OBJECTIVE: To explore the effect of dichloromethane extraction phase of ethanol extract from stem of Patrinia scabiosaefolia Fisch.(DPSS) on proliferation and differentiation of K562 cells and its related mechanism. METHODS: MTT assay was used to detect the effects of DPSS at 0, 25, 50, 100 and 200 μg/ml on the proliferation of K562 cells at 24, 48 and 72 hours. Flow cytometry was used to analyze the changes of cell cycle and apoptosis at 24 and 48 hours. Wright-Giemsa staining was used to observe the morphological changes of K562 cells. The cell surface antigens CD33 and CD11b were detected by flow cytometry. RESULTS: The proliferation of K562 cells treated with different concentrations of DPSS was inhibited in a time-dose dependent manner (r=-0.96). Cell cycle analysis showed that with the increase of DPSS concentration, cells in G₂/M phase increased (r=0.88), and cells were blocked in G₂/M phase. Flow cytometry results showed that with the apoptosis rate of K562 cells was the highest when treated with 200 μg/ml DPSS for 48 h. Morphological observation showed that the K562 cell body increased, the amount of cytoplasm increased, the ratio of nucleus to cytoplasm decreased, and the nuclear chromatin was rough after DPSS treatment. Cell differentiation antigen, CD33 and CD11b, were positively expressed after treated with DPSS. CONCLUSION DPSS can induce apoptosis through cell cycle arrest, inhibit the proliferation of K562 cells, and induce K562 cells to differentiate into monocytes, which has a potential anti-leukemia effect.
Humans ; K562 Cells ; Patrinia ; Methylene Chloride/pharmacology* ; Apoptosis ; Cell Proliferation ; Cell Differentiation

Inula helenium L. is rich source of eudesmane-type sesquiterpene lactones, mainly alantolactone and isoalantolactone, which have the various pharmacological functions. In this study, we examined the inhibitory effects of nitric oxide (NO) production of hexane, methylene chloride, ethyl acetate, butanol, and water fractions from I. helenium and investigated the anti-inflammatory effect of hexane fraction of I. helenium (HFIH) in LPS-induced RAW 264.7 cells. Quantification of alantolactone and isoalantolactone from HFIH was carried out for the standardization by multiple reaction monitoring using triple quadrupole mass spectrometer. HFIH significantly inhibited inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) protein as well as their downstream products NO and prostaglandin E₂ (PGE₂) in LPS-stimulated RAW 264.7 cells. Moreover, HFIH suppressed NF-κB transcriptional activity by decreasing the translocation of p65 to the nucleus. The in vivo study further confirmed that HFIH attenuated the paw edema induced by carrageenan in an acute inflammation model. These findings suggest that HFIH may be useful as a promising phytomedicine for inflammatory-associated diseases.
Carrageenan ; Cyclooxygenase 2 ; Edema ; Inflammation ; Inula ; Lactones ; Methylene Chloride ; Nitric Oxide ; Nitric Oxide Synthase ; RAW 264.7 Cells ; Staphylococcal Protein A ; Water

Lauraceae is a family medicinal plant whose tubers possesses antimicrobial, and cytotoxic, such as antiparasitic and anti-inflammatory special effects and has been used for the medicine in the cure of hepatitis and rheumatism. The antimicrobial activities of bioactive compounds including one neolignan; kunstlerone (1) and two alkaloids include isocaryachine (2) and noratherosperminine (3) as well as crude hexane, methanol and dichloromethane extracts were evaluated. Additionally, the effect of compounds 1, 2 and 3 were evaluated on A549, PC-3, A375, HT-29 and WRL-68 cell lines. In conclusion, kunstlerone 1 showed moderate cytotoxicity against various cancer cell lines such as A549, PC-3, A375, HT-29 and WRL-68, respectively with EC₅₀ values of 28.02, 26.78, 33.78, 33.65 and 16.46 µg/mL. The crude methanol extract showed antigrowth activity against S. pyogenes II and B. cereus, with MICs of 256 µg/mL. The compounds kunstlerone (1), isocaryachine (2) and noratherosperminine (3) showed complete inhibition against P. shigelloides, with MIC ≤60 µg/mL compare to ampicillin, as a positive control, which showed antigrowth activity against P. shigelloides at MIC 10 µg/mL.
Alkaloids ; Ampicillin ; Cell Line ; Hepatitis ; Humans ; Lauraceae ; Methanol ; Methylene Chloride ; Plants, Medicinal ; Plesiomonas ; Rheumatic Diseases

Piper nigrum L. (Piperaceae), which is a well-known food seasoning, has been used as a traditional medicine for the treatment of vomiting, abdominal pain, diarrhea and anorexia in Korea, China and Japan. Methanol extract from the fruit of P. nigrum was successively partitioned as n-hexane, methylene chloride, ethyl acetate, n-butanol and H₂O soluble fractions. Among those fractions the ethyl acetate soluble fraction showed the most potent DPPH radical scavenging activity, and piperine was isolated from the ethyl acetate fraction. To know the antioxidant activity of piperine, we tested the activities of superoxide dismutase (SOD) and catalase together with oxidative stress tolerance and intracellular ROS level in Caenorhabditis elegans. To investigate whether piperine-mediated increased stress tolerance was due to regulation of stress-response gene, we quantified SOD-3 expression using transgenic strain including CF1553. Consequently, piperine enhanced SOD and catalase activities of C. elegans, and reduced intracellular ROS accumulation in a dose–dependent manner. Moreover, piperine-treated CF1553 worms exhibited significantly higher SOD-3::GFP intensity.
1-Butanol ; Abdominal Pain ; Anorexia ; Caenorhabditis elegans ; Caenorhabditis ; Catalase ; China ; Diarrhea ; Fruit ; Japan ; Korea ; Medicine, Traditional ; Methanol ; Methylene Chloride ; Oxidative Stress ; Piper nigrum ; Piper ; Seasons ; Superoxide Dismutase ; Vomiting

Burns ; Burns, Chemical ; Hand ; Humans ; Inhalation ; Male ; Methylene Chloride ; Paint ; Protective Clothing ; Skin ; Skin Transplantation ; Transplants

BACKGROUND: Rice bran is the outer layer of the rice grain, and contains high amounts of bioactive phytochemicals. Here, we investigated and compared chemopreventive properties of purple and white rice bran extracts. METHODS: Rice bran was extracted with dichloromethane and methanol. Chemical constituents in the extracts were analyzed by colorimetric assay and high performance liquid chromatography. The mutagenicity and antimutagenicity of the extracts were determined via the Salmonella mutation assay. The anticarcinogenic enzyme induction and antioxidant activities of the extracts were examined using Hepa1c1c7 cells and 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay, respectively. RESULTS: The methanol extracts of rice bran contained high amounts of phenolic acids, flavonoids, anthocyanins, and phytic acid, whereas large amounts of γ-oryzanol and vitamin E were presented in the dichloromethane extract. None of the extracts were mutagenic to Salmonella typhimurium. All rice bran extracts had strong antimutagenic effects against aflatoxin B1- and 2-amino-3,4-dimethylimidazo [4,5-f]quinoline-induced mutagenesis. The inhibitory effect against 2-aminofluorene-induced mutagenesis was found in the dichloromethane extract, while only the methanol extract of purple rice bran exhibited antimutagenic effects against benzo(a)pyrene. None of the extracts induced quinone reductase activity in Hepa1c1c7 cells. Additionally, the greatest antioxidant capacity was found in the methanol extract of purple rice bran. CONCLUSIONS: The methanol extract of purple rice bran containing high amount of phenolic acids, flavonoids, anthocyanins, and phytic acid showed the most effective antioxidant and antimutagenic activities by inhibiting mutagenic metabolizing enzymes and/or scavenging free radicals. These results demonstrate the nutritional and medical value of Thai rice for cancer prevention.
Aflatoxins ; Anthocyanins ; Antimutagenic Agents ; Asian Continental Ancestry Group* ; Benzo(a)pyrene ; Chromatography, Liquid ; Enzyme Induction ; Flavonoids ; Free Radicals ; Humans ; Methanol ; Methylene Chloride ; Mutagenesis ; NAD(P)H Dehydrogenase (Quinone) ; Phenol ; Phytic Acid ; Phytochemicals ; Salmonella ; Salmonella typhimurium ; Vitamin E ; Vitamins

OBJECTIVES: Aflatoxin M1 (AFM1)-contaminated dairy products pose serious human health risks, causing liver and renal failure if consumed. They are also related to decreased milk and egg production in infected animals. This study investigated the AFM1 contamination levels in cheeses sold in Isfahan province, Iran, by enzyme-linked immunosorbent assay (ELISA). METHODS: A total of 100 white cheese samples were randomly collected from supermarkets in Isfahan province and after extraction using dichloromethane were prepared for the ELISA. RESULTS: Of the 100 samples, 52 (52%) were contaminated by AFM1, at levels ranging from 50.2 to 424.4 ng/kg. The remaining 48% of the samples had undetectable AFM1 levels (< 50 ng/kg). Based on the standard limit set by the European Commission and Iran, 8% (8/100) of the AFM1-positive samples (with concentrations between 250.2 and 424.4 ng/kg) had levels higher than the permissible value of 250 ng/kg. CONCLUSION: Although the percentage of cheese samples in Isfahan province with AFM1 levels exceeding the national permissible limit was low, the examination of cheeses and the milk used for their production is nevertheless important for ensuring public health. Furthermore, optimum storage conditions of animal feed should be ensured, and livestock nutrition must be monitored for the presence of AFM1 and other aflatoxins.
Aflatoxin M1* ; Aflatoxins* ; Animal Feed ; Animals ; Cheese* ; Dairy Products ; Enzyme-Linked Immunosorbent Assay ; Humans ; Iran* ; Liver ; Livestock ; Methylene Chloride ; Milk ; Ovum ; Public Health ; Renal Insufficiency

This study describes the anti-inflammatory, anti-oxidant, and melanogenesis inhibition activities of methanol extract and various organic solvent fractions of Arecae Pericarpium. We examined the inhibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 cells, 1,1-diphenyl-2-picrylhydrazine (DPPH) scavenging activity, mushroom tyrosinase inhibition activity and melanin contents. The study showed that, among all tested fractions, methylene chloride fraction showed the strongest inhibition of LPS-induced NO production in RAW 264.7 cells (IC₅₀ value 8.89 µg/mL) and DPPH radical scavenging activity (EC₅₀ value 21.39 µg/mL). Methylene chloride and ethyl acetate fractions similarly inhibited mushroom tyrosinase activity. Methanol extract exhibited strongest reduction of melanin content in B16F10 melanoma cells. Based on the bioactivity assay results, methylene chloride and ethyl acetate fractions were further separated. Eight phenolic compounds were isolated, which are dimeric syringol (1), catechol (2), 4-hydroxybenzaldehyde (3), vanillin (4), 4-hydroxyacetophenone (5), apocynin (6), protocatechuic acid (7) and 4-hydroxybenzoic acid (8). Among the isolated compounds tested, catechol showed the strongest inhibition of LPS-induced NO production in RAW 264.7 cells. Catechol also showed the concentration-dependent NF-κB inhibition activity. Arecae Pericarpium might have potentials to be developed as anti-inflammatory agent or dermatological product for skin-whitening agent.
Agaricales ; Areca* ; Melanins ; Melanoma ; Methanol ; Methylene Chloride ; Monophenol Monooxygenase ; Nitric Oxide ; Phenol ; RAW 264.7 Cells

BACKGROUND: IgA nephropathy (IgAN) is the most common form of glomerulonephritis, a principal cause of end-stage renal disease (ESRD) worldwide. The mechanisms of onset and progression of IgAN have not been fully revealed, and epidemiologic studies have yielded diverging opinions as to the role of occupational exposure to organic solvents in the initiation or worsening of IgAN. As the authors encountered a laboratory worker with IgAN that progressed to ESRD, we present a case report of IgAN progression due to dichloromethane exposure along with a review of literature. CASE PRESENTATION: A 41-year-old male laboratory worker began to experience gross painless hematuria after two years of occupational exposure to toluene. Although clinical follow-up was initiated under the impression of IgAN based on clinical findings, the patient continued to work for four more years in the same laboratory, during which he was in charge of laboratory analysis with direct exposure to a high concentration of dichloromethane without proper protective equipment. During that time, his renal function rapidly worsened and finally progressed to ESRD 10 years after the first clinical symptoms. The result of exposure assessment through reenactment of his work exceeded the occupational exposure limit for dichloromethane to a considerable degree. CONCLUSIONS: The causal association between occupational solvent exposure and IgAN is still unclear; therefore, this case report could be used as a basis to support the relevance of occupational solvent exposure to IgAN and/or its progression. Early intervention as well as close monitoring of laboratory workers exposed to various organic solvents is important to prevent or delay the progression of glomerulonephritis to ESRD in the occupational setting.
Adult ; Early Intervention (Education) ; Epidemiologic Studies ; Follow-Up Studies ; Glomerulonephritis ; Glomerulonephritis, IGA* ; Hematuria ; Humans ; Immunoglobulin A* ; Kidney Failure, Chronic* ; Male ; Methylene Chloride ; Occupational Exposure ; Solvents ; Toluene

Seven phenolic compounds including p-coumaric acid (1), 4-hydroxybenzoic acid (2), 4-hydroxybenzaldehyde (3), vanillic acid (4), luteolin (5), acacetin (6), and tricin (7), were isolated from the methylene chloride and ethyl acetate fractions of Echinochloa utilis grains. Compounds (1 - 4, 6) were isolated for the first time from this plant. These compounds were tested for inhibitory activities against LPS-induced NO production in RAW 264.7 cells. Compounds 5 and 6 displayed significant inhibitory effects, with IC₅₀ values of 27.9 ± 2.6 and 14.0 ± 1.1 µM, respectively. The results suggested that E. utilis ethanolic extract may be used as a potential source of anti-inflammatory agents and functional foods for the treatment of allergic diseases.
Anti-Inflammatory Agents ; Echinochloa* ; Ethanol ; Functional Food ; Luteolin ; Methylene Chloride ; Phenol* ; Plants ; RAW 264.7 Cells ; Vanillic Acid