Does hypericin boost the efficacy of high-power laser? A preliminary experimental study on rats / A hipericina aumenta a eficácia do laser de alta potência? Um estudo preliminar e experimental em ratos

CONTEXT AND OBJECTIVE: Lasers are widely used in treating symptomatic benign prostatic hyperplasia. In current practice, potassium titanyl phosphate (KTP) lasers are the most common type of laser systems used. The aim here was to evaluate the rapid effect of high-power laser systems after application of hypericin. DESIGN AND SETTING: Experimental animal study conducted in the Department of Urology, Gülhane Military Medical Academy, Ankara, Turkey, in 2012. METHODS: Sixteen rats were randomized into four groups: 120 W KTP laser + hypericin; 120 W KTP laser alone; 80 W KTP laser + hypericin; and 80 W KTP laser alone. Hypericin was given intraperitoneally two hours prior to laser applications. The laser incisions were made through the quadriceps muscle of the rats. The depth and the width of the laser incisions were evaluated histologically and recorded. RESULTS: To standardize the effects of the laser, we used the ratio of depth to width. These new values showed us the depth of the laser application per unit width. The new values acquired were evaluated statistically. Mean depth/width values were 231.6, 173.6, 214.1 and 178.9 in groups 1, 2, 3 and 4, respectively. The most notable result was that higher degrees of tissue penetration were achieved in the groups with hypericin (P < 0.05). CONCLUSIONS: The encouraging results from our preliminary study demonstrated that hypericin may improve the effects of KTP laser applications. .

CONTEXTO E OBJETIVO: Lasers são amplamente utilizados no tratamento de hiperplasia benigna de próstata sintomática. Na prática atual, lasers de fosfato de titanilo de potássio (KTP) são os tipos mais comuns usados dos sistemas. O objetivo foi avaliar o efeito rápido do sistema laser de alta potência após a aplicação de hipericina. TIPO DE ESTUDO E LOCAL: Estudo experimental animal, realizado no Departamento de Urologia, Academia de Medicina Militar de Gülhane, Ancara, Turquia, em 2012. MÉTODOS: 16 ratos foram divididos aleatoriamente em 4 grupos: 120W KTP laser + hipericina; 120W KTP laser somente; 80W KTP laser + hipericina; 80W KTP laser somente. Hipericina foi dada intraperitonealmente duas horas antes da aplicação do laser. As incisões a laser foram feitas através do músculo quadríceps dos ratos. A profundidade e a largura das incisões a laser foram avaliadas histologicamente e registradas. RESULTADOS: Para padronizar o efeito do laser foi utilizada a razão entre profundidade e largura. Estes novos valores nos mostraram a profundidade da aplicação do laser de largura por unidade. Os novos valores adquiridos foram avaliados estatisticamente. Os valores da média de profundidade/largura foram 231,6, 173,6, 214,1 e 178,9 nos grupos 1, 2, 3 e 4, respectivamente. O resultado mais notável foi atingir altos graus de penetração tecidual nos grupos com hipericina (P < 0,05). CONCLUSÕES: Os resultados promissores do nosso estudo preliminar mostraram que hipericina pode melhorar os efeitos das aplicações do laser KTP. .

Hypericin: chemical synthesis and biosynthesis / 中国天然药物

Hypericin is one of the most important phenanthoperylene quinones extracted mainly from plants of the genus Hypericum belonging to the sections Euhypericum and Campylosporus of Keller's classification. Widespread attention to the antiviral and anti-tumor properties of hypericin has spurred investigations of the chemical synthesis and biosynthesis of this unique compound. However, the synthetic strategies are challenging for organic and biological chemists. In this review, specific significant advances in total synthesis, semi-synthesis, and biosynthesis in the past decades are summarized.

A new perylenequinone from a halotolerant fungus, Alternaria sp. M6 / 中国天然药物

AIM@#To study the metabolites of a halotolerant fungus Alternaria sp. M6.@*METHODS@#The metabolites were isolated and purified by various chromatographic techniques. Their structures were determined on the basis of physical properties and spectroscopic data.@*RESULTS@#Nine compounds were isolated and identified as 8β-chloro-3, 6aα, 7β, 9β, 10-pentahydroxy-9, 8, 7, 6a-tetrahydroperylen-4(6aH)-one (1), alterperylenol (2), dihydroalterperylenol (3), adenine (4), adenosine (5), deoxyadenosine (6), guanosine (7), tryptophan (8), and hexadecanoic acid (9).@*CONCLUSION@#Compound 1 is a new perylenequinone.

Simultaneous determination of hypocrellin A, hypocrellin B, and hypocrellin C by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To develop a high-performance liquid chromatography (HPLC) method for simultaneous determination of hypocrellin A, hypocrellin B, and hypocrellin C.</p><p><b>METHOD</b>The separation was carried out on a Kromasil C18 (4.6 mm x 250 mm, 5 micrm) colum eluted with in mobile phases of water containing 0.5% glacial acetic acid and acetonitrile. The column temperature was 35 degrees C, and the flow rate was 1.0 mL x min(-1). The detection wavelength was set at 265 nm.</p><p><b>RESULT</b>The three compounds were well separated. Calibration curves of hypocrellin A, hypocrellin B, and hypocrellin C showed good linear relationship RSD > 2.0%. The average recoveries of the hypocrellin A, hypocrellin B, and hypocrellin C were 101.8%, 102.3%, 100.0%, respectively.</p><p><b>CONCLUSION</b>The developed method is simple, accurate, and repeatable, and can be readily used as valid tool for the quality control of Hypocrella bambusae.</p>

Rapid cloning and functional characterization of hypericin synthase gene / 药学学报

Hypericin, a red-colored naphtodianthrone, is a natural product synthesized in the medicinal plant Hypericum perforatum, commonly known as St. John's wort. Hypericin has attracted a growing attention of the pharmaceutical industry because of its potential application to various therapies, including the treatment of depression and remarkable antiviral and photodynamic activities, hyp-1 gene encodes for phenolic coupling protein which catalyzes in vitro direct and specific conversion of emodin to hypericin which, however, has not formed common opinion so far. Six pairs of primers specific to hyp-1 gene were synthesized. The rapid cloning of hyp-1 gene was performed based on step-by-step extension of a short region of the gene through a series of PCR reactions. All cloned sequences were confirmed by DNA sequencing. A vector named pET32ahyp containing hyp-1 gene was constructed and was transformed into E. coli to induce heterologous expression. SDS-PAGE and Western blot results showed the recombinant Hyp-1 protein was expressed successfully in E. coli. The soluble fraction was used to test the function of the recombinant Hyp-1. Hypericin was detected by LC-MS/MS with emodin as a substrate under in vitro conditions. The above results corroborated the Hyp-1 function, a confusing question, which lay a material foundation for the synthesis of hypericin by synthetic biotechnology.

Immunomodulatory Effects of Hypocrellin A on MHC-restricted Antigen Processing

Hypocrellin A has gained much attention in recent years due to its light-induced antitumor, antifungal and antiviral activities. Here we report that hypocrellin A exerts immunomodulatory effects on MHC-restricted presentation of antigen. Hypocrellin A inhibited class II-MHC restricted presentation of exogenous antigen, but not class I MHC-restricted presentation of exogenous antigen, in dendritic cells. Hypocrellin A also inhibited the cytosolic pathway of endogenous antigen presentation. However, hypocrellin A did not inhibit the expression of class I and class II MHC molecules on dendritic cells (DCs), the phagocytic activity of DCs, or the H-2K(b)-restricted presentation of a synthetic peptide, SIINFEKL. These results show that hypocrellin A differentially modulates the MHC-restricted antigen presentation pathways.

Cluster analysis on hypocrellin A content in anamorphs of Shiraia bambusicola / 中国中药杂志

<p><b>OBJECTIVE</b>By examining the colony growth rate, colony texture, substrate mycelium color and separation method of 40 anamorph strains of Shiraia bambusicola to investigate the relationship between these indexes and content of hypocrellin A.</p><p><b>METHOD</b>In UV-spectrophotometry and microscopic observation were applied in the experiments. The results were analyzed by the clustering and variance analysis.</p><p><b>RESULT</b>SPSS clustering analysis showed that the colony characters of S. bambusicola anamorph strains had significant differences with hypocrellin A content.</p><p><b>CONCLUSION</b>In the tested conditions, substrate mycelium color of S. bambusicola anamophs has the most distinct correlation with the yield of hypocrellin A.</p>

Estudo experimental da inibição da proliferação vitreorretiniana pelo uso da hiperecina / Experimental study of vitreoretinal proliferation inhibition by the use of hypericin

OBJETIVOS: Induzir a produção de membranas vitreorretinianas em modelo de trauma ocular animal. Avaliar a inibição do desenvolvimento da proliferação vitreorretiniana (PVR) com o uso de hiperecina. MÉTODOS: Estudo Experimental. Foram utilizados 19 coelhos machos pigmentados adultos com peso entre 2.000 e 3.000 gramas. Todos submetidos a modelo de trauma com dispase associada à diatermia da retina para indução de membranas de PVR. Separados randomicamente para receberem hiperecina (10 µM em 0,1 ml) ou solução salina (0,1 ml) como placebo. Avaliados clinicamente no sétimo, décimo quarto, vigésimo primeiro e vigésimo oitavo dias de pós-operatório com oftalmoscopia indireta e retinografia colorida digitalizada. O grau de PVR foi classificado em estágios (de 0 a 7) segundo Hida e colaboradores. RESULTADOS: A formação de membranas esteve presente em 79 por cento dos olhos, sendo 100 por cento nos olhos do grupo placebo e 60 por cento nos olhos do grupo tratamento (hiperecina). A comparação entre as médias dos estágios de PVR entre os grupos mostrou diferença estatisticamente significativa, com valor p=0,0321 pelo teste Wilcoxon. CONCLUSÕES: O modelo de trauma com uso de dispase e diatermia da retina produz membranas vitreorretinianas. A hiperecina mostrou-se eficaz na diminuição do aparecimento e progressão do PVR.

PURPOSE: To produce proliferative vitreoretinopathy (PVR) in an animal ocular trauma model. To evaluate the inhibition of (PVR) emergence and progression by hypericin. METHODS: Experimental Study. Nineteen pigmented male adult rabbits weighing between 2,000 and 3,000 grams were used in this study. All of them were submitted to trauma model with dispase and retinal diathermy to induce PVR membranes formation. They were randomly assigned to receive hypericin (10 µM in 0.1 ml) or saline solution (0.1 ml) as placebo. They were evaluated clinically in the seventh, fourteenth, twenty-first and twenty-eighth postoperative days with indirect ophthalmoscopy and digital color retinography. The PVR degree was classified according to Hida (0 to 7). RESULTS: Membranes formation was present in 79 percent of the eyes; being 100 percent in the eyes of placebo group and 60 percent in the eyes of treatment group (hypericin). The comparison between PVR phases averages within the groups showed a statistically significant difference between the two groups, with a p value of 0.0321 for Wilcoxon test. CONCLUSIONS: The trauma model with dispase and retinal diathermy produces vitreoretinal membranes. Hypericin was considered effective in PVR emergence and progression decrease.

Dereplication of bioactive constituents of the genus hypericum using LC-[+,-]-ESI-MS and LC-PDA techniques: Hypericum triquterifolium as case study

Utilizing liquid chromatography-electro spray ionization-mass spectrometry [LC-[+,-]-ESI-MS] and liquid chromatography-photodiode array detection [LC-PDA] techniques, a dereplication strategy for the analysis of the secondary metabolites constituents of the genus Hypericum has been developed. From the crude methanolic extract of the aerial parts of H. triquetrifolium [leaves, stems, and flowers] and on the basis of their UV- profiles, chromatographic retention times and [+,-]-ESI-MS [TIC and SIM] mass spectral data, seven known [1-7] compounds were dereplicated fairly rapidly. The compounds were classified into three structural classes: phloroglucinols: hyperfirin and adhyperfirin; naphthodianthrones: hypericin, pseudo-hypericin, proto-hypericin, and protopseudo-hypericin; and the flavonoid rutin

Morphological identification of 20 medicinal species in Hypericum / 中国中药杂志

<p><b>OBJECTIVE</b>To provide anatomical evidences for the morphological and histological identification of 20 medicinal species in Hypericum.</p><p><b>METHOD</b>Morphological and anatomical study on the organs of 20 medicinal species in Hypericum using tissue clearing, paraffin sectioning and thin sectioning.</p><p><b>RESULT</b>According to their anatomical characteristics, the secretory structures can be divided into nodules, secretory cavities (canals) and tiny secretory tubes of 20 medicinal species in Hypericum. Hypericin was produced and stored in the nodules, while the volatile oil was produced and stored in the secretory cavities (canals) and tiny secretory tubes. The types differed markedly from each other in location, diameter and distributional density of leaf, and the anatomical structures differed from each other of stem, calyx, petal, anther and fruit among the 20 species in Hypericum.</p><p><b>CONCLUSION</b>The secretory structures may be as anatomical evidences for the morphological and histological identification of 20 medicinal species in Hypericum.</p>

Evaluation of preparing process of extract of hypericum perforatum using HPLC fingerprint analysis / 中国中药杂志

<p><b>OBJECTIVE</b>To establish the quality control method of the extract enriched hypericins and flavonoids (HFEE) prepared from Hypericum perforatum by means of HPLC fingerprint analysis, and to evaluate the validity, stability of within-day and between-day of the preparing process of HFEE as well as the influence of antioxidant and the herbal source to the quality of HFEE.</p><p><b>METHOD</b>HPLC-UV-MS was employed to the qualitative and quantitative analyses of HFEE.</p><p><b>RESULT</b>Ten peaks on the HPLC fingerprint of HFEE were indicated, and eight compounds of them had been identified. With respect of the preparing process of HFEE, the validity and stability were significantly observed. There was almost no effect observed on the quality of HFEE whether or not adding the antioxidant during the process. However, different parts of the plant collected as the materials could significantly affect the quality of HFEE.</p><p><b>CONCLUSION</b>The method established in the present study is convenient, reliable, and could be used for the quality control of the extract of H. perforatum and for the monitor and control of the preparing process of the extract.</p>

Relationship between apoptosis induced by 2-butylamino-2-demethoxy-hypocrellin B in human pancreatic cancer cells Capan-1 and photosensitization of mitochondria / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the possible mechanism of apoptosis induced by photodynamic therapy (PDT) in human pancreatic cancer cells Capan-1 with 2-butylamino-2-demethoxy-hypocrellin B (BAHB) as photosensitizer.</p><p><b>METHODS</b>The localization of BAHB in Capan-1 cells was studied, apoptosis was determined by DNA gel electrophoresis after PDT. The mitochondria membrane potential (DYm) and cytochrome C release were observed by laser scan confocal microscopy and Western blotting.</p><p><b>RESULTS</b>The low concentration photosensitizer was mainly localized in mitochondria and also in lysosomes when the concentration is high. DNA ladder analysis showed characteristic of apoptosis. The mitochondria membrane potential (DYm) showed a loss of 30% around, after 6 hours by PDT under laser scan confocal microscopy, which is caused by a sudden increase in the permeability of mitochondria membrane accompanied with apoptosis. In Western blotting, cytochrome C release was observed from the mitochondria into the cytoplasm during BAHB-induced apoptosis.</p><p><b>CONCLUSION</b>The research suggests that BAHB-induced apoptosis is related to photosensitization of mitochondria.</p>

analysis of force between attractive magnets used in orthodontics

To compare the results of hard and soft attractive magnet combinations to the gold standard Neodymium Iron Boron Magnets used for extrusion of impacted maxillary canines. Experimental and comparative study. Eastman Dental Institute and Hospital, London, UK in collaboration with Department of Physics and Astronomy University College of London [UCL] during 1996-1998. In vitro experiments with combinations of hard and soft magnets of different morphologies. Load Cell Transducer was used to record the force levels in grams with magnets placed in different vertical, horizontal and angular offsets. Three-dimensional mesh plots were created to visualize force level decay with increasing distance. The results were compared with previous study on hard magnet combinations only and observed for "edge effect". The present study force levels were very low albeit the edge effect was negligible. Although the edge effect phenomenon present with powerful hard magnets was minimal or nearly absent with the hard and soft magnet combination - the overall force levels were too low to be of any use in clinical scenario for extrusion of deep impacted canines

Enhancement of hypericin production and cell growth of Hypericum perforatum L. suspension cultures by nitric oxide / 生物工程学报

Nitric oxide has emerged as a key signaling molecule in plants recently. The role of nitric oxide in elicitor-induced defense responses of plants has been extensively investigated. In this work, sodium nitroprusside was utilized as the donor of nitric oxide to investigate the effects of exogenous nitric oxide on hypericin production and cell growth of suspension cell cultures of Hypericum perforatum L.. Compared with the untreated Hypericum perforatum L. suspension cells, external application of 0.5 and 15.0 mmol/L sodium nitroprusside induced 1.4 and 0.5-fold dry cell weight, and 0.9 and 2.1-fold hypericin content respectively. The results showed that low concentration of sodium nitroprusside promoted the growth of Hypericum perforatum L. suspension cells, while high concentration of sodium nitroprusside enhanced hypericin biosynthesis in Hypericum perforatum L. suspension cells. The maximum hypericin production was achieved by adding 0.5 mmol/L and 15.0 mmol/L sodium nitroprusside to the culture at day 0 and day 14 respectively, increasing the total hypericin yield by nearly 3.2-fold. The effects of sodium nitroprusside on hypericin content and growth of Hypericum perforatum L. suspension cells were abolished by nitric oxide specific scavenger 2-4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, which indicated that the effects of the application of sodium nitroprusside were caused by nitric oxide released from sodium nitroprusside rather than sodium nitroprusside itself. The results also showed that 15.0 mmol/L sodium nitroprusside stimulated the activities of phenylalanine ammonia-lyase (PAL), one of the key enzymes of phenylpropanoid pathway, in suspension cells of Hypericum perforatum L., which suggested that the synthetic pathway of hypericin might be activated by NO through triggering the defense responses of Hypericum perforatum L. suspension cells.

Effects of hypericin associated with light irradiation on human laryngeal squamous cell carcinoma strain Hep-2 / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study effects of hypericin associated with light irradiation on human laryngeal squamous cell carcinoma strain Hep-2.</p><p><b>METHODS</b>Using techniques of tumor cells culture in vitro, Hep-2 cells were exposed to different concentration hypericin as 0.5, 1.0, 2.0, 3.0, 5.0 microg/ml, then 10 minutes 7.5 J/cm2 light irradiation was given after an hour. Other groups Hep-2 culture cells were also exposed to different concentration hypericin as 5.0, 10.0, 20.0, 25.0 microg/ml, without light irradiation. In all groups, contrast groups were set up. And 48 hours later, growth characteristics of Hep-2 cells were studied by morphological observation, fluorescence microscope, MTT assay and flow cytometry.</p><p><b>RESULTS</b>In normal contrast group, Hep-2 cells grew intensively and contacted with each other. However, cells which were treated with hypericin, combination with light irradiation were declined greatly. In higher dose hypericin group, necrosis could be found. MTT assay showed that hypericin associated with light irradiation inhibited growth of laryngeal cell with dose dependence manner. Flow cytometry showed that hypericin with light irradiation could block cell growth at G0/G1 phase, inducing apoptosis of laryngeal cell. Under fluorescence microscope, some sings of cell apoptosis including coagulation of chromatin, fragmentation of nuclei and apoptotic body could be found.</p><p><b>CONCLUSION</b>Human laryngeal squamous cell carcinoma strain Hep-2 can be inhibited and induced into apoptosis by treated with hypericin combination with light irradiation.</p>

Interaction of Hypericin Embedded in Liposomes with the ocular tissue of new Zealand rabbits for application in photodynamic therapy

The potent photodynamic properties of hypericin [HY] elicit a range of light-dependant virucidal and tumoricidal activities. This work highlights the interaction of liposomes loaded with HY with the aqueous humor and crystalline lens of the rabbits' eye in comparison with those treated with HY only. Six group of New Zealand rabbits of six rabbits each were used in this study. The rabbits received 30 micro l of HY or liposomes loaded with hypericin topically once a day for one week as an eye drop of HY concentration 1-9 micro M. The eyes of rabbits were exposed to He-Ne laser for 5 min after 30 min from administration of HY or liposomes loaded with HY for the studied groups. Total soluble protein, molecular weight distribution, electrophoretic mobility and refractive index for both aqueous humor and soluble lens crystallins were investigated for the studied samples. The obtained results showed an increase in the solubility of lens protein and aqueous humor, an increase in the molecular weights of beta and gamma proteins accompanied by fluctuations in he electrophoretic mobility and slight decrease in the refractive index at high concentration of HY. The changes were higher in the samples treated with HY more than those treated with liposomes loaded with HY. It is concluded from the obtained data that hypericin embedded in liposomes may be used as a hotosensitizer for photodynamic therapy at low fractionated doses with minimal side effects

Determining the quantity of hypericin in medicinal materials and asepsis seedings of Hypericum perforatum by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To establish the method of determining the quantity of hypericin in Hypericum perforatum and determine the quantity of the hypericin in defferent medicinal materials and asepsis seedings which grow in defferent environment.</p><p><b>METHOD</b>The specimen is extracted with methanol--Pyridine (9:1) ultrasound extraction. Chromatographic assay is performed on a hypersily ODS2 (4.6 mm x 150 mm, 5 microm) column. The mobile phase is composed of methanol -1.56% dihydric natrium phosphate hydrogen natrium solution (shift solution's acidity to 2.1 with phosphoric acid)--ethyl acetate (4:1.9:1), velocity of flow is 1 mL x min(-1); column temperature is 35 degrees C; the detection wavelength is 590 nm.</p><p><b>RESULT</b>A satisfactory seperaration between hypericin and impurity. The calibration curve is linear over the range of 0.0524-0.2620 microg for hypericin (r = 0.9998). The average recovery of hypericin is 97.50%.</p><p><b>CONCLUSION</b>The quantity of hypericin in Hypericum perforatum has something to do with the genetic factor, environment factor, growing period and dry means. The method of determining the quantity of hypericin can be regarded as the method of controling the quantity of medicinal materials.</p>

Effect of factors on callus biomass and synthetic mass of hypericin in Hypericum perforatum / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of several factors on the quantity of hypericin in H. perforatum callus.</p><p><b>METHOD</b>High efficiency liquid phase chromatography and plant tissue culture were applied.</p><p><b>RESULT AND CONCLUSION</b>When the ratio of nitro-nitrogen to amina-nitrogen is 3:1, the callus biomass is 1.6-fold and the synthetic mass of hypericin rises. 0.1-0.20 mg x L(-1) mannose improves the content of total hypericin. The addition of PVP or PVPP can promote improvement of the growth and biosynthesis of callus.</p>

Studies on the chemical constituents of a fungus producing perylenequinones / 药学学报

<p><b>AIM</b>To study the chemical constituents in the mycelia of Hypomyces sp..</p><p><b>METHODS</b>Silica gel column chromatography was employed for the isolation and purification. Chemical and spectral methods were used to determine the structures of the isolated compounds.</p><p><b>RESULTS</b>Two compounds were isolated and identified as: hypomycin C (I) and hypomycin D (II).</p><p><b>CONCLUSION</b>Compounds I and II are new compounds.</p>