Enhanced inhibition of protein disulfide isomerase and anti-thrombotic activity of a rutin derivative: rutin:Zn complex.

Rutin is a flavonoid that exists in plants and in commonly consumed foods. In recent years, rutin has been demonstrated to have anti-thrombotic efficacy through its inhibition of protein disulfide isomerase. However, the low aqueous solubility and high dose limit the therapeutic applications of rutin. In this study, we found that the chelation of zinc ions increased rutin aqueous solubility by 4-fold. More importantly, the thus-formed rutin:Zn complex inhibited PDI activity more potently than rutin itself. In a murine model with electric current-induced arterial thrombosis, the rutin:Zn complex slowed mouse arterial occlusion compared to rutin without increasing bleeding risk. Thus, the zinc chelation not only improved rutin aqueous solubility but achieved stronger inhibition of PDI. Furthermore, zinc chelation of a selected list of flavonoids containing the adjacent keto and phenoxy groups also increased their inhibition of PDI. Hence, our study provides a strategy to promote flavonoids' anti-thrombotic properties.

Chemical Stability of a Chinese Herbal Spirit Using LC-MS-Based Metabolomics and Accelerated Tests.

As a prevalent medicinal liquor among Chinese people, a type of Chinese herbal spirit from Jing Brand Co., Ltd (CHS-J) is a newly developed health beverage with the health functions of anti-fatigue and immune enhancement. The researchers from the enterprise found that the contents of several components in CHS-J samples have been significantly decreasing during the stated storage period, as detected by the HPLC-UV method, which would make a great challenge for quality control of CHS-J. Furthermore, the chemical stability of CHS-J during the storage period is greatly challenged affected, especially in the environment of high temperature and light exposure. To systematically reveal the unstable components and promote the quality control of CHS-J, the chemical stability of CHS-J during the shelving storage period was characterized by the UPLC/Q-TOFMS-based metabolomics approach. First, the targeted and untargeted metabolomics approaches discovered the significantly changed components in CHS-J samples produced in different years. Furthermore, the accelerated tests of newly produced CHS samples and several authorized standards were conducted to validate the above results and elucidate the possible mechanisms underlying these chemical changes. Moreover, these chemical changes during the storage period had little influence on the anti-fatigue effect of CHS-J samples. These findings will offer new insight into the understanding of the chemical stability of CHS-J and will facilitate the quality control of CHS-J.

Simultaneous quantification of bioactive components in Chinese herbal spirits by ultra-high performance liquid chromatography coupled to triple-quadrupole mass spectrometry (UHPLC-QQQ-MS/MS).

BACKGROUND: The Chinese medicinal wine made from herbal medicines became prevalent among Chinese people. The Chinese herbal spirit is composed of several herbal extracts, and has the certain health functions, such as anti-fatigue and immune regulation. The quality evaluation of Chinese herbal spirit is greatly challenged by the enormous and complex components with great structural diversity and wide range of concentration distribution. METHODS: An ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS) with multiple reaction monitoring (MRM) method was developed to simultaneously determine forty-three bioactive components in the Chinese herbal spirits produced by year 2014 and 2018. RESULTS: Quantitative results showed that 11 components, i.e.., puerarin (5), purpureaside C (7), daidzin (8), echinacoside (9), acteoside (15), epimedin B (22), epimedin C (23), icariin (24), eugenol (27), chikusetsusaponin iva (30) and Z-ligustilide (40), significantly decreased along with the increasing years of storage, while 5 compounds, i.e.., geniposidic acid (1), protocatechuic acid (2), crustecdysone (14), daidzein (18) and icariside I (35), were basically stable in all samples across the years. CONCUSION: The established method allowing to simultaneously determined 43 components with wide structural diversity and trace amounts will facilitate the quality control research of Chinese herbal spirits.

A strategy for rapid discovery of traceable chemical markers in herbal products using MZmine 2 data processing toolbox: A case of Jing Liqueur.

Objective: The quality evaluation of herbal products remains a big challenge. Traceable markers are the core concept of the authentication of herbal products. However, the discovery of traceable markers is labor-intensive and time-consuming. The aim of this study is to develop a convenient approach to rapidly screen the traceable markers for herbal product authentication. Methods: Commercial Jing Liqueur and its 22 species of herbal ingredients were analyzed using HPLC-QTOF-MS and GC-MS to characterize nonvolatile and volatile chemicals. The acquired data were imported into MZmine 2 software for mass detection, chromatogram building, deconvolution and alignment. The aligned data were exported into a csv file and then traceable markers were selected using the built-in filter function in Excel. Finally, the traceable markers were identified by searching against online databases or publications, some of which were confirmed by reference standards. Results: A total of 288 chemical features transferred from herbal materials to Jing Liqueur product were rapidly screened out. Among them, 52 markers detected by HPLC-QTOF-MS were annotated, while nine volatile markers detected by GC-MS were annotated. Moreover, 30 of these markers were confirmed by comparing with reference standards. A chemical fingerprint consisting of traceable markers was finally generated to ensure the authentication and quality consistency of Jing Liqueur. Conclusion: A strategy for rapid discovery of traceable markers in herbal products using MZmine 2 software was developed.

A strategy for rapid discovery of traceable chemical markers in herbal products using MZmine 2 data processing toolbox: A case of Jing Liqueur / 中草药·英文版

Objective: The quality evaluation of herbal products remains a big challenge. Traceable markers are the core concept of the authentication of herbal products. However, the discovery of traceable markers is labor-intensive and time-consuming. The aim of this study is to develop a convenient approach to rapidly screen the traceable markers for herbal product authentication. Methods: Commercial Jing Liqueur and its 22 species of herbal ingredients were analyzed using HPLC-QTOF-MS and GC–MS to characterize nonvolatile and volatile chemicals. The acquired data were imported into MZmine 2 software for mass detection, chromatogram building, deconvolution and alignment. The aligned data were exported into a csv file and then traceable markers were selected using the built-in filter function in Excel. Finally, the traceable markers were identified by searching against online databases or publications, some of which were confirmed by reference standards. Results: A total of 288 chemical features transferred from herbal materials to Jing Liqueur product were rapidly screened out. Among them, 52 markers detected by HPLC-QTOF-MS were annotated, while nine volatile markers detected by GC–MS were annotated. Moreover, 30 of these markers were confirmed by comparing with reference standards. A chemical fingerprint consisting of traceable markers was finally generated to ensure the authentication and quality consistency of Jing Liqueur. Conclusion: A strategy for rapid discovery of traceable markers in herbal products using MZmine 2 software was developed.

Inhibition of the Citrus Canker Pathogen Using a Photosensitizer Assisted by Sunlight Irradiation.

Citrus canker, induced by bacterial infection, seriously affects the growth and productivity of citrus around the world and has attracted strong research interest. The current treatment for this disease uses copper salts to inactivate the pathogenic bacteria: Xanthomonas citri subsp. citri (Xcc) strain. However, copper salts may have a negative impact on the environment or plant. In this work, we identify a chemical compound, 2,6-diiodo-1,3,5,7-tetramethyl-8-(P-benzoic acid)-4,4'-difluoroboradiazaindacene (DIBDP), to inactivate the pathogenic Xcc strain (29-1). DIBDP is activated by sunlight and generates reactive oxygen species to kill the bacteria. In order to overcome the degradation of DIBDP under sunlight, an adjuvant agent was identified to limit the photodegradation of DIBDP by forming a photosensitizer complex (PSC). This complex demonstrated significant antimicrobial activity to Xcc 29-1, which was 64-fold more potent than the copper biocides. The antimicrobial efficacy of PSC on citrus leaves infected by Xcc 29-1 also was much stronger than copper agent and, at the same time, the PSC was safe to the host exposed to sunlight. Thus, this PSC is a promising antibacterial agent to control citrus canker disease.

Research progress of P450 in the biosynthesis of bioactive compound of medicinal plants / 药学学报

Most of the active ingredients of herbs are secondary metabolites of plants. Cytochrome P450s (P450s) are hemoglobin-containing monooxygenases encoded by a super-gene family, which play important roles in the metabolic network of plants. This review focuses on the role of P450s on biosynthesis of secondary metabolites such as terpenoids, alkaloids, flavonoids and phenylpropanoids. This will provide references for biosynthesis and regulation of secondary metabolites in medicinal plants.

Screening of long non-coding RNA related to CYP450s involved in biosynthesis of tanshinones / 中国中药杂志

Tanshinones are abietane-type norditerpenoid quinones that make up the main bioactive ingredients of traditional Chinese medicine Salvia miltiorrhiza. Cytochrome CYP450 plays an important role in the post-structural modification of tanshinone biosynthesis pathway. Long non-coding RNA( lncRNA) have been defined as transcripts longer than 200 nucleotides,which have been functionally characterized in regulating the growth and development,secondary metabolism and stress of medicinal plants. In this study,we perform a comprehensive identification of lncRNAs in response to tanshinone metabolism induced by yeast extract( YE) and Ag~+ S. miltiorrhiza hairy roots. Deep RNA sequencing was used to identify a set of different 8 942 lncRNAs,of which 6 755 were intergenic lncRNAs. We predicted a total of 1 115 814 lncRNA-coding gene pairs,including 122 lncRNA-coding gene as cis pairs. The correlation analysis between lncRNA and CYP450 related to tanshinone biosynthesis was carried out and a total of 16 249 lncRNA-CYP450 target gene pairs were identified. Further analysis with functional known CYP76 AH1,CYP76 AH3 and CYP76 AK1 involved in tanshinone biosynthesis,we also identified a set of 216 target genes. These candidate genes will be the important target in the downstream regulation mechanism analysis of the tanshinone biosynthesis pathway.

Construction of Saccharomyces cerevisiae whole-cell biocatalyst system for conversion miltiradiene / 药学学报

Tanshinones are the bioactive components of the Chinese medicinal herb Salvia miltiorrhiza, while its biosynthetic pathway remains to be characterized. Rapid identification and characterization of the genes correlated to tanshinones biosynthesis is very important. As one of the intermediates of tanshinones biosynthesis, the ferruginol content is relative low in both root and engineered bacteria. It is urgent to construct an efficient system for conversion of miltiradiene to ferruginol to obtain large amount of ferruginol as the substrates for further identifying other downstream genes involved in tanshinones biosynthesis. In this study, we constructed the whole-cell yeast biocatalysts co-expressing miltiradiene oxidase CYP76AH1 and cytochrome P450 reductases (SmCPR1) from Salvia miltiorrhiza, and then characterized it with RT-PCR. After permeabilization, the yeast whole-cell could catalyze turnover of miltiradiene to ferruginol efficiently through single-step biotransformation with a conversion efficiency up to 69.9%. The yeast whole-cell biocatalyst described here not only provide an efficient platform for producing ferruginol in recombinant yeast but also an alternative strategy for identifying other CYP genes involved in tanshinones biosynthesis.

Construction of Saccharomyces cerevisiae whole-cell biocatalyst system for conversion miltiradiene / 药学学报

Tanshinones are the bioactive components of the Chinese medicinal herb Salvia miltiorrhiza, while its biosynthetic pathway remains to be characterized. Rapid identification and characterization of the genes correlated to tanshinones biosynthesis is very important. As one of the intermediates of tanshinones biosynthesis, the ferruginol content is relative low in both root and engineered bacteria. It is urgent to construct an efficient system for conversion of miltiradiene to ferruginol to obtain large amount of ferruginol as the substrates for further identifying other downstream genes involved in tanshinones biosynthesis. In this study, we constructed the whole-cell yeast biocatalysts co-expressing miltiradiene oxidase CYP76AH1 and cytochrome P450 reductases (SmCPR1) from Salvia miltiorrhiza, and then characterized it with RT-PCR. After permeabilization, the yeast whole-cell could catalyze turnover of miltiradiene to ferruginol efficiently through single-step biotransformation with a conversion efficiency up to 69.9%. The yeast whole-cell biocatalyst described here not only provide an efficient platform for producing ferruginol in recombinant yeast but also an alternative strategy for identifying other CYP genes involved in tanshinones biosynthesis.

Effect of roots of Ficus hirta on cocaine-induced hepatotoxicity and active components / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of the roots of F. hirta against the cocaine-induced hepatotoxicity and it's active components.</p><p><b>METHOD</b>Cocaine hydrochloride was subcutaneously injected to make male ICR mice liver wounded. Male ICR mice were randomly ig administered with the F. hirta decoction. The dose groups are 100, 200, 300 g x kg(-1) herb materials per body weight. Cocaine hydrochloride was subcutaneously injected into the mice after the administration. The serum ALT, AST activity and the activity of CAT in liver homogenate were assayed, and liver change of pathomorphism was evaluated to prove the effect of the F. hirta decoction on cocaine-induced hepatotoxicity. And the activity of psoralean which was separated from the F. hirta decoction by bioassay-guided fractionation, was proofed in the same method.</p><p><b>RESULT</b>We find that the F. hirta decoction shows a distinct effect on reducing serum transferase. The serum transferase and the content CAT in liver homogenate were dose-related reduced, and the histopathological examination found a significantly change of the liver tissues. And the psoralean, qua the mainly component, shows the same effect.</p><p><b>CONCLUSION</b>F. hirta has the protective effect against the cocaine-induced hepatotoxicity. Psoralean is the basis.</p>

Effects of Lanthanum on Excitatory Amino Acids and Calcium Homeostasis of Cerebral Cortex in Rats / 环境与健康杂志

Objective To observe the effects of lanthanum on learning and memory,and explore the corresponding mechanism on the basis of the levels of excitatory amino acids and calcium homeostasis of cerebral cortex in rats.Methods Forty female Wistar rats were randomly divided into control,low-,moderate-and high-dose lanthanum groups(ten rats in each group).The offspring of lanthanum exposure groups were administrated with lanthanum from weaning for one month.The capability of learning and memory was measured by neuroethology testing.The levels of excitatory amino acids,intracellular calcium ion and Nissl body expression of cerebral cortex were also detected.Results The capabilities of learning and memory in lanthanum exposure groups were significantly lower compared with the control,with a dose-dependent manner.The levels of glutamate and intracellular calcium ion of cerebral cortex in low-and moderate-dose group were significantly higher than those in controls,and the levels of glutamate,aspartate and intracellular calcium ion of cerebral cortex in high-dose group were increased significantly as compared with the control group,low dose group and moderate dose lanthanum group.Besides,Nissl body expression of cerebral cortex in low-dose group was lower than that in control group,and the levels of Nissl body expression of cerebral cortex in middle dose lanthanum group and high dose groups decreased further.Conclusion The increased levels of excitatory amino acids,calcium homeostasis unbalance and cellular injuries in cerebral cortex may be involved in lower capacity of learning and memory in the rats exposed to lanthanum.

Preliminary study on treatment of partial androgen deficiency in aging males with Jingui Shenqi Pill / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the efficacy and safety of Jingui Shengqi Pill in treating partial androgen deficiency in aging males (PADAM), and to explore the new approach in improving the quality of life in PADAM patients.</p><p><b>METHODS</b>Forty patients with PADAM were treated with JSP, the efficacy was evaluated with international index of erectile function (IIEF) scoring, PADAM questionnaire scoring, hormone, prostatic specific antigen (PSA), etc., and the data before treatment were compared with those after treatment in the same group.</p><p><b>RESULTS</b>After 3 months of treatment, PADAM scoring and IIEF scoring were all significantly improved. Symptoms regarding physical ability, vasomotion, and psychical and mental condition all got improved more markedly than symptoms regarding sexual hypofunction. The serum level of testosterone was 3.85 +/- 0.36 before treatment and 5.02 +/- 0.83 after treatment (P < 0.05); luteinizing hormone of 7.33 +/- 2.14 and 4.84 +/- 1.43 (P < 0.01), follicle-stimulating hormone of 10.22 +/- 4.48 and 6.47 +/- 3.28 (P < 0.01), respectively. The level of PSA failed to change significantly (1.94 +/- 0.55 and 2.06 +/- 0.47, P > 0.05).</p><p><b>CONCLUSION</b>JSP is effective and safe in treating PADAM, the mechanism of it is different from supplementing extrinsic androgen. It may have produced the effect by means of favorably regulating the condition of sex hormone to improve the balance of pituitary-sex gland axis, so it has more extensive clinical application.</p>

[Study on the spectral properties of caffeic acid and its derivatives].

Caffeic acid, chlorogenic acid and forsythiaside are active components of Chinese herbs and have diverse biological activities. As the similar compounds, they all include 3, 4-dihydroxy-phenyl in their structures. In this paper, the fluorescence and UV spectra of the caffeic acid, chlorogenic acid and forsythiaside were studied in different pH. The spectra properties of fluorescence and UV were further discussed in view of the phenol hydroxyl states, free, dissociative and protonated. It is found that the luminescence intensity, the peak shape and peak wavelength change with pH. Experimental results also indicate that caffeic acid, chlorogenic acid and forsythiaside can emit fluorescence in wide range of pH (2-12), and fluorescent intensity is enhanced as the dissociation of 4-phenol hydroxyl. On the other hand, fluorescent intensity may be quenched as the dissociation of 3-phenol hydroxyl. Furthermore, in strong acidic or in strong basic media, the fluorescence is quenched partly, and the shape of UV spectra changed greatly in strong basic media, meaning that their molecular structures are changed considerable.