Four new steroidal glycosides from Lilium lancifolium Thunb. and their antitumor activity.

Four new steroidal glycosides (1-4), including two steroidal saponins named lililancifoloside B and C (1-2), one pregnane glycoside named lililancifoloside D (3), and one C22-steroidal lactone glycoside named lililancifoloside E (4), together with five known ones (5-9), were isolated from the bulbs of Lilium lancifolium Thunb. By using spectroscopic analysis, including 1D, 2D NMR, and HR-ESI-MS, the structures of 1-4 were elucidated. All isolates were tested for their cytotoxic potential against the MCF-7, MDA-MB-231, HepG2, and A549 cell lines. Compound 6 distinguished out among them, IC50 values of 3.31, 5.23, 1.78, and 1.49 µM against the four cell lines, respectively. Other compounds such as compound 3, 5, and 9 have also shown specific cytotoxic activity. Next, studies showed that compound 6 might cause HepG2 cells to undergo a cell cycle arrest during the G2/M phase and apoptosis.

Modified Acupotomy versus Percutaneous Release for Trigger Thumb: A Retrospective Study.

Background: Acupotomy is now increasingly used for trigger thumb, while recent evidence showed it increased the risk of nerve injury. Based on the close proximity of the neurovascular bundles and the A1 pulley, we designed a modified acupotomy. Given that percutaneous release is the common surgical treatment, this retrospective study aimed to compare the effect and safety of modified acupotomy versus percutaneous release for trigger thumb. Methods: This is a retrospective study. All patients with trigger thumb were retrieved in the electronic records of the Department of Pain Medicine at the Beijing Hospital of Traditional Chinese Medicine from January 2016 to September 2018. Both short-term (3 months) and long-term (2 years) outcomes were evaluated using the criteria established through Gilberts et al.'s questionnaire, including triggering, residual pain, stiffness, digital nerve injury, scar, infection and satisfaction. Chi-square test or Fisher's exact test was used to compare differences between two groups. Results: A total of 305 patients with 334 trigger thumbs treated with either modified acupotomy (n = 194 thumbs) or percutaneous release (n = 140 thumbs) were included. Of them, 221 (72.5%) were female, and the mean age was 56.2 ± 10.0 years. The mean duration of trigger thumb lasted for 7.5 ± 3.6 months. At 3 months, all triggering were alleviated by both therapies. Although more digital nerve injury occurred in the percutaneous release (0 [0%] thumbs vs 5 [3.6%] thumbs, P = 0.012), more residual pain (30 [15.5%] thumbs vs 6 [4.3%] thumbs; rate ratio, 3.61; 95% confidence interval [CI]: 1.54-8.43; P = 0.001) and less satisfaction occurred in the modified acupotomy group. At 2 years, more recurrent triggering, residual pain and digital nerve injury occurred in the percutaneous release group (2 [1.0%] thumbs vs 12 [8.6%] thumbs; rate ratio, 0.12; 95% CI: 0.03-0.53; P = 0.001; 9 [4.6%] thumbs vs 22 [15.7%] thumbs; rate ratio, 0.30; 95% CI: 0.14-0.62 P < 0.001; 0 [0%] thumbs vs 4 [2.9%] thumbs, P = 0.030, respectively). Moreover, satisfaction was significantly better in the modified acupotomy group. Conclusion: The modified acupotomy had better long-term outcomes and satisfaction than the percutaneous release for trigger thumb, although percutaneous release has less residual pain and better satisfaction in the short term. The modified acupotomy is a treatment option for trigger thumb.

Effectiveness and safety of acupotomy for knee osteoarthritis: study protocol for a randomized controlled trial.

BACKGROUND: Knee osteoarthritis (KOA) is one of the most common musculoskeletal disorders. Acupotomy may be effective for KOA, but the evidence is limited. This trial aims to determine the effectiveness and safety of acupotomy for KOA. METHODS/DESIGN: This is a parallel-group, assessor-blinded randomized controlled trial. Two hundred patients with KOA will be recruited and randomly assigned to two groups (group A or group D) in a 1:1 ratio. Patients in group A will receive acupotomy and topical diclofenac diethylamine for 4 weeks, while patients in group D will receive topical diclofenac diethylamine alone for 4 weeks. The primary outcome will be the response rate-the proportion of patients who achieve the minimal clinically important improvement in pain and function at week 4 compared with baseline. Secondary outcomes will include pain, function, quality of life, the use of rescue medicine (loxoprofen sodium), and adverse events at weeks 4, 8, and 24 after randomization. Besides, joint fluid and serum will be collected to assess the level of inflammatory cytokines, like TNF-α, IL-1ß, and MMP-3. DISCUSSION: This study will contribute to a better understanding of the effectiveness and safety of acupotomy in combination with topical nonsteroidal anti-inflammatory drugs. If the hypothesis is confirmed, acupotomy may be recommended as adjunctive therapy for patients with KOA. Results of the study will be of great importance for the guidelines of clinical therapy. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR2100043005 Registered on 4 February 2021.

Acupotomy Alleviates Energy Crisis at Rat Myofascial Trigger Points.

The aim of this study was to determine the effects of acupotomy on energy crises in rat trigger points (TrPs) by measuring mechanical pain thresholds (MPTs) and levels of acetylcholinesterase (AChE), free sarcoplasmic calcium (Ca2+), adenosine 5'-triphosphate (ATP), adenosine 5'-monophosphate (AMP), substance P (SP), and calcitonin gene-related peptide (CGRP) in rat muscle TrP tissue. Male Sprague Dawley rats (n = 32) were randomly divided into four groups: control, TrP, acupotomy, and lidocaine injection. Enzyme-linked immunosorbent assays were used to measure AChE, and free sarcoplasmic Ca2+ concentrations were determined by fluorescent staining with Fura-2 AM; high-performance liquid chromatography was used to measure ATP and AMP, and SP and CGRP were evaluated by immunohistochemistry. Compared with the control group, free sarcoplasmic Ca2+, AMP, SP, and CGRP were higher in the model group, while MPT, AChE, and ATP were lower. Treatment with acupotomy or lidocaine injection reduced free sarcoplasmic Ca2+, SP, and CGRP and increased MPTs and AChE levels compared with the model group. However, only acupotomy also led to decreased AMP and increased ATP levels relative to the model group. We conclude that acupotomy can alleviate energy crises at TrPs.

New phenolic acids from Salvia yunnanensis C.H.Wright.

Two new phenolic acids, ethyl pro-lithospermate (1), n-butyl pro-lithospermate (2) were isolated from Salvia yunnanensis C.H.Wright, along with nineteen known compounds (3-21). The structures of the isolated compounds were elucidated on the basis of extensive spectrometry and by comparing their physical and spectroscopic data to the literature. Among them, compounds 11, 12 and 14-16 were firstly isolated from S. yunnanensis C.H.Wright. Some of the isolated compounds were evaluated for their neuroprotection. Compounds 10-12 showed significant neuroprotective effects in PC12 cells and compounds 1, 4-7 displayed moderate neuroprotective effects.

Salvianolic acids T and U: a pair of atropisomeric trimeric caffeic acids derivatives from root of Salvia miltiorrhiza.

Two new trimeric caffeic acids, named salvianolic acids T and U (1 and 2), were isolated from the underground part of Salvia miltiorrhiza. Their structures, consisting of three caffeic acid units, were determined based on extensive 1D- and 2D-spectroscopic analyses and electronic circular dichroism (ECD) calculations.

Simultaneous determination of five phenolic components and paeoniflorin in rat plasma by liquid chromatography-tandem mass spectrometry and pharmacokinetic study after oral administration of Cerebralcare granule(®).

A simple, sensitive and selective high-performance liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) method was developed for simultaneous determination and pharmacokinetic study of six active components, protocatechuic acid, chlorogenic acid, caffeic acid, ferulic acid rosmarinic acid and paeoniflorin in rat plasma after oral administration of Cerebralcare granule(®) for the first time. The method involves a simple liquid-liquid extraction with ethyl acetate. The separation was performed on a Luna C18 column (2.0×100mm i.d., 3.0µm, particle, Phenomenex, USA) with gradient elution using a mobile phase composed of acetonitrile and water (containing 0.1% formic acid) at a flow rate of 0.2ml/min. Electrospray ionization (ESI) in negative ion mode and selective reaction monitoring (SRM) was used for the quantification of six active components and internal standard (IS, Chloroamphenicol). The method was linear for all analytes over investigated range with all correlation coefficients greater than 0.9914. The lower limits of quantification (LLOQ) were 1.0ng/ml for protocatechuic acid, 1.0ng/ml for chlorogenic acid, 1.0ng/ml for caffeic acid, 5.0ng/ml for ferulic acid, 1.5ng/ml for rosmarinic acid and 6.0ng/ml for paeoniflorin, respectively. The intra- and inter-day precisions (R.S.D.%) were less than 6.60% and 11.68%, and accuracy (RE %) between -3.26% and 1.13% (n=6). The developed method was applied for the first time to the pharmacokinetic study of protocatechuic acid, chlorogenic acid, caffeic acid, ferulic acid, rosmarinic acid and paeoniflorin in rat plasma after oral administration of Cerebralcare granule(®).

Antioxidant phenolic glycosides from the bark of Populus ussuriensis Kom.

Study on the EtOAc soluble fraction from the bark of Populus ussuriensis Kom. resulted in the isolation of three phenolic glycosides, including populoside (1), 7-O-ρ-coumaroylsalirepin (2) and 7-O-caffeoylsalirepin (3), among which 3 is a new compound. The structures of these compounds were elucidated on the basis of spectroscopic evidence. Phenolic glycosides 1-3 exhibited excellent antioxidant activity, evaluated in the ABTS⁺ radical scavenging assay.

Phenolic compounds in the leaves of Populus ussuriensis and their antioxidant activities.

Two new phenolic glucosides [isograndidentatin A (1) isograndidentatin B (2)], 5 known phenolic glucosides [grandidentatin (3), salireposide (4), populoside (5), populoside A (6), and salicortin (7)], and 2 known phenolic acids [P-coumaric acid (8) and caffeic acid (9)] were isolated from the leaves of Populus ussuriensis. Structure elucidation of 1 and 2 was achieved through extensive spectroscopic techniques. Compounds 1-6 and 9 showed significant antioxidant activities, which were evaluated by the DPPH radical-scavenging method (IC(50) values of 6.68, 6.61, 6.75, 6.84, 6.76, 6.79, and 5.92 microM, respectively) and the ABTS .+ radical-scavenging system (TEAC values of 1.21, 1.28, 1.26, 1.05, 1.69, 1.60, and 2.00 mM, respectively).

Post-genome research on the biosynthesis of ergot alkaloids.

Genome sequencing provides new opportunities and challenges for identifying genes for the biosynthesis of secondary metabolites. A putative biosynthetic gene cluster of fumigaclavine C, an ergot alkaloid of the clavine type, was identified in the genome sequence of ASPERGILLUS FUMIGATUS by a bioinformatic approach. This cluster spans 22 kb of genomic DNA and comprises at least 11 open reading frames (ORFs). Seven of them are orthologous to genes from the biosynthetic gene cluster of ergot alkaloids in CLAVICEPS PURPUREA. Experimental evidence of the identified cluster was provided by heterologous expression and biochemical characterization of two ORFs, FgaPT1 and FgaPT2, in the cluster of A. FUMIGATUS, which show remarkable similarities to dimethylallyltryptophan synthase from C. PURPUREA and function as prenyltransferases. FgaPT2 converts L-tryptophan to dimethylallyltryptophan and thereby catalyzes the first step of ergot alkaloid biosynthesis, whilst FgaPT1 catalyzes the last step of the fumigaclavine C biosynthesis, i. e., the prenylation of fumigaclavine A at C-2 position of the indole nucleus. In addition to information obtained from the gene cluster of ergot alkaloids from C. PURPUREA, the identification of the biosynthetic gene cluster of fumigaclavine C in A. FUMIGATUS opens an alternative way to study the biosynthesis of ergot alkaloids in fungi.

The biosynthetic gene clusters of aminocoumarin antibiotics.

Plants and microorganisms are the most important sources of secondary metabolites in nature. For research in the functional genomics of secondary metabolism, and for the biotechnological application of such research by genetic engineering and combinatorial biosynthesis, most microorganisms offer a unique advantage to the researcher: the biosynthetic genes for a specific secondary metabolite are not scattered over the genome, but rather are clustered in a well-defined, contiguous region - the biosynthetic gene cluster of that metabolite. This is exemplified in this review for the biosynthetic gene clusters of the aminocoumarin antibiotics novobiocin, clorobiocin and coumermycin A (1), which are potent inhibitors of DNA gyrase. Cloning, sequencing and analysis of the biosynthetic gene clusters of these three antibiotics revealed that the structural differences and similarities of the compounds are perfectly reflected by the genetic organisation of the biosynthetic gene clusters. The function of most biosynthetic genes could be identified by gene inactivation experiments as well as by heterologous expression and biochemical investigation. The prenylated benzoic acid moiety of novobiocin and clorobiocin, involved in the interaction with gyrase, is structurally similar to metabolites found in plants. However, detailed investigations of the biosynthesis revealed that the biosynthetic pathway and the enzymes involved are totally different from those identified in plants.

Pharmacokinetic study of artemisinin after oral intake of a traditional preparation of Artemisia annua L. (annual wormwood).

Artemisia annua L. (annual wormwood) contains the antimalarial artemisinin. Aqueous preparations of the dried herb are included in the pharmacopoeia of the People's Republic of China for treatment of fever and malaria. Fourteen healthy male volunteers received one liter of tea prepared from nine grams of Artemisia annua leaves. Blood samples were taken and artemisinin was detected by reversed phase high-performance liquid chromatography. The mean +/- SD maximum plasma concentration of artemisinin was 240 +/- 75 ng/mL and the mean +/- SD area under the plasma concentration-time curve was 336 +/- 71 ng/mL x hr. Artemisinin was absorbed faster from herbal tea preparations than from oral solid dosage forms, but bioavailability was similar. One liter of an aqueous preparation of nine grams of Artemisia annua contained 94.5 milligrams of artemisinin (approximately 19% of the usually recommended daily dose). Artemisinin plasma concentrations after intake of this herbal tea are sufficient for clinical effects, but insufficient to recommend such preparations as equivalent substitutes for modern artemisinin drugs in malaria therapy.