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Epimedium is widely used in traditional Chinese medicine and contains rich bioactive compounds. These compounds often have a methyl group at their 4'-OH position catalyzed by methyltransferases. Therefore, studying methyltransferases in Epimedium plants is of great significance. In this study, a flavonol methyltransferase, EpOMT4, was isolated from Epimedium pseudowushanense B.L. Guo. The recombinant enzyme regiospecifically transferred a methyl group to the 4'-OH position of 8-prenylkaempferol forming icaritin. The study demonstrates that enzymatic methylation of flavonoids in Epimedium plants holds significant potential and could provide a promising alternative method for the biosynthetic production of bioactive methylated prenylflavonoids.

Short-chain dehydrogenase/reductases (SDRs) belong to the NAD(P)(H)-dependent oxidoreductase superfamily, which have various functions of catalyzing oxidation/reduction reactions and have been generally used as powerful biocatalysts in the production of pharmaceuticals. In this study, ScSDR1 and ScSDR2, two new SDRs have been identified and characterized from Stachybotrys chartarum 3.5365. Substrate scope investigation revealed that both of the enzymes possessed the ability to oxidize β -OH to ketone specifically, and exhibited substrate promiscuity and high stereo-selectivity for efficiently catalyzing the structurally different prochiral ketones to chiral alcohols. These findings not only suggest that ScSDR1 and ScSDR2 might be potent synthetic tools in drug research and development, but also provide good examples for further engineered enzymes with higher efficiency and stereo-selectivity.

Anhydroicaritin ( 1a ), baohuoside ( 1b ) and icariin ( 1c ) were recognized as major pharmacologically active ingredients of Epimedium plants. Their primary means of acquisition were chemical isolation from plants. However, it suffers from low yield, environmental pollution and shortage of plants. Herein, to remedy these problems, biosynthesis was explored to obtain the three active ingredients. Fortunately, with SfFPT as 8-prenyltransferase, EpPF3RT and Ep7GT as glycosyltransferases, kaempferide ( 1 ) was transferred to 1a , 1b and 1c enzymatically. Thus, we report the details of this method. This approach represents a promising environmental friendly alternative for the production of these compounds from an abundant analogue.

In this work, a novel isophorone-based fluorescent probe H-1 was designed and synthesized. The probe H-1 could achieve highly selective detection of Al 3+ through forming a 1:1 complex, with a recognition mechanism based on intramolecular charge transfer (ICT). The detection limit of the probe H-1 for Al 3+ is as low as 8.25 × 10 -8 M which was determined by fluorescent titration. It is confirmed that H-1 could be used not only for fluorescence spectrometry to detect Al 3+ ions in actual water samples, but also for biological imaging to detect Al 3+ ions in cells and plants., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Peridecalins C and D ( 1 and 2 ), one decalin and one oxygen-decalin containing polyketide-amino acid hybrids with 5/6/6 ring system, was isolated from a genetic mutant of Periconia sp. F-31. Their structures were elucidated through extensive spectroscopic data analysis, including 1 D/2D NMR and HR-MS spectra. Biosynthetically, two proposed Diels-Alder reactions are supposed to be involved in the skeleton construction of 1 and 2 .

One new eremophilane sesquiterpene periconianone L ( 1 ), together with four known guaiane-type sesquiterpenes 4,10,11-trihydroxyguaiane ( 2 ), (-)-guai-1(10)-ene-4 α ,11-diolhydroxymecuration ( 3 ), guaidiol A ( 4 ), and epi-guaidiol A ( 5 ) were isolated from the endophytic fungus Periconia sp. F-31. The structure of the new compound was established by spectroscopic methods, including UV, IR, HRESIMS, and extensive NMR techniques. Compound 3 was isolated as natural product for the first time.

Objective: To explore clinical efficicacy of closed manipulative reduction and external fixation with cardboard splint in treating Monteggia fracture., Methods: Fifty-eight children with Monteggia fracture were underwent closed manipulative reduction and external fixation with cardboard splint from January 2010 to Junuary 2018. Among them, including 37 males and 21 females, aged from 3.5 to 12 years old with an average of (8.48±2.29) years old;the courses of disease ranged from 0.5 hours to 9 days with an average of (4.21±1.46) days. Broberg and Morrey scores before treatment, 1, 3 and 6 months after treatment were used to evaluate clinical effects., Results: All children were followed up from 1 to 6 months with an average of (3.35±2.12) months. Broberg and Morrey score (7.24±2.81) before treatment, (32.06 ±8.33) at 1 month after treatment, (73.18±5.56) at 3 months after treatment and (95.87±6.75) at 6 months after treatment; there were statistical differences at each time points after treatment with before treatment ( P <0.05);37 children got excellent results, 19 good and 1 moderate., Conclusion: Treatment of Monteggia fractures with closed manipulative reduction and external fixation with cardboard splint could reach integration of motion and quietness, also could remove external fixation at early stage, and get obvious short-term and medium-term therapeutic results.

Objective: In order to obtain new glycosyltransferases with highly efficient catalysis, the glycosyltransferases from Carthamus tinctorius which contains diverse types of glycosides were mined. Methods: A new glycosyltransferase gene (UGT88B2) with full length was obtained by PCR and further transformed into Escherichia coli for heterologous expression. The catalytic activity of recombinant UGT88B2 was determined by HPLC-MS n . The structures of representative catalytic products were elucidated by MS and NMR., Results: UGT88B2 exhibited catalytic promiscuity and various patterns in glycosylation of flavonoids with high efficiency., Conclusion: A new glycosyltransferase named UGT88B2 was successfully mined and can be employed as enzymatic tools in glycosylation of flavonoids., Competing Interests: The authors declare no conflict of interest., (© 2020 Tianjin Press of Chinese Herbal Medicines. Published by ELSEVIER B.V.)

Various bioactive polyketides have been found in Aloe barbadensis. However, the polyketide synthases (PKSs), which participate in biosynthesis of polyketides in A. barbadensis remain unknown. In this study, two type III PKSs (AbPKS1 and AbPKS2) were identified from A. barbadensis . AbPKS1 and AbPKS2 were able to utilize malonyl-CoA to yield heptaketides (TW93a and aloesone) and octaketides (SEK4 and SEK4b), respectively. AbPKS1 also exhibited catalytic promiscuity in recognizing CoA thioesters of aromatics to produce unusual polyketides. What Is more, a whole cell biocatalysis system with the capability of producing 26.4 mg/L of SEK4/SEK4b and 2.1 mg/L of aloesone was successfully established.

Bistachybotrysin K ( 1 ), one new phenylspirodrimane dimer with a central 6/7 oxygen heterocycle core, was isolated from the fungus Stachybotrys chartarum CGMCC 3.5365. Its structure was elucidated by extensive spectroscopic data and single-crystal X-ray diffraction. Compound 1 showed significant cytotoxicity against human tumor cell lines HCT116, NCI-H460, BGC823, Daoy, and HepG2 with IC 50 values in the range of 1.1-4.7 µM.

Three new phenylspirodrimanes derivatives named stachybotrysins H and I ( 1 and 2 ) and stachybotrin E ( 3 ), together with one known compound stachybotrylactam ( 4 ), were isolated from Stachybotrys chartarum CGMCC 3.5365. Their structures were determined by extensive NMR data and mass spectroscopic analysis. Compounds 1 and 2 showed inhibitory effect towards potassium channel Kv1.3 with IC 50 values of 13.4 and 10.9 μM, respectively.

Two new lanostane triterpenoids (1 and 2), two new ergostane-type steroids (3 and 4) together with two known lanostane triterpenoids (5 and 6) and one known steroid (7) were isolated from the cultured mycelia of Ganoderma capense (CGMCC 5.71). Their structures were determined on the basis of extensive spectroscopic (HRESIMS, 1D NMR, 2D NMR) data analyses. Compound 1 exhibited moderate cytotoxic activity against the human cancer cell line NCI-H1650 with an IC 50 value of 22.3 μM, and 7 displayed cytotoxic activity against the human cancer cell line HCT116 with an IC 50 value of 17.4 μM. In addition, compounds 2, 3, 5, and 6 displayed weak anti-HIV activity with IC 50 values of 23.5, 46.7, 21.6, and 30.1 μM, respectively.

Glucuronidation is an important and popular metabolic reaction in vivo of drugs. The further evaluation of biological activity and toxicity of glucuronides is necessary in the course of the drug research and development. However, the synthesis of glucuronides is limited by the lack of efficient approach. Herein, we have developed a new glucuronide synthesis method using plant uridine diphosphate-dependent glucuronosyltransferases (UGTs), UGT88D4, UGT88D7, and EpGT8, enabling the convenient preparation for corresponding O-glucuronide metabolites (1a, 2a, 3a, and 3b) in milligram scale of two neurological active agents, IMM-H004 (1) and FLZ (2). Their structures were characterized by spectroscopic data analyses.

A new steroid glucoside (1), along with nine known steroids (2-10) and four known sorbicillinoids (11-14), were isolated from the endophytic fungus Trichoderma sp. Xy24. Their structures were elucidated on the basis of spectroscopic data analyses and by comparison with reported data. Compounds 3, 5-7, 9, 10, and 13 exhibited significant inhibitory effects on HIV-1 virus with IC 50 values ranging 1.9-9.3 μM; compounds 10, 13, and 14 showed potent inhibitory activity on LPS-induced NO production in BV2 microglia cells with inhibitory rates of 108.2, 100, and 75.1% at 10 μM, respectively. In addition, compound 10 displayed moderate cytotoxicity against BCG823 and HePG2 cell lines with IC 50 values of 11.1 and 17.7 μM, respectively.

Three new sesquiterpenoids trichoacorenols B-C and cyclonerodiol B (1-3), along with three known ones (4-6), were isolated from the mangrove plant endophytic fungus Trichoderma sp. Xy24 using various column chromatography techniques. The structures of these compounds were determined on the basis of extensive spectroscopic data analyses. Compounds 1, 2, 4, and 5 were four acorane sesquiterpenes, 3 and 6 were two monocyclic sesquiterpenediols. Compounds 3 and 5 exhibited significant neural anti-inflammatory activity by inhibiting LPS-induced NO production in BV2 cells with the inhibitory rates of 75.0% and 39.2% at 0.1 μM, respectively, which are more potent than curcumin, a positive control with the inhibitory rate of 21.1% at 0.1 μM.

The glycosides of 4'-demethylepipodophyllotoxin (DMEP) possess various pharmacological activities; however, the chemical synthesis of these glycosides faces challenges in regioselectivity, stereoselectivity, and the protection and de-protection of functional groups. In this work, a novel glycosyltransferase (GT) gene AbGT5 from Aloe barbadensis was successfully cloned, heterogeneously expressed and purified. Recombinant AbGT5 was able to catalyze the glycosylation of DMEP and the glycosylated product, which was separated from the preparative scale reaction, was characterized as DMEP 4'-O-β-D-glucoside via MS, 1H-NMR, 13C-NMR, HSQC and HMBC. According to the investigations of enzyme properties, AbGT5 show the highest activity around 20 ℃ in the buffer of pH 9.0, and it was independent of divalent metal ions. Under the optimum conditions, the conversion rate of DMEP can reach 80%. Above all, in this work the enzymatic glycosylation of DMEP was achieved with high efficiency by the novel GT AbGT5., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Five monoterpenoids were isolated from the endophytic fungus Periconia sp. F-31, including three new carene-type monoterpenoids, 2-carene-5,8-diol (1), 2-carene-8,10-diol (2), 2-carene-8-acetamide (3), one new menthene-type monoterpenoid 8-hydroxy-1,7-expoxy-2-menthene (4), and one known monoterpenoid anethofuran (5). The structures of all compounds were elucidated based on a comprehensive spectroscopic data analysis, electronic circular dichroism (ECD), and calculated ECD.

Secoisolariciresinol dehydrogenase (SDH) is a key enzyme involved in the biosynthetic pathway of podophyllotoxin.In this study, two SDH candidate genes,SO282 and SO1223, were cloned from callus of Dysosma versipellis by homology-based PCR and rapid amplification of cDNA end (RACE).The SDH candidate genes were expressed in Escherichia coli and the subsequent enzyme assay in vitro showed that recombinant SO282 had the SDH activity. These results pave the way to the follow-up investigation of the biosynthetic of podophyllotoxin., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Objective: To investigate the therapeutic effects of manipulative reduction for calcaneal fractures., Methods: From January 2009 to June 2012, 53 cases (60 affected feet) of Sanders type I-IV calcaneal fractures were treated by manipulative reduction and external fixation with cardboard splint,including 45 males and 8 females with an average age of (33.5 +/- 1.54) years old ranging from 18 to 65. The course of disease ranged 0.5 h to 7 d. Before treatment the feet were swelling, ache and activity limitation in evidence, some feet were wide flat deformity, the cortical bone was broken in the imaging examination. All selected cases were evaluated using Creighton-Nebraska health foundation assessment scale for fractures of calcaneus., Results: All cases were followed up at 12 months after treatment, 13 feet got excellent curative effect, 34 good, 11 moderate and 2 poor., Conclusion: Manipulative reduction for Sanders type I-IV calcaneal fractures could get excellent curative effect. Manipulative reduction for calcaneal fractures could avoid surgical trauma while assure high curative effects. Manipulative reduction is not only economical and easy therapy, but also can restore maximum function of the calcaneus with few complications and facilitate early rehabilitation of ankle and joint function.

Naringenin (1) was transformed to three metabolites (2-4) by Mucor sp. Based on LCMS(n)-IT-TOF and NMR spectroscopic data, 2-4 were identified as naringenin-7-O-sulphate, naringenin-4'-O-sulphate, and naringenin-5-O-sulphate, respectively. These results might provide hints to the mammalian/human metabolism of naringenin.

Objective: To explore clinical effects of manipulative reduction with minimally invasive percutaneous plate osteosynthesis in treating distal tibiofibular fractures., Methods: From 2009 to 2011, 60 patients with distal tibiofibular fractures were treated by manipulative reduction with minimally invasive percutaneous plate osteosynthesis. Among them, there were 32 males and 28 females aged from 14 to 70 years old with an average of 41.22 +/- 2.06. According to AO classification of fractures,5 cases were type A1, 22 cases were type A2,21 cases were type A3 and 12 cases were type C1. Operation time, blood loss,time of callus and fracture healing were observed, Mazur scoring of ankle joint were used to evaluate therapeutic., Results: Fifty-eight incisions were healed at stage I ,and 2 cases were infected at distal tibial. Operation time was with an average of (62.34 +/- 5.66) min ranged 45 to 90 min;blood loss was 30 to 150 ml with an average of (80.57 +/- 5.59) ml;formation of callus appeared from 4 to 12 weeks,with an average of (8.24 +/- 2.06) weeks, and fracture healing time was from 3 to 6 months, with an average of (4.50 +/- 1.13) months. According to Mazur scoring of ankle joint 40 cases got excellent results, 18 good, and 2 fair., Conclusion: Manipulative reduction with minimally invasive percutaneous plate osteosynthesis can obtain reliable fixation. It is a good choice of treating distal tibiofibular fractures by protecting blood supply of fractures.

In the present study, Potamogeton crispus L. plants exposed to various concentrations of silver (Ag) (5, 10, 15, and 20 microM) for 5d were investigated to determine the accumulating potential of Ag and its influence on nutrient elements, chlorophyll pigments and fluorescence, various antioxidant enzymes and compounds, adenosine triphosphate (ATP), protein content and ultrastructure. The accumulation of Ag was found to increase in a concentration dependent manner with a maximum of 29.3 microg g(-1) at 20 microM. The nutrient elements (except Ca), photosynthetic pigments, chlorophyll a fluorescence parameters (Fo, Fv, Fv/Fm, Fv/Fo), malondialdehyde (MDA), ATP, peroxidase (POD) activity, ascorbate (AsA), reduced glutathione (GSH) and protein contents decreased significantly as concentration of Ag augmented. In contrast, an induction in SOD activity was recorded, while an initial rise in Ca content and CAT activity was followed by subsequent decline. Morphological symptoms of senescence phenomena such as chlorosis and damage of chloroplasts and mitochondria were observed even at the lowest concentration of Ag, which suggested that Ag hastened the senescence of the tested plants. The loss of nutrients and chlorophyll content and damage of chloroplasts were associated with disturbances in photosynthetic capacity as indicated by the quenching of chlorophyll a fluorescence. Decreased chlorophyll and protein contents suggest oxidative stress induced by Ag. In addition, both the reduction of ATP and the damage to the ultrastructure of organelles were indicative of general disarray in the cellular functions exerted by Ag.

On the basis of a JA-inducible EST, the full length cDNA of a putative protein kinase gene, OsSJMKJ, was isolated from rice (Oryza sativa L.). The predicted protein of this gene, consisting of 498 amino acids with a predicted PI value of 8.43, contains a mitogen-activated protein kinase (MAPK) domain with a dual-phosphorylation activation motif TDY (Thr-Asp-Tyr) and a C-terminus domain that feature a typical MAPK belonging to subgroup V. OsSJMK1 showed 81% sequence identity to the reported BWMK1, a blast fungus and wounding inducible protein, in the N-terminus kinase domain but had low sequence identity in the C-terminus domain. Despite the high similarity of gene structure between OsSJMK1 and BWMK1, the expression of OsSJMK1 was weakly induced by blast fungus but not wounding, rather it was transiently induced by jasmonic acid (JA) and salicylic acid (SA) at early stages of treatment. This gene was not induced by other hormones (such as ABA) or abiotic stresses, such as drought and salinity. The low level of transcript of this gene was detected in the various tissues of rice. The results suggest that OsSJMK1 might be activated specifically by JA and SA and involved in defense signaling pathways.