Scopoletin is a coumarin compound with various biological activities including detumescence and analgesic, insecticidal, antibacterial and acaricidal effects. However, interference with scopolin and other components often leads to difficulties in purification of scopoletin with low extraction rates from plant resource. In this paper, heterologous expression of the gene encoding β-glucosidase An-bgl3 derived from Aspergillus niger were carried out. The expression product was purified and characterized with further structure-activity relationship between it and β-glucosidase analyzed. Subsequently, its ability for transforming scopolin from plant extract was studied. The results showed that the specific activity of the purified β-glucosidase An-bgl3 was 15.22 IU/mg, the apparent molecular weight was about 120 kDa. The optimum reaction temperature and pH were 55 ℃ and 4.0, respectively. Moreover, 10 mmol/L metal ions Fe²⁺ and Mn²⁺ increased the enzyme activity by 1.74-fold and 1.20-fold, respectively. A 10 mmol/L solution containing Tween-20, Tween-80 and Triton X-100 all inhibited the enzyme activity by 30%. The enzyme showed affinity towards scopolin and tolerated 10% methanol and 10% ethanol solution, respectively. The enzyme specifically hydrolyzed scopolin into scopoletin from the extract of Erycibe obtusifolia Benth with a 47.8% increase of scopoletin. This demonstrated that the β-glucosidase An-bgl3 from A. niger shows specificity on scopolin with good activities, thus providing an alternative method for increasing the extraction efficiency of scopoletin from plant material.
Aspergillus niger/genetics* ; beta-Glucosidase/chemistry* ; Scopoletin ; Polysorbates ; Coumarins

This study compared the ameliorating effects of L-borneol, natural borneol, and synthetic borneol on the injury of different brain regions in the rat model of acute phase of cerebral ischemia/reperfusion(I/R) for the first time, which provides a reference for guiding the rational application of borneol in the early treatment of ischemic stroke and has important academic and application values. Healthy specific pathogen-free(SPF)-grade SD male rats were randomly assigned into 13 groups: a sham-operation group, a model group, a Tween model group, a positive drug(nimodipine) group, and high-, medium-, and low-dose(0.2, 0.1, and 0.05 g·kg~(-1), respectively) groups of L-borneol, natural borneol, and synthetic borneol according to body weight. After 3 days of pre-administration, the rat model of I/R was established by suture-occluded method and confirmed by laser speckle imaging. The corresponding agents in different groups were then administered for 1 day. The body temperature was monitored regularly before pre-administration, days 1, 2, and 3 of pre-administration, 2 h after model awakening, and 1 d after model establishment. Neurological function was evaluated based on Zea-Longa score and modified neurological severity score(mNSS) 2 h and next day after awakening. The rats were anesthetized 30 min after the last administration, and blood was collected from the abdominal aorta. Enzyme-linked immunoassay assay(ELISA) was employed to determine the serum levels of tumor necrosis factor-alpha(TNF-α), interleukin-6(IL-6), IL-4, and transforming growth factor-beta1(TGF-β1). The brain tissues were stained with triphenyltetrazolium chloride(TTC) for the calculation of cerebral infarction rate, and hematoxylin-eosin(HE) staining was used for observing and semi-quantitatively evaluating the pathological damage in different brain regions. Immunohistochemistry was employed to detect the expression of ionized calcium binding adapter molecule 1(IBA1) in microglia. q-PCR was carried out to determine the mRNA levels of iNOS and arginase 1(Arg1), markers of polarization phenotype M1 and M2 in microglia. Compared with the sham-operation group, the model group and the Tween model group showed significantly elevated body temperature, Zea-Longa score, mNSS, and cerebral infarction rate, severely damaged cortex, hippocampus, and striatum, increased serum levels of IL-6 and TNF-α, and decreased serum levels of IL-4 and TGF-β1. The three borneol products had a tendency to reduce the body temperature of rats 1 day after modeling. Synthetic borneol at the doses of 0.2 and 0.05 g·kg~(-1), as well as L-borneol of 0.1 g·kg~(-1), significantly reduced Zea-Longa score and mNSS. The three borneol products at the dose of 0.2 g·kg~(-1) significantly reduced the cerebral infarction rate. L-borneol at the doses of 0.2 and 0.1 g·kg~(-1) and natural borneol at the dose of 0.1 g·kg~(-1) significantly reduced the pathological damage of the cortex. L-borneol and natural borneol at the dose of 0.1 g·kg~(-1) attenuated the pathological damage of hippocampus, and 0.2 g·kg~(-1) L-borneol attenuated the damage of striatum. The 0.2 g·kg~(-1) L-borneol and the three doses of natural borneol and synthetic borneol significantly reduced the serum level of TNF-α, and the 0.1 g·kg~(-1) synthetic borneol reduced the level of IL-6. L-borneol and synthetic borneol at the dose of 0.2 g·kg~(-1) significantly inhibited the activation of cortical microglia, and 0.2 g·kg~(-1) L-borneol up-regulated the expression of Arg1 and down-regulated the expression level of iNOS. In conclusion, the three borneol products may alleviate inflammation to ameliorate the pathological damage of brain regions of rats in the acute phase of I/R by inhibiting the activation of microglia and promoting the polarization of microglia from M1 type to M2 type. The protective effect on brain followed a trend of L-borneol > synthetic borneol > natural borneol. We suggest L-borneol the first choice for the treatment of I/R in the acute phase.
Rats ; Male ; Animals ; Transforming Growth Factor beta1/metabolism* ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha/metabolism* ; Interleukin-6/metabolism* ; Interleukin-4/metabolism* ; Polysorbates ; Brain ; Brain Ischemia/metabolism* ; Reperfusion Injury/metabolism* ; Cerebral Infarction ; Reperfusion

The effective components of flavonoids in the "Pueraria lobata-Hovenia dulcis" drug pair have low bioavailability in vivo due to their unstable characteristics. This study used microemulsions with amphoteric carrier properties to solve this problem. The study drew pseudo-ternary phase diagrams through titration compatibility experiments of the oil phase with emulsifiers and co-emulsifiers and screened the prescription composition of blank microemulsions. The study used average particle size and PDI as evaluation indicators, and the central composite design-response surface method(CCD-RSM) was used to optimize the prescription; high-dosage drug-loaded microemulsions were obtained, and their physicochemical properties, appearance, and stability were evaluated. The results showed that when ethyl butyrate was used as the oil phase, polysorbate 80(tween 80) as the surfactant, and anhydrous ethanol as the cosurfactant, the maximum microemulsion area was obtained. When the difference in results was small, K_(m )of 1∶4 was chosen to ensure the safety of the prescription. The prescription composition optimized by the CCD-RSM was ethyl butyrate(16.28%), tween 80(9.59%), and anhydrous ethanol(38.34%). When the dosage reached 3% of the system mass, the total flavonoid microemulsion prepared had a clear and transparent appearance, with average particle size, PDI, and potential of(74.25±1.58)nm, 0.277±0.043, and(-0.08±0.07) mV, respectively. The microemulsion was spherical and evenly distributed under transmission electron microscopy. The centrifugal stability and temperature stability were good, and there was no layering or demulsification phenomenon, which significantly improved the in vitro dissolution of total flavonoids.
Polysorbates/chemistry* ; Flavonoids ; Pueraria ; Surface-Active Agents/chemistry* ; Ethanol ; Emulsions ; Particle Size ; Solubility

BACKGROUND: Microsporum gypseum has been isolated from South Korea since 1966. However, the incidence of M. gypseum infection is very low. OBJECTIVE: The aim of this study is to add massive data on M. gypseum to the literature and to provide useful information on clinical and mycological characteristics of M. gypseum. METHODS: We retrospectively analyzed data of 198 cases infected with M. gypseum from 1979 to 2016. The identification of M. gypseum was done with mycological examination using 15% KOH preparation and potato corn meal tween 80 agar culture media. RESULTS: The incidence of M. gypseum infection was very low in South Korea, showing a tendency to decrease. Out of 198 cases, men were 94 cases (47.5%) and women were 104 cases (52.5%). Mean age of all patients was 29.83 years old: 24.97 years old in men and 34.22 years old in women. M. gypseum infection occurred most frequently in September (16.7%) and August (16.2%). The most common clinical type of M. gypseum infection was tinea corporis (38.4%). CONCLUSION: M. gypseum infection shows very low incidence but still remains around us until recent years. We should keep in mind the characteristics of M. gypseum.
Agar ; Culture Media ; Epidemiology ; Female ; Humans ; Incidence ; Korea* ; Male ; Meals ; Microsporum* ; Polysorbates ; Retrospective Studies ; Solanum tuberosum ; Tinea ; Zea mays

OBJECTIVES: The main objectives of this study were to verify the antibacterial activity of two essential oils, lavender and peppermint, against dental caries and to review their synergistic effect when used in combination. Our results provide basic data for the evaluation of the use of these two substances towards the prevention and cure of dental caries. METHODS: The sample solutions of lavender and peppermint oils were prepared in three different concentrations (30%, 50%, and 70% (v/v)) by diluting them with third-distilled water and Tween 20. Streptococcus mutans was selected as the bacterial species for testing. The disk diffusion method was used to measure the antibacterial activity of the sample solutions. For generating growth curves and measuring the number of clusters of the bacterial, the liquid medium-dilution method was used; the absorbance of the medium was measured at 600 nm after 3, 6, 12 and 24 hours. RESULTS: When the antibacterial activity of the oils was tested via the disk diffusion method, the activity improved with increasing concentrations of all the sample solutions of peppermint, lavender, and the blend, but there was no significant difference between them with respect to the type of oil. In the growth curves of S. mutans, growth inhibition was observed after 12 hours. The inhibitory effect of 30% lavender oil on growth was 64.9% and 80.1% after 12 and 24 hours of treatment, respectively whereas that of peppermint oil was 71.3% and 80.1% after 12 and 24 hours of treatment, respectively. The inhibitory effect of the blended oil was 71.9% and 81.0% after 12 and 24 hours of treatment, respectively. CONCLUSIONS: Further research is still required in order to determine the efficacy of lavender and peppermint oils, as well as other essential oils, for wider use in preventing dental caries.
Dental Caries ; Diffusion ; Lavandula ; Mentha piperita ; Methods ; Oils ; Oils, Volatile ; Polysorbates ; Streptococcus mutans ; Streptococcus ; Water

Based on the fact that chromogenic reaction of blue complex, a reaction product which can be dissolved in organic solvents, can be realized by polyethoxy and ammonium thiocyanate in tween 80, a rapid and accurate way for the determination for tween 80 in pharmaceutical adjuvant was established in this study, providing reliable technical means for quality control of traditional Chinese medicine injections. Based on the study of reaction kinetics, chromogenic reaction temperature and time, as well as extraction of organic solvents and other key conditions were optimized, and Kumu injection was used as the test material for method validation and applicability investigation. It was finally determined that 3 mL ammonium thiocyanate solution was added in the sample solution, and the reaction was carried out in a boiling water bath for 2 h. After cooling to room temperature, 5 mL of dichloromethane was added to extract the chromogenic product. The absorbance value was measured at the wavelength of 623 nm to calculate the tween 80 content in the sample. Under optimized conditions, tween 80 solution showed a good linear relationship with the absorbance in the range from 0.8 mg to 3.0 mg. The linear regression equation was ＝0.258-0.047. The correlation coefficient was 0.999 6. Under the experimental conditions, the average recovery was 99.66%, and the precision RSD was less than 2.0%. The results showed that this method can quickly and accurately determine the content of tween 80 in Kumu injection, and it could be applicable to the quality control of traditional Chinese medicine injections.
Adjuvants, Pharmaceutic ; chemistry ; Medicine, Chinese Traditional ; Polysorbates ; chemistry ; Quality Control ; Solvents ; Temperature

Paclitaxel (PTX) is a effectively chemotherapeutic agent which is extensively able to treat the non-small cell lung, pancreatic, breast and other cancers. But it is a practically insoluble drug with water solubility less than 1 μg/mL, which restricts its therapeutic application. To overcome the problem, hyaluronic acid-complexed paclitaxel nanoemulsions (HPNs) were prepared by ionic complexation of paclitaxel (PTX) nanoemulsions and hyaluronic acid (HA) to specifically target non-small cell lung cancer. HPNs were composed of dl-α-tocopheryl acetate, soybean oil, polysorbate 80, ferric chloride, and HA and fabricated by high-pressure homogenization. The HPNs were 85.2 ± 7.55 nm in diameter and had a zeta potential of −35.7 ± 0.25 mV. The encapsulation efficiency was almost 100%, and the PTX content was 3.0 mg/mL. We assessed the in vivo antitumor efficacy of the HPNs by measuring changes in tumor volume and body weight in nude mice transplanted with CD44-overexpressing NCI-H460 xenografts and treated with a bolus dose of saline, Taxol®, PTX nanoemulsions (PNs), or HPNs at a dose of 25 mg/kg. Suppression of cancer cell growth was higher in the PN- and HPN-treated groups than in the Taxol® group. In particular, HPN treatment dramatically inhibited tumor growth, likely because of the specific tumor-targeting affinity of HA for CD44-overexpressed cancer cells. The loss of body weight and organ weight did not vary significantly between the groups. It is suggest that HPNs should be used to effective nanocarrier system for targeting delivery of non-small cell lung cancer overexpressing CD44 and high solubilization of poorly soluble drug.
Animals ; Body Weight ; Breast ; Carcinoma, Non-Small-Cell Lung* ; Heterografts ; High Pressure Neurological Syndrome ; Hyaluronic Acid ; Lung ; Mice ; Mice, Nude ; Organ Size ; Paclitaxel* ; Polysorbates ; Solubility ; Soybean Oil ; Tumor Burden ; Water

BACKGROUND: There are several media for culture of Malassezia spp., such as Leeming & Notman (LN) medium, modified Leeming & Notman (mLN) medium, Dixon's medium and modified Dixon's medium etc. It is known that Malassezia spp. grow well in these media in general, but the kind and amounts of their ingredients are various and un-uniform according to researchers. Author propose the new and transparent BHI based medium for the optimal growth of Malassezia spp. OBJECTIVES: The purpose of this study was to design the simple and transparent BHI based medium and find essential ingredients for the growth of M. globosa and M. obtusa. METHODS: The colony size of eight standard strains (M. dermatis, M. furfur, M. globosa, M. japonica, M. obtusa, M. sloofiae, M. sympodialis, M. yamatoensis) on the modified BHI (mBHI) agar media with different ingredients was observed by naked eye after seven day culture. The compositions of mBHI medium were as follows; mBHI-1 was supplemented with 0.7% dextrose, 1.5% Tween 80, 1% glycerol to BHI medium, mBHI-2 was supplemented with 1.5% Tween 40 to mBHI-1 instead of Tween 80, mBHI-3 was supplemented with 1.5% Tween 60 to mBHI-1 instead of Tween 80, mBHI-4 was added with 0.8% bile salts to mBHI-1. mBHI-5 was supplemented with 1.5% Tween 60 to mBHI-4 instead of Tween 80, and mBHI-6 was supplemented with 1.5% Tween 40 to mBHI-4 instead of Tween 80. pH of six mBHI media was all adjusted to 6.5. RESULTS: M. furfur & M. japonica were grown well on mBHI-1 agar, but M. globosa & M. obtusa were not grown and others grown poorly. M. globosa & M. obtusa were not grown on mBHI-1 & mBHI-4 containing Tween 80 as lipid source, but others grown on all mBHI media. The media that all eight Malassezia strains grew well were slightly turbid mBHI-5 & transparent mBHI-6 medium. CONCLUSIONS: M. globosa & M. obtusa need glycerol and bile salts as well as Tween 60 or 40 instead of Tween 80 for growth. M. furfur & M. japonica need not bile salts for growth. Author proposes the transparent modified BHI medium supplemented with 0.7% dextrose, 1.5% Tween 40, 1% glycerol and 0.8% bile salts (mBHI-6) as new standard medium for culture of eight Malassezia species.
Agar ; Bile Acids and Salts ; Glucose ; Glycerol ; Hydrogen-Ion Concentration ; Malassezia* ; Polysorbates

A 71-year-old man presented with pain in the left eye that revealed a 3x3 mm deep corneal stromal infiltrate, with a 2x2 mm epithelial defect. The patient started topical moxifloxacin, voriconazole 2%, and natamycin for 2 weeks. However, the treatment was not effective and the corneal infiltration worsened. Subsequently, the patient underwent therapeutic penetrating keratoplasty. Thick brown/gray mold colonies on Potato Corn Meal Tween 80 agar was isolated from excised corneal tissue and on slide culture many septated, and club-shaped ascospores were revealed. Histological findings also showed numerous hyphae scattered in corneal tissue. A. alternata colonies were confirmed by 18S rRNA sequencing. Intracameral voriconazole was injected every other day for 2 weeks to eliminate remaining fungi on the deep corneal stroma. The remaining corneal infiltration was improved one month after the injection. During 5 months postoperative follow up, the infection did not recurred. In conclusion, deep corneal infection of A. alternata was effectively treated with intracameral voriconazole injection.
Agar ; Aged ; Alternaria* ; Corneal Stroma ; Follow-Up Studies ; Fungi ; Humans ; Hyphae ; Keratitis* ; Keratoplasty, Penetrating ; Meals ; Natamycin ; Polysorbates ; Solanum tuberosum ; Zea mays

Tacrolimus and cyclosporine have been used in various formulations, but their hypersensitivity reactions are rare in practice. Castor oil derivatives are nonionic surfactants used in aqueous preparations of hydrophobic active pharmaceutical ingredients. Castor oil derivatives that can be used as additives to tacrolimus and cyclosporine may play a role in the development of hypersensitivity reactions, especially anaphylaxis. Various immunologic and nonimmunologic mechanisms have been implicated in hypersensitivity reactions induced by castor oil derivatives. Physicians should be aware that not only the drug itself, but also its additives or metabolites could induce hypersensitivity reactions. We report a case of anaphylaxis caused by vitamin K (phytonadine), serotonin antagonist (granisetron), intravenous tacrolimus, and cyclosporine. Interestingly, the patient tolerated oral cyclosporine, which did not contain Cremophor EL or polysorbate 80.
Anaphylaxis ; Castor Oil ; Cyclosporine ; Humans ; Hypersensitivity ; Polysorbates ; Serotonin ; Surface-Active Agents ; Tacrolimus ; Vitamin K