The medicines targeted at α-glucosidase played an important role in anti-diabetes and anti-HIV therapy. Unfortunately, the method based on fluorescent assay strategy for α-glucosidase inhibitor screening remains poorly investigated. In this study, a novel "Turn On" fluorescence sensor platform has been developed for trace α-glucosidase inhibitor screening from natural medicines. Firstly, carbon dots were prepared by one-pot synthesis and used as the signal output. Combining with the carbon dots, cobalt oxyhydroxide (CoOOH) nanoflakes were employed to build the fluorescence resonance energy transfer (FRET) based sensor platform. Secondly, L-ascorbic acid-2-O-α-D-glucopyranosyl (AAG) was innovatively introduced as α-glucosidase substrate. With hydrolysis of AAG by α-glucosidase, ascorbic acids (AA) were released that can rapidly reduce CoOOH nanoflakes to Co(2+), and then FRET was stopped accompanying with the fluorescence recovery of CDs. The sensor platform was ultrasensitive to AA with a detection limit of 5 nM, ensuring the sensitive monitoring of enzyme activity. Acarbose was used as the inhibitor model and its inhibition rate is proportional to the logarithm of concentration in range of 10(-9)-10(-3)M with the correlation coefficient of R(2)=0.996, and an ultralow limit of detection of ~1×10(-9)M was obtained. The inhibiting ability of seven compounds isolated from natural medicines was also evaluated. The constructed sensor platform was proven to be sensitive and selective as well as cost-effective, facile and reliable, making it promising as a candidate for trace α-glucosidase inhibitor screening.
<b>Rheum tanguticum</b> (Polygonaceae), a plant species used in both traditional Chinese and Tibetan medicine, has been listed as endangered species because its distribution in the Tibetan Plateau has rapidly decreased in recent years. We estimated the genetic diversity of <b>R. tanguticum</b> using the cpDNA <b>trn</b>L-F region for 95 individuals from nine populations. The results demonstrated high genetic diversity in this species (Ht = 0.632), primarily due to variation among populations (68.4%, GST = 0.6839), as opposed to variation within populations (31.6%, Hs = 0.2). AMOVA analyses indicated that genetic differentiation among populations was very high (FST = 0.7009), and gene flow among populations was low (Nm = 0.11). The reason for high genetic differentiation among populations might be due to the geography of the alpine environment of the region and human activity. Furthermore, genetic structure in <b>R. tanguticum</b> implies that wild populations with high genetic diversity should be protected, and collecting seeds from populations with high genetic diversity is necessary for conservation breeding programs.<br>• High genetic diversity and large genetic differentiation among populations were observed in <b>Rheum tanguticum.</b> • The main reason for the observed genetic structure might be the geography of the alpine environment and human activity. • Conservation plans for <b>in situ</b> and <b>ex situ</b> protection were proposed.
To achieve a high yield of tropane alkaloids (TA) and exploit the alpine plant sustainably, an optimized protocol for induction and establishment of hairy roots culture of <i>Prezwalskia tangutica</i> Maxim was developed through selection of appropriate <i>Agrobacterium</i> strain and the explant type. The hypocotyl is more readily facile to induce the HR than the cotyledon is when infected with the three different agrobacterium strains. MUS440 has an efficiency (of up to 20%), whereas the ATCC10060 (A4) can induce HR on both types of explants with the highest frequency (33.33%), root length (21.17 ± 2.84 cm), and root number (10.83 ± 1.43) per explant than the other strains. The highest HR production resulted from using hypocotyl as explants. Independent transformed HR was able to grow vigorously and to propagate on a no-hormone 1/2MS liquid medium. The presence of pRi <i>rol</i>B gene in transformation of HR was confirmed by PCR amplification. In the liquid medium, the HR growth curve appeared to be “S” shaped, and ADB had increased to 4.633 g/l. Moreover, HPLC analysis showed that HR lines have an extraordinary ability to produce atropine (229.88 mg/100 g), anisodine (4.09 mg/100 g), anisodamine (12.85 mg/100 g), and scopolamine (10.69 mg/100 g), which were all more significant than the control roots. In conclusion, our study optimized the culture condition and established a feasible genetics reactor for <i>P. tangutica</i> green exploration and biological study in the alpine region.
<i>Rheum tanguticum</i> is a widely used Chinese medicinal plant. Recently, because of the great demand, the wild populations have been declining rapidly. In this study, the levels of genetic variation of 11 wild and five cultivated populations of <i>R. tanguticum</i> were investigated by ISSR markers. The 13 selected ISSR primers amplified 306 polymorphic bands out of a total of 326 (93.87 %). Based on Nei’s gene diversity and Shannon’s index, the genetic diversity in cultivated populations of <i>R. tanguticum</i> (<i>H</i> = 0.2490; <i>I</i> = 0.3812; <i>H</i> <sub>B</sub> = 0.3033) was relatively lower than that of wild populations (<i>H</i> = 0.2666; <i>I</i> = 0.4124; <i>H</i> <sub>B</sub> = 0.3115), although no significant differences were identified. Assignment was performed with AFLPOP program, and XGM was the most likely source population of HM. The origins of the rest cultivated populations were admixture. UPGMA and PCoA analyses showed that wild and cultivated populations were not separated into two groups, indicating that a large number of wild genotypes were maintained in the cultivated gene pool. The coefficient of genetic differentiation between wild and cultivated populations was 0.0305 (<i>G</i> <sub>st</sub>), which was in good agreement with the results of analysis of molecular variance (AMOVA), in which, only 1.85 % of the total variance existed between groups of wild and cultivated populations, while 70.91 % occurred within populations and 27.24 % among populations. Together, these results indicated that cultivated populations were not genetically differentiated from wild populations. On the basis of this study, we have made some suggestions for the conservation and efficient management of the genetic resources of this important medicinal herb.
The genus Adonis L. (Ranunculaceae), native to Europe and Asia, comprises 32 annual or perennial herbaceous species. Due to their cardiac-enhancing effects, Adonis spp. have long been used in European and Chinese folk medicine. These plants have been widely investigated since the late 19th century, when the cardiovascular activity of Adonis vernalis L. was noted in Europe. The present paper provides a review of the phytochemistry, biological activities and toxicology in order to highlight the future prospects of the genus. More than 120 chemical compounds have been isolated, with the most important components being cardiac glycosides as well as flavones, carotenoids, coumarins and other structural types. Plants of the genus, especially A. vernalis L. and A. amurensis Regel & Radde, their extracts and their active constituents possess broad pharmacological properties, including cardiovascular, antiangiogenic, antibacterial, antioxidant, anti-inflammatory and acaricidal activities, and exhibit both diuretic effects and effects on the central nervous system. However, most plants within the 32 species have not been comprehensively studied, and further clinical evaluation of their cardiovascular activity and toxicity should be conducted after addressing the problem of the rapidly decreasing resources. This review provides new insight into the genus and lays a solid foundation for further development of Adonis.
Abstract Ethnopharmacological relevance Herpetospermum caudigerum Wall. (HCW) is a traditional Tibetan medicine, which has been used to ameliorate liver injuries in the folk. Aim of the study Liver fibrosis has been recognized as a major lesion of the liver that leads to liver cirrhosis/hepatocarcinoma and even to death in the end. This study aims to demonstrate the protective effect of HCW against CCl 4 -induced liver injury in rats and to explore the underlying mechanisms. Materials and methods Hepatic fibrosis was induced by intraperitoneal injection of CCl 4. Liver function markers, fibrosis markers, serum anti-oxidation enzymes as well as elements levels were determined. Serum and liver tissues were subjected to NMR-based metabolomics and multivariate statistical analysis. Results HCW could significantly reduce the elevated levels of fibrosis markers such as hyaluronidase, laminin, Type III procollagen and Type IV collagen in the serum, improve the activities of the antioxidant enzymes, and effectively reverse the abnormal levels of elements in liver fibrosis rats. Correlation network analysis revealed that HCW could treat liver fibrosis by ameliorating oxidative stress, repairing the impaired energy metabolisms and reversing the disturbed amino acids and nucleic acids metabolisms. Conclusion This integrated metabolomics approach confirmed the validity of the traditional use of HCW in the treatment of liber fibrosis, providing new insights into the underlying mechanisms. Graphical abstract fx1 [ABSTRACT FROM AUTHOR]
ETHNOPHARMACOLOGICAL RELEVANCE: Herpetospermum caudigerum Wall. (HCW) is a traditional Tibetan medicine, which has been used to ameliorate liver injuries in the folk. AIM OF THE STUDY: Liver fibrosis has been recognized as a major lesion of the liver that leads to liver cirrhosis/hepatocarcinoma and even to death in the end. This study aims to demonstrate the protective effect of HCW against CCl4-induced liver injury in rats and to explore the underlying mechanisms. MATERIALS AND METHODS: Hepatic fibrosis was induced by intraperitoneal injection of CCl4. Liver function markers, fibrosis markers, serum anti-oxidation enzymes as well as elements levels were determined. Serum and liver tissues were subjected to NMR-based metabolomics and multivariate statistical analysis. RESULTS: HCW could significantly reduce the elevated levels of fibrosis markers such as hyaluronidase, laminin, Type III procollagen and Type IV collagen in the serum, improve the activities of the antioxidant enzymes, and effectively reverse the abnormal levels of elements in liver fibrosis rats. Correlation network analysis revealed that HCW could treat liver fibrosis by ameliorating oxidative stress, repairing the impaired energy metabolisms and reversing the disturbed amino acids and nucleic acids metabolisms. CONCLUSION: This integrated metabolomics approach confirmed the validity of the traditional use of HCW in the treatment of liber fibrosis, providing new insights into the underlying mechanisms.
Our previous study isolated a natural high-methoxyl homogalacturonan (HRWP-A) from Hippophae rhamnoides and showed antitumor activity in vivo. In this study, the immunomodulatory activity and mechanisms of action of HRWP-A were further investigated. Results showed that HRWP-A could recover the body condition and activated macrophage in Cyclophosphamide (CTX)-induced immunosuppressed mice. Further, we investigated the possible mechanism underlying the effects of HRWP-A on mouse peritoneal macrophages. qPCR and western blot revealed that HRWP-A upregulated the expression of TLR4 mRNA in vitro. This process was accompanied by a clear increase in MyD88 expression and p-IκB-α, but these effects were largely abrogated by pretreatment with anti-TLR4 antibodies. The effects of HRWP-A on macrophage NO, IL-1β and IL-6 production were also inhibited by anti-TLR4 antibodies and were greatly influenced by the NF-κB inhibitor PDTC. Moreover, HRWP-A failed to induce the production of NO, IL-1β and IL-6 in peritoneal macrophages prepared from C3H/HeJ mice, which have a point mutation in the Tlr4 gene, suggesting the involvement of the TLR4 molecule in HRWP-A-mediated macrophage activation. These results may have important implications for our understanding of the structure-activity relationship of immunopotentiating polysaccharides from medicinal herbs.
Our previous study isolated a natural high-methoxyl homogalacturonan (HRWP-A) from Hippophae rhamnoides and showed antitumor activity in vivo. In this study, the immunomodulatory activity and mechanisms of action of HRWP-A were further investigated. Results showed that HRWP-A could recover the body condition and activated macrophage in Cyclophosphamide (CTX)-induced immunosuppressed mice. Further, we investigated the possible mechanism underlying the effects of HRWP-A on mouse peritoneal macrophages. qPCR and western blot revealed that HRWP-A upregulated the expression of TLR4 mRNA in vitro. This process was accompanied by a clear increase in MyD88 expression and p-IκB-α, but these effects were largely abrogated by pretreatment with anti-TLR4 antibodies. The effects of HRWP-A on macrophage NO, IL-1β and IL-6 production were also inhibited by anti-TLR4 antibodies and were greatly influenced by the NF-κB inhibitor PDTC. Moreover, HRWP-A failed to induce the production of NO, IL-1β and IL-6 in peritoneal macrophages prepared from C3H/HeJ mice, which have a point mutation in the Tlr4 gene, suggesting the involvement of the TLR4 molecule in HRWP-A-mediated macrophage activation. These results may have important implications for our understanding of the structure-activity relationship of immunopotentiating polysaccharides from medicinal herbs.
Fatty acids in Herpetospermum seed oil from supercritical CO2 extraction were analyzed by HPLC fluorescence detection (HPLC-FLD) with pre-column derivatization and GC-MS. After derivatizing 39 kinds of saturated and unsaturated fatty acids used 1-[ 2- ( p-toluenesulfonate ) ethyl]-2-phenylimidazole [ 4,5-f] 9,10-phenanthrene ( TSPP ) as pre-column derivatization reagent. All the fatty acid derivatives were separated with a good baseline resolution in conjunction with a gradient elution. The external standard method for the simultaneous quantitative determination of 39 fatty acids was developed and applied for the determination of the free fatty acid contents in Herpetospermum seed oil samples obtained from supercritical CO2 extraction coupled with orthogonal tests, ultrasound-assisted extraction and microwave-assisted reflux extraction. The mass percent of oleic acid, linoleic acid and linolenic acid and the ratio of unsaturated fatty acids and all fatty acids in 9 orthogonal test samples were contrasted. The results indicated that the mass percent of oleic acid, linoleic acid and linolenic acid in Herpetospermum seed oil are up to 34.65% ( 147. 14 mg/g), 22.85% (97.03 mg/g), 20. 86% ( 88. 56 rag/g), respectively, and ratio of unsaturated fatty acids and all fatty acids is 79%. Furthermore, by GC-MS method, the acid catalysis and alkaline catalysis of the methyl esterifying reaction were discussed for the analysis fatty acids in Herpetospermum seed oil, and the optimum GC-MS conditions were obtained. Simultaneously, the characteristics of HPLC and GC-MS methods were discussed about analyzing fatty acids.
<p>A simple and sensitive method for the determination of free fatty acids (FFAs) using acridoné9́ethyĺṕtoluenesulfonate (AETS) as a fluorescence derivatization reagent by high performance liquid chromatography (HPLC) has been developed. Free fatty acid derivatives were separated on an Eclipse XDB́C<sub>8</sub> column with a good baseline resolution and detected with the fluorescence of which excitation and emission wavelengths of derivatives were set at <sub>ex</sub>=404 and <sub>em</sub>=440 nm, respectively. Identification of 19 fatty acid derivatives was carried out by online post́column mass spectrometry with an atmospheric pressure chemical ionization (APCI) source under positivéion detection mode. Nineteen FFAs from the extract of <i>Lomatogonium rotatum</i> are sensitively determined. The results indicate that the plant <i>Lomatogonium rotatum</i> is enriched with an abundance of FFAs and FFAs of higher contents, which mainly focus on even carbon atoms, C<sub>14</sub>, C<sub>16</sub>, and C<sub>18</sub>. The validation of the method including linearity, repeatability, and detection limits was examined. Most linear correlation coefficients for fatty acid derivatives are >0.9989, and detection limits (at signaĺtónoise of 3:1) are 12.3-43.7 fmol. The relative standard deviations (RSDs) of the peak areas and retention times for 19 FFAs standards are <2.24% and 0.45%, respectively. The established method is rapid and reproducible for the separation determination of FFAs from the extract of <i>Lomatogonium rotatum</i> with satisfactory results.</p>
During the screening of a traditional Chinese folk herb library against HepG2 and Hep3B cell lines, the EtOAc extract from the Tibetan medicine, Caragana tibetica (CT-EtOAc) exhibited potential anti-hepatocellular carcinoma (anti-HCC) activity. HPLC-based activity profiling was performed for targeted identification of anti-HCC activity from CT-EtOAc by MS-directed purification method. CT-EtOAc was separated by time-based fractionation for further anti-HCC bioassay by a semipreparative HPLC column (150 mm × 10 mm i.d., 5 μm) with a single injection of 5 mg. Bioassay-guided and ESIMS-directed large scale purification was performed with a single injection of 400 mg of CT-EtOAc by peak-based fractionation. A 1.4-mm heavy wall micro NMR tube with z-gradient was used to measure one and two dimensional NMR spectra for the minor or trace amounts of components of the extract. Two active compounds could be elucidated as naringenin chalcone (CT-1) and 3-hydroxy-8, 9-dimethoxypterocarpan (CT-2) relevant to anti-HCC effects for the EtOAc extract of C. tibetica rapidly and unambiguously by this protocol.
During the screening of a traditional Chinese folk herb library against HepG2 and Hep3B cell lines, the EtOAc extract from the Tibetan medicine, Caragana tibetica (CT-EtOAc) exhibited potential anti-hepatocellular carcinoma (anti-HCC) activity. HPLC-based activity profiling was performed for targeted identification of anti-HCC activity from CT-EtOAc by MS-directed purification method. CT-EtOAc was separated by time-based fractionation for further anti-HCC bioassay by a semipreparative HPLC column (150 mm × 10 mm i.d., 5 μm) with a single injection of 5 mg. Bioassay-guided and ESIMS-directed large scale purification was performed with a single injection of 400 mg of CT-EtOAc by peak-based fractionation. A 1.4-mm heavy wall micro NMR tube with z-gradient was used to measure one and two dimensional NMR spectra for the minor or trace amounts of components of the extract. Two active compounds could be elucidated as naringenin chalcone (CT-1) and 3-hydroxy-8, 9-dimethoxypterocarpan (CT-2) relevant to anti-HCC effects for the EtOAc extract of C. tibetica rapidly and unambiguously by this protocol.
This study established an HPLC fingerprint of Tibetan medicine Shaji Gao from different habitats and lay a foundation for Shaji Gao varieties identification and preparation process. The chromatographic condition was as follow: Agilent zorbax SB-C18 (4.6 mm x 250 mm, 5 μm) eluted with the mobile phases of acetonitrile and 0.4% phosphoric acid water in gradient mode. The flow rate was 1.0 mL x min(-1), and the detection wavelength was set at 360 nm. The fingerprints of 15 batches Shaji Gao were carried out by similarity comparation, 7 chromatographic peaks were extracted as the common peaks of fingerprint, 3 peaks were identified, which were quercetin, kaempferol and isorhamnetin. The similarity degrees of 14 batches of samples were above 0.9 and 1 batch of samples was below 0.9. This is the first established fingerprint of Shaji Gao by using HPLC. This method has good precision, stability and repeatability that it could provide basis for quality control and evaluation of Shaji Gao.
Obesity, a major health problem worldwide, is a complex multifactorial chronic disease that increases the risk for insulin resistance, type 2 diabetes, coronary heart disease, and hypertension. In this study, we assessed methods to isolate hypaphorine, a potent drug candidate for obesity and insulin resistance. Semi-preparative reversed-phase liquid chromatography (semi-preparative RPLC) was established as a method to separate three compounds, adenosine, l-tryptophan, and hypaphorine, from the crude extracts of <i>Caragana korshinskii </i>Kom. Due to its specific chemical structure, the effect of hypaphorine on differentiation and dexamethasone (DXM) induced insulin resistance of 3T3-L1 cells was investigated. The structures of the three compounds were confirmed by UV, ¹H-NMR, and <sup>13</sup>C-NMR analysis and compared with published data. The activity results indicated that hypaphorine prevented the differentiation of 3T3-L1 preadipocytes into adipocytes by down-regulating hormone-stimulated protein expression of peroxisome proliferator activated receptor <i>γ</i> (PPAR<i>γ</i>) and CCAAT/enhancer binding protein (C/EBP<i>α</i>), and their downstream targets, sterol regulatory element binding protein 1 c (SREBP1c) and fatty acid synthase (FAS). Hypaphorine also alleviated DXM-induced insulin resistance in differentiated 3T3-L1 adipocytes <i>via</i> increasing the phosphorylation level of Akt2, a key protein in the insulin signaling pathway. Taken together, we suggest that the method can be applied to large-scale extraction and large-quantity preparation of hypaphorine for treatment of obesity and insulin resistance.
Presents an interview with Qamba Chilai, a Tibetan medical master in Tibet, China. Opinion on the date of the formation of Tibetan medicine; Comment on the relationship between Tibetan medicine and traditional Chinese and Western medicine; Insights on the challenges facing Tibetan medicine. INSET: PROFILE..
ETHNOPHARMACOLOGICAL RELEVANCE: 'Ershiwuwei Shanhu' pill (ESP), a classical and famous prescription of traditional Tibetan medicine, has a long history of empirical clinical use for the treatment of cerebrovascular and neurological diseases, but the absence of scientific evidence for its effect restricted its clinical application and further development.MATERIALS AND METHODS: The methodology of plasma pharmacochemistry was adopted to analyze the potentially bioactive components in ESP extracts. A method based on UPLC-DAD/Q-TOF-MS was established to identify herb components in ESP extracts and analyze the absorbed components of ESP and their metabolites in rat plasma, brain, heart, liver and kidney samples after oral administration of ESP extracts.
RESULTS: A total of 61 herb components were detected and identified in ESP extracts, while 35 absorbed components-including 19 prototype compounds and 16 metabolites-were discovered as potentially bioactive components in rat plasma and tissues by comparative analysis of the UV and MS chromatograms of ESP extracts, blank biosamples and dosed biosamples.
CONCLUSIONS: The potentially bioactive components of ESP extracts identified from rat plasma and tissues provide useful information for further study of the pharmacology and mechanism of action of ESP.
‘Ershiwuwei Shanhu’ pill (ESP), a classical and famous prescription of traditional Tibetan medicine, has a long history of empirical clinical use for the treatment of cerebrovascular and neurological diseases, but the absence of scientific evidence for its effect restricted its clinical application and further development. The methodology of plasma pharmacochemistry was adopted to analyze the potentially bioactive components in ESP extracts. A method based on UPLC-DAD/Q-TOF-MS was established to identify herb components in ESP extracts and analyze the absorbed components of ESP and their metabolites in rat plasma, brain, heart, liver and kidney samples after oral administration of ESP extracts. A total of 61 herb components were detected and identified in ESP extracts, while 35 absorbed components—including 19 prototype compounds and 16 metabolites—were discovered as potentially bioactive components in rat plasma and tissues by comparative analysis of the UV and MS chromatograms of ESP extracts, blank biosamples and dosed biosamples. The potentially bioactive components of ESP extracts identified from rat plasma and tissues provide useful information for further study of the pharmacology and mechanism of action of ESP.
• HPLC-DAD-APCI/MS was set up for analysis of flavonoid aglycones in the RBP. • The method is capable of providing higher sensitivity and repeatability. • Four methods were applied and assessed for extraction of flavonoids from RBP. • The highest extraction efficiency of flavonoids from RBP was achieved by MAE. • MAE is of short extraction time, low solvent consumption and homogeneous conditions.<br>For identification and quantification of flavonoid aglycones in rape bee pollen (RBP) collected from the Qinghai-Tibetan Plateau, a high-performance liquid chromatography (HPLC) separation method with diode array detector (DAD) and atmospheric pressure chemical ionization/mass spectrometric (APCI/MS) detection and four extraction methods (i.e. microwave-assisted extraction, Soxhlet extraction, cold-soaked extraction, and heat reflux extraction) were developed in this study. The identification of flavonoid aglycones was based on retention time and mass spectra by comparison with standards. Results demonstrated that this method showed excellent reproducibility and correlation coefficient, and offered the detection limits of 0.77-15.50 pmol at signal-to-noise ratio of 3. Quercetin and kaempferol were presented in RBP, and microwave-assisted extraction (MAE) was superior to the other three methods in terms of efficiency, convenience and high content of quercetin (1.37 ± 0.059 mg/g) and kaempferol (23.44 ± 0.544 mg/g). Our work indicated that: 1) the proposed HPLC-DAD-APCI/MS was an accurate and precise analysis method to identify and quantify the flavonoid aglycones in RBP; and 2) MAE was efficient to extract flavonoids from RBP with short extraction time, low solvent consumption, and homogeneous extraction conditions.
AIM OF THE STUDY: Based on the authors' collection of specimens used as jie-ji in local Tibetan areas, China, and taxonomic determination, this paper aims to give a list of medicinal plants as jie-ji, formally identify the ones recognized as jie-ji ga-bao or jie-ji na-bao and to offer basic data for further studies on these Tibetan herbs.MATERIALS AND METHODS: Local herbalists were visited in Tibetan areas, China to observe which plants were being used as jie-ji. Samples of the indigenous plants were collected at the same time. Also, the medicinal plants as jie-ji were taxonomically identified.
RESULTS: A list of medicinal plants including 10 species of jie-ji in local Tibetan areas is given, including their morphological pictures used for identification.
CONCLUSIONS: The origin of jie-ji is from 10 species of the Section Cruciata, Genus Gentiana (Gentianaceae). five species with dark blue flowers are used as jie-ji na-bao, the other five with white flowers are used as jie-ji ga-bao. Also, Gentiana macrophylla Pall. with dark blue flowers in the Section Cruciata, Genus Gentiana is not the original plant of jie-ji na-bao. The species endemic to the province are used as the original plants of jie-ji only in local Tibetan area of the province. Finally, the drug use of jie-ji in Traditional Tibetan Medicine is reasonable and it is efficacious.
AIM OF THE STUDY: Based on the authors' collection of specimens used as jie-ji in local Tibetan areas, China, and taxonomic determination, this paper aims to give a list of medicinal plants as jie-ji, formally identify the ones recognized as jie-ji ga-bao or jie-ji na-bao and to offer basic data for further studies on these Tibetan herbs. MATERIALS AND METHODS: Local herbalists were visited in Tibetan areas, China to observe which plants were being used as jie-ji. Samples of the indigenous plants were collected at the same time. Also, the medicinal plants as jie-ji were taxonomically identified. RESULTS: A list of medicinal plants including 10 species of jie-ji in local Tibetan areas is given, including their morphological pictures used for identification. CONCLUSIONS: The origin of jie-ji is from 10 species of the Section Cruciata, Genus Gentiana (Gentianaceae). five species with dark blue flowers are used as jie-ji na-bao, the other five with white flowers are used as jie-ji ga-bao. Also, Gentiana macrophylla Pall. with dark blue flowers in the Section Cruciata, Genus Gentiana is not the original plant of jie-ji na-bao. The species endemic to the province are used as the original plants of jie-ji only in local Tibetan area of the province. Finally, the drug use of jie-ji in Traditional Tibetan Medicine is reasonable and it is efficacious.
An HPLC-UV-MS method for simultaneous identification of predominant phenolics and minor nucleoside derivatives in<i> Gastrodia elata</i> was developed, which was based on their UV and MS characteristics summarized through a series of homemade reference standard experiments. Phenolics showed characteristic UV λ<sub>max</sub> at 267 nm, [M + NH₄]⁺ base peak in positive mode and [M-H]⁻ base peak in negative mode while nucleosides exhibited UV λ<sub>max</sub> at 255 nm, [M + H]⁺, [M-H + 2H₂O]⁻ or [M-H + CH₃COOH]⁻. Phenolics conjugates mainly underwent the consecutive loss of gastrodin residue (-268 U) and the combined loss of H₂O and CO<sub>2 </sub>from the citric acid unit under negative MS/MS conditions whereas nucleosides simply lost the ribose (-132 U) under positive MS/MS conditions. According to these characteristics, a special pattern under MS/MS conditions and reported compound data for<i> G. elata</i> in the literature, not only 15 phenolics were identified but also 6 nucleoside derivatives were identified. Among these compounds, seven phenolics and three nucleoside derivatives have not been reported yet from<i> G. elata</i>.
The preparation of biocatalysts based on immobilized trypsin is of great importance for both proteomic research and industrial applications. Here, we have developed a facile method to immobilize trypsin on hydrophobic cellulose-coated silica nanoparticles by surface adsorption. The immobilization conditions for the trypsin enzyme were optimized. The as-prepared biocatalyst was characterized by Fourier transform infrared spectroscopy, transmission electron microscopy, and elemental analysis. In comparison with free enzyme, the immobilized trypsin exhibited greater resistances against thermal inactivation and denaturants. In addition, the immobilized trypsin showed good durability for multiple recycling. The general applicability of the immobilized trypsin for proteomic studies was confirmed by enzymatic digestion of two widely used protein substrates: bovine serum albumin (BSA) and cytochrome c. The surface adsorption protocols for trypsin immobilization may provide a promising strategy for enzyme immobilization in general, with great potential for a range of applications in proteomic studies.
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