Efficacy and safety comparison of infrared laser moxibustion and traditional moxibustion in knee osteoarthritis: study protocol for a Zelen-design randomized controlled non-inferiority clinical trial.

BACKGROUND: Knee osteoarthritis (KOA) is the most common chronic degenerative joint disease and places a substantial burden on the public health resources in China. The purpose of this study is to preliminarily evaluate whether infrared laser moxibustion (ILM) is non-inferior to traditional moxibustion (TM) in the treatment of KOA. MATERIALS AND METHODS: In the designed Zelen-design randomized controlled non-inferiority clinical trial, a total of 74 patients with KOA will be randomly allocated to one of two interventions: ILM treatment or TM treatment. All participants will receive a 6-week treatment and a follow-up 4 weeks after treatment. The primary outcomes will be the mean change in pain scores on the numeric rating scale (NRS) measured at baseline and the end of last treatment at week 6. The secondary outcomes will be the pain scores on the NRS from weeks 1 to 5 after the start of treatment and the changes from baseline to endpoints (weeks 6 and 10) in the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), SF-36, knee circumference, and 6-min walking test. In addition, safety assessment will be performed throughout the trial. CONCLUSION: The results of our study will help determine whether a 6-week treatment with ILM is non-inferior to TM in patients with KOA, therefore providing evidence to verify if ILM can become a safer alternative for TM in clinical applications in the future. TRIAL REGISTRATION: Clinical Trial Registration Platform (ChiCTR2200065264); Pre-results. Registered on 1 November 2022.

Identification of the high-yield monacolin K strain from Monascus spp. and its submerged fermentation using different medicinal plants.

BACKGROUND: Medical plants confer various benefits to human health and their bioconversion through microbial fermentation can increase efficacy, reduce toxicity, conserve resources and produce new chemical components. In this study, the cholesterol-lowering monacolin K genes and content produced by Monascus species were identified. The high-yield monacolin K strain further fermented with various medicinal plants. The antioxidant and anti-inflammatory activities, red pigment and monacolin K content, total phenolic content, and metabolites in the fermented products were analyzed. RESULTS: Monacolin K was detected in Monascus pilosus (BCRC 38072), and Monascus ruber (BCRC 31533, 31523, 31534, 31535, and 33323). It responded to the highly homologous mokA and mokE genes encoding polyketide synthase and dehydrogenase. The high-yield monacolin K strain, M. ruber BCRC 31535, was used for fermentation with various medicinal plants. A positive relationship between the antioxidant capacity and total phenol content of the fermented products was observed after 60 days of fermentation, and both declined after 120 days of fermentation. By contrast, red pigment and monacolin K accumulated over time during fermentation, and the highest monacolin K content was observed in the fermentation of Glycyrrhiza uralensis, as confirmed by RT-qPCR. Moreover, Monascus-fermented medicinal plants including Paeonia lactiflora, Alpinia oxyphylla, G. uralensis, and rice were not cytotoxic. Only the product of Monascus-fermented G. uralensis significantly exhibited the anti-inflammatory capacity in a dose-dependent manner in lipopolysaccharide-induced Raw264.7 cells. The metabolites of G. uralensis with and without fermentation (60 days) were compared by LC/MS. 2,3-Dihydroxybenzoic acid, 3,4-dihydroxyphenylglycol, and 3-amino-4-hydroxybenzoate were considered to enhance the antioxidant and anti-inflammatory ability. CONCLUSIONS: Given that highly homologous monacolin K and citrinin genes can be observed in Monascus spp., monacolin K produced by Monascus species without citrinin genes can be detected through the complementary methods of PCR and HPLC. In addition, the optimal fermentation time was important to the acquisition of antioxidants, red pigment and monacolin K. These bioactive substances were significantly affected by medicinal plants over fermentation time. Consequently, Monascus-fermented G. uralensis had a broad spectrum of biological activities.

Tao-Hong-Si-Wu Decoction promotes angiogenesis after cerebral ischaemia in rats via platelet microparticles.

Platelet microparticles (PMPs) are membrane particles derived from the platelet membrane that enter into the blood circulation. We sought to explore the therapeutic effects of Tao-Hong-Si-Wu Decoction (THSWD) on angiogenesis in a rat model of cerebral ischaemia-reperfusion (I/R). The protective effect of THSWD on I/R rats was observed morphologically by immunohistochemical expression of VEGF and CD34, along with immunofluorescence results of co-expression of BrdU and vWF. Then, PMPs from different groups of rats were extracted, and cytokine array analysis was used to screen for angiogenesis associated proteins. The results showed that THSWD can promote the expression of VEGF, CD34, BrdU and vWF. Cytokine array analysis revealed the changes in the expression of 29 related angiogenic proteins in the total protein of PMPs, which involved the Notch signalling pathway. Compared with model group, the expression levels of NICD and Hes-1 in the THSWD group were significantly increased. In the context of I/R, the angiogenesis-related proteins of PMPs are different. THSWD may involve the promotion of activation of the Notch signalling pathway to achieve therapeutic effects on cerebral ischaemia.

Benzophenones from Anemarrhena asphodeloides Bge. Exhibit Anticancer Activity in HepG2 Cells via the NF-κB Signaling Pathway.

A chemical investigation of the fibrous roots of Anemarrhena asphodeloides Bge. led to the isolation of four benzophenones, including one new compound (1) and three known ones (2-4). Comprehensive 1D, 2D NMR and HRESIMS data established the structures of the isolated compounds. The absolute configurations were determined by comparison of the calculated optical rotation (OR) with experimental data. All the isolates were evaluated for their cytotoxicities on hepatocellular carcinoma cell lines (HepG2 and Hep3B). Compound 1 showed strong cytotoxicity against HepG2 and Hep3B cells, with IC50 values at 153.1 and 180.6 nM. Through MTT assay, flow cytometry and Western blot analysis, compound 1 demonstrated the ability to stimulate apoptosis via the NF-κB signaling pathway in HepG2 cells. These benzophenones are potential lead compounds for the development of better treatments for hepatocellular carcinoma.

Monoterpene indole alkaloids from Rhazya stricta.

Twenty-seven monoterpene indole alkaloids (MIAs) including three new ones were isolated from the plant of Rhazya stricta. Their structures were elucidated by analyses of HRMS and NMR data. Secopleiocarpamine A (1) represents a novel 2,3-seco pleiocarpamine type MIA possessing a cyano group. A possible biosynthetic pathway for 1 was postulated. All compounds were evaluated for their inhibitory activities against six Candida strains, and the results showed that 2, 5, 12, 21, 23, and 27 exhibited moderate inhibitory activities with MIC values ranging from 3.125 to 50 µg/mL.

Two new steroidal saponins isolated from Anemarrhena asphodeloides.

Two new steroidal saponins, named timosaponin P (1) and timosaponin Q (2), were isolated from the rhizome parts of Anemarrhena asphodeloides Bunge using various chromatographic methods. Their structures and absolute configurations were elucidated by a combination of spectroscopic and spectrometric data, including 1D, 2D NMR, HR-ESI-MS and ECD calculations, and this is the first time the absolute configuration of C-23 of steroidal saponin was confirmed by ECD calculations.

Perceived criticism predicts outcome of psychotherapy for panic disorder: Replication and extension.

OBJECTIVE: We tested the relation of perceived criticism (PC) from a parent or spouse/romantic partner to outcome of psychotherapy for panic disorder (PD). METHOD: Participants were 130 patients with PD (79% with agoraphobia) who received 24 twice-weekly sessions of panic-focused psychodynamic psychotherapy, cognitive-behavioral therapy, or applied relaxation therapy. Patients were predominantly White (75%), female (64%), and non-Hispanic (85%). At baseline, Week 5 of treatment, termination, and at 6- and 12-month follow-up, patients rated PC from the relative with whom they lived. Independent evaluators assessed the severity of PD at baseline, Weeks 1, 5, and 9 of treatment, termination, and the 2 follow-up points. Data were analyzed with piecewise (treatment phase, follow-up phase) latent growth curve modeling. RESULTS: The latent intercept for PC at baseline predicted the latent slope for panic severity in the follow-up (p = .04) but not the active treatment phase (p = .50). In contrast, the latent intercept for PD severity at baseline did not predict the latent slope on PC in either phase (ps ≥ .29). Nor did the slopes of PC and PD severity covary across treatment (p = .31) or follow-up (p = .13). Indeed, PC did not change significantly across treatment (p = .45), showing the stability of this perception regardless of significant change in severity of patients' PD (p < .001). CONCLUSIONS: Because PC predicts worse long-term treatment outcome for PD, study findings argue for interventions to address perceived criticism in treatment. (PsycINFO Database Record

[Steroidal saponins and their pharmacological activities in Solanum plants].

The genus Solanum which is the largest genus in Solanaceae consists of more than 2 000 species, most of them are distributed in tropics and subtropics areas, and only a small amount in temperate regions throughout the world. Steroidal saponins could be found in many species of the Solanum, and they are an important group of natural products exhbiting a number of potent beneficent properties, such as anti-tumor, antibacterial, antifungal, anti-inflammatory, antiviral, antioxidant, hypoglycemic, and antihyperlipidemic effects. As supplement, this paper gives a review of different structural categories of steroidal saponins and their pharmacological effects from the solanum plants over the past decade, and it is intended to provide a point of reference for further research on steroidal saponins in Solanum plants.

Magnetic molecularly imprinted polymer for the selective extraction of sildenafil, vardenafil and their analogs from herbal medicines.

The successfully developed magnetic molecularly imprinted polymers (MMIPs) toward six synthetic phosphodiesterase type-5 (PDE-5) inhibitors were described. Sildenafil was used as template for the preparation of MMIPs using superparamagnetic core-shell nanoparticle as supporter. The obtained MMIPs were characterized using transmission electron microscope, Fourier transform infrared, X-ray diffraction, and vibrating sample magnetometer. High performance liquid chromatography (HPLC) with diode array detector (DAD) was used for the analysis of target analytes. The application of MMIPs as selective sorbent in the cleanup of herbal medicine samples prior to HPLC offered simple sample preparation. The adsorption capacity and selectivity of prepared MMIPs and magnetic non-molecularly imprinted polymers were investigated. The binding isotherms were obtained for sildenafil and fitted by Freundlich isotherm model. Structurally similar compound of sildenafil and a reference compound protocatechuic acid were used for investing the selective recognition of MMIPs.

[Effect of Evn-50 on cell growth and apoptosis in tamoxifen-resistance human breast cancer cell line MCF-7/TAM-R].

OBJECTIVE: To investigate the effect of Evn-50 extracted from Vitex negundo on human breast cancer cell line MCF-7 and MCF-7/TAM-R cells in vitro. METHODS: MCF-7 and tamoxifen-resistant MCF-7/TAM-R cells were treated with Evn-50,tamoxifen or combination of Evn-50 and tamoxifen. Cell proliferation inhibition rates were determined by MTT assay. The apoptosis rate and the change of cell cycle were detected by PI staining flow cytometry. Protein expression of phospho-MAPK 44/42 (Thr202/Tyr204),MAPK P44/42, phospho-AKT (Ser473) and AKT were detected with Western blotting. RESULTS: The viability of MCF-7 cells was decreased in combination group [(28.65 ±11.43)%] and Evn-50 group [(53.02 ±15.14)%] compared with TAM group (P<0.01). The cell viability of MCF-7/TAM-R in combination group [(42.11 ±14.30)%] was significantly lower than that in TAM group [(92.18 ±13.16)%] (P<0.01). The cell apoptosis rate was dependent on the time of treatment in all groups,the effects on apoptosis and G2/M phase cells were most prominent at 72 h (P<0.01). Western blotting revealed that protein levels of phosphorylated AKT and p-MAPK44/42 decreased,while the expression of total AKT and MAPK44/42 was stable. In MCF-7/TAM-R cells,the expression of phosphorylation of AKT and MAPK44/42 protein was not changed in Evn-50 or TAM alone group,but significantly inhibited in the combination group at 72 h. CONCLUSION: Evn-50 can inhibit cell growth and induce apoptosis in MCF-7 and MCF-7/TAM-R cells,it can reverse tamoxifen-resistance of MCF-7/TAM-R cells.The mechanisms may be related to the down-regulation of phosphorylated ERK1/2 in MAPK signal pathway and phosphorylated AKT in AKT signal pathway.

Magnetic molecularly imprinted polymer for the detection of rhaponticin in Chinese patent medicines.

The magnetic molecularly imprinted polymers (MMIPs) had been synthesized for the selective extraction and clean-up of rhaponticin in Chinese patent medicines (CPM). The MMIPs were prepared via suspension polymerization, using Fe(3)O(4) as magnetically component, rhaponticin as template molecule, acrylamide (AM) as functional monomers, styrene (St) as copolymer monomer and ethylene glycol dimethacrylate (EGDMA) as a cross-linker in dimethyl sulfoxide porogen. The obtained MMIPs were characterized using scanning electron microscopy (SEM) and Fourier transform infrared (FT-IR), X-ray diffraction (XRD) and vibrating sample magnetometer (VSM). High performance liquid chromatography (HPLC) was used for analysis the target analytes. The polymers were evaluated further by batch rebinding experiments. From the derived Freundlich isotherm (FI) equation their binding capacity and binding strength were determined. Structurally similar compound resveratrol and a reference compound kirenol were used for investing the selective recognition capability of MMIPs. The MMIPs were used as sorbents for selective pre-concentration of rhaponticin from CPM samples. The contents of rhaponticin for the four CPM samples were found to be 11.84, 3.35, 4.47, and 7.57µg g(-1), respectively, and the recoveries of spiked samples ranged from 77.82% to 91.00%. The prepared MMIPs could be applied to selectively pre-concentrate and determine rhaponticin in CPM samples.

Molecularly imprinted polymer for the specific solid-phase extraction of kirenol from Siegesbeckia pubescens herbal extract.

Molecular imprinted polymers (MIPs) were prepared by thermal polymerization using a non-covalent molecularly imprinting strategy with kirenol as the template, acrylamide (AM) as the functional monomer and ethylene glycol dimethacrylamide (EGDMA) as the cross-linker in the porogen of tetrahydrofuran (THF). The synthesized MIPs were characterized by scanning electron microscopy (SEM) and Fourier transform infrared (FT-IR). Its molecular recognition property was investigated by UV spectrogram. High-pressure liquid chromatography (HPLC) was used for analysis of target analytes. The polymers were evaluated further by batch rebinding experiments, and from the derived isotherms their binding capacity and binding strength were determined. Then the selectivity of the MIPs was checked toward the selected structurally related compounds and the recognition coefficients for kirenol, darutigenol, and ent-2-oxo-15, 16, 19-trihydroxypimar-8(14)-ene (TD) were 2.47, 3.43 and 3.40, respectively. The properties of MIPs for SPE were also evaluated. The results obtained demonstrate that the good imprinting effect and the excellent selectivity of MIPs were obtained. The optimized molecular imprinted SPE procedure was applied to extract kirenol directly from the extracts of the aerial part of Siegesbeckia pubescens herb. A selective extraction of kirenol from traditional Chinese medicine (TCM) was achieved with extraction yield of 80.9%.

Ultra-performance LC-photodiode array-eλ-ESI-MS/MS screening method for the detection of radical-scavenging natural antioxidants from radix et rhizoma Rhei.

A novel ultra-performance LC-photodiode array-el-ESI-MS/MS screening method was developed for the detection and identification of natural antioxidants from radix et rhizoma Rhei. Nine compounds were found to possess a potential antioxidant activity, and their free radical-scavenging capacities were investigated in detail. The nine compounds were identified as 1-O-galloyl-2-O-cinnamoylglucose, 6-hydroxymusizin-8-O-ß-D-glucopyranoside, (+)-catechin, gallic acid 3-O-ß-D-glucopyranoside, trans-3,5,4'-trihydroxystilbene- 4'-O-ß-D-(2"-O-galloyl)-glucopyranoside, sennoside A, 4-(4'-hydroxyphenyl)-2- butanone-4'-O-ß-D-(2"-O-galloyl-6"-O-p-coumaroyl) glucopyranoside, emodin-8-O-(6'-Omalonyl) glucopyranoside, and physcion-8-O-ß-D-glucopyranoside. The reactivity and SC(50) values of those compounds were investigated, respectively. 1-O-Galloyl-2-O-cinnamoylglucose showed the strongest capability for scavenging 1,1-diphenyl-2-picrylhydrazylfree radical; trans-3,5,4'-trihydroxystilbene-4'-O-ß-D-(2"-O-galloyl) glucopyranoside showed the strongest capability for scavenging superoxide radical; 4-(4'-hydroxyphenyl)-2-butanone- 4'-O-2-D-(2"-O-galloyl-6"-O-p-coumaroyl) glucopyranoside exhibited the highest reactivity in the lipid peroxidation processes. The use of the analytical screening method based on ultra-performance LC-photodiode array-el-ESI-MS/MS would provide a new way for rapid detection of radical-scavenging natural compounds from radix et rhizoma Rhei or complex matrices.

Expression and localization of activin receptor-interacting protein 2 in mouse tissues.

Activin plays important roles in reproductive tissues as a stimulator of follicle-stimulating hormone (FSH) secretion. Activin receptor-interacting protein 2 (ARIP2) has been recently identified in mouse tissues as a regulatory protein of activin signal transduction. However, the localization and function of ARIP2 are not well characterized. In this study, we found that ARIP2 mRNA and protein were widely expressed in mouse tissues by reverse transcription-PCR (RT-PCR) and Western blotting. The immunoreactivities of ARIP2 were mainly localized at myocardial cells of heart, Leydig cells in testis, macrophages and epithelial cells of bronchus in lung, renal tubule and collecting tubule, pancreatic islet, adrenal gland, adenohypophysis and hypothalamus by immunohistochemical staining. Furthermore, ARIP2 overexpression down-regulated signal transduction induced by activin A in pituitary gonadotroph LbetaT2 cells and inhibited FSH secretion from primary cultured anterior pituitary cells induced by activin A. These findings suggest that ARIP2 is widely distributed in various tissues and may be a negative regulator of activin action in pituitary cells.