Study on the Compositional Analysis, Extraction Process, and Hemostatic and Anti-Inflammatory Activities of Cirsium japonicum Fisch. ex DC.-Cirsium setosum (Willd.) MB Extracts.

Cirsium japonicum Fisch. ex DC. (CF) and Cirsium setosum (Willd.) MB (CS) are commonly used clinically to stop bleeding and eliminate carbuncles. Still, CF is mainly used for treating inflammation, while CS favors hemostasis. Therefore, the present study used UHPLC-MS to analyze the main chemical constituents in CF-CS extract. We optimized the extraction process using single-factor experiments and response surface methodology. Afterward, the hemostatic and anti-inflammatory effects of CF-CS extract were investigated by determining the clotting time in vitro, the bleeding time of rabbit trauma, and the induction of rabbit inflammation using xylene and lipopolysaccharide. The study of hemostatic and anti-inflammatory effects showed that the CF-CS, CF, and CS extract groups could significantly shorten the coagulation time and bleeding time of rabbits compared with the blank group (p < 0.01); compared with the model group, it could dramatically inhibit xylene-induced ear swelling in rabbits and the content of TNF-α, IL-6, and IL-1ß in the serum of rabbits (p < 0.01). The results showed that combined CF and CS synergistically increased efficacy. CF-CS solved the problem of the single hemostatic and anti-inflammatory efficacy of a single drug, which provided a new idea for the research and development of natural hemostatic and anti-inflammatory medicines.

Characterization of Three Polysaccharide-Based Hydrogels Derived from Laminaria japonica and Their Hemostatic Properties.

Three Laminaria japonica polysaccharides (LJPs) extracted via water extraction (LJP-W), acid extraction (LJP-A), and enzymatic extraction (LJP-E) were used as raw materials to be cross-linked with chitosan and polyvinyl alcohol to prepare hydrogels. Compared with conventional hydrogel systems, all three types of LJP-based polysaccharide hydrogels exhibited better swelling properties (14 times their original weight) and the absorption ability of simulated body fluid (first 2 h: 6-10%). They also demonstrated better rigidity and mechanical strength. Young's modulus of LJP-E was 4 times that of the blank. In terms of hemostatic properties, all three polysaccharide hydrogels did not show significant cytotoxic and hemolytic properties. The enzyme- and acid-extracted hydrogels (LJP-Gel-A and LJP-Gel-E) demonstrated better whole-blood coagulant ability compared with the water-extracted hydrogel (LJP-Gel-W), as evidenced by the whole blood coagulation index being half that of LJP-Gel-W. Additionally, the lactate dehydrogenase viabilities of LJP-Gel-A and LJP-Gel-E were significantly higher, at about four and three times those of water extraction, respectively. The above results suggested that LJP-Gel-A and LJP-Gel-E exhibited better blood coagulation capabilities than LJP-Gel-W, due to their enhanced platelet enrichment and adhesion properties. Consequently, these hydrogels are more conducive to promoting coagulation and have good potential for wound hemostasis.

Antioxidant and hemostatic properties of preparations from Asteraceae family and their chemical composition - Comparative studies.

Asteraceae, known as sunflower family, is one of the largest flowering plants family around the world. Sunflower family contains numerous phytochemical compounds. The aim of this study was to describe phytochemical characteristics and investigate the effect of four sunflower vegetable preparations (extracts): chicory leaves (Cichorium intybus), green lettuce leaves (Lactuca sativa), red lettuce leaves (Lactuca sativa var. crispa) and sunchoke roots (Helianthus tuberosus) on different biomarkers of oxidative stress in human plasma in in vitro model. The antioxidant potential was also tested using the DPPH method. The phytochemical composition of the tested preparations was determined by UPLC-ESI-QTOF-MS. All the tested extracts demonstrated antioxidant activity in human plasma. We have observed chicory's and sunchoke's extracts had strongest antioxidant properties in the used models with human plasma. None of the tested vegetables changed ORAC and TAC in vitro. The obtained results suggest that sunflower vegetables might help to prevent oxidative stress related with cardiovascular diseases.

Comparisons of the anti-inflammatory, antiviral, and hemostatic activities and chemical profiles of raw and charred Schizonepetae Spica.

ETHNOPHARMACOLOGICAL RELEVANCE: A common view in traditional Chinese medicine (TCM) theory is that "processing can alter the efficacy of crude drugs". The clinical usage of some processed products may have already changed greatly over time during the development of modern scientific analysis. Therefore, the view of "processing can alter the efficacy of crude drugs" should be confirmed by comparative studies. Schizonepetae Spica (SS), a Chinese medicinal herb, is the dried spike of Schizonepeta tenuifolia Briq. It is available in two forms: raw products and charred products (Schizonepetae Spica Carbonisata, SSC; raw SS processed by stir-frying until carbonization). Raw SS is commonly used to treat TCM symptoms that resemble common cold, fever, respiratory tract infection and allergic dermatitis, while SSC has long been used as a remedy for TCM symptoms that resemble bloody stool and metrorrhagia. AIM OF THE STUDY: We aimed to examine whether stir-fry processing alters the anti-inflammatory, antiviral and hemostatic activities of SS and explore the chemical profile behind the potential changes in medicinal properties caused by stir-fry processing. MATERIALS AND METHODS: We used cell models to examine the anti-inflammatory and antiviral effects of raw SS and SSC. The bleeding time of the tail bleeding model and clotting time of the capillary method in mice were used to compare the hemostasis properties of raw SS and SSC. The chemical profiles of SS and SSC were compared using a method combining gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (HPLC/Q-TOF-MS) analysis. RESULTS: The anti-inflammatory effects of SSC were less potent than those of raw SS. Both raw SS and SSC effectively inhibited viral infection in a dose-dependent manner, with IC50 values of 96.30 and 9.73 µg/mL and selectivity index (SI) values of >1.56 and 7.78, respectively. Interestingly, SSC showed more potent antiviral activities than raw SS. Intragastric administration of raw SS and SSC to mice demonstrated that the hemostatic effects of SSC were more potent than those of raw SS. By comparing the volatile chemical profiles of SSC, we found that twenty-nine constituents disappeared and that fifty-four new constituents were formed while the relative contents of five other components decreased and three other components increased. Additionally, the nonvolatile chemical profiles of raw SS and SSC differed, with thirty-two lower peaks and seven higher peaks in SSC than in SS. CONCLUSION: Our study showed that raw SS and SSC support traditional practice for the clinical applications of these two products except for raw SS used for the treatment of viral infection. It is a fascinating challenge to form SSCs with both traditional hemostatic activities and antiviral properties after stir-fry processing. In addition, the volatile and nonvolatile chemical constituents of raw SS changed dramatically during processing. Further studies are warranted to explore whether the change in chemical constituents is in accordance with the purpose of processing.

Rapid Hemostatic Biomaterial from a Natural Bath Sponge Skeleton.

Uncontrolled bleeding is the main cause of mortality from trauma. Collagen has been developed as an important hemostatic material due to its platelet affinity function. A bath sponge skeleton is rich in collagen, also known as spongin. To understand the hemostatic effect of spongin, spongin materials, SX, SFM and SR were prepared from the bath sponge Spongia officinalis, and hemostatic experiments were performed. The SX, SFM and SR were significantly better than the positive control, type I collagen, in shortening the whole blood clotting time in vitro and hemostasis upon rat tail amputation. In a hemostatic experiment of rabbit common carotid artery injury, the hemostatic time and 3 h survival rate of the SFM group were 3.00 ± 1.53 min and 100%, respectively, which are significantly better than those of the commercial hemostat CELOX-A (10.33 ± 1.37 min and 67%, respectively). Additionally, the SFM showed good coagulation effects in platelet-deficient blood and defibrinated blood, while also showing good biocompatibility. Through a variety of tests, we speculated that the hemostatic activity of the SFM is mainly caused by its hyperabsorbency, high affinity to platelets and high effective concentration. Overall, the SFM and spongin derivates could be potential hemostatic agents for uncontrolled bleeding and hemorrhagic diseases caused by deficiency or dysfunction of coagulation factors.

Anti-inflammatory and hemostatic effects of Linaria reflexa Desf.

The work presented here was aimed to investigate the in vivo anti-inflammatory and in vitro hemostatic activities of Linaria reflexa extract and to establish the relationship between its bioactivity and chemical composition. Twenty-three secondary metabolites were identified, most of them are good anti-inflammatory agents, in line with data by carrageenin-induced rat paw edema assays of the n-butanol extract showing high anti-inflammatory inhibition (63.90%) of edema swelling in the rat paw at the dose 200 mg/kg after 4 h. Furthermore, both extent of inflammatory response and tissue injury were prevented keeping the levels of rate myeloperoxidase (60.16%) and of malondialdehyde, which is the final product of lipid peroxidation generated by free radicals (58.58%). The same extract showed also a remarkable hemostatic effect established by measuring the coagulation time of decalcified plasma (45 s), related to its flavonoid glycosides content.

Evaluation of VEGF mediated pro-angiogenic and hemostatic effects and chemical marker investigation for Typhae Pollen and its processed product.

ETHNOPHARMACOLOGICAL RELEVANCE: Typhae Pollen (TP) is a well-known Traditional Chinese Medicine (TCM) to remove blood stasis. Carbonized Typhae Pollen (CTP), a processed product of TP after being stir-fried, has been widely applied to clinical practice with its capability of hemostasis. However, the underlying mechanism of TP and CTP are still not fully elucidated and discrimination against TP and CTP remains a challenge. AIM OF STUDY: The aim of this study is to investigate whether TP could remove blood stasis by promoting angiogenesis and the process of carbonizing it could enhance hemostatic effect. Meanwhile, some chemical markers for quality control of CTP had better to be found. MATERIAL AND METHODS: The changes of constituents between TP and CTP were analyzed by UPLC-QTOF-MS/MS. We investigated pro-angiogenic and hemostatic effects of TP and CTP in two zebrafish models: VRI-induced ISV insufficiency model and Ator-induced cerebral hemorrhage model. Subsequently, quantitative real-time PCR (qRT-PCR) was applied to investigate the mechanism of pharmacological effects. Finally, chemometric method was applied to find chemical markers. RESULTS: A total of 19 compounds were identified in qualitative analysis. The loss rate of each compound was calculated and compared. Two compounds (huaicarbon A/B) could only be detected in CTP and the content of flavonoid glycosides in CTP was significantly decreased compared with TP. The average content of the three identified flavonoid aglycones (quercetin, isorhamnetin and kaempferol) was increased about 30 percent in CTP. TP promoted pro-angiogenesis by up-regulating the expression of VEGFA, flt1 and kdr. After heating process, the pro-angiogenic activity was reduced and hemostatic activity was enhanced in CTP. Then qRT-PCR analysis found that CTP could significantly up-regulate the expression of VEGFA and vWF. In the discovery of markers, 6 chemical markers for discrimination of TP and CTP were obtained by chemometric method. CONCLUSION: Our research indicated that the pro-angiogenic activity of TP was involved in VEGF signaling pathway. After processing, hemostatic activity of CTP has been enhanced by up-regulating the expression of VEGFA and vWF. A chemical marker database was established to provide a scientific evidence for quality control, mechanism and the clinical application of TP and CTP.

Elucidation of the active ingredients of Lamiophlomis herba against hemorrhage based on network pharmacology and tail snipping model in mice.

This study aimed to elucidate the active ingredients of Lamiophlomis herba (LH), the overground part of Lamiophlomis rotata (Benth.) Kudo, against hemorrhage based on network pharmacology and tail snipping model in mice. A total of 118 hemorrhage-related target genes were identified by retrieving public databases, and 39 genes were identified as the hub genes of hemorrhage based on protein-protein interaction and module analyses. The interactions between 67 potentially active ingredients in LH and 7 genes in the 39 hub genes were established and analyzed through molecular docking and Cytoscape. A total of 21 ingredients were involved in the interactions, and were divided into three categories: iridoid (15 ingredients), flavonoid (2 ingredients) and other category (4 ingredients). Based on the "multi-ingredient, multi-target" characteristic of traditional Chinese medicines (TCMs), the results of network pharmacology indicated that iridoid might be the key active ingredient group of LH against hemorrhage. The contribution of iridoid to the hemostatic effect of LH was investigated by the tail snipping model in mice. The results showed that iridoid was the key active ingredient group of LH against hemorrhage, which confirmed the prediction in network pharmacology. Additionally, the previous reports also supported this prediction. In conclusion, the finding of the present study indicates that iridoid is the key hemostatic ingredient group of LH. This work provides valuable references for investigation of the hemostatic ingredients of LH based on the holistic theory of TCMs. Meanwhile, this work also provides further insight into the development of hemostatic drugs based on LH.

Hemostatic activity of aqueous extract of Myrtus communis L. leaf in topical formulation: In vivo and in vitro evaluations.

ETHNOPHARMACOLOGICAL RELEVANCE: Myrtus communis L. (MC) is a well-known medicinal plant in traditional Persian medicine, which contains a large amount of phenolic compounds (mainly hydrolyzable tannins). As mentioned in ancient literature, MC was widely used to control bleeding in every part of the body. Nevertheless, there is no pharmacological study on the anti-hemorrhagic activity of this plant till now. AIM OF THE STUDY: The current in vivo and in vitro study aimed at evaluating the hemostatic activity of M. communis aqueous leaf extract (MCE) in topical formulation. MATERIALS AND METHODS: Two parameters of bleeding time and amount in tail bleeding model were measured in vivo in rats treated with MCE (1%, 2.5%, 5%, 7.5%, 10%, 15%, and 20% w/v), 5% M. communis aqueous leaf extract gel (G), tannic acid (TA) (1%, 2.5%, 5%, 7.5%, and 10%), normal saline (NS), and the Monsel's solution (MS), a commercial hemostatic agent. Also, the effect of 5% MCE and 5% TA on PT (prothrombin time) and aPTT (activated partial thromboplastin time) as well as protein precipitation and platelet aggregation were assessed in vitro. RESULTS: In the rat-tail bleeding model, bleeding time and amount significantly (P < 0.001) reduced by the application of 5% MCE solution on the cut tail compared with the NS group. The bleeding time and amount in the MS group were not significantly different from those of the 5% MCE group. Platelet microaggregates were detected by fluorescent microscope. PT and aPTT values increased >120 s and >180 s by 5% MCE, respectively. Also, protein precipitation and significant reduction in serum proteins were observed in the 5% MCE group. CONCLUSION: The current study provided new insights into the hemostatic effect of MCE, which may be partially mediated by platelet aggregation activity. Hence, it could be evaluated as the resource of new plant origin hemostatic agent.

Fabrication of an expandable keratin sponge for improved hemostasis in a penetrating trauma.

Effective hemostasis improvements for penetrating traumas remain a research priority for civilian and noncivilian applications. Herein, we fabricated an expandable keratin sponge (EKS) for the hemostatic treatment of a penetrating trauma based on the excellent hemostatic ability of keratin and the expandable property of polyacrylamide (PAM). EKSs with semi-interpenetrating networks were fabricated by radical polymerization of keratin and PAM, and the EKS showed rapid expansion upon blood absorption. This sponge exhibited effective hemostasis on a rat penetrating liver hemorrhage, and the expansion of the EKS was dependent on the bleeding volume. In addition, the results of a shear wave elastography analysis showed that the elasticity of the liver tissue increased from 12.5 kPa to 21.2 kPa after the penetrating liver trauma treated by the EKS, and the mechanical strength of the liver tissue was maintained after 1 h of the EKS application. Further in vivo tests indicated the effectiveness of the EKS for hemostasis in a swine femoral artery transection hemorrhage model. This EKS is promising for hemostatic applications.

Chemical structure and snake antivenom properties of sulfated agarans obtained from Laurencia dendroidea (Ceramiales, Rhodophyta).

Aqueous and KCl-soluble polysaccharides were extracted from Laurencia dendroidea (Rhodomelaceae, Ceramiales) and their chemical profile was accessed by anion-exchange chromatography, chemical and spectroscopic analyses. The homogeneous agaran DHS-4 (181.3 × 103 g. mol-1, 21.3% of NaSO3) presents A units mostly 2-sulfated (18.9 mol%), nonsubstituted (15.3 mol%) and 6-O-methylated (10.1 mol%), while B units are l-sugars composed predominantly by galactose 6-sulfate precursor units (19.2 mol%) and 3,6-anhydrogalactose (13.8 mol%), besides non-precursor galactose 6-sulfate units bearing d-xylose substituents on C-3 (8.1 mol%). The crude KCl-soluble DHS agaran (20.5% of NaSO3) inhibited proteolysis and hemolysis induced by Lachesis muta and Bothrops jararaca venoms. DHS was able to inhibit up to 75% the L. muta venom hemorrhagic effect and to reduce the lethality displayed by B. jararaca venom, increasing the mice survival time up to 3 times. Therefore, this agaran has high potential to be used as an additional tool to treat snakebite envenomation.

Identification of certain Panax species to be potential substitutes for Panax notoginseng in hemostatic treatments.

Panax notoginseng (Burkill) F. H. Chen ex C. H. Chow (P. notoginseng) is a highly valued Chinese materia medica having a hemostatic effect and mainly used for the treatment of trauma and ischemic cardiovascular diseases. Stringent growth requirements, weak resistance to insect pests and plant diseases, arsenic contamination and continuous cropping constitute hurdles to further increases in the agricultural production of P. notoginseng. This review focuses on the traditional uses (based on traditional Chinese medicine theory), major chemical components, biological activities, pharmacological properties, geographical distributions and historical development of taxonomy of P. notoginseng and its related species in Panax genus, including Panax japonicus C. A. Meyer (P. japonicus), Panax japonicus C. A. Meyer var. major (Burkill) C. Y. Wu et K. M. Feng (P. japonicus var. major) and Panax japonicus C. A. Meyer var. bipinnatifidus (Seem.) C. Y. Wu et K. M. Feng (P. japonicus var. bipinnatifidus) are reviewed. This review sheds light on the origin herbs of Zhujieshen (ZJS) and Zhuzishen (ZZS), e.g., P. japonicas var japonicas, P. japonicus var. major and P. japonicus var. bipinnatifidus could be used as a substitute for P. notoginseng as hemostatic herbs.

Comparing coagulation activity of Selaginella tamariscina before and after stir-frying process and determining the possible active constituents based on compositional variation.

CONTEXT: Selaginella tamariscina (P. Beauv.) Spring (Selaginellaceae) (ST) has been widely used in China as a medicine for improving blood circulation. However, its processed product, S. tamariscina carbonisatus (STC), possesses opposite haemostatic activity. OBJECTIVE: To comprehensively evaluate the activity of ST and STC on physiological coagulation system of rats, and seek potential active substances accounting for the activity transformation of ST during processing. MATERIALS AND METHODS: The 75% methanol extracts of the whole grass (fine powder) of ST and STC were prepared, respectively. Male Sprague-Dawley rats were randomly divided into five groups: control group, model group, model + ST group, model + STC group and positive control group (model + Yunnanbaiyao). The duration of intragastric administration was 72 h at 12 h intervals. Haemorheology parameters were measured using an LB-2 A cone-plate viscometer and the existed classic methods, respectively. SC40 semi-automatic coagulation analyzer was employed to determine coagulation indices. Meanwhile, HPLC and LC-MS were applied for chemical analyses of ST and STC extracts. RESULTS: STC shortened tail-bleeding time, increased whole blood viscosity (WBV) and plasma viscosity (PV), decreased erythrocyte sedimentation rate blood (ESR), reduced activated partial thromboplastin time (APTT) and increased the fibrinogen (FIB) content in the plasma of bleeding model rats. Although ST could shorten APTT and TT, the FIB content was significantly decreased by ST. Dihydrocaffeic acid with increased content in STC vs. ST showed haemostatic activity for promoting the platelet aggregation induced by collagen and trap-6, and reducing APTT and PT significantly with a concentration of 171.7 µM in vitro. Amentoflavone with reduced content in STC vs. ST inhibited ADP and AA-induced platelet aggregation significantly with a concentration of 40.7 µM. DISCUSSION AND CONCLUSIONS: As the processed product of ST, STC showed strong haemostatic activity on bleeding rat through regulating the parameters involved in haemorheology and plasma coagulation system. Two active compounds, dihydrocaffeic acid and amentoflavone, might be partially responsible for the haemostatic and anticoagulant activity of STC and ST, respectively.

Evaluation of Artemisia scoparia for hemostasis promotion activity.

Excessive hemorrhage through any reasons is a life threatening process. Artemisia scoparia of family Asteraceae has been used in local system of medicine to stop bleeding from wounds and in injuries, antiseptic, in healing urticarial and for removal of worms from the body. Aerial parts of A. scoparia was extracted with 95% methanol (ASM) and fractionated through liquid-liquid partition in ascending order of n-hexane (ASH), chloroform (ASC), ethyl acetate (ASE), and the remaining as the aqueous fraction (ASA). Phytochemical classes of the extract/fractions were determined by qualitative assays. Prothrombin time (PT) was estimated on the plasma of human blood by Owren method. Capillary tube method was applied to determine the hemostasis activity in Sprague-Dawley rat. Tannins, saponins, terpenoids, quinones, betacyanins and flavonoids were present whereas phlobatannins, anthraquinones and alkaloids were established absent in ASM, ASC, ASE and ASA. Prothrombin time was significantly decreased by mixing (10 µg) of ASM (16.67±1.15 sec), ASH (12.33±0.57 sec), ASC (15.33±0.57 sec) and ASA (9.0±1.0 sec) to that of vehicle (20.0±1.0 sec). Administration (200 mg/kg) of all the extract/fractions showed significantly less (26.00±11.79 sec - 41.00±7.21 sec) hemostasis time as compared to the (242.67±39.67 sec) control rats. The results suggested the therapeutics importance of A. scoparia use in bleeding pathologies.

Isolation, Characterization and Evaluation of Collagen from Jellyfish Rhopilema esculentum Kishinouye for Use in Hemostatic Applications.

Hemostat has been a crucial focus since human body is unable to control massive blood loss, and collagen proves to be an effective hemostat in previous studies. In this study, collagen was isolated from the mesoglea of jellyfish Rhopilema esculentum Kishinouye and its hemostatic property was studied. The yields of acid-soluble collagen (ASC) and pepsin-soluble (PSC) were 0.12% and 0.28% respectively. The SDS-PAGE patterns indicated that the collagen extracted from jellyfish mesoglea was type I collagen. The lyophilized jellyfish collagen sponges were cross-linked with EDC and interconnected networks in the sponges were revealed by scanning electron microscope (SEM). Collagen sponges exhibited higher water absorption rates than medical gauze and EDC/NHS cross-linking method could improve the stability of the collagen sponges. Compared with medical gauze groups, the blood clotting indexes (BCIs) of collagen sponges were significantly decreased (P < 0.05) and the concentration of collagen also had an influence on the hemostatic property (P < 0.05). Collagen sponges had an improved hemostatic ability compared to the gauze control in tail amputation rat models. Hemostatic mechanism studies showed that hemocytes and platelets could adhere and aggregate on the surface of collagen sponge. All properties make jellyfish collagen sponge to be a suitable candidate used as hemostatic material and for wound healing applications.

Exploring hemostatic and thrombolytic potential of heynein - A cysteine protease from Ervatamia heyneana latex.

ETHNOPHARMACOLOGICAL RELEVANCE: The latex of Ervatamia heyneana (Wall.) T. Cooke plant has been used for wound healing and various skin diseases by Indian tribes and folklore. AIM OF THE STUDY: To validate the scientific basis of heynein - a key protease of Ervatamia heyneana, in hemostasis and wound healing process. MATERIALS AND METHODS: The latex from E. heyneana was processed and subjected to two step purification. The purified heynein was assayed for proteolytic activity using casein as substrate and also attested by zymography. The inhibition studies confirmed the nature of heynein. Pure fibrinogen was used for fibrinogenolytic activity and citrated plasma was used for coagulant and fibrinolytic activities. The edema inducing action and hemorrhagic activity of heynein were assessed on mice model. RESULTS: The purified heynein exhibited proteolytic activity, which was confirmed by caseinolytic assay and zymography. The inhibition studies confirmed heynein to be a cysteine protease. Heynein showed complete hydrolysis of all the three subunits of human fibrinogen (Aα, Bß, γ). It exhibited strong pro-coagulant activity by reducing plasma clotting time from 248 to 39s at 40µg concentration. Heynein cleaved α polymer subunit in fibrin clot and did not induce edema and hemorrhage in mice models. The non-hemorrhagic nature was supported with histopathological studies of skin samples. CONCLUSION: Heynein displays strong pro-coagulant action associated with fibrin(ogen)olytic activity. This provides basis for the observed pharmacological action of Ervatamia heyneana and thereby justifies its use in folk medicine.

Bletilla striata: Medicinal uses, phytochemistry and pharmacological activities.

ETHNOPHARMACOLOGICAL RELEVANCE: Bletilla striata (Thunb.) Reichb. f. (Orchidaceae), also known as Hyacinth Orchid and Baiji (Simplified Chinese: ), not only has been widely used for the treatment of hematemesis, hemoptysis, and traumatic bleeding due to the efficacy of arresting bleeding with astringent action, but also has been applied topically to overcome ulcers, sores, swellings, and chapped skin due to the efficacy of dispersing swelling and promoting tissue regeneration. Additional medical applications include the treatment of tuberculosis, malignant ulcers, hemorrhoids, anthrax, eye diseases, and silicosis. AIM OF THIS REVIEW: This review aims to provide up-to-date information on the botanical characterization, medicinal uses, chemical constituents, pharmacological activities, and toxicity of B. striata. In addition, this paper also focuses on the possible exploitation of this plant for the treatment of different diseases, and uncovers opportunities for future research. MATERIALS AND METHODS: The relevant information on B. striata was gathered from worldwide accepted scientific databases via an electronic search (Google Scholar, Web of Science, ScienceDirect, ACS Publications, PubMed, Wiley Online Library, SciFinder, CNKI). Information was also obtained from The Plant List, Chinese pharmacopoeia, Chinese herbal classics books, PhD and MSc dissertations, etc. RESULTS: A comprehensive analysis of the literature obtained through the above-mentioned sources confirmed that the ethnomedical usages of B. striata have been recorded in Mongolia, Korea, Japan, and China. Phytochemical investigations revealed that the major chemical constituents of B. striata are polysaccharides, bibenzyls, phenanthrenes, triterpenoids and its saponins, steroids and its saponins, which also have been proven to be the main bioactive substances capable of exhibiting numerous pharmacological activities including wound healing, antiulcer, hemostasis, cytotoxicity, antimicrobial, anti-inflammation, anti-oxidation, immunomodulation, anti-fibrosis, antiaging, anti-allergy, and anti-itch. CONCLUSIONS: Preliminary investigations on pharmacological properties of B. striata have shown that B. striata is an outstanding astringent hemostatic medicinal, B. striata polysaccharides (BSP) as the major bioactive components not only capable of promoting wound healing, but also show good performance as a kind of promising natural biomaterial. More importantly, BSP are also reported to be excellent embolic material. However, further investigations need to be carried out to fully clarify its efficacy of dispersing swelling and promoting tissue regeneration. Moreover, this plant also needs a lot more investigations to clarify the pathways of absorption, distribution, metabolism and excretion, and to evaluate its long-term in vivo chronic toxicity before proceeding to the development of pharmaceutical formulation.

Spirostanol glycosides with hemostatic and antimicrobial activities from Trillium kamtschaticum.

Ten spirostanol glycosides, trillikamtosides A-J, together with eleven known analogues, were isolated from the hemostatic fraction of the 75% aqueous EtOH extract of the whole herbs of Trillium kamtschaticum. Their structures were established by extensive spectroscopic data analysis and chemical methods. The aglycones of three of these compounds had unique 3ß,17α-dihydroxy-spirostanes featuring a double bond between C-4 and C-5, while two others represent a rare class of spirostanol glycosides which possess a 5(6 â†’ 7) abeo-steroidal aglycone. All the compounds were evaluated for their hemostatic and antimicrobial activities. Three of the spirostanol glycosides exhibited induced-platelet aggregation at a concentration of 300 µg/mL with maximal induced-platelet aggregation rates of 72%, 71%, and 62% in rabbits, respectively, and their EC50 values were 492.7, 203.3, and 109.8 µM. Five of the spirostanol glycosides showed an anti-Candida albicans effect with MIC values of 21.1, 10.6, 8.8, 21.6, and 11.0 µM, respectively.

Orthogonal optimization of prokaryotic expression of a natural snake venom phospholipase A2 inhibitor from Sinonatrix annularis.

Phospholipase A2 (PLA2) is a calcium-dependent enzyme that is involved in inflammatory processes such as the liberation of free arachidonic acid from the membrane pool for the biosynthesis of eicosanoids. Snake venom are known containing PLA2s (svPLA2s) which exhibit a wide variety of pharmacological effects including neurotoxicity, cardiotoxicity, myotoxicity and hemorrhage. Therefore, inhibition of svPLA2 would be advantageous to successful envenomation treatment. A gamma type PLI (PLA2 inhibitor) has been extracted from the serum of Sinonatrix annularis, a non-venomous snake indigenous to China. This showed strong inhibition of Deinagkistrrodon acutus PLA2, however, the PLIγ level in the serum and snake resource are not sufficiently sustainable for further research. To overcome these limitations, we constructed a His6-PLIγ pET28 fusion expression vector and transformed Escherichia coli BL21. To improve the expression of PLIγ, an orthogonal experiment [L16(4)(5)] was performed to optimize induction parameters. The optimized condition was determined to be: induction by 0.4 mM isopropyl-ß-D-thiogalactoside (IPTG) for 6 h to the recombinant BL21 after its OD600 was 0.8, with continuous shaking cultivation at 190 rpm and 35 °C. Under these conditions, the amount of expressed protein could reach 57 mg/L. The His6-PLIγ was purified by nickel affinity chromatography and renatured by On-column refolding. The resulting PLIγ showed a good inhibitory effect of enzymatic activities to venom PLA2 isolated from D. acutus. Moreover, the PLIγ had a wide anti-hemorrhage activities to D. acutus, Naja atra and Agkistrodon halys venom.