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We grew cucumber in pots in greenhouse for 9-successive cropping cycles and analyzed the rhizosphere Pseudomonas spp. community structure and abundance by PCR-denaturing gradient gel electrophoresis and quantitative PCR. Results showed that continuous monocropping changed the cucumber rhizosphere Pseudomonas spp. community. The number of DGGE bands, Shannon-Wiener index and Evenness index decreased during the 3rd cropping and thereafter, increased up to the 7th cropping, however, however, afterwards they decreased again. The abundance of Pseudomonas spp. increased up to the 5th successive cropping and then decreased gradually. These findings indicated that the structure and abundance of Pseudomonas spp. community changed with long-term cucumber monocropping, which might be linked to soil sickness caused by its continuous monocropping.

Millettia speciosa, a traditional Chinese folk medicine, used as immunity enhancer and anti-bronchitis. The study was proposed to investigate phytocomponents and pharmacological activities (antityrosinase, sunscreen and anticancer) of the different fractions of M. speciosa leaves. The sample was extracted with solvents of increasing polarity i.e. petroleum ether (PE), ethyl acetate (EtOAc), methanol (MeOH) and the total flavonoid content for each fraction was determined. The GC-MS analysis revealed various important bioactive compounds. The identified major compounds included beta sitosterol (12.039%), (Z)-9-octadecenamide (6.299%) and gamma sitosterol (4.910%). The highest flavonoid content was recorded for the PE extract i.e. 47.50 ± 0.40%. The PE extract showed significant antityrosinase activity (70.30 ± 0.90 %) with IC50 0.035 mg/ml followed by the MeOH extract i.e. 66.50 ± 1.20% with IC50 0.039 mg/ml. The highest sunscreen activity was noted for the EtOAc extract by observing maximum absorbance value (1.518 ± 0.49) in UVA region followed by 1.214 ± 0.46 in UVB zone. The highest cytotoxicity (29.45 ±1.17% viability) towards MCF-7 breast cancer cells line was noted for the PE extract with IC50 6.73 µg/ml. Overall, our study revealed the presence of potential bioactive compounds for each extract that exhibited significant therapeutic activities and therefore validate the importance of M. speciosa leaves as a source of natural medicine.

Introduction: The root of Millettia speciosa Champ. (Leguminosae) is one of the wellknown traditional Chinese medicines abundant in phenolic compounds and plays important roles in the treatment of pain or numbness of the joints, blood deficiency sallow, chronic bronchitis and chronic hepatitis. Objective: An ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) based chemical profiling approach was established for the separation and characterization of isoflavonoids in the roots of Millettia speciosa Champ. Methods: The roots of Millettia speciosa Champ. were prepared by ethanol extraction followed by further extraction with chloroform. The extracts were analyzed by UPLC-Q-TOF-MS in positive and negative electrospray ion modes at different Collision Energy (CE) values. Results: A total of 48 components were simultaneously detected, of which 38 components, including 21 isoflavones, 4 phenolic acids, 4 isoflavanones, 2 phenolic aldehydes, 2 flavanones, 1 aliphatic acid, 1 chalcone, 1 flavonol, 1 isoflavane and 1 pterocarpan were unambiguously identified or tentatively assigned based on the retention time, UV spectra, characteristic molecular ions, MS/MS fragmentation data and reference standards. The isomers of isoflavonoids were distinguished using accurate mass, the diagnostic fragmentations of C-ring and UV spectra. The major diagnostic fragment ions of isoflavonoids were observed and the corresponding fragmentation pathways were proposed. Conclusion: This investigation provides important analysis data for further quality control, pharmacological and toxicological research on Millettia speciosa Champ.

The root of Millettia speciosa Champ. (MSCP) is used in folk medicine and is popular as a soup ingredient. The root is composed of the rhizome and radix, but only the radix has been used as a food. Thus, it is very important to compare the chemical components and antioxidant activities between the rhizome and radix. The extracts were analyzed by UHPLC-Q-Exactive Orbitrap-MS and multivariate analysis, and the antioxidant activities were evaluated by 2,20-azinobis (3-ethylbenzothiazo-line-6-sulfonic acid) diammonium salt (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays. Ninety-one compounds were detected simultaneously and temporarily identified. Ten compounds were identified as chemical markers to distinguish the rhizome from the radix. The antioxidant activities of the radix were higher than the rhizome. Correlation analysis showed that uvaol-3-caffeate, 3-O-caffeoyloleanolic acid, and khrinone E were the main active markers for antioxidant activity, which allowed for the rapid differentiation of rhizomes and the radix. Therefore, it could be helpful for future exploration of its material base and bioactive mechanism. In addition, it would be considered to be used as a new method for the quality control of M. speciosa.

Ultrasonic-assisted extraction (UAE) and microwave-assisted extraction (MAE) were developed to extract flavonoids antioxidants from the root of Millettia speciosa (Millettia speciosa Champ.). The optimal experimental parameters for antioxidant extraction from M. speciosa Champ. were measured using single-factor experimentation combined with response surface methodology (RSM). Final extraction rate (FER) of flavonoids assay was used to quantify antioxidant compounds. Then, 2,2ˊ-diphenyl-1-picryhydrazyl (DPPH) scavenging capacity, 2,2ˊ-azino-bis(3-ethylbenzothiazoline-6-sulphonicacid) (ABTS) scavenging capacity and ferric reducing antioxidant power (FRAP) were used to determine the antioxidant capacities of extract. Besides, UAE and MAE were compared with conventional Soxhlet extraction (CSE) and M. speciosa Champ. powders were observed by scanning electronic microscopy to analyze the damage of cell structure caused by different extraction methods. The results showed that UAE was the optimal extraction method to extract flavonoids compounds from M. speciosa Champ. and the optimized UAE conditions of flavonoids was as follows: ethanol, 80%, temperature, 40 °C and ultrasonic power, 500 W. This paper provided useful information to utilize the resource of Chinese herbal medicine and manifest the great antioxidant activities of M. speciosa Champ., and the optimized UAE conditions of flavonoids from M. speciosa Champ. could be used in industrial production.

Objective To study the chemical constituents from the roots of Millettia speciosa. Methods The chemical constituents were isolated and purified by silica gel, Sephadex LH-20, and ODS column chromatography. The structures were identified by means of spectral data. Results Fifteen compounds were isolated and identified as naringenin ( 1 ), liquiritigenin ( 2 ), garbanzol ( 3 ), 7-hydroxy-6,4′- dimethoxyisoflavone ( 4 ), calycosin ( 5 ), 2′,5′,7-trihydroxy-4′-methoxyisoflavone ( 6 ), 2′-hydroxybiochanin A ( 7 ), 6-methoxycalopogonium isoflavone A ( 8 ), demethylmedicarpin ( 9 ), 4,4′-dihydroxy-2′-methoxychalcone ( 10 ), 2′,4′-dihydroxy-4-methoxychalcone ( 11 ), rhododendrol ( 12 ), secoisolariciresinol ( 13 ), bisdihydrosiringenin ( 14 ), and polystachyol ( 15 ). Conclusion All compounds are obtained from this plant for the first time.

Two new rotenoids, named millettiaosas A–B (1–2), together with four known compounds were isolated from the roots of Millettia speciosa. Their structures were elucidated on the basis of spectroscopic analysis including 1D and 2D NMR techniques and HRESIMS. Evaluation of the two new compounds for cytotoxicity against four human cancer cell lines (MCF-7, HCT-116, A549 and HepG-2) showed moderate activities (10 μM

In Chinese medicine Suberect Spatholobus Stem is used to treat menoxenia, blood deficiency, numb paralyses, and so on. In folk, Millettia speciosa is often used as a substitute for Suberect Spatholobus Stem in some areas but it has not been reported whether M. speciosa is the eligible substitute for Suberect Spatholobus Stem or not till now. In this study, a capillary zone electrophoretic method was applied to determinate the interactions between natural products isolated from M. speciosa Champ. and thrombin for the first time. Both qualitative and quantitative characterizations of the molecule-enzyme binding were determined. Twenty ingredients were isolated from M. speciosa Champ. and the results showed that compared with positive and negative control, the compounds YT-1, YT-2, YT-3, YT-8, YT-9, YT-10, YT-11, YT-12, YT-14, YT-15, YT-16, and YT-20 interacted with thrombin while the other eight had no binding to thrombin. The binding constants of the interaction between compounds and thrombin were calculated by the Scatchard analysis formula. Because M. speciosa contains these compounds which have different levels of anticoagulant activity, it may be the eligible substitute for Suberect Spatholobus Stem.

Three new phenolic glycosides, millettiaspecosides A--C (1-3), were isolated from the caulis of Millettia speciosa along with three known phenolic glycosides, khaephuoside B (4), seguinoside K (5), albibrissinoside B (6). Their structures were established by spectroscopic methods. The spectral analysis permitted correct assignments for (1)H and (13)C NMR data of compounds 4 and 5.

A new flavonol triglycoside, millettiaspecoside D, was isolated from the caulis of Millettia speciosa Champ. Its structure was elucidated on the basis of spectroscopic analysis.

Purpose: To evaluate whether Millettiae Speciosae Champ. (Leguminosae) can enhance exercise performance as well as ascertain if it a potential functional food material. Methods : The extract of Millettia speciosa Champ. (MSE) was orally administered to mice in 500, 1000, 2000 mg/kg doses to investigate its anti-fatigue effect in both forced swimming and climbing tests. Glycogen, triglyceride (TG), blood urea nitrogen (BUN) and creatine phosphokinase (CK) levels in plasma which can indicate alterations in energy utilization during exercise performance, were determined to analyze the operating exercise mechanisms. Results : The results showed that swimming time to exhaustion was longer in all treated groups (41.06 ± 1.92, 47.84 ± 1.60, 54.00 ± 2.45 min for 500, 10000 and 2000 mg/kg doses, respectively) than for control (19.45 ± 0.62 min, p < 0.05). The middle and high doses of MSE-treated groups significantly prolonged the climbing time compared with control (p < 0.05). Furthermore, MSE reduced the content of TG significantly by increasing fat utilization, delayed the accumulation of BUN and decreased the level of CK (p < 0.05). In addition, administration of MSE significantly protected the depletion of muscle glycogen when compared with control (p < 0.05). Conclusion : The results show for the first time that Millettia speciosa Champ. (Leguminosae) has significant anti-fatigue activity, and also suggest that it is a potential functional food material. Keywords : Radix millettiae speciosae, Anti-fatigue activity, Exercise performance, Serum urea nitrogen, Gastrocnemius muscle glycogen, Triglyceride, Functional food

The 70% ethanol extract of the roots of Millettia speciosa gave two new oleanane-type saponins along with two known pterocarpans, medicarpin and maackiain. The structures of new compounds were established on the basis of chemical and spectroscopic evidences. Medicarpin and maackiain showed cytotoxicity against leukemia cells (HL-60) and inhibited leukotrine secretion from RBL-2H3 cells.