Multi-omics analysis reveals promiscuous O-glycosyltransferases involved in the diversity of flavonoid glycosides in Periploca forrestii (Apocynaceae).

Flavonoid glycosides are widespread in plants, and are of great interest owing to their diverse biological activities and effectiveness in preventing chronic diseases. Periploca forrestii, a renowned medicinal plant of the Apocynaceae family, contains diverse flavonoid glycosides and is clinically used to treat rheumatoid arthritis and traumatic injuries. However, the mechanisms underlying the biosynthesis of these flavonoid glycosides have not yet been elucidated. In this study, we used widely targeted metabolomics and full-length transcriptome sequencing to identify flavonoid diversity and biosynthetic genes in P. forrestii. A total of 120 flavonoid glycosides, including 21 C-, 96 O-, and 3 C/O-glycosides, were identified and annotated. Based on 24,123 full-length coding sequences, 99 uridine diphosphate sugar-utilizing glycosyltransferases (UGTs) were identified and classified into 14 groups. Biochemical assays revealed that four UGTs exhibited O-glycosyltransferase activity toward apigenin and luteolin. Among them, PfUGT74B4 and PfUGT92A8 were highly promiscuous and exhibited multisite O-glycosylation or consecutive glycosylation activities toward various flavonoid aglycones. These four glycosyltransferases may significantly contribute to the diversity of flavonoid glycosides in P. forrestii. Our findings provide a valuable genetic resource for further studies on P. forrestii and insights into the metabolic engineering of bioactive flavonoid glycosides.

Synergistic Effect of Rhodiola rosea and Caffeine Supplementation on the Improvement of Muscle Strength and Muscular Endurance: A Pilot Study for Rats, Resistance Exercise-Untrained and -Trained Volunteers.

Multi-level studies have shown that Rhodiola rosea (RHO) and Caffeine (CAF) have the potential to be nutritional supplements to enhance physical performance in resistance exercise-untrained and -trained subjects. This study examined the synergistic effects of RHO (262.7 mg/kg for rats and 2.4 g for volunteers) and CAF (19.7 mg/kg for rats and 3 mg/kg for volunteers) supplementation on improving physical performance in rats, resistance exercise-untrained volunteers and resistance exercise-trained volunteers. Rats and volunteers were randomly grouped into placebo, CAF, RHO and CAF+RHO and administered accordingly with the nutrients during the training procedure, and pre- and post-measures were collected. We found that RHO+CAF was effective in improving forelimb grip strength (13.75%), erythropoietin (23.85%), dopamine (12.65%) and oxygen consumption rate (9.29%) in the rat model. Furthermore, the current results also indicated that the combination of RHO+CAF significantly increased the bench press one-repetition maximum (1RM) (16.59%), deep squat 1RM (15.75%), maximum voluntary isometric contraction (MVIC) (14.72%) and maximum repetitions of 60% 1RM bench press (22.15%) in resistance exercise-untrained volunteers. Additionally, despite the excellent base level of the resistance exercise-trained volunteers, their deep squat 1RM and MVIC increased substantially through the synergistic effect of RHO and CAF. In conclusion, combined supplementation of RHO+CAF is more beneficial in improving the resistance exercise performance for both resistance exercise-untrained and -trained volunteers. The present results provide practical evidence that the synergies of RHO and CAF could serve as potential supplementary for individuals, especially resistance exercise-trained subjects, to ameliorate their physical performances effectively and safely.

Strengths, Microstructure and Nanomechanical Properties of Concrete Containing High Volume of Zeolite Powder.

In order to save resources and reduce the carbon footprint of concrete, the addition of high volumes of supplementary cementitious materials (SCMs) to replace cement is one of the most effective and promising methods. Zeolite powder (ZP), with a high specific surface area, exhibits high pozzolanic reactivity in cement-based materials. This paper investigates the effects of ZP addition used to replace cement at the levels of 20%, 40% and 60% on the strength development and microstructure evolution of concrete, and the nanomechanical properties are analyzed using nanoindentation technique. The results show that the replacement of ZP for cement generally has a dilution effect on the concrete, leading to a detrimental effect on the strength development. However, the 20% ZP replacement for cement slightly enhances the 90-day compressive strength. The pore structure analysis shows that the sample with 20% ZP content has a lower total porosity than the control sample. The hydration of ZP goes against the dilution effect and reduces the total porosity of concrete to compact the microstructure. Nanoindentation investigation of the matrix shows that 20% ZP decreases the content of portlandite but increases the content of high density calcium silicate hydrate (C-S-H). This is beneficial for improving the nanomechanical properties of interface transition zone. However, further increases in the content of ZP (40% and 60%) decrease the total volume of C-S-H and increase the porosity to degrade the microstructure.

Integrated metabolomic and transcriptomic profiling reveals the tissue-specific flavonoid compositions and their biosynthesis pathways in Ziziphora bungeana.

BACKGROUND: Ziziphora bungeana Juz. is a folk medicine from the Xinjiang Uygur Autonomous Region. The herb or the aerial parts of it have been used to medicinally treat cardiovascular diseases. Flavonoids are the main pharmacologically active ingredients in Z. bungeana. Identification of the tissue-specific distribution of flavonoids in Z. bungeana is crucial for effective and sustainable medicinal use of the plant. Furthermore, understanding of the biosynthesis pathways of these flavonoids in Z. bungeana is of great biological significance. METHODS: The flavonoids from different tissues of Z. bungeana were identified using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The full-length transcriptome of Z. bungeana was determined using a strategy based on a combination of Illumina and PacBio sequencing techniques. The functions of differentially expressed unigenes were predicted using bioinformatics methods and further investigated by real-time quantitative PCR and phylogenetic relationship analysis. RESULTS: Among the 12 major flavonoid components identified from Z. bungeana extracts, linarin was the most abundant component. Nine flavonoids were identified as characteristic components of specific tissues. Transcriptome profiling and bioinformatic analysis revealed that 18 genes were putatively involved in flavonoid biosynthesis. The gene expression and phylogenetic analysis results indicated that ZbPALs, Zb4CL3, ZbCHS1, and ZbCHI1 may be involved in the biosynthesis of the main flavonoid intermediate. ZbFNSII, ZbANS, and ZbFLS may be involved in the biosynthesis of flavones, anthocyanins, and flavonols, respectively. A map of the biosynthesis pathways of the 12 major flavonoids in Z. bungeana is proposed. CONCLUSIONS: The chemical constituent analysis revealed the compositions of 9 characteristic flavonoids in different tissues of Z. bungeana. Linarin can be hydrolysed into acacetin to exert a pharmaceutical role. Apigenin-7-O-rutinoside is hypothesised to be the precursor of linarin in Z. bungeana. There was greater content of linarin in the aerial parts of the plant than in the whole herb, which provides a theoretical basis for using the aerial parts of Z. bungeana for medicine. These results provide a valuable reference for further research on the flavonoid biosynthesis pathways of Z. bungeana and will be significant for the effective utilisation and ecological protection of Z. bungeana.

[Transcriptome analysis reveals candidate genes involved in flavonoid biosynthesis in Ziziphora bungeana].

Ziziphora bungeana is a kind of medicinal plants belongs to Labiatae,and it also a kind of geoherbs in Xinjiang. The main active ingredient linarin has a higher content in inflorescence than in other parts. In this study,high-throughput sequencing technology was used to reveal the transcriptome of the inflorescence of Z. bungeana,77 366 unigenes were acquired,of which 56 375 unigenes were annotated based on search of the database and classification. Through the analysis of metabolic pathways,sixty unigenes were probably encoding some enzymes involved in the flavonoid biosynthesis pathways. The contents of linarin in different parts were determined and the key genes were verified by qRT-PCR. The discovery provides the research basis for further analysis of the enzyme genes involved in the biosynthesis of the major flavonoid components in Z. bungeana.

Preparation of Magnetic Nanoliposomes of Sesquiterpene-Rich Fraction from Cichorium glandulosum and Its Tissue Distribution in Mice.

OBJECTIVE: To prepare and study the distribution of magnetic nanoliposomes of Sesquiterpene-Rich Fraction from Cichorium glandulosum (SRF-MLN) (under magnetic field) in the main organs of mice. METHODS: The SRF-MLN was prepared by ethanol injection-sonication precipitation method. The quality of its pharmaceutical properties was investigated by the active ingredient lactucin. The drug concentration and distribution of lactucin in different tissues and organs including whole blood, liver, heart, spleen, lung, and kidney were evaluated with Sesquiterpene-Rich Fraction from Cichorium glandulosum (SRF-LP) as a control. RESULTS: The prepared SRF-MLN was spherical and monodispersed with an average particle diameter of 65 ± 1 nm, encapsulation efficiency of 91.3% ± 1.5% (RSD% = 1.5), and drug loading rate of 4.7% ± 0.3% (RSD% = 0.7). Dispersion coefficient PDI was 0.157 and Zeta potential was -17.5 mV. The lactucin concentration of SRF-MLN after 10 min of intravenous injection in heart, liver, spleen, lung, and kidney was significantly higher than that of SRF-LP group. The AUC0-12h values of liver tissue were obviously higher than other tissues and MRT was significantly prolonged under the action of the magnetic field (p<0.01). CONCLUSION: The prepared SRF-MLN can change the distribution of drugs in different tissues and organs, prolonging the action time of the drugs in the body, and it has certain specificity under the action of applied magnetic field.

[Classification, distribution and change of Uighur medicinal materials resources].

With the development and change of Uygur medicine, The sources,medicinal parts and producing area of some Uygur Medicine have changed. It is more important to master the classification, distribution and change of Uygur medicinal materials. These were more than 1 200 kinds Uygur medicine in history were resaerched by field investigation, philological research, and textual research, which main source of original plant were 140 families, 510 genera, 840 species; and source of original animal were 76 families, 107 genera, the original animal 141 species; 55 kinds of original mineral, which main producer were Xinjiang and Central Asia, West Asia, the Mediterranean, and North Africa, Southeast Asia and other provinces in China, there are individual medicinal materials from the Americas, Europe and other places. Through this study the classification, distribution, source and evolution of specific families and genera of Uygur medicine resources have mastered.It is hoped to provide theoretical basis for further research and development of Uygur medicinal materials.

[Determination of 3 components in Xinjiang Ziziphora bungeana by HPLC].

This study aimed to develop an HPLC method for simultaneous determination of the 3 components in Ziziphora bungeana. The optimum HPLC condition was as follows：ZORBAX SB-C18 column（4.6 mm×250 mm, 5.0 µm）with gradient elution of methanol (A)-0.2% glacial acetic acid (B), detection wavelength 340 nm,column temperature 30 °C, flow rate 1 mL·min⁻¹. There were good linearity between peak areas and injection quantity of caffeic acid, rosmarinic acid, and linarin in the range of 2-40 mg·L⁻¹ï¼ˆr=0.999 9）, 3-60 mg·L⁻¹ï¼ˆr=1）, 7-140 mg·L⁻¹ï¼ˆr=0.999 9）, respectively. The average recoveries were 100.9%（RSD 1.3%）,98.25%（RSD 2.0%）,and 98.73%（RSD 1.5%）, respectively. The HPLC method was stable and accurate, which could be used to detect caffeic acid,rosmarinic acid, and linarin in Ziziphora bungeana.

Ziziphora clinopodioides Flavonoids Protect Myocardial Cell Damage from Myocardial Ischemia-Reperfusion Injury.

To investigate effects of Ziziphora clinopodioides Lam. flavonoids on ischemia-reperfusion injury of myocardial cells. After application of 6.25, 25, and 100 µg/mL Ziziphora clinopodioides Lam. flavonoids to H9C2 myocardial cells for 24H, they were treated for 4 hours with hydrogen peroxide to induce oxidative damage, whereas controls were cells without treatment and cells only incubated with hydrogen peroxide. Cell viability, lactate dehydrogenase release and mitochondrial membrane potential, intracellular Na+/K+-ATPase activity and ATP content, and reactive oxygen species formation were monitored. An ischemia-reperfusion injury rat model was established by left anterior descending coronary artery ligature in 48 Sprague-Dawley rats, which were divided into positive control with isosorbide mononitrate (10 mg/kg) injection (n=8), model (ischemia-reperfusion, n=8), sham-operated (n=8), and Ziziphora clinopodioides Lam. flavonoids low (75 mg/kg, n=8), medium (150 mg/kg, n=8), and high concentration (300 mg/kg, n=8) groups. Superoxide dismutase activity and malondialdehyde content in homogenized hearts were measured and ischemic and infarction areas were triphenyl tetrazolium chloride and H&E stained for pathological and morphological examinations. Ziziphora clinopodioides Lam. flavonoids preconditioning improved cell viability (P<0.01), intracellular Na/K ATPase activity (P<0.001), and intracellular ATP content (P<0.001) and maintained mitochondrial membrane potential (P<0.05) in hydrogen peroxide treated H9C2 cells as well as rescued superoxide dismutase activity (P<0.01), decreased the malondialdehyde content (P<0.001), and reduced myocardial damage in the ischemia-reperfusion rat model (P<0.001) compared to the controls. Ziziphora clinopodioides Lam. flavonoids may be an effective drug for protecting myocardial tissue from ischemia-reperfusion injury by reducing reactive oxygen species related damage.

[The present situation and countermeasures of Uighur medicine compound].

Uighur medicine compound, which created and used by Uighur nationality, is under the guidance of the Uighur medical theory system of herbal formula and dialectical use of minority nationality conventional medicines. In recent years, Uighur medicine attracted more and more attention of people which have used and were using it. Combining the history of Uighur medicine, this article summarizes the Uighur resources, medicinal materials, drugs preparation, ancient documents, and the establishment of the clinical evaluation system and so on, and then analyzes the status quo and the existing problems in Uighur medicine compound research and industry. On this basis, we put forward countermeasures and suggestions for the development of Uighur medicine.

Developmental changes in carbon and nitrogen metabolism affect tea quality in different leaf position.

Leaf position represents a specific developmental stage that influences both photosynthesis and respiration. However, the precise relationships between photosynthesis and respiration in different leaf position that affect tea quality are largely unknown. Here, we show that the effective quantum yield of photosystem II [ΦPSⅡ] as well as total chlorophyll concentration (TChl) of tea leaves increased gradually with leaf maturity. Moreover, respiration rate (RR) together with total nitrogen concentration (TN) decreased persistently, but total carbon remained unchanged during leaf maturation. Analyses of major N-based organic compounds revealed that decrease in TN was attributed to a significant decrease in the concentration of caffeine and amino acids (AA) in mature leaves. Furthermore, soluble sugar (SS) decreased, but starch concentration increased with leaf maturity, indicating that source-sink relationship was altered during tea leaf development. Detailed correlation analysis showed that ΦPSⅡ was negatively correlated with RR, SS, starch, tea polyphenol (TP), total catechins and TN, but positively correlated with TChl; while RR was positively correlated with TN, SS, TP and caffeine, but negatively correlated with TChl and starch concentrations. Our results suggest that biosynthesis of chlorophyll, catechins and polyphenols is closely associated with photosynthesis and respiration in different leaf position that greatly influences the relationship between primary and secondary metabolism in tea plants.

[Influence on Strychni Semen's Analgesic Effect and Toxicity of Milk-Impregnated- Processing-Technology of Traditional Uighur Medicine].

OBJECTIVE: To investigate the effects of different processing methods on analgesic effect and toxicity of Strychni Semen, and to provide the scientific basis of milk-impregnated-processing-technology of traditional Uighur medicine for toxic medicines. METHODS: Three different methods, frying with sand of traditional Chinese medicine (1), cooled milk-impregnated-processing-technology of traditional Uighur medicine (2), and heated milk-impregnated-processing-technology of traditional Uighur medicine (3) were used to process raw Strychni Semen and the processed products were compared with those in the raw drug (4). Karber method was used to caculate the LD50. RESULTS: The LD50 of (4), (1), (2) and (3)were 273.0, 289.3, 289.3 and 339.2 mg/kg,successively. Low dose group of heated milk-impregnated-processing-technology, low and high dose groups of cooled milk-impregnated-processing-technology had significant inhibition on the acetic acid-induced writhing reaction in mice (P < 0.05). CONCLUSION: Strychni Semen processed by cooled and heated milk can enhance the analgesic effect in mice while improving the LD50 of mouse, which illustrates milk-impregnated-processing- technology's pharmacological mechanism on efficacy enhancing and toxicity reducing of Strychni Semen.

The mechanism of hepatoprotective effect of sesquiterpene rich fraction from Cichorum glandulosum Boiss. et Huet on immune reaction-induced liver injury in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Cichorum glandulosum Boiss. et Huet is a traditional Uygur herbal medicine that has been used as a cholagogic and diuretic agent to improve liver function. However, the mechanism is not known for the liver-protective function. We investigated the antioxidant effects of plant extraction (CGE60) in vitro and in vivo, and find the mechanism of liver protection in Bacille Calmette-Guerin vaccine (BCG)+Lipopolysaccharides (LPS) induced liver injury in mice. MATERIALS AND METHODS: CGE60 was made, and the antioxidant activity was investigated by comparing the ability of scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH), and 2,2-azinobis(3-ehtylbenzothiazolin-6-sulfnicAcid) diammonium salt (ABTS) free radicals in vitro. Then, CGE60 was administrated in mice of liver damage model which was induced in mice using the BCG+LPS protocol. The CGE 60 extract was tested at three dosages: 50 mg/kg, 100 mg/kg, and 200 mg/kg. Product of lipid peroxidation (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX,), nitric oxide (NO), nitric oxide synthetase (NOS), hydroxyproline and alkaline phosphatase (ALP) contents were evaluated in liver to determine the CGE60 activity in the hepatic injury model. Tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and transforming growth factor-ß (TGF-ß) proteins were determined in the liver tissues using ELSIA. The signaling activities were evaluated in Western blot. RESULTS: CGE60 exhibited strong antioxidant ability in vitro. With oral administration, CGE60 significantly increased the activity of CAT, SOD, GSH-PX, and decreased the level of NO, NO synthase, hydroxyproline, ALP and lipid peroxidation liver of in the BDG+ LPS model. CGE60 attenuated hepatic inflammation via down- regulation of TNF-α, IL-6 and TGF-ß. CGE60 blocked protein expression of cytochrome P450 2E1 (CYP2E1), nuclear factor kappa-B (NF-κB), phosphorylation of extracellular signal-regulated kinase (p-ERK1/2), and cyclooxygenase-2 (COX-2),but activated the expression of p-P38 MAPK. CONCLUSION: This study suggests that CGE60 possesses antioxidant activity and this activity associates with hepatoprotective effect in the mice of BCG +LPS model, and the mechanisms underlying these effects may involve antioxidant actions and anti-inflammation activities.

[Pharmacognostical studies on imported and cultivated Citrullus colocynthis].

OBJECTIVE: In order to use Citrullus colocynthis safety and correctly,The pharmacognosy of imported and cultivated Citrullus colocynthis was systematically studied. METHODS: The morphological characters, microscopical identification, and thin layer Chromatograpy (TLC) were studied. RESULTS: The characters of import and cultivated Citrullus colocynthis were approximately identical, but there were still some differences between them, such as: the fruit size of cultivated Citrullus colocynthis was largish, granulose were more, trachea were thicker, collenchyma were longer, the colour of spots in TCL were more clear. CONCLUSION: There were seldon difference between import and cultivated Citrullus colocynthis, and cultivated Citrullus colocynthis could take the place of import's if planted on a large scale.

Response of metallothionein gene-1 to laboratory exposure to heavy metals and thermal stress in the freshwater prawn Macrobrachium rosenbergii.

Metallothioneins, metal-inducible proteins, are being characterized from different organisms and shown as potential biomarkers of exposure to pollution by certain heavy metals. Here we report the identification of a new metallothionein cDNA (433bp) from the shrimp Macrobrachium rosenbergii, putatively encoding a 61 residue polypeptide. Tissue specific analysis indicated that Mar-MT-I (M. rosenbergii Metallothionein Gene-1) is expressed with the highest levels in the hepatopancreas and lowest in the thoracic ganglia, and none in the gills or muscles. In addition, our data showed that Mar-MT-I is differentially regulated in the hepatopancreas by certain heavy metals and thermal stress: Cd and Cu produce somewhat similar expression profile patterns, Zn has a reductional effect and thermal stress alone entirely stops its expression. These results show that Mar-MT-I mRNA levels can potentially be used as biomarkers for Cd, Cu or Zn pollution individually. However, in the case of combined metal treatment, different combinations of these metals have quite different effect on Mar-MT-I expression. Therefore, factors of such differential behaviors should be kept as a priority for further biomonitoring studies.

[Purification technology of Rhizoma et Radix Notopterygii and Radix Angelicae Pubescentis in Fufang Xuelian dropping pills by macroporous resin].

OBJECTIVE: To study the purification technology of Rhizoma et Radix Notopterygii and Radix Angelicae Pubescentis in Fufang Xuelian dropping pills by macroporous resin. METHOD: Taking osthole, isomperatorin as index ingredients, the type of resin sampling amount and elution solvent were decided, and the influence of sample concentration pH of sample and ratio of diameter to height of column to adsorption were studied. RESULT: HPD400A was chosen to purify, the suitable sampling ratio of resin volume to raw material was 1:2; pH 3.5 (crude drug) and ratio of diameter to height was 1:7; 95% ethanol of the elution solvent was satisfactory eluant for desorption. CONCLUSION: HPD400A macroporous resin can be used to purify Rhizoma et Radix Notopterygii and Radix Angelicae Pubescentis.