, the physicochemical structures and antioxidant activities of KPS extracted by different techniques were investigated. The optimal extraction conditions of UAE and MAE for the extraction of KPS were obtained by response surface methodology. Different extraction techniques significantly affected the contents of uronic acids, molecular weights, molar ratios of constituent monosaccharides, and the degree of esterification of KPS. Results showed that KPS exhibited remarkable DPPH and ABTS radical scavenging activities, and reducing power. The high antioxidant activities observed in KPS extracted by the MAE method (KPS-M) might be partially attributed to its low molecular weight and high content of unmethylated galacturonic acid. Results suggested that the MAE method could be a good potential technique for the extraction of KPS with high antioxidant activity, and KPS could be further explored as functional food ingredients.

Phenolic Profiles, Antioxidant Capacities, and Inhibitory Effects on Digestive Enzymes of Different Kiwifruits.

In order to obtain high-quality kiwifruits with health-promoting characteristics, physicochemical properties, phenolic profiles, antioxidant capacities, and inhibitory effects on digestive enzymes (pancreatic lipase and α-glucosidase), of fourteen different types of kiwifruit obtained from China were systematically investigated and compared. Noticeable variations in the fruits' physicochemical properties and phenolic profiles were observed among them. The total phenolic content of Actinidia chinensis cv. Hongshi, A. chinensis cv. Jinshi, and A. chinensis cv. Jinlong were 16.52 ± 0.26 mg GAE/g DW (dry weight), 13.38 ± 0.20 mg GAE/g DW, and 11.02 ± 0.05 mg GAE/g DW, respectively, which were much higher than those of the other tested kiwifruits. According to high performance liquid chromatography (HPLC) analysis, phenolic compounds, including procyanidin B1, procyanidin B2, (-)-epicatechin, chlorogenic acid, gallic acid, and quercetin-3-rhamnoside, were found to be the major compounds in kiwifruits, while procyanidin B1, procyanidin B2, and chlorogenic acid were the most abundant phenolic compounds. Furthermore, all the tested kiwifruits exerted remarkable antioxidant capacities and inhibitory effects on pancreatic lipase and α-glucosidase. Indeed, A. chinensis cv. Hongshi, Actinidia chinensis cv. Jinshi, and Actinidia chinensis cv. Jinlong exhibited much better antioxidant capacities and inhibitory effects on digestive enzymes than those of the other tested kiwifruits. Particularly, A. polygama showed the highest inhibitory activity on α-glucosidase. Therefore, Actinidia chinensis cv. Hongshi, Actinidia chinensis cv. Jinshi, and Actinidia chinensis cv. Jinlong, as well as A. polygama could be important dietary sources of natural antioxidants and natural inhibitors against pancreatic lipase and α-glucosidase, which is helpful for meeting the growing demand for high-quality kiwifruits with health-promoting characteristics in China.

Kinsenoside screening with a microfluidic chip attenuates gouty arthritis through inactivating NF-κB signaling in macrophages and protecting endothelial cells.

Gouty arthritis is a rheumatic disease that is characterized by the deposition of monosodium urate (MSU) in synovial joints cause by the increased serum hyperuricemia. This study used a three-dimensional (3D) flowing microfluidic chip to screen the effective candidate against MSU-stimulated human umbilical vein endothelial cell (HUVEC) damage, and found kinsenoside (Kin) to be the leading active component of Anoectochilus roxburghi, one of the Chinese medicinal plant widely used in the treatment of gouty arthritis clinically. Cell viability and apoptosis of HUVECs were evaluated, indicating that direct Kin stimulation and conditioned medium (CM) from Kin-treated macrophages both negatively modulated with MSU crystals. Additionally, Kin was capable of attenuating MSU-induced activation of nuclear factor-κB/mitogen-activated protein kinase (NF-κB/MAPK) signaling, targeting IκB kinase-α (IKKα) and IKKß kinases of macrophages and influencing the expressions of NF-κB downstream cytokines and subsequent HUVEC bioactivity. Inflammasome NLR pyrin domain-containing 3 (NALP3) and toll-like receptor 2 (TLR2) were also inhibited after Kin treatment. Also, Kin downregulated CD14-mediated MSU crystals uptake in macrophages. In vivo study with MSU-injected ankle joints further revealed the significant suppression of inflammatory infiltration and endothelia impairment coupled with alleviation of ankle swelling and nociceptive response via Kin treatments. Taken together, these data implicated that Kin was the most effective candidate from Anoectochilus roxburghi to treat gouty arthritis clinically.

A renal-targeted triptolide aminoglycoside (TPAG) conjugate for lowering systemic toxicities of triptolide.

Triptolide (TP), a naturally derived compound, is proven effective in the treatment of nephritis and chronic allograft nephropathy. However, the severe multiorgan toxicity greatly limited it from further clinic use. 2-Glucosamine was demonstrated as a potential targeting ligand that could specifically interact with megalin receptors highly expressed in renal proximal tubules. In this study, 2-glucosamine was employed as a glycosyl donor while triptolide the acceptor to afford a nonhydrolyzable triptolide derivative-triptolide aminoglycoside (TPAG). The kidney-targeting efficiency, pharmacodynamic properties and safety of TPAG were thus evaluated. TPAG displayed 6.94-fold of AUC(0-t, kidney) and 13.96-fold of MRT(0-t, kidney) compared to TP. Additionally, TPAG presented improved protective effect against renal ischemia/reperfusion injury. Compared to TP's multiorgan toxicity, TPAG showed minimum toxicity toward the kidney and genital systems, and greatly lowered toxicity in the liver and immune systems. In sum, our study presented an alternative structure modification of triptolide with improved safety and efficacy profiles.

Dysregulation of cystathionine γ-lyase (CSE)/hydrogen sulfide pathway contributes to ox-LDL-induced inflammation in macrophage.

Hydrogen sulfide (H2S), mainly produced by cystathionine γ-lyase (CSE) in vascular system, emerges as a novel gasotransmitter exerting anti-inflammatory and anti-atherosclerotic effects. Alterations of CSE/H2S pathway may thus be involved in atherosclerosis pathogenesis. However, the underlying mechanisms are poorly understood. The present study showed that the levels of CSE mRNA and protein expression, as well as H2S production were decreased in ox-LDL-treated macrophage. CSE overexpression reduced the ox-LDL-stimulated tumor necrosis factor-α (TNF-α) generation in Raw264.7 and primary macrophage while CSE knockdown enhanced it. Exogenous supplementation of H2S with NaHS and Na2S also decreased the production of TNF-α and intercellular adhesion molecule-1 (ICAM-1) in ox-LDL-stimulated macrophage, and alleviated the adhesion of macrophage to endothelial monolayer. Cysteine, a CSE preferential substrate for H2S biosynthesis, produced similar effects on the pro-inflammatory cytokine generation, which were reversed by CSE inhibitors PAG and BCA, respectively. Moreover, NaHS and Na2S attenuated the phosphorylation and degradation of IκBα and p65 nuclear translocation, as well as JNK activation caused by ox-LDL. The JNK inhibitor suppressed the NF-κB transcription activity in ox-LDL-treated cells. Furthermore, inhibitors of NF-κB (PDTC), ERK (U0126 and PD98059) and JNK (SP600125) partially blocked the suppression by ox-LDL on the CSE mRNA levels. Taken together, the findings demonstrate that ox-LDL may down-regulate the CSE/H2S pathway, which plays an anti-inflammatory role in ox-LDL-stimulated macrophage by suppressing JNK/NF-κB signaling. The study reveals new therapeutic strategies for atherosclerosis, based on modulating CSE/H2S pathway.

Diversity and taxonomy of endophytic xylariaceous fungi from medicinal plants of Dendrobium (Orchidaceae).

Dendrobium spp. are traditional Chinese medicinal plants, and the main effective ingredients (polysaccharides and alkaloids) have pharmacologic effects on gastritis infection, cancer, and anti-aging. Previously, we confirmed endophytic xylariaceous fungi as the dominant fungi in several Dendrobium species of tropical regions from China. In the present study, the diversity, taxonomy, and distribution of culturable endophytic xylariaceous fungi associated with seven medicinal species of Dendrobium (Orchidaceae) were investigated. Among the 961 endophytes newly isolated, 217 xylariaceous fungi (morphotaxa) were identified using morphological and molecular methods. The phylogenetic tree constructed using nuclear ribosomal internal transcribed spacer (ITS), large subunit of ribosomal DNA (LSU), and beta-tubulin sequences divided these anamorphic xylariaceous isolates into at least 18 operational taxonomic units (OTUs). The diversity of the endophytic xylariaceous fungi in these seven Dendrobium species was estimated using Shannon and evenness indices, with the results indicating that the dominant Xylariaceae taxa in each Dendrobium species were greatly different, though common xylariaceous fungi were found in several Dendrobium species. These findings implied that different host plants in the same habitats exhibit a preference and selectivity for their fungal partners. Using culture-dependent approaches, these xylariaceous isolates may be important sources for the future screening of new natural products and drug discovery.

Sclerotial formation of Polyporus umbellatus by low temperature treatment under artificial conditions.

BACKGROUND: Polyporus umbellatus sclerotia have been used as a diuretic agent in China for over two thousand years. A shortage of the natural P. umbellatus has prompted researchers to induce sclerotial formation in the laboratory. METHODOLOGY/PRINCIPAL FINDING: P. umbellatus cultivation in a sawdust-based substrate was investigated to evaluate the effect of low temperature conditions on sclerotial formation. A phenol-sulfuric acid method was employed to determine the polysaccharide content of wild P. umbellatus sclerotia and mycelia and sclerotia grown in low-temperature treatments. In addition, reactive oxygen species (ROS) content, expressed as the fluorescence intensity of mycelia during sclerotial differentiation was determined. Analysis of ROS generation and sclerotial formation in mycelia after treatment with the antioxidants such as diphenyleneiodonium chloride (DPI), apocynin (Apo), or vitamin C were studied. Furthermore, macroscopic and microscopic characteristics of sclerotial differentiation were observed. Sclerotia were not induced by continuous cultivation at 25°C. The polysaccharide content of the artificial sclerotia is 78% of that of wild sclerotia. In the low-temperature treatment group, the fluorescent intensity of ROS was higher than that of the room temperature (25°C) group which did not induce sclerotial formation all through the cultivation. The antioxidants DPI and Apo reduced ROS levels and did not induce sclerotial formation. Although the concentration-dependent effects of vitamin C (5-15 mg mL(-1)) also reduced ROS generation and inhibited sclerotial formation, using a low concentration of vitamin C (1 mg mL(-1)) successfully induced sclerotial differentiation and increased ROS production. CONCLUSIONS/SIGNIFICANCE: Exposure to low temperatures induced P. umbellatus sclerotial morphogenesis during cultivation. Low temperature treatment enhanced ROS in mycelia, which may be important in triggering sclerotial differentiation in P. umbellatus. Moreover, the application of antioxidants impaired ROS generation and inhibited sclerotial formation. Our findings may help to provide new insights into the biological mechanisms underlying sclerotial morphogenesis in P. umbellatus.

HIV-1 integrase inhibitory activity of endophytic fungi from Aquilaria sinensis (Lour.) Gilg / 中国药学杂志

OBJECTIVE: To investigate the HIV-1 integrase inhibitory activity of endophytic fungi from medicinal plant Aquilaria sinensis (Lour.) Gilg. METHODS: HIV-1 integrase inhibitory activity of the isolated endophytic fungi was determined by HIV-1 integrase strand transfer inhibitory activity assay, and the active metabolites of the endophytic fungi with the greatest potential were studied by activity tracking. RESULTS: Among 78 strains of endophytic fungi isolated from A. sinensis, nine strains (11.54%) showed strong inhibitory activity against HIV-1 integrase. Three compounds were obtained from the fermentation of strain HN-AS-8 which was identified to be Chaetomium globosum by HIV-1 integrase strand reaction. Compound 1 inhibited HIV-1 integrase with IC50 value of 35.4 μmol · L-1, and compounds 1 and 3 were for the first time isolated from Chaetomium sp. CONCLUSION: Potential anti-HIV-1 metabolites exist in the endophytic fungi from A. sinensis, which can be new resources for new anti-AIDS drugs.