Leaf Spot caused by Alternaria tenuissima on Rhamnella franguloides in China.

The small tree species Rhamnella franguloides, belonging to the genus Rhamnella in the tribe Rhamneae Hook. f. of Rhamnaceae (Hauenschild et al. 2016), is an important medicinal plant commonly used for making tea in China. In August 2023, leaf spot symptoms were observed on R. franguloides in Shangyao county, Yantai, Shandong, China, with a disease incidence of 45-65%. Initially appearing as small dark brown spots on the tip lesions, they later expanded and merged into irregular-shaped brown necrotic lesions with yellowish halos. To isolate pathogen, 20 symptomatic tissue fragments (5 × 5 mm) from ten sampling randomly plants were surface sterilized, placed on potato dextrose agar (PDA) plates, and incubated at 25°C in darkness for 3 days to obtain colonies.10 purified isolates with similar morphological characteristics were obtained by single-spore isolation from the colonies. The representative isolate MR13 was chosen for morphological and molecular analysis. The colonies On PDA medium initially appear as a circular yellow-brown ring with white round margins, ultimately turning into olive green with fluffy aerial hyphae. The conidiophores displayed brown pigmentation, solitary or branched, producing abundant short chains of conidia. The conidia were typically obclavate to obpyriform or ellipsoid in shape, 22.5-64.5 × 12.5-23.6µm in size, with a short conical beak at the apex, zero to three longitudinal septa and one to five transverse septa. Based on cultural and morphological characteristics, the fungus was identified as Alternaria spp (Simmons 2007). Due to morphological traits, five genes (the internal transcribed spacer [ITS], actin [ACT], plasma membrane ATPase [ATP], Alternaria major allergen [Alt a1] and histone 3 [H3]) form MR13 were amplified using primer pairs ITSI / ITS4, ACTDF1/R1, ATPDF1 / ATPDRI, Alt-for / Alt-rev, and H3-1a/1b, respectively (Hong et al. 2005; Lawrence et al. 2013; Lousie and Donaldson 1995). BLASTn analysis failed to confirm the identification of MR13 species based on ITS, ACT, ATP and Alt a1(ITS, OR668512; ACT, OR676918; ATP, OR676917; Alt a1, OR676919). The phylogenetic tree showed that it was closely related to Alternaria alternate, A. tenuissima, and A. destruens. The H3 sequence (OR676920) exhibited 100% similarity to A. tenuissima (OR485421). The phylogenetic tree constructed solely with H3 further confirmed MR13 as A. tenuissima. Pathogenicity tests were conducted by introducing the fungus onto healthy R. franguloides leaves in the field. Fifty leaves (five per plant) were treated with a 20ml suspension containing around 1x10^4 spores/ml, while an equal number of control samples were sprayed with distilled water. Transparent plastic bags were used to cover the treated leaves for 48 hours and maintain humidity. After fourteen days of inoculation, consistent leaf spotting symptoms were observed. In contrast, the control leaves showed no sign of infection. The fungal pathogen was successfully reisolated and identified as A. tenuissima through morphological and sequence analysis, fulfilling Koch's postulates. To our knowledge, this is the first report of A. tenuissima causing leaf spot disease on R. franguloides in China. Identifying the disease's causal agent is crucial for developing effective management strategies.

[Common diseases and drug use characteristics of Chinese herbal medicines and suggestions].

Based on the data of 56 kinds of diseases and drug use in 100 kinds of cultivated Chinese herbal medicines, this paper used frequency analysis method to count the types of diseases and their drug use characteristics, and systematically analyzed the status of drug registration and monitoring standards for disease prevention and control of Chinese herbal medicines. The results showed that 14 diseases such as root rot, powdery mildew, and drooping disease were common in the production of Chinese herbal medicines. Among the 99 pesticides reported, 67.68% were chemically synthesized, 23.23% were biological pesticides, and 9.09% were mineral pesticides. Among the reported pesticides, 92.93% of them were low toxic, with relative safety. However, 70% of the production drugs were not registered in Chinese herbal medicines, and the phenomenon of overdose was serious. The current pesticide residue monitoring standards does not match well with production drugs in China. Although the matching degree between Maximum Residue Limit of Pesticide in Food Safety National Standard(GB 2763-2021) and production drugs is more than 50%, there are few varieties of Chinese herbal medicines covered. The matching degree between Chinese Pharmacopoeia(2020 edition), Green Industry Standard of Medicinal Plants and Preparations(WM/T2-2004), and production drugs is only 1.28%. It is suggested to speed up the research and registration of Chinese herbal medicine production and further improve the pesticide residue limit standard combined with the actual production, so as to promote the high-quality development of Chinese herbal medicine industry.

[Common diseases and drug use of Pseudostellaria heterophylla].

Pseudostellaria heterophylla in large-scale cultivation needs to apply pesticides to control diseases, and non-standard use of pesticide may cause excessive pesticide residues in medicinal materials, increasing the risk of clinical medication. To accurately monitor the residual pesticides, this paper investigated the drug use during the process of P. heterophylla disease prevention in 25 P. he-terophylla planting enterprises or individual households in Guizhou province. It was found that there were 8 common diseases in P. he-terophylla planting, including leaf spot, downy mildew, virus disease, root rot, dropping disease, purple feather disease, white silk disease, and damping-off disease. Twenty-three kinds of pesticides were used in disease control, mainly chemical synthetic pesticides, accounting for 78.3%, followed by biological pesticides and mineral pesticides, accounting for 13.0% and 8.7%, respectively. The disease prevention and control drugs were all low-toxic pesticides, and there were no varieties banned in the Chinese Pharmacopoeia(2020 edition). However, the pesticides used have not been registered on P. heterophylla, and the excessive use of drugs was serious. The present monitoring of pesticide residues in P. heterophylla is mainly based on traditional pesticides such as organochlorine, organophosphorus, and carbamate, which does not effectively cover the production of drugs and had certain safety risks. It is suggested to speed up the research and registration of drug use in the production of P. heterophylla, increase the use of biological pesticides, and further improve the monitoring indicators of pesticide residues in combination with the actual production of drugs, so as to promote the high-quality development of P. heterophylla industry.

[Contamination status and exposure risk of mycotoxins in Coicis Semen].

By investigating the contamination status and predicting the exposure risk of mycotoxin in Coicis Semen, we aim to provide guidance for the safety supervision of Chinese medicinal materials and the formulation(revision) of mycotoxin limit standards. The content of 14 mycotoxins in the 100 Coicis Semen samples collected from five major markets of Chinese medicinal materials in China was determined by UPLC-MS/MS. The probability evaluation model based on Monte Carlo simulation method was established after Chi-square test and One-way ANOVA of the sample contamination data. Health risk assessment was performed on the basis of margin of exposure(MOE) and margin of safety(MOS). The results showed that zearalenone(ZEN), aflatoxin B_1(AFB_1), deoxynivalenol(DON), sterigmatocystin(ST), and aflatoxin B_2(AFB_2) in the Coicis Semen samples had the detection rates of 84%, 75%, 36%, 19%, and 18%, and the mean contamination levels of 117.42, 4.78, 61.16, 6.61, and 2.13 µg·kg~(-1), respectively. According to the limit standards in the Chinese Pharmacopoeia(2020 edition), AFB_1, AFs and ZEN exceeded the standards to certain extents, with the over-standard rates of 12.0%, 9.0%, and 6.0%, respectively. The exposure risks of Coicis Semen to AFB_1, AFB2, ST, DON, and ZEN were low, while 86% of the samples were contaminated with two or more toxins, which needs more attention. It is suggested that the research on the combined toxicity of different mycotoxins should be strengthened to accelerate the cumulative exposure assessment of mixed contaminations and the formulation(revision) of toxin limit standards.

[Effect of Danlou Tablets on alleviating hepatic adipogenesis, inflammatory response, and insulin resistance in db/db mice].

This study explored the effect and potential mechanism of Danlou Tablets(DLT) on insulin resistance in db/db mice with type 2 diabetic mellitus(T2 DM). The db/db male mice were randomly assigned into model control(MC) group, metformin(MET, tablet, 100 mg·kg~(-1)) group, and DLT(1 g·kg~(-1)) group, and C57 BL/6 J mice were taken as normal control(NC) group. The mice in the MET group and DLT group were given corresponding drugs by gavage once a day for 16 weeks. The fasting blood glucose, glucose tolerance, and insulin tolerance were measured to evaluate the effect of DLT on blood glucose and insulin resistance in diabetic mice. The serum free fatty acid, triacylglycerol, and total cholesterol levels were determined to evaluate the effect of DLT on blood lipids in diabetic mice. The liver index and perirenal fat index were calculated to measure the effect of DLT on lipid accumulation in non-adipose tissue and adipose tissue. Western blot was performed to determine the protein levels of insulin receptor-ß(IRß), phospho-IRß(p-IRß), phosphatidylinositol 3-kinase(PI3 K), and insulin receptor substrate-1(IRS-1) involved in IRS-1/PI3 K/Akt signaling pathway in the livers of mice to reveal the mechanism of DLT in alleviating insulin resistance in diabetic mice. The protein levels of sterol regulatory element binding protein-1(SREBP-1) and the mRNA levels of sterol regulatory element binding protein-1 c(SREBP-1 c), fatty acid synthase(FAS), acetyl-CoA carboxylase(ACC), diacylglycerol acyltransferase-1(Dgat1), and diacylglycerol acyltransferase-2(Dgat2) involved in the SREBP-1/FAS signaling pathway were detected to evaluate the effect of DLT on lipid metabolism in diabetic mice. Real-time quantitative PCR was employed to determine the mRNA levels of galectin-3(Gal-3), interleukin-6(IL-6), and monocyte chemoattractant protein-1(MCP-1) in mouse liver to evaluate the effect of DLT on inflammatory response in diabetic mice. The results showed that DLT significantly reduced the blood glucose and lipid levels and alleviated the insulin resistance in diabetic mice. Compared with the MC group, DLT significantly up-regulated the protein levels of p-IRß, PI3 K, and IRS-1(P<0.05 or P<0.01), and down-regulated the protein level of SREBP-1 in liver tissues of diabetic mice(P<0.05). DLT lowered the mRNA levels of SREBP-1 c, FAS, ACC, Dgat1, and Dgat2 related to lipid metabolism as well as those of Gal-3, IL-6, and MCP-1 associated with inflammation in the livers of diabetic mice(P<0.05 or P<0.01). The findings of this study suggest that DLT may alleviate insulin resistance in diabetic mice by regulating IRS-1/PI3 K/Akt signaling pathway and SREBP-1/FAS signaling pathway to reduce lipid production and inhibit inflammatory response.

Sensory-guided isolation and identification of new sweet-tasting dammarane-type saponins from Jiaogulan (Gynostemma pentaphyllum) herbal tea.

The purpose of this study was to elucidate the chemical basis for the sweet property produced by Gynostemma pentaphyllum and find new natural high-potency (HP) sweeteners. Sixteen new compounds (gypenosides YN 1-16) were obtained by sensory-guided isolation and identification, in which fifteen of them were sweet-tasting constituents with sweetness intensities 10-100 times higher than that of sucrose evaluated by human sensory panel test. Their structures were established by 1D and 2D nuclear magnetic resonance spectra, mass spectroscopy, infrared spectroscopy, UV-visible spectroscopy, and chemical method. Gypenoside YN 4 was the sweetest compound with a concentration of 15.504 ± 1.343 mg/kg, while gypenoside YN 12 has the highest concentration (1397.674 ± 12.948 mg/kg), as shown by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Structure-activity relationship analysis implied that the compounds' sweetness intensity was associated with side-chain substitutions at C-20 or the number of glucosyl groups at C-3. These new plant-derived natural products may be potential natural sweeteners.

[Methods for synchronous detection of 14 mycotoxins in Pseudostellariae Radix and investigation on its contamination].

This study aimed to establish a method for synchronous detection of 14 mycotoxins in Pseudostellariae Radix and investigate its contamination with mycotoxins, so as to provide technical guidance for monitoring the quality of Chinese medicinal materials and medication safety. The sample was extracted with 80% acetonitrile in an oscillator for 1 h, purified using the modified QuEChERS purifying agent(0.1 g PSA + 0.3 g C_(18) + 0.3 g MgSO_4), and separated on a Waters HSS T3 chromatographic column(2.1 mm×100 mm, 1.8 µm). The gradient elution was carried out with 0.1% formic acid in water and acetonitrile, followed by the scanning in the multi-reaction monitoring(MRM) mode and the analysis of mycotoxin contamination in 26 Pseudostellariae Radix samples. The recovery rates of the established method were within the range of 82.17%-113.6%, with the RSD values less than 7% and the limits of quantification(LOQ) being 0.019-0.976 µg·kg~(-1). The detection rate of 14 mycotoxins in 26 batches of medicinal materials was 53.85%. The detection rate of sterigmatocystin(ST) was the highest, followed by those of zearalenone(ZEN), aflatoxin G_2(AFG_2), fumonisin B_1(FB_1), HT-2 toxin, and nivalenol(NIV). Their respective detection rates were 38.46%, 26.92%, 23.08%, 11.54%, 11.54%, and 7.69%, with the pollution ranges being 1.48-69.65, 0.11-31.05, 0.11-0.66, 0.28-0.83, 20.86-42.56, and 0.46-1.84 µg·kg~(-1), respectively. The established method for the detection of 14 mycotoxins is accurate, fast and reliable. The research results have very important practical significance for guiding the monitoring and prevention and control of exogenous fungal contamination of Chinese medicinal materials.

Repellent, larvicidal and adulticidal activities of essential oil from Dai medicinal plant Zingiber cassumunar against Aedes albopictus.

Zingiber cassumunar is an important plant used in traditional medicine and as a natural mosquito repellent. However, the compounds responsible for the repellent activity of the plant are still unknown. The aim of the study is to identify the components of Z. cassumunar essential oil that show repellent activity against Aedes albopictus. We also evaluated the larvicidal and adulticidal activities of Z. cassumunar essential oil against Ae. albopictus. In-cage mosquito repellent experiments showed that Z. cassumunar essential oil possessed moderate repellent activity with a minimum effective dose (MED) of 0.16 ± 0.01 mg/cm2, compared to reference standard N,N-diethyl-3-methylbenzamide (DEET, 0.03 ± 0.01 mg/cm2). Bioassay-guided fractionation identified the major active compound of Z. cassumunar essential oil as (-)-terpinen-4-ol (1) (MED: 0.19 ± 0 mg/cm2). We also found that Z. cassumunar essential oil showed moderate larvicidal activity against first instar larvae of Ae. albopictus with a LC50 (50% lethal concentration) of 44.9 µg/L after 24 h. Fumigation bioassays showed that Z. cassumunar essential oil exhibits moderate adulticidal activity against Ae. albopictus with a LC50 of 5.44%, while (-)-terpinen-4-ol showed significant adulticidal activity with a LC50 of 2.10% after 24 h. This study verifies that the Z. cassumunar essential oil has mosquito repellent activity, and that (-)-terpinen-4-ol is mainly responsible for this activity. Furthermore, this study provides scientific support for the folk usage of Z. cassumunar essential oil as mosquito repellent and indicates that Z. cassumunar essential oil and (-)-terpinen-4-ol can be used as plant-derived repellents and insecticides for mosquito control.

[Preventive and therapeutic effects of safflower water extract on systemic scleroderma in mice and its mechanism].

OBJECTIVE: To study the preventive and therapeutic effects of safflower water extract on systemic scleroderma (SSc) in mice and its mechanism. METHODS: Sixty BALB/C mice were randomly divided into the control group, model group, prednisone group and safflower low, middle, high dose groups, 10 mice in each group.The control group was injected with normal saline, and the other five groups were subcutaneously injected with bleomycin hydrochloride with 100 µl at the concentration of 200 µg /ml on the back, once a day for 28 days to establish the SSc models.At the same time, the control group and model group were treated with normal saline (10 ml/kg), the prednisone group was treated with prednisone 4.5 mg/kg (10 ml/kg), and the low, middle, and high dose safflower groups were treated with safflower at the doses of 1.5, 3, 6 g/kg (10 ml/kg), and all groups were treated for 28 days.After 28 days, all mice were decapitated. The blood samples and back skin of the BLM injection part were collected.After that, all the tissue slices were taken to measure the dermal thickness, and the content of hydroxyproline (HYP) in the skin tissues was detected by hydrolysis method.The contents of tissue growth factor (CTGF) and transforming growth factor-ß (TGF-ß ) in the skin tissues and the levels of interleukin-6 (IL-6) and interleukin-17 (IL-17) in serum were determined by ELISA. RESULTS: Compared with the control group, the dermal thickness of the model group was increased(P＜0.05), the contents of CTGF, TGF-ß and HYP in the skin tissues and the levels of IL-6 and IL-17 in the serum of the model group were increased(P＜0.05); compared with the model group, the dermal thickness in the prednisone group and safflower groups was decreased (P＜0.05), the levels of CTGF, TGF-ß and HYP in the skin tissues and the serum levels of IL-6 and IL-17 in the prednisone group and safflower groups were decreased (P＜0.05). CONCLUSION: Safflower water extract can improve skin condition (or dermal thickness) in SSc mice, and its mechanism may be related to reducing immune inflammatory response.

Effect of Ginkgo biloba extract-761 on motor functions in permanent middle cerebral artery occlusion rats.

BACKGROUND: Ginkgo biloba extract (EGb-761) has been in use to treat variety of ailments including memory loss and emotional disorders usually experienced after ischemic stroke. However, data regarding its protective role in stroke associated motor dysfunction is scarce. PURPOSE: The present work was designed to investigate the long-term effects of EGb-761 on the motor dysfunctions associated with permanent middle cerebral artery occlusion (pMCAO) in rats. STUDY DESIGN/METHODS: Focal ischemic stroke was induced in male Sprague-Dawley rats by pMCAO. These rats were orally administered with EGb-761 (25, 50, 100 mg/kg) and positive control butylphthalide (50 mg/kg) for up to 28 consecutive days. The motor function was evaluated by assessing neurological scores, rotarod performance and gait analysis after 7, 14, 21 and 28 days. After 28 days, the histological examination of in frontal cortex and hippocampus was also carried out. RESULTS: EGb-761 treatment significantly improved motor function with better outcome in coordination and gait impairment rats. EGb-761 (25, 50, 100 mg/kg) treatment for 28 days significantly decreased the neurological scores. After 28 days of treatment EGb-761 (50 and 100 mg/kg) significantly increased the latency in rotarod test, walk speed, and the body rotation, whereas, decreased the stride time and the left posterior swing length in gait were observed. EGb-761 (50, 100 mg/kg). EGb-761 (50, 100 mg/kg) significantly improved the pathological changes related to pMCAO. CONCLUSIONS: EGb 761 could improve motor function especially gait impairments among pMCAO rat model related to the decreased neuronal damage. Therefore, it might be the potential to be explored further as an effective therapeutic drug to treat post stroke motor dysfunctions.

New Sweet-Tasting C21 Pregnane Glycosides from the Roots of Myriopteron extensum.

To investigate the sweet-tasting components in the roots of Myriopteron extensum, the phytochemical study of its roots was conducted, which led to the discovery of 12 new C21 pregnane glycosides (extensumside M-X, 1-12) and two known ones (extensumside C and extensumside E, 13-14). Their chemical structure elucidation was accomplished by means of spectroscopic methods: IR, UV, ESI-MS, and NMR (1H NMR, 13C NMR, HSQC, 1H-1H COSY, HMBC, HSQC-TOCSY, and ROESY), as well as the chemical evidence. Sensory analysis of these compounds revealed that nine of them (1, 3, 4, 5, 6, 7, 8, 13, and 14) are highly sweet-tasting compounds. Their sweetness intensities are 25 to 400 times greater than that of sucrose. Analysis of the structure-activity relationship (SAR) indicated that the sweet intensities of the isolated compounds are closely related to the aglycone 3ß,16α-dihydroxy-pregn-5-en-20-one, the number and type of the monosaccharide in the sugar chain linked to C-3 and C-16 and the position of the mBe group.

Antidepressant effects of dammarane sapogenins in chronic unpredictable mild stress-induced depressive mice.

Depression is a common, dysthymic, and psychiatric disorder, resulting in enormous social and economic burden. Dammarane sapogenins (DS), an active fraction from oriental ginseng, has shown antidepressant-like effects in chronic restraint rats and sleep interruption-induced mice, and the present study aimed to further confirm the antidepressant effects of DS in a model of chronic unpredictable mild stress (CUMS) and to explore the underlying mechanism. Oral administration of DS (20, 40, and 80 mg/kg) markedly improved depressant-like behaviors, increasing the sucrose intake in the sucrose preference test and reducing the latency in the novelty-suppressed feeding test, and decreasing the immobility time in both the tail suspension and forced swimming tests, compared with the CUMS mice. Biochemical analysis of brain tissue and serum showed that DS treatment restored the decreased hippocampal neurotransmitter concentrations of serotonin, dopamine, norepinephrine (noradrenaline), and gamma-aminobutyric acid, and decreased the elevated of serum hormone levels (corticotrophin releasing factor, adrenocorticotrophic hormone, and corticosterone) induced by CUMS. Our findings confirm that DS exerts an antidepressant-like effect in the CUMS model of depression in mice, and suggest it may be mediated by regulation of neurotransmitters and hypothalamic-pituitary-adrenal axis.

A history of standardization in the English translation of traditional Chinese medicine terminology.

In order to facilitate and propose further international standardization of traditional Chinese medicine (TCM) terminology, this article applies methods of historiography, philology and descriptive study to divide the history of TCM into three phases, based on representative experts and social events; to illustrate different aspects of these experts and their translation principles and standards and to discuss associated factors and inherent problems. The authors find that the development of a terminology standard for TCM has generally progressed from early approaches that were ill-suited to the contemporary needs to culturally and professionally referenced approaches, from uncoordinated research to systematic studies, and from individual works to collaborative endeavors. The present international standards of TCM terminology have been attained through the work of numerous scholars and experts in the history of the field. The authors are optimistic that a more comprehensive and recognized standard will come out soon.

Epidemiological investigation of risk factors of the pregnant women with early spontaneous abortion in Beijing.

OBJECTIVE: To determine the risk factors of the pregnant women with early spontaneous abortion in Beijing. METHODS: A total of 34,417 cases of pregnant women were participated in the survey from January 2000 to December 2013. A questionnaire was informed to each woman. The content of questionnaire includes four parts: general condition, obstetrical history, past history and family history, and living environment and habits. The mental condition was evaluated with Self-Rating Anxiety Scale (SAS) and Self-Rating Depression Scale (SDS). RESULTS: A total of 32,296 questionnaires were collected. The spontaneous abortion rate in the total sample was 3.0%. There was no significant difference between the normal pregnancy group and spontaneous abortion group in terms of general condition, obstetrical and past history (P>0.05). Significant differences between the two groups were found in terms of decoration during pregnancy, keeping pets, near mobile communication base station within 100 m around the residence, drinking during pregnancy, having a cold during pregnancy and SAS (P<0.05). Having a cold during pregnancy, decoration during pregnancy, near mobile communication base station within 100 m around the residence, keeping pets and high SAS were determined the independent risk factors of spontaneous abortion by Logistic regression analysis. CONCLUSIONS: Having a cold during pregnancy, decoration, keeping pets, near mobile communication base station within 100 m around the residence and high SAS are the independent risk factors of spontaneous abortion in Beijing.

New Sweet-Tasting C21-Pregnane Glycosides from Pericarps of Myriopteron extensum.

Ten novel C21 pregnane glycosides, extensumside C-L (1-10), were isolated as highly sweet-tasting substances from the edible pericarps of Myriopteron extensum (Wight) K. Schum by sensory-guided fractionation and purification. Their structures were determined through 1D and 2D NMR, such as HSQC, HMBC, 1H-1H COSY, HSQC-TOCSY, and ROESY, as well as other spectroscopic analysis combined with chemical evidence. These compounds shared the same aglycone, 3ß,16α-dihydroxy-pregn-5-en-20-one, and contained the deoxysugar chain and the glucose chain which were linked to C-3 and C-16 of the aglycone, respectively. The sweetness potency was evaluated by a human sensory panel test and preliminary structure-taste relationship was discussed. The sweetness intensities of these compounds are between 50 and 400 times greater than that of sucrose. Furthermore, quantitation analyses of compounds 1, 3, 4, and 6 in different parts of M. extensum indicated that the concentrations of these sweet components in the pericarps are obviously higher than those in stems and roots.

Asiatic acid exerts anticancer potential in human ovarian cancer cells via suppression of PI3K/Akt/mTOR signalling.

Context Asiatic acid, a triterpenoid compound extracted from the tropical medicinal plant Centella asiatica (Family: Apiaceae), has exhibited various biological activities. Objective This study was performed to investigate the cytotoxic effects of asiatic acid on human ovarian cancer cells. Materials and methods SKOV3 and OVCAR-3 ovarian cancer cells were exposed to different concentrations of asiatic acid (10-100 µg/mL) for 72 or 48 h. Cell viability, colony formation, cell cycle distribution, apoptotic response were examined. Involvement of the phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathway was tested. Results At the concentration of 40 µg/mL, asiatic acid caused about 50% reduction in the viability of ovarian cancer cells, but had little effect on the viability of normal human ovarian epithelial cells. Asiatic acid at 10 µg/mL reduced colony formation of ovarian cancer cells by 25-30%. Asiatic acid-treated cells showed a cell cycle arrest at the G0/G1 phase and 7- to 10-fold increase in apoptosis. The phosphorylation levels of PI3K, Akt and mTOR were remarkably lower in asiatic acid-treated cells. Overexpression of constitutively active Akt partially reversed the cytotoxic effects of asiatic acid, as evidenced by increased cell viability and colony formation. Furthermore, knockdown of Akt mimicked the growth-suppressive activity of asiatic acid. Discussion and conclusion These results provide first the evidence for the anticancer potential of asiatic acid in ovarian cancer cells, partially via inactivation of the PI3K/Akt/mTOR pathway. Asiatic acid may represent a potential therapeutic agent for ovarian cancer.

Flavonoids and stilbenoids from Derris eriocarpa.

One new resveratrol analogue, 1-(3',4',5'-trimethoxyphenyl)-2-methoxy-2-(4″-methoxyphenyl)-ethane-1-ol (1), and two new prenylisoflavones, 4'-hydroxy-5,7-dimethoxy-6-(3-methyl-2-butenyl)-isoflavone (2), and derrubon 5-methyl ether (3), together with 17 known compounds including one new natural product, 5,7-dihydroxy-3-[4'-O-(3-methyl-2-butenyl)-phenyl]-isoflavone (4), were isolated from the stems of ethnomedicinal plant Derris eriocarpa How. (Leguminosae). Their structures were elucidated based on chemical evidence and spectroscopic techniques including two-dimensional NMR methods. All compounds are reported from this species for the first time. Antimicrobial activities of the new compounds were evaluated. Compound 2 exhibited good inhibitory activities against Candida guilliermondii, C. albicans and Microsporium gypseum with the minimal inhibitory concentration (MIC) values of 12.5 µg/ml.

[Effects of applying pig manure on lettuce yield and nitrate content and soil nutrients].

A pot experiment with two representative soils (purple soil and yellow soil) in Southwest China was conducted to study the effects of applying pig manure on the lettuce (Lactuca sativa L. var. capitata L. ) yield and nitrate content and the soil nutrients. Applying pig manure increased the lettuce yield significantly, and the increment was higher for yellow soil than for purple soil. The nitrate and total nitrogen, phosphorus, and potassium contents in lettuce plants were closely related to soil type and pig manure application rate. According to the evaluation standards of the nitrate pollution level of vegetables formulated by the Chinese Academy of Agricultural Sciences, the nitrate content in lettuce plants growing on purple soil was lower than the grade I (< or = 432 mg x kg(-1), slight pollution) in treatments CK (no pig manure application) and M1 ( applying 200 kg N x hm(-2) of pig manure), but generally higher than the grade II (< or = 758 mg x kg(-1), moderate pollution) while not exceeded the grade III (< or = 1440 mg x kg(-1), heavy pollution) in other treatments. The nitrate content in lettuce plants growing on yellow soil was lower than the grade I, except that in the treatments of chemical fertilizations and of M8 (applying 1600 kg N x hm(-2) of pig manure) where the plant nitrate content was exceeded the grade II. The critical value of Olsen-P characterizing the apparent leaching risk level of phosphorous in yellow soil and purple soil was 96.3 and 107.7 mg x kg(-1), respectively. The environmental safety capacity of pig manure was higher for yellow soil than for purple soil. Applying pig manure increased the organic carbon and total nitrogen contents of the two soils significantly.

[Rapid identification of 14 bile acids contained in Qingkailing injection by HPLC-ESI-MS/MS].

OBJECTIVE: To investigate ESI-MS/MS fragmentation pattern of bile acids contained in Qingkailing injection, and the rapid identification method. METHOD: HPLC-ESI-MS/MS technology was adopted to investigate ESI-MS/MS fragmentation pattern of five bile acids contained in Qingkailing injection, and rapidly identify bile acids contained in Qingkailing injection from eight different manufacturers. RESULT: 5 bile acids showed similar ESI-MS/MS fragmentation patterns, based on which 14 bile acids were rapidly separated from Qingkailing injection. Among them, 11 were found in injections of all of the manufacturers, and the rest three were found in individual manufacturers. CONCLUSION: HPLC-ESI-MS/MS is used to rapidly identify bile acids from complex material systems, which provides an effective method for interpreting the complex material base of compound traditional Chinese medicine preparations and enhance the quality standards.

Characterization of polymethoxylated flavonoids (PMFs) in the peels of 'Shatangju' mandarin ( Citrus reticulata Blanco) by online high-performance liquid chromatography coupled to photodiode array detection and electrospray tandem mass spectrometry.

A sensitive HPLC-DAD-ESI-MS/MS method was established to screen and identify the polymethoxylated flavonoids (PMFs) in the peels of 'Shatangju' mandarin ( Citrus reticulata Blanco). Eight PMF standards, including four polymethoxylated flavones, two polymethoxylated flavanones, and two polymethoxylated chalcones, were first to be analyzed in positive mode by CID-MS/MS. On the basis of the ESI-MS(n) characteristics of PMFs and the results of EIC-MS/MS experiment, 32 PMFs including 24 flavones and 8 flavanones or chalcones were screened from the complex extract of the peels of 'Shatangju' mandarin. Among them, 10 PMFs were hydroxylated polymethoxyflavonoids (OH-PMFs), and the rest were all permethoxylated PMFs. This was the first systematic report of the presence of PMFs in the peels of 'Shatangju' mandarin, especially for polymethoxylated flavanones and chalcones. Meanwhile, the contents of the three main PMFs and total flavonoids in the peels of 'Shatangju' were determined by HPLC and UV spectrophotometry, respectively. The results indicated that the developed analytical method could be employed as an effective technique for the characterization of PMFs.