Smithii Var Research Articles

Identification of Bupleurum (Apiaceae) seeds by allele-specific PCR based on ITS sequences.

The traditional Chinese medicine Bupleuri radix (chaihu) is the dried roots of Bupleurum chinense and Bupleurum scorzonerifolium and many adulterants exist because of the differences in traditional understanding, medication habits and seed resources. Therefore, rapid and accurate identification methods for Bupleurum (Apiaceae) seeds are required. We analyzed the internal transcribed spacer (ITS) sequences of five common Bupleurum species to detect variations in them, including B. chinense, B. scorzonerifolium, B. marginatum var. stenophyllum, B. falcatum and B. smithii var.parvifolium. Based on single nucleotide polymorphisms (SNPs) in the ITS region, we designed five specific primer pairs and used these primers in an allele-specific PCR technique to establish a robust molecular identification method. The neighbor-joining (NJ) tree of ITS sequences showed that five Bupleurum species formed their own monophyly. Five specific primer pairs were designed and integrated into a specific PCR master mix and cycling conditions. The primer pair of BCF/R8 for B. chinense could amplify a specific identification band of 429bp and the minimum detection limit of the 5g mixture was about 5%; BSF/R11 for B. scorzonerifolium could amplify a specific 464bp band and the minimum detection limit was about 5%; BMSF/R1 for B. marginatum var. stenophyllum could amplify a specific 344bp band and the minimum detection limit was about 1%; BFF/R7 for B. falcatum could amplify a specific 137bp band and the minimum detection limit was about 1%; BSPF/R1 for B. smithii var. parvifolium could amplify a specific 390bp band. Subsequent analysis proved the reliable accuracy and good practicability of the five specific identification primers, indicating that the allele-specific primer PCR identification method can quickly identify Bupleurum seeds. The method elaborated in this study has the advantages of simple operation, good accuracy and high efficiency.

Bupleurum polysaccharides ameliorated renal injury in diabetic mice associated with suppression of HMGB1-TLR4 signaling.

Bupleurum polysaccharides (BPs) is isolated from Bupleurum smithii var. parvifolium, a key traditional Chinese medicine. The study was to investigate the effects of BPs on diabetic kidney injury. After two intraperitoneal injections of streptozotozin (STZ) 100 mg·kg-1, renal injury in diabetic mice was induced and BPs was orally administrated at dosages of 30 and 60 mg·kg-1·d-1. The STZ injected mice developed renal function damage, renal inflammation and fibrosis known as diabetic kidney disease (DKD). BPs significantly reduced serum creatinine level and urinary albumin excretion rate, with the attenuated swelling of kidneys. BPs treatment obviously alleviated the pathological damage of renal tissue. The progression of renal injury in BPs treated mice was inhibited with less expression of type IV collagen (Col IV), fibronectin (FN) and α-smooth muscle actin (α-SMA). The inhibition of inflammation in kidney was associated with the reduced level of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). BPs administration suppressed the over-expression of toll like receptor 4 (TLR4) and high-mobility group box 1 (HMGB1) with lowered activity of nuclear factor kappa B (NF-κB) in renal tissue of diabetic mice. Oral administration of BPs effectively prevented the development ofrenal injury in diabetic mice. This study suggested that the protection provided by BPs might affect through the interruption of HMGB1-TLR4 pathway, leading to the inhibition of renal inflammation and fibrotic process.

Comparative analysis on chemical constituents in Bupleurum chinense,B. marginatum,B. marginatum var. stenophyllum and B. smithii var. parvifolium

UPLC-Q-TOF-MS was used to analyze the chemical differences in Bupleurum. chinense,B. marginatum,B. marginatum var. stenophyllum and B. smithii var. parvifolium. Chromatographic separation was carried out on an Acquity HSS T3 C_(18) column( 2. 1 mm ×100 mm,1. 8 μm,Waters) with the mobile phase composed of 0. 1% formic acid in water-acetonitrile in the gradient elution. A hybrid quadrupole time-of-flight tandem mass spectrometry( Q-TOF-MS~E) was used for mass spectrometric analysis. Finally,25 peaks were identified based on their exact mass data and fragmentation characteristics. B.chinense,B.marginatum,B. marginatum var. stenophyllum and B. smithii var. parvifolium were obviously clustered into 3 types through processing by principal component analysis( PCA). There was almost no difference between B. chinense and B. marginatum. However,the compounds existed in B. chinense were different from those in B. marginatum var. stenophyllum,and B. smithii var. parvifolium.

Comparative analysis on seedling growth dynamics and physiological indices of Bupleurum smithii var. parvifolium and B. scorzonerifolium

With Bupleurum smithii var. parvifolium and B. scorzonerifolium as test objects, in order to provide a theoretical basis for the introduction and domestication of B. smithii var. parvifolium, the growth and development dynamics of seedlings, biomass accumulation, the content of malonaldehyde(MDA), the activity of antioxidase such as SOD, POD, CAT and APX between them were comparatively analyzed by direct sowing culture in the open field. The results indicated that the morphological index and the biomass accumulation of B. smithii var. parvifolium such as root diameter, root length, plant height and leaf number were inferior to B. scorzonerifolium, the antioxidase SOD and POD activity of B. smithii var. parvifolium was significantly inferior to B. scorzonerifolium (P<0.05), the antioxidase CAT and APX activity of B. smithii var. parvifolium was inferior to B. scorzonerifolium but the difference wasn't significant, while MDA content was superior to B. scorzonerifolium(P<0.05). Thus, compared with cultivated B. scorzoneri folium, the plant growth velocity of wild B. smithii var. parvifolium was relatively slower and its resistance was relatively weaker after introduction and domestication.

Freshwater Mastogloia (Bacillariophyceae) taxa from Macedonia, with a description of the epizoic M. sterijovskii sp. nov.

Mastogloia is one of the few species-rich diatom genera the majority of whose taxa inhabit the marine environment with only a small number of taxa in freshwater habitats. An ongoing revision of the Macedonian diatom flora, mainly over the last decade, has revealed many previously misidentified taxa. Currently, seven freshwater Mastogloia taxa have been recorded from Macedonia: M. elliptica (C. Agardh) Cleve, M. dansei (Thwaites) W. Smith, M. grevillei W. Smith, M. lacustris (Grunow) Grunow, M. ohridana Cvetkoska & Levkov, M. albertii nom. nov. and M. sterijovskii sp. nov. The main morphological features have been analysed with light and scanning electron microscopy. These taxa inhabit a variety of freshwater habitats, including oligotrophic rivers, eutrophic ponds, mine lakes, thermo-mineral springs, with one species from fossil sediments of ancient Lake Ohrid. Mastogloia albertii nom. nov., which we also lectotypify, is a new name for M. smithii var. amphicephala Grunow, since the latter is clearly distinct from the lectotype of M. smithii Thwaites. In addition, one of the observed taxa does not fit within any previously described Mastogloia taxa and is hereby described as a new species, M. sterijovskii. Mastogloia sterijovskii was found as epizoic on the dorsal carapace of the European pond turtle, Emys orbicularis Linnaeus, from the Mladost reservoir near Veles, central Macedonia. This species resembles M. albertii in valve outline, but can be distinguished by the shape of the valve apices and its size.

Therapeutic Effects of Bupleurum Polysaccharides in Streptozotocin Induced Diabetic Mice.

Diabetes mellitus is related to low-grade chronic inflammation and oxidative stress. Bupleurum Polysaccharides (BPs), isolated from Bupleurum smithii var. parvifolium has anti-inflammatory and anti-oxidative properties. However, little is known about its therapeutic effects on diabetes. In this experiment, the effects of BPs on alleviation of diabetes and the underlying mechanisms were investigated. Diabetic mice model was established via successive intraperitoneal injections of streptozotocin (100 mg/kg body weight) for two days. Mice with blood glucose levels higher than 16.8mmol/L were selected for experiments. The diabetic mice were orally administered with BPs (30 and 60 mg/kg) once a day for 35 days. BPs not only significantly decreased levels of blood glucose, but also increased those of serum insulin and liver glycogen in diabetic mice compared to model mice. Additionally, BPs adminstration improved the insulin expression and suppressed the apoptosis in pancreas of the diabetic mice. Histopathological observations further demonstrated that BPs protected the pancreas and liver from oxidative and inflammatory damages. These results suggest that BPs protect pancreatic β cells and liver hepatocytes and ameliorate diabetes, which is associated with its anti-oxidative and anti-inflammatory properties.

Development of a C3c-based ELISA method for the determination of anti-complementary potency of Bupleurum polysaccharides

Traditionally, determination of inhibitory potency of complement inhibitors is performed by the hemolytic assay. However, this assay is not applicable to the lectin pathway, thus impeding the understanding of complement inhibitors against the overall function of the complement system. The main objective of our study was to develop a specific enzyme-linked immunosorbent assay (ELISA) as an alternative method to assess the anti-complement activity, particularly against the lectin pathway. By using respective coating substrates against different activation pathways, followed by capturing the stable C3c fragments, our ELISA method can be used to screen complement inhibitors against the classical pathway and the lectin pathway. The inhibitory effect of suramin on the classical pathway, as measured by our hemolytic assay is consistent with previous reports. Further assessment of suramin and Bupleurum polysaccharides against the lectin pathway showed a good reproducibility of the method. Comparison of the lectin pathway IC50 between Bupleurum smithii var. parvifolium polysaccharides (1.055mg/mL) and Bupleurum chinense polysaccharides (0.98mg/mL) showed that, similar to the classical and alterative pathway, these two Bupleurum polysaccharides had comparable anti-complementary properties against the lectin pathway. The results demonstrate that the described ELISA assay can compensate for the shortcomings of the hemolytic assay in lectin pathway.

Morphology and typification of Mastogloia smithii and M. lacustris, with descriptions of two new species from the Florida Everglades and the Caribbean region

The names Mastogloia smithii Thwaites ex Smith and M. smithii var. lacustris Grunow have been attributed to a variety of related diatom morphologies, partly due to the poor availability of type material and complicated nomenclatural history. The history is detailed, clarifying the type morphologies of M. smithii and reconfirming a neglected elevation of M. smithii var. lacustris to M. lacustris (Grunow) Grunow. Populations reported as M. smithii and M. lacustris from the temperate zone (Ontario, Canada and Iowa and Michigan, USA), karstic wetlands of the subtropical Everglades (Florida, USA) and the tropics (Jamaica, Mexico and Belize) are compared with each other. Based on morphological differences including density of partecta, striae and areolae, M. calcarea sp. nov. and M. pseudosmithii sp. nov. are described from the Everglades and the Caribbean region, and a lectotype of M. smithii and a neotype of M. lacustris are designated.

Bupleurum Polysaccharides Attenuates Lipopolysaccharide-Induced Inflammation via Modulating Toll-Like Receptor 4 Signaling

Background Bupleurum polysaccharides (BPs), isolated from Bupleurum smithii var. parvifolium, possesses immunomodulatory activity, particularly on inflammation. Bacterial endotoxin lipopolysaccharide (LPS) triggers innate immune responses through Toll-like receptor 4 (TLR4) on host cell membrane. The present study was performed to evaluate whether the therapeutic efficacy of BPs on suppression of LPS’s pathogenecity could be associated with the modulating of TLR4 signaling pathway.Methodology/Principal FindingsLPS stimulated expression and activation of factors in the TLR4 signaling system, including TLR4, CD14, IRAK4, TRAF6, NF-κB, and JNK, determined using immunocytochemical and/or Western blot assays. BPs significantly inhibited these effects of LPS. LPS increased pro-inflammatory cytokines (TNF-α, IL-6, IL-1β, IL-12p40, and IFN-β) and NO production, evaluated using ELISA and Griess reaction assays, respectively. BPs antagonized these effects of LPS. Interestingly, BPs alone augmented secretion of some pro-inflammatory cytokines of non-LPS stimulated macrophages and enhanced phagocytic activity towards fluorescent E.coli bioparticles. In a rat model of acute lung injury (ALI) with pulmonary hemorrhage and inflammation, BPs ameliorated lung injuries and suppressed TLR4 expression.SignificanceThe therapeutic properties of BPs in alleviating inflammatory diseases could be attributed to its inhibitory effect on LPS-mediated TLR4 signaling.

Beneficial Effect of the Polysaccharides from Bupleurum smithii var. parvifolium on “Two-Hit” Acute Lung Injury in Rats

Radix Bupleuri is a traditional Chinese herb frequently used in the prescriptions for the treatment of inflammatory diseases. This study was to determine whether the crude polysaccharides isolated from the roots of Bupleurum smithii var. parvifolium (BPs) had beneficial effects on a "two-hit" acute lung injury model in rats. A two-hit lung injury model characterized by hemorrhagic shock followed by reperfusion and intratracheal lipopolysaccharide (1mgkg(-1)) administration was established in Wistar rats. Drugs and saline were administered 30min after the start of resuscitation. The two-hit acute lung injury model was successfully established. After oral administration, all BPs groups ameliorated pathological injury with lessened complement C3c deposit in lung. BPs 10 and 20mgkg(-1) diminished the ratio of wet-to-dry weight. In bronchoalveolar lavage (BAL) fluids, the protein concentration, the leukocytes counts, and the lung myeloperoxidase were significantly reduced. BPs also mediated the decreases in interleukin-6 and tumor necrosis factor-α in BAL fluids and in sera. Furthermore, BPs 20mgkg(-1) decreased the total complement hemolytic activity. BPs had beneficial effects on two-hit acute lung injury. The mechanisms of BPs on inflammatory disease might relate to its inhibitory effect on increased production of proinflammatory mediators and on overactivation of complement.

Mastogloia smithiivarlacustrisGrun.: A Structural Engineer of Calcareous Mats in Karstic Subtropical Wetlands

Mastogloia smithii var. lacustris Grun. is the dominant diatom in periphyton mats of the calcareous, freshwater to brackish wetlands of Caribbean coasts. Despite oligotrophy, frequent desiccation, high irradiance and temperatures, and occasional fire, periphyton communities in these wetlands can produce over 2000 g m-2 of organic biomass, prompting studies that examine stress resistance and maintenance of algal mats under extreme conditions. The diatom flora inhabiting periphyton mats from over 500 sites in the Florida Everglades and similar wetlands in Belize, Jamaica and Mexico was examined, and M. smithii var. lacustris was a persistent component, present in 97% of samples and comprising up to 80% of a diverse diatom assemblage. Valves at various stages of division were observed encased in extracellular polysaccharide that exceeded the cell volume; SEM observations confirm issuance from mantle pores resulting in suspension of the cell in a matrix dominated by cyanobacterial filaments. Using corresponding biophysical data from the collection sites, we define the optima for M. smithii var. lacustris along salinity, pH, phosphorus, and water depth gradients. Experiments revealed a collapse of M. smithii var. lacustris populations in the presence of above-ambient phosphorus concentrations and a rapid resurgence upon reflooding of desiccated mats. This widespread diatom taxon appears to play a critical role similar to that of cyanobacteria in microbial mats, and its disappearance in the presence of enrichment threatens biodiversity and the natural function in these systems that are increasingly influenced by urbanization.

Determination of flavones in different origin and parts of Bupleurum smithii var.parvifoliaum by UPLC-PDA

An ultra performance liquid chromatography method has been developed for determination of flavones in different origin and different parts from Bupleurum smithii var. parvifoliaum. The separation was performed at 30 degrees C on an Acquity UPLC BEH C18 column eluted with methanol and 0.1% phosphoric acid solution as mobile phases in gradient elution. The detection wavelength was set at 256 nm and the flow rate was 0.4 mL x min(-1). There flavones of rutin, quercetin and isorhamnetin were separated well, the linear calibration curves were obtained over the ranges of 0.106-1.06 microg for rutin (r = 0.999 8, n = 6), 0.004 87-0.048 7 microg for quercetin (r = 0. 999 7, n = 6), 0.022 0-0.220 microg for isorhamnetin (r = 0.999 8, n = 6). The mean recoveries of the three compounds were 99.3%, 97.8%, 98.9% with RSD of 2.4%, 3.0%, 3.2%. The result showed that the method is convenient and feasible for determination of the flavone content in B. smithii var. parvifoliaum.

Macrophage immunomodulatory activity of the polysaccharides from the roots of Bupleurum smithii var. parvifolium

Aim of the study Radix Bupleuri, one of the most frequently prescribed crude herbs in traditional Chinese medicine, has been used for centuries to treat inflammatory diseases. However, little is known about the therapeutic mechanisms of crude polysaccharides (BPs) isolated from the roots of Bupleurum smithii var. parvifolium. Macrophages play important roles in inflammatory diseases such as systemic lupus erythematosus (SLE). The purpose of the present work was to investigate immunomodulative effects of Bupleurum polysaccharides on murine peritoneal macrophages. Materials and methods BALB/c mice were administered intragastrically with Bupleurum polysaccharides 20, 40, and 80 mg kg −1 day −1, or prednisone 3 mg kg −1 day −1 or levamisole 25 mg kg −1 day −1 from day 0 to day 6. Peritoneal macrophages were isolated 5 days after intraperitoneal injection of 1 mL 5% sodium thioglycollate. Phagocytic functions of macrophages were studied; cytokines concentrations in the culture supernatants were determined by enzyme-linked immunosorbent assay and the secretion of nitric oxide (NO) was quantified by Griess reaction. Results Treatment with BPs enhanced phagocytic functions of macrophages (phagocytosis of apoptotic thymocytes, IgG-opsonized sheep red blood cells and chicken red blood cells) and inhibited LPS-induced productions of NO and proinflammatory cytokines (interleukin-1β, interleukin-6 and tumor necrosis factor-α). Conclusions Bupleurum polysaccharides up-regulated phagocytic activities but inhibited LPS-induced productions of proinflammatory mediators. These data suggested that at least part of the traditional beneficial effects of Bupleurum on inflammatory diseases could be ascribed to the immunomodulatory effects of Bupleurum polysaccharides on macrophages.

Beneficial effect of Bupleurum polysaccharides on autoimmune disease induced by Campylobacter jejuni in BALB/c mice

Aim of the study Radix Bupleuri, is one of the most frequently prescribed crude herbs in the prescriptions of traditional Chinese medicine for the treatment of inflammatory diseases and auto-immune diseases. This study was to determine whether the crude polysaccharides (BPs) isolated from the roots of Bupleurum smithii var. parvifolium, had beneficial effects on autoimmune disease. Materials and methods BALB/c mice were immunized with CJ-S 131 in Freund's complete adjuvant on day 0, and then boosted on day 14. BPs 15 or 30 mg kg −1 day −1, or prednisone 5 mg kg −1 day −1 was given to BALB/c mice intragastrically from day 0 to day 34. Results Treatment with BPs 15 or 30 mg kg −1 day −1 for 35 days protected kidney from glomerular injury with reduced immunoglobulin deposition and lowered proteinuria. The increased production of serum autoantibodies and total immunoglobulin G (IgG) was also inhibited. BPs 30 mg kg −1 day −1 improved weight loss and spleen swelling when compared with vehicle-treated group. Conclusions These findings suggested that Bupleurum polysaccharides had a beneficial effect on systemic lupus erythematosus-like syndroma induced by CJ-S 131 in BALB/c mice.

Relationships between Epipelic Diatom Taxa (Bacillariophyceae) and Nutrients in Two Karstic Lakes in Central Anatolia, Turkey

ABSTRACT The relationship between epipelic diatom communities and 16 environmental variables from two gypsum karstic lakes in central Anatolia was explored using canonical correspondence analysis. Temperature, ammonium (NH4 +), sulphate (SO4 2-) and calcium (Ca2+) concentrations were the most important variables related to species distribution. The genera Mastogloia, Gomphonema, Navicula, Cymbella, and Nitzschia were the richest in taxa. Diatom species associated with temperature, NH4 +, SO4 2-, and Ca2+ gradients were Mastogloia braunii, Mastogloia smithii var. lacustris, Campylodiscus clypeus, Navicula salinarum and Amphora coffeaeformis.

Desmids (Chlorophyta), including two new species and three new varieties, in two swamps, a lake, and a tarn in the South Island of New Zealand

Swamps at Okarito (Westland), Lake Hill (near Lake Coleridge), and Red Tarn and Blue Lake (in Mount Cook National Park) contained 24, 19, 20, and 8 desmid taxa, respectively. Genera represented by the largest number of taxa were Cosmarium with 17 taxa (7 of them in Okarito Swamp), Closterium with 9 taxa (6 of them in Lake Hill Swamp), and Staurastrum with 9 taxa (4 of them in Red Tarn). New species are Closterium okaritoense and Cosmarium paludicola in Okarito Swamp. New varieties are Pleurotaenium ehrenbergii var. contractum and Cosmarium crassipelle var. ornatum (both in Lake Hill Swamp), and Actinotaenium adelochondrum var. parvum (in Red Tarn). New records for the New Zealand flora are Euastrum verrucosum (in Lake Hill Swamp), Micrasterias suboblonga var. tecta (in Okarito Swamp), Cosmarium retusiforme var. incrassatum and Xanthidium smithii var. maius (both in Red Tarn), and Staurastrum neglectum and S. subarmigerum (in Blue Lake). Six other taxa previously recorded in the North Island were found in the South Island, including the endemic Cosmarium subquadratum var. genuosum and Xanthidium intermedium.

New Combinations within North American Schoenoplectus smithii and S. purshianus (Sect. Actaeogeton, Cyperaceae) and Comparison with Eastern Asian Allies

The new combinations Schoenoplectus purshianus (Fernald) M. T. Strong var. williamsii (Fernald) S. G. Smith, S. smithii (A. Gray) Sojaik var. setosus (Fernald) S. G. Smith, and S. smithii var. levisetus (Fassett) S. G. Smith are made. Diagnostic features, mainly from spikelet dimensions, perianth bristles, and achenes, are provided that distinguish the eastern North American S. smithii and S. purshianus, and the eastern Asian and Pacific island S. hotarui (Ohwi) Holub, S. juncoides (Roxburgh) Palla, and S. komarovii (Roshevitz) Sojaik, all five of which have been confused with each other. A description of Schoenoplectus sect. Actaeogeton (Reichenbach) J. Raynal is provided based mostly on North American and eastern Asian species.

The Diatom History of an Urban Lake: Summit Lake, Akron, Ohio

ABSTRACT Sedimentary diatoms from a 609 cm core indicated that Summit Lake, Akron, Ohio, originated as a shallow alkaline pool following late Pleistocene glaciation. The lake gradually developed a limnetic zone and became a peat bog. In recent years, the lake has become a slightly alkaline hypertrophic waterbody probably because of increased soil erosion and domestic sewage pollution. One hundred thirty (130) taxa (species and varieties) of diatoms were identified from the sediments of Summit Lake. Species most frequently encountered in the oldest sediments were Fragilaria brevistriata, F. construens, and F. pinnata. Those common in younger peat sediments were Achnanthes exigua, Cyclotella compta, Cymbella diluviana, Fragilaria brevistriata, F. construens, F. pinnata, Mastogloia smithii var. lacustris, and Tabellaria quadrisepta. The most abundant diatoms in recent sediments (since 1800 A.D.) were Asterionella formosa, Cocconeis placentula, Cyclotella meneghiniana, Fragilaria crotonensis, Melosira granula...

THE OCCURRENCE OF ANOPHELES SMITHII VAR. RAGEAUI MATTINGLY AND ADAM IN GHANA, WITH A NOTE ON ITS POSSIBLE IMPLICATION AS A VECTOR OF NON-HUMAN MALARIA.

THE OCCURRENCE OF ANOPHELES SMITHII VAR. RAGEAUI MATTINGLY AND ADAM IN GHANA, WITH A NOTE ON ITS POSSIBLE IMPLICATION AS A VECTOR OF NON-HUMAN MALARIA.

EXPERIMENTS WITH SOME NATIVE AND INTRODUCED PLANTS AS FOODS FOR CAMNULA PELLUCIDA (SCUDD.) (ORTHOPTERA: ACRIDIDAE) IN WESTERN CANADA

In field cage experiments, 17 native and introduced grasses and a native sedge, grown in pure culture, differed in their effect on the growth, survival and rate of population increase of the clear-winged grasshopper, Camnula pellucida. In terms of eggs deposited, Festuca rubra permitted a 24-lold population increase in a 1-year test; in tests repeated in 2 consecutive years, Agropyron elongatum, A. cristatum, Elymus junceus, Poa pratensis and P. compressa produced average annual increases of 10- to 18-fold; A. intermedium, A. dasystachyum, A. trachycaulum var. typicum, A. riparium, Poa ampla, and Bromus inermis, 3- to 8-fold. Agropyron smithii, Elymus canadensis, Stipa spartea, S. viridula, Bouteloua gracilis and Carex stenophylla var. enervis (1 year’s results), 2-fold or less. Percentage survival to the adult stage, rate of development, and apparently, robustness, were all positively correlated with population increase, but not strongly.