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Alcoholic liver disease (ALD) results from continuous and heavy alcohol consumption. The current treatment strategy for ALD is based on alcohol withdrawal coupled with antioxidant drug intervention, which is a long process with poor efficacy and low patient compliance. Alcohol-induced CYP2E1 upregulation has been demonstrated as a key regulator of ALD, but CYP2E1 knockdown in humans was impractical, and pharmacological inhibition of CYP2E1 by a clinically relevant approach for treating ALD was not shown. In this study, we developed a RNAi therapeutics delivered by lipid nanoparticle, and treated mice fed on Lieber-DeCarli ethanol liquid diet weekly for up to 12 weeks. This RNAi-based inhibition of Cyp2e1 expression reduced reactive oxygen species and oxidative stress in mouse livers, and contributed to improved ALD symptoms in mice. The liver fat accumulation, hepatocyte inflammation, and fibrosis were reduced in ALD models. Therefore, this study suggested the feasibility of RNAi targeting to CYP2E1 as a potential therapeutic tool to the development of ALD.
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Based on ITS sequences, the molecular identification of Cordyceps cicadae and Tolypocladium dujiaolongae was carried out, and high-performance liquid chromatography(HPLC) fingerprint combined with chemical pattern recognition method was established to differentiate C. cicadae from its adulterant T. dujiaolongae. The genomic DNA from 10 batches of C. cicadae and five batches of T. dujiaolongae was extracted, and ITS sequences were amplified by PCR and sequenced. The stable differential sites of these two species were compared and the phylogenetic tree was constructed via MEGA 7.0. HPLC was used to establish the fingerprints of C. cicadae and T. dujiaolongae, and similarity evaluation, cluster analysis(CA), principal component analysis(PCA), and partial least squares discriminant analysis(PLS-DA) were applied to investigate the chemical pattern recognition. The result showed that the sources of these two species were different, and there were 115 stable differential sites in ITS sequences of C. cicadae and T. dujiao-longae. The phylogenetic tree could distinguish them effectively. HPLC fingerprints of 18 batches of C. cicadae and 5 batches of T. dujiaolongae were established. The results of CA, PCA, and PLS-DA were consistent, which could distinguish them well, indicating that there were great differences in chemical components between C. cicadae and T. dujiaolongae. The results of PLS-DA showed that six components such as uridine, guanosine, adenosine, and N~6-(2-hydroxyethyl) adenosine were the main differential markers of the two species. ITS sequences and HPLC fingerprint combined with the chemical pattern recognition method can serve as the identification and differentiation methods for C. cicadae and T. dujiaolongae.
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Cromatografía Líquida de Alta Presión/métodos , Cordyceps/genética , Hypocreales , FilogeniaRESUMEN
Tuber rot has become a serious problem in the large-scale cultivation of Gastrodia elata. In this study, we compared the resistance of different ecotypes of G. elata to tuber rot by field experiments on the basis of the investigation of G. elata diseases. The histological observation and transcriptome analysis were conducted to reveal the resistance differences and the underlying mechanisms among different ecotypes. In the field, G. elata f. glauca had the highest incidence of tuber rot, followed by G. elata f. viridis, and G. elata f. elata and G. elata f. glauca×G. elata f. elata showed the lowest incidence. Tuber rot showcased obvious plant source specificity and mainly occurred in the buds and bottom of G. elata plants. After infection, the pathogen spread hyphae in host cortex cells, which can change the endophytic fungal community structure in the cortex and parenchyma of G. elata. G. elata f. glauca had thinner lytic layer and more sugar lumps in the parenchyma than G. elata f. elata. The transcription of genes involved in immune defense, enzyme synthesis, polysaccharide synthesis, carbohydrate transport and metabolism, hydroxylase activity, and aromatic compound synthesis had significant differences between G. elata f. glauca and G. elata f. elata. These findings suggested that the differences in resis-tance to tuber rot among different ecotypes of G. elata may be related to the varied gene expression patterns and secondary metabolites. This study provides basic data for the prevention and control of tuber rot and the improvement of planting technology for G. elata.
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Ecotipo , Gastrodia/microbiología , Perfilación de la Expresión Génica , Tubérculos de la Planta/genéticaRESUMEN
The continuous cropping obstacle of Gastrodia elata is outstanding, but its mechanism is still unclear. In this study, microbial changes in soils after G. elata planting were investigated to explore the mechanism correlated with continuous cropping obstacle. The changes of species and abundance of fungi and bacteria in soils planted with G. elata after 1, 2, and 3 years were compared. The pathogenic fungi that might cause continuous cropping diseases of G. elata were isolated. Finally, the prevention and control measures of soil-borne fungal diseases of G. elata were investigated with the rotation planting pattern of "G. elata-Phallus impudicus". The results showed that G. elata planting resulted in the decrease in bacterial and fungal community stability and the increase in harmful fungus species and abundance in soils. This change was most obvious in the second year after G. elata planting, and the soil microbial community structure could not return to the normal level even if it was left idle for another two years. After G. elata planting in soils, the most significant change was observed in Ilyonectria cyclaminicola. The richness of the Ilyonectria fungus in soils was significantly positively correlated with the incidence of G. elata diseases. When I. cyclaminicola was inoculated in the sterile soil, the rot rate of G. elata was also significantly increased. After planting one crop of G. elata and one to three crops of P. impudicus, the fungus community structure in soils gradually recovered, and the abundance of I. cyclaminicola decreased year by year. Furthermore, the disease rate of G. elata decreased. The results showed that the cultivation of G. elata made the Ilyonectria fungi the dominant flora in soils, and I. cyclaminicola served as the main pathogen of continuous cropping diseases of G. elata, which could be reduced by rotation planting with P. impudicus.
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Bacterias , Hongos , Gastrodia/microbiología , Micobioma , Suelo , Microbiología del SueloRESUMEN
This study aims to explore the resource utilization of used fungus-growing materials produced in the cultivation of Gastrodia elata. To be specific, based on the production practice, this study investigated the recycling mechanism of used fungus-growing materials of G. elata by Phallus inpudicus. To screen edible fungi with wide adaptability, this study examined the allelopathic effects of Armillaria mellea secretions on P. impudicus and 6 kinds of large edible fungi and the activities of enzymes related to degradation of the used fungus-growing materials of G. elata. The results showed that P. impudicus can effectively degrade cellulose, hemicellulose, and lignin in used fungus-growing materials of G. elata. The cellulase activity of A. mellea was significantly higher than that of P. impudicus, and the activities of lignin peroxidase, polyphenol oxidase, and xylanase of P. impudicus were significantly higher than those of A. mellea, which was the important reason why A. mellea and P. impudicus used different parts and components of the used fungus-growing materials to absorb carbon sources and develop ecological niche differences. The growth of P. impudicus was significantly inhibited on the used fungus-growing materials of G. elata. The secretions of A. mellea had allelopathic effects on P. impudicus and other edible fungi, and the allelopathic effects were related to the concentration of allelopathy substances. The screening result showed that the growth and development of L. edodes and A. auricular were not significantly affected by 30% of A. mellea liquid, indicating that they had high resistance to the allelopathy of A. mellea. The results showed that the activities of extracellular lignin peroxidase, polyphenol oxidase, and xylanase of the two edible fungi were similar to those of P. impudicus, and the cellulase activity was higher than that of P. impudicus. This experiment can be further verified by small-scale production tests.
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Agaricales , Ascomicetos , Basidiomycota , Catecol Oxidasa , Celulasas , GastrodiaRESUMEN
Through consulting the ancient herbs, medical books and modern literature, this paper made textual research on the name, origin, producing area, quality evaluation, collection and processing of medicinal materials of Sang (Mori Folium, Mori Cortex, Mori Ramulus, Mori Fructus) in famous classical formulas, in order to provide a basis for the development of famous classical formulas containing medicinal materials of Sang. According to the research, Mori Folium and Mori Cortex were first used as medicines in Shengnong Bencaojing , Mori Ramulus was first used as medicine in Jinxiaofang, and Mori Fructus was first used as medicine in Xinxiu Bencao. Before the Tang dynasty, there were Nyusang and Shansang. Since Tang dynasty, there were many sources of medicinal materials of Sang, including Baisang (Morus alba), Jisang (M. australis), Shansang (M. mongolica), etc. According to textual research, the mainstream varieties were M. australis, M. alba and their cultivated varieties. In modern times, according to the relevant information and the Chinese Pharmacopoeia, M. alba is the original base. In ancient times, the origin of mulberry changed with the development of sericulture, mulberry has been widely planted since the Song dynasty. In the Ming and Qing dynasties, mulberry has been planted most in Jiangsu and Zhejiang. In modern times, they are mainly produced in Jiangsu, Zhejiang, Anhui, Hunan and other places. In recent years, due to the related policies and strategies such as "moving silkworms from east to west", the center of silkworm breeding has gradually transferred to the west. As for the quality evaluation and harvesting and processing of mulberry medicinal materials, Most of the ancient and modern records of Mori Folium are the same. They are harvested after frost, and dried after removing impurities. The quality is better when the leaves are large and thick, yellowish green, holding prickly hands and undergoing frost. The harvesting period of Mori Cortex is slightly different in ancient and modern records. Ancient books record that it can be harvested all the year round, but in modern times, it is mostly harvested from late autumn to the next spring. The processing methods include removing soil and fibrous roots, scraping off yellow-brown rough skin, peeling off white skin and drying in the sun. The quality is better when they are white, thick, flexible, free of rough skin and full of powder. There are few records about the collection, processing and quality evaluation of Mori Ramulus and Mori Fructus in ancient Chinese herbal books. According to modern literature, Mori Ramulus is usually collected in late spring and early summer, with leaves removed, slightly dried, sliced while fresh, and dried in the sun. The best quality of Mori Ramulus is fine and tender with the yellow and white section. Mori Fructus is harvested from April to June when the fruit turns red, and dried in the sun, or slightly steamed and dried in the sun, and it is better to be big, dark purple, oily and thick. There are many processing methods of mulberry medicinal materials. Ancient books record stir frying, baking, burning and steaming of Mori Folium, in modern times, there is honey-roasted method, but most of them are used as raw products. In ancient materia medica, Mori Cortex has firing method, baking method, stir-frying method, honey-fried method, etc. In modern times, there are stir-fried and honey-fried methods, and most of them are used as raw products. Ancient books record that Mori Ramulus has cutting and frying methods, while modern ones have cutting, frying, wine-processed and bran-processed methods. Processing methods of Mori Fructus are consistent in ancient and modern times, and they are mostly dried after being cleaned or steamed. Based on the research results, it is suggested that M. alba should be selected as mulberry medicinal materials in the famous classical formulas, and appropriate medicinal parts and processing methods can be selected according to the indications of the famous classical formulas.
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In this paper, the name, origin, producing area and other aspects of Menthae Haplocalycis Herba in the famous classical formulas were carried out by consulting herbal literature, medical books, prescription books in the past dynasties and related modern documents. Through the textual research, it can be seen that the name of Bohe was used as the correct name in the mainstream of the past dynasties, and there were still multiple synonyms, most of which originated from the false transmission of dialectal accent, producing area and efficacy. There are many varieties recorded in the literature of the past dynasties such as Bohe, Longnao Bohe, Hubohe and Shibohe. According to the textual research, Bohe, Longnao Bohe and Yebohe are consistent with Mentha haplocalyx, whcih is the mainstream variety. Longnao Bohe is named for its form of producing area, Shibohe is Mosla chinensis, Daye Bohe is Agastache rugosa, and Nanbohe is M. crispata. Menthae Haplocalycis Herba has been widely planted since Tang dynasty. It was mainly grown in Jiangsu, Zhejiang, Jiangxi and Sichuan in Ming and Qing dynasties, and Jiangsu is the genuine production area. Its quality is best if it has dry body, no roots, many leaves, green color and strong smell. In ancient times, the stems and leaves of Menthae Haplocalycis Herba were often picked and dried in summer and autumn, which is basically the same as the records of modern times when the stems and leaves are luxuriant in summer and autumn, or when the flowers bloom to three rounds, they are picked in sunny days and cut in different times, and then dried in the sun or in the shade, and the raw products was often used as medicine in ancient and modern times. Before the Song dynasty, Menthae Haplocalycis Herba was recorded as pungent and warm. Until the Song dynasty, it was written as “extremely cool” in Lyuchanyan Bencao. It may have been thought in the early stage that it was similar to several warm herbs, such as Perilla frutescens, Stachys japonica, Elsholtzia ciliata and M. chinensis in appearance, all of which have the function of Xinsan, so it was recorded as warm. Since the Qing dynasty, Menthae Haplocalycis Herba has been recorded as cool property in the mainstream materia medica, Menthae Haplocalycis Herba recorded as pungent and cool in the 2020 edition of Chinese Pharmacopoeia, and its effect is to dissipate wind heat, clear the head, relieve the pharynx and so on, the records of efficacy in ancient and modern times are basically the same. Based on the research results, it is suggested that raw products of M. haplocalyx should be selected when developing the famous classical formulas containing Menthae Haplocalycis Herba.
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Visci Herba, a commonly used Chinese medicinal, was often mistaken as Taxilli Herba in ancient Chinese materia medica. The two Chinese medicinals have often been confused even in present clinical practice, and their origins are difficult to be identified. Hence, it is necessary to carry out systematic and in-depth textual research. This paper explored the origin, producing area, quality evaluation, harvesting and processing, property, flavor, and efficacy of Visci Herba based on ancient Chinese materia medica of the past dynasties and modern plant morphology, so as to provide evidence for the development and utilization of Visci Herba. The findings demonstrated that Visci Herba was mostly recorded as Taxilli Herba until the name of Visci Herba appeared in the Tang Dynasty. The records of the two Chinese medicinals could be traced back to the Song Dynasty. Visci Herba and Taxilli Herba were officially listed as two different Chinese medicinals in the 1977 edition Chinese Pharmacopoeia for the first time, where the origin of Visci Herba was determined to be Viscum coloratum(Komar.)Nakai. According to the ancient Chinese materia medica, V. coloratum was mainly distributed in Lingbao, Henan province and Xuzhou, Jiangsu province and now it mostly grows in Northeast China and North China. In ancient times, Visci Herba with deep yellow cross-section and sticky fruit juice on the tree was preferred, which was often harvested on 3, March in spring, dried in the shade, grinded together with the roots, branches, stems, and leaves using the copper file, and preserved away from fire. By contrast, it is now usually harvested from winter to the next spring and then cut into sections for drying after the removal of thick stems, or dried after being steamed. As described in ancient Chinese materia medica, Visci Herba, bitter and sweet in flavor, neutral in property, possessed the effects of dispelling rheumatism, tonifying kidney, strengthening sinews and bones, benefiting blood vessels, and preventing abortion, basically consistent with its modern functions of dispelling rheumatism, tonifying liver and kidney, strengthening sinews and bones, and preventing abortion.
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The differences of the active ingredients in Dendrobium huoshanense of different growth years and their protective effects on acute liver injury were studied to provide evidence for optimizing harvest time. The contents of polysaccharides, total flavonoids and total alkaloids in D. huoshanense of different growth years were determined by UV spectrophotometry, and the contents of gigantol in D. huoshanense were determined by HPLC. C57 BL/6 mice were randomly divided into blank control group(saline), modeling group(saline), high-dose(7.5 g·kg~(-1)) and low-dose(1.25 g·kg~(-1)) groups of D. huoshanense of different growth years. Each group was intragastrically administered every day for 2 weeks. 500 mg·kg~(-1) paracetamol was injected intraperitoneally 2 h after last treatment except the control group. After 12 hours, the serum and liver tissues were collected to detect the activities of ALT and AST, and the levels of SOD and MDA. The hepatic histopathological examination was performed. The results showed that the chemical constituents of D. huo-shanense of different growth years were significantly different(P<0.05). The contents of polysaccharide and gigantol of D. huoshanense of 2 growth years were the highest. The contents of flavonoids and alkaloids of D. huoshanense of 3 growth years were the hig-hest, followed by the D. huoshanense of 2 growth years, and the lowest were that of 1 growth year. Compared with the modeling group, D. huoshanense of different growth years could decrease the activities of ALT and AST in serum. Meanwhile, the levels of MDA reduced significantly, while those of SOD increased markedly. Histopathological results suggested that all D. huoshanense samples were effective in the reduction of the necrosis of hepatocytes in different degrees. The results of the multi-component SPSS paired tests showed that polysaccharide and gigantol probably played a leading role in the liver protection effects, while D. huoshanense of 2 growth years showed the best efficacy. The optimal harvesting time of D. huoshanense is 2 growth years.
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Animales , Ratones , Alcaloides , Cromatografía Líquida de Alta Presión , Dendrobium , Hígado , PolisacáridosRESUMEN
Rubi Fructus is a commonly used traditional Chinese medicine. The origin of Rubi Fructus is the dried fruit of Rubus chingii, a plant of the family Rosaceae, according to the 2015 edition of Chinese pharmacopoeia. There are some differences in the plant origin of Rubi Fructus in ancient herbal literature, to trace back its sources, we conducted a textual research on its origin, producing areas, quality evaluation, processing and concocting, properties, tastes and efficacy etc. based on the records of ancient herbal literatures and combined with plant morphology and related investigation. RESULTS:: showed that the variety of Rubi Fructus was more complex among ancient herbal literature, including R. coreanus, R. hirsutus, R. corchorifolius, R. foliolosus and other mixed varieties. Most scholars believe that the R. chingii has not been recorded in ancient herbal literature, while R. chingii was recorded as early as the Ming Dynasty in Compendium of materia medica through our textual research. Ancient Chinese herbs recorded that Rubi Fructus was mostly produced in Hubei, Shandong, Shanxi and Jiangsu provinces, while R. chingii mainly produced in Anhui, Jiangsu, Zhejiang, Jiangxi, Fujian and other provinces nowadays. Also, it was recorded that Rubi Fructus harvested in wheat field during May were the best. Besides, R. chingii with big, full, grain integrate, firm, yellow and green color, sour taste and impurity free possess the best quality in the contemporary. The ancient records of processing and concocting, properties, tastes and efficacy were basically the same as modern ones.These results provide the basis for the correct utilization and further development of Rubi Fructus.
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Humanos , China , Medicamentos Herbarios Chinos , Frutas , Materia Medica , Medicina Tradicional China , RubusRESUMEN
In this study, HPLC-ESI-MS and HPLC methods were established to explore the differences in the main chemical components and content of Mori Cortex with(mulberry root bark) and without(Mori Cortex) the phellem layer from both qualitative and quantitative aspects. The HPLC-ESI-MS method was used for quality analysis in positive and negative ion modes, and 33 compounds were identified in mulberry root bark, 22 compounds in Mori Cortex, and 26 compounds in phellem layer; mulberry root bark and Mori Cortex shared 22 components, and mulberry root bark has 11 unique compounds; Mori Cortex and its phellem layer shared 15 components, while Mori Cortex has 7 unique compounds. HPLC method was used to simultaneously determine 7 major constituents, including mulberroside A, chlorogenic acid, dihydromorin, oxyresveratrol, moracin O, kuwanon G, and kuwanon H, and the developed method showed good linearity(r>0.998 9) within the concentration range and the recoveries varied from 99.88% to 103.0%, and the RSD was 1.7%-2.9%. The HPLC results showed that the contents of the 7 compounds have great differences in 13 batches samples, compared with mulberry root bark, the contents of mulberroside A, chlorogenic acid, dihydromorin and moracin O of Mori Cortex were increased, while the contents of oxyresveratrol, kuwanon G and kuwanon H were decreased after peeling process. These results can provide a basis for the rationality and quality control of Mori Cortex required to remove the phellem layer.
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Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos , Espectrometría de Masas , Morus , Corteza de la PlantaRESUMEN
A highly sensitive monoclonal antibody against aflatoxin B_1(AFB_1) was prepared and an indirect competition enzyme-linked immunosorbent assay(ic-ELISA) was established based on the antibody which was used for high-throughput and rapid screening of AFB_1 contamination in Chinese herbal medicines to ensure the safety of medication. In this study, the structure of AFB_1 was modified by improved oxime method, and the carrier protein was coupled by EDC-NHS method to obtain the complete antigen of AFB_1, which was more convenient and environmental friendly. The Balb/c female mice were immunized using increasing the immunization dose and various ways of injection, and finally the AFB_1 monoclonal antibody was prepared. The AFB_1 monoclonal antibody belongs to IgG_(2 b) immunoglobulin by identifying its immunological characteristics, and its sensitivity(IC_(50)) can reach 0.15 μg·L~(-1), and the affi-nity is 2.81×10~8 L·mol~(-1). The cross-reaction rates of AFB_2, AFG_1, and AFG_2 were 35.07%, 8.75%, and 1.15%, respectively, and there was almost no cross-reactivity with other mycotoxins. Based on the high sensitivity and specificity of the antibody, an ic-ELISA method was established and applied to the determination of AFB_1 contamination in Ziziphi Spinosae Semen. According to the matrix matching standard curve, the linear concentration range for AFB_1 was 0.05-0.58 μg·L~(-1)(R~2=0.992), the recoveries were 88.00%-119.0%, and the detection limit was 1.69 μg·kg~(-1). The AFB_1 in 33 batches of Ziziphi Spinosae Semen samples was determined by ic-ELISA, and the contamination level was 3.62-206.58 μg·kg~(-1). The linear correlation coefficient between the detection results of ic-ELISA and UHPLC-MS/MS was 0.996, and there were no false positive and false negative cases. It indicates that the established ic-ELISA is accurate and reliable, and could provide a simple and effective technique for fast screening of AFB_1 contamination in Ziziphi Spinosae Semen, and also could be considered as the reference for the detection and monitoring of AFB_1 contamination in other Chinese herbal medicines.
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Animales , Femenino , Ratones , Aflatoxina B1 , Anticuerpos Monoclonales , Contaminación de Medicamentos , Ensayo de Inmunoadsorción Enzimática , Semen , Química , Espectrometría de Masas en TándemRESUMEN
OBJECTIVE@#To explore the clinical effect of knee denervation combined with micro-fracture under arthroscope in the treatment of patellofemoral arthritis.@*METHODS@#From May 2015 to May 2018, 60 patients with knee joint degenerative patellofemoral arthritis were treated, including 28 males and 32 females, aged from 24 to 56(40.5±3.35) years old. Among them, 30 cases underwent arthroscopic debridement alone(control group), 30 cases underwent arthroscopic debridement, peripatellar denervation combined with cartilage cone and patellofemoral articular surface microfracture treatment (treatment group). VAS, Lysholm and Kujala scores were used to evaluate the therapeutic effect.@*RESULTS@#No complications such as wound infection, vascular nerve injury and deep venous thrombosis occurred in all patients. The patients were followed up for 7 to 36 months with an average of 12.5 months. The VAS scores of the two groups were improved 4 weeks after operation, and the improvement of the treatment group was significantly better than that of the control group, the difference was statistically significant (<0.05). Lysholm and Kujala scores were compared at the last follow-up of the two groups, the improvement of the treatment group was significantly better than that of the control group.@*CONCLUSIONS@#The treatment of patellofemoral arthritis with patellar denervation combined with microfracture can better relieve pain and improve knee function.
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Adulto , Anciano , Femenino , Humanos , Masculino , Adulto Joven , Artritis , Desnervación , Fémur , Fracturas por Estrés , Articulación de la Rodilla , Rótula , Articulación Patelofemoral , Resultado del TratamientoRESUMEN
This study aimed to analyze the residues of aflatoxin B_1( AFB_1) in Ziziphi Spinosae Semen from different producing areas and to assess the health risk of aflatoxin B_1 residue based on the obtained data. A total of 72 samples of Ziziphi Spinosae Semen from different areas were detected by IAC-HPLC-FLD. Based on the data of AFB_1 pollution,a probabilistic assessment model with Monte Carlo simulation was developed. Then,the risk assessment of AFB_1 exposure by Ziziphi Spinosae Semen intake was carried out by MOE( margin of exposure). The results showed that 32 out of 72 of samples( 44. 4%) were found to be contaminated with AFB_1,and the average and maximum concentration of AFB_1 in samples was 5. 42 μg·kg~(-1) and 55. 09 μg·kg~(-1),respectively. After health risk assessment,the average and 97. 5%( 90% confidence interval) exposure level of daily exposure of AFB_1 by Ziziphi Spinosae Semen intake were 0. 008 6( 0. 008 1-0. 009 2) and 0. 057 3( 0. 053 2-0. 061 4) μg·kg~(-1)·d~(-1),respectively. The results showed common use of Ziziphi Spinosae Semen had low level of risk associated with AFB_1. However,the high consumption of Ziziphi Spinosae Semen showed a higher risk than common intake,requiring attention. This study laid a foundation for clinical safe prescription of Ziziphi Spinosae Semen.
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Aflatoxina B1/análisis , Cromatografía Líquida de Alta Presión , Contaminación de Medicamentos , Preparaciones de Plantas/análisis , Medición de Riesgo , Ziziphus/químicaRESUMEN
OBJECTIVE: Osteosarcoma is an aggressive malignant neoplasm, which commonly afflicts patients of 20-30 years of age, and its morbidity has increased markedly in recent years. Certain genes and signal pathways have been identified to exert key roles in osteosarcoma progression. Here, we set out to characterize in more detail of the role of HIF-1/AKT/Cyclin D1 pathway in the progression of osteosarcoma. METHODS: Immunohistochemistry, western blot and qPCR were used to test the protein or mRNA levels of HIF-1 in osteosarcoma tissues or adjacent nontumor tissues. MTT, clone formation, wound healing, Transwell, in vivo tumorigenesis, flow cytometry and western blot analysis were used to determine cell proliferation, clone formation ability, migration, invasion, tumorigenesis, and cell apoptosis in MG63 and U2OS cells, respectively. Immunoprecipitation and immunofluorescence assays were performed to investigate the protein-protein interaction between HIF-1α and proteins related to signal pathways. RESULTS: HIF-1 was overexpressed in osteosarcoma tissues and cell lines, which promoted cell proliferation, clone formation, migration, invasion and inhibited cell apoptosis. Results also demonstrated that HIF-1 combined with AKT, and there might be a positive loop between the two proteins of HIF-1 and AKT, then the protein-protein interaction up-regulated the expression of Cyclin D1 in protein level, but not mRNA level, made Cyclin D1 protein more stable, triggered cell proliferation, clone formation, tumorigenesis, but inhibited cell apoptosis. CONCLUSIONS: The present study showed that HIF-1 modulated Cyclin D1 expression might through shaping a positive loop with AKT proteins. Additionally, HIF-1α promoted the tumor cells growth, migration and invasion in osteosarcoma through the activation of the AKT/Cyclin D1 signal cascade. We proposed that HIF-1 could be served as a marker for distinguishing osteosarcoma and an effective therapeutic target for osteosarcoma.
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Neoplasias Óseas/genética , Ciclina D1/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Osteosarcoma/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Animales , Apoptosis/genética , Neoplasias Óseas/patología , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Humanos , Ratones Endogámicos BALB C , Ratones Desnudos , Invasividad Neoplásica , Osteosarcoma/patología , Transducción de SeñalRESUMEN
As an important part of traditional medicine in China, traditional Chinese medicine(TCM) plays a significant role because of its unique medical efficiency, less adverse reactions and extensive resources. However, in recent years, the aflatoxins in medicinal herbs have been detected excessive both at home and abroad, seriously affecting the reputation and credibility of traditional Chinese medicine. In this paper, the current status of aflatoxins contamination in medicinal herbs was analyzed, and the internal and external factors of aflatoxins contamination in traditional Chinese medicine were also summarized. In view of the high toxicity of aflatoxins, it is proposed to strengthen the mildew prevention and control from the early planting to storage stage, and the reasonable detoxification mode should also be considered. This review aims to provide a reference in guaranteeing the clinical safe administration of medicinal herbs and reducing the risk of being poisoned by aflatoxins.
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Aflatoxinas , China , Contaminación de Medicamentos , Medicamentos Herbarios Chinos , Química , Medicina Tradicional China , Plantas MedicinalesRESUMEN
This study was carried out to investigate the effect of oral administration of Dendrobium huoshanense on the expressions and activities of hepatic microsomal cytochrome P450s in mice, and to provide a reference for the evaluation of drug-drug interactions between D. huoshanense and clinical drugs. The C57BL/6 mice were randomly divided into blank control group, D. huoshanense low dose group (crude drug 1.25 g·kg⁻¹), D. huoshanense high dose group (crude drug 7.5 g·kg⁻¹), and phenobarbital positive control group (0.08 g·kg⁻¹). Each group was intragastrically administered with drugs for 2 weeks. The mice were sacrificed and their liver microsomes were prepared. The expressions of major subtypes of P450 enzyme were determined by Western blot and the probe drugs were used to detect the enzyme activities of P450 subtypes with protein expression changes. Western blot analysis showed that the protein expressions of CYP1A1, CYP1A2 and CYP2B in liver tissues were up-regulated in D. huoshanense-treated group. In vitro enzyme activity tests showed that there were no significant difference in metabolism of 7-ethoxyresorufin (a probe drug for CYP1A1) and bupropion (a probe drug for CYP2B) between D. huoshanense group and control group. The metabolism of phenacetin (a probe drug for CYP1A2) showed a statistical difference in rate Vmax, and it was significantly increased by approximately 20% in D. huoshanense group as compared with the blank control group, and the clearance CLint in treated group was also increased by about 32%. Therefore, oral administration of D. huoshanense had no effects on the activities of most hepatic P450 enzymes in mice, with no drug-drug interaction related to the P450 enzyme system in most clinical drugs theoretically. However, oral administration of D. huoshanense may accelerate the metabolism of CYP1A2-catalyzed drugs, which needs to be considered in clinical practice.
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Animales , Ratones , Citocromo P-450 CYP1A1 , Metabolismo , Citocromo P-450 CYP1A2 , Metabolismo , Sistema Enzimático del Citocromo P-450 , Metabolismo , Dendrobium , Química , Medicamentos Herbarios Chinos , Farmacología , Ratones Endogámicos C57BL , Microsomas Hepáticos , Distribución AleatoriaRESUMEN
AIM To investigate the effects of imperatorin and isoimperatorin on the expression of mouse liver cytochrome P450s and hepatic toxicity in mice.METHODS C57BL/6 mice were randomly divided into control and administration groups,which were treated with imperatorin or isoimperatorin by intragastric administration for two weeks.The effects of two compounds on mRNA expressions of major P450s isoforms were analyzed by RT-PCR.The P450 expression was determined by Western blot.The serum levels of glutamicpyruvic transaminase (GPT),glutamic-oxalacetic transaminase (GOT) and blood total bilirubin (TBIL) were detected by kits.The change of liver tissue was observed with HE staining.RESULTS The Cyp1a2 mRNA expression was significantly induced by 40 mg/kg imperatorin as compared with the control group.For isoimperatorin,the Cyp2c37 mRNA expression was significantly induced.Western blot results showed that CYP1 A2 expression was significantly induced by imperatorin.For isoimperatorin,the CYP2C and CYP2E1 expressions were significantly induced.Blood biochemical indices showed that 40 mg/kg isoimperatorin led to increased serum GOT and TBIL levels.Pathological analysis showed that both compounds (at the doses of 20 mg/kg and 40 mg/kg) could cause liver edema to a certain degree.CONCLUSION Imperatorin is the inducer of CYP1A2,while isoimperatorin is the inducer of CYP2C and CYP2E1.These two compounds (at the doses of 20 mg/kg and 40 mg/kg) can lead to damage for mouse liver.The toxicity of isoimperatorin is stronger than that of imperatorin.
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AIM To observe the effects of Sangtongjian Mixture (STJ) on glucose and lipid metabolism,insulin resistance and fat cytokines in type 2 diabetic rats,and their mechanisms of action.METHODS One hundred and forty rats fed on the combination of STZ and high fat diet were established as the type 2 diabetic models.Fasting blood glucose (FBG) level reached more than 16.7 mmol/L and then the rats were randomly divided into model group,metformin (180 mg/kg) group,STJ (73.5,147 and 294 mg/kg) groups.Ten rats were set as the blank group.Each treatment group was intragastrically given the corresponding agents for twelve weeks.The fasting blood glucose levels of rats were measured once every two weeks after the administration.After a 12-week administration period,glycosylated serum protein (GSP),glycosylated hemoglobin (GHb) and lipid profile indices (TC,TG,HDL-C and LDL-C) were determined.The serum insulin level was measured by radioimmunoassay,and homeostasis model assessment of insulin resistance (HOMA-IR) and insulin sensitivity index (ISI) were calculated.The levels of serum adiponectin and leptin were detected by ELISA.RESULTS STJ remarkably decreased the levels of FBG,GSP,GHb,TC,TG,LDL-C,leptin and HOMR-IR in type 2 diabetic rats.Furthermore,STJ also significantly increased the levels of HDL-C,adiponectin and ISI.CONCLUSION STJ can improve glucose and lipid metabolism in type 2 diabetic rats by ameliorating insulin resistance and regulating fat cytokine levels.
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This paper investigated the diversity of the silkworm excrement bacterial communities in different ages before and after drying, aiming to clarify the differences of bacterial communities in composition and bacterial abundance and the influences of drying treatment, and provide scientific basis for the efficacy of scientific connotation and utilization of silkworm excrement. High-throughput sequencing technique was used to measure the sequence of 16S rDNA-V4 variable region of bacteria in silkworm excrement. QIIME, Mothur and PICRUSt software programs were employed to sort and calculate the number of sequences and operational taxonomic units (OTUs) for each sample. Thereafter, the abundance, distribution, alpha diversity index of species, beta diversity and bacterial communities diversity among different sample groups and predicted the bacterial gene functions were analyzed. In this study, the numbers of effective sequences for six samples were 259 250; the rarefaction curves showed a sufficient sequencing depth, and the number of OTUs was close to saturation. The bacteria in silkworm excrement belonged to the following five phylums: Proteobacteria (89.3%), Actinobacteria (5.0%), Firmicutes (4.4%), Bacteroidetes (1.1%) and Cyanobacteria (0.2%). The dominant specie was Cyanobacteria of the total bacteria identified, respectively. The abundances and diversities of the silkworm excrement bacterial communities have been reduced after drying treatment, especially the silkworm excrement of the fifth instar. PICRUSt analysis was performed to show that abundance of the functional genes such as membrane transport, carbohydrate metabolism, amino acid metabolism, lipid metabolism, nucleotide metabolism, cellular processes and signaling were relatively high. The result showed that the drying treatment could decreased the species and numbers of pathogenic bacteria in silkworm excrement obviously and improve the quality of medicinal materials. Compared with the lower ages, silkworm excrement of fifth instar seems like to be more suitable for use in medicine. Illumina MiSeq high-throughput sequencing system provides a more accurate and scientific data resource for the study of bacteria in silkworm excrement.