[Clinical therapeutic strategies of refracture after bone transport for tibial bone defect].

OBJECTIVE: To explore the clinical therapeutic strategies of refracture after Ilizarov bone transport technique in the treatment of tibial bone defect. METHODS: A retrospective study was performed on 19 patients with infected tibial bone defect treated by Ilizarov bone transport technique and then refracture from August 2010 to January 2020, including 18 males and 1 female with an average age of (37.7±13.0) years old ranging from 15 to 66 years old. Cause of injury invlved falling injury in 4 cases, crashing injury 1 case, crushing injury in 1 case and without obvious injury history in 13 cases. The ipsilateral distal femoral fracture in 2 cases occurred before the external fixator of tibia was removed, and the other 17 cases had a minimum of 1 day and a maximum of 30 months after the external fixator had been removed. The site of refracture was at regenerative zone in 8 cases, at docking site in 9 cases, at ipsilateral femoral shaft in 2 cases. According to the modified Simpson classification proposed by the author, the refracture was classified. The treatment of refracture include plaster splint, traction or external fixation. Bone healing and function were evaluated according to the standards of the Association for the Study and Application of the Method of Ilizarov(ASAMI). RESULTS: All patients were followed up, and the duration ranged from 9 to 17 months with an average of (12.3±3.2) months. According to the modified Simpson classification, there were 3 cases of type Ⅰa, 1 case of type Ⅰb, 3 cases of type Ⅰc, 1 case of type Ⅱ, 9 cases of type Ⅲ and 2 cases of type Ⅴ. All the refractures healed without infection or malunion. The fracture healing time of conservative treatment for 6 cases were 3, 5, 3, 2, 2, 2 months fespectively;and the healing time of fracture treated by surgery for 13 cases was 2 to 6 months, with an average of(4.4±1.4) months. According to ASAMI evaluation criteria, bony results showed all patients obtained excellent results, and functional results showed 6 patients got excellent results, 13 good beacause of ankle or knee stiffness. CONCLUSION: The modified Simpson classification could contain most clinical types of refracture after bone transport, and the external fixation is a simple and effective method for refracture.

Restless abdomen: a spectrum or a phenotype variant of restless legs syndrome?

BACKGROUND: With the growing awareness of restless legs syndrome (RLS), sensory disorders similar to RLS but initially confined to the arms, abdomen, and perineum have been reported. One of them is restless abdomen, which refers to a restless sensation in abdomen. Our study is designed to evaluate the clinical phenotype of restless abdomen and investigate its relationship with RLS. METHODS: We enrolled 10 patients with restless abdomen according to RLS diagnostic criteria, excluding the requiring of leg involvement. Laboratory examinations were performed to exclude mimics and notable comorbidities. RESULTS: All 10 patients had RLS like symptoms in the abdomen and otherwise satisfied all other RLS diagnostic criteria, and responded to dopaminergic therapy. CONCLUSIONS: Neurologists and gastroenterologists should be aware that RLS-related restlessness can occur in extra-leg anatomy in the absence of episodes of worsening or augmentation of restlessness.

Improving the antioxidant and anti-tyrosinase activities of Stemonae Radix by solid-state fermentation with Mucor circinelloides T2-12.

Stemonae Radix, a medicinal and edible herb, has been reported to possess various pharmacological effects. In the present study, Stemonae Radix was fermented by fungi to improve the antioxidant and anti-tyrosinase activities. The results showed that Stemonae Radix fermented by Mucor circinelloides T2-12 exhibited two-folds more antioxidant and anti-tyrosinase activities than non-fermented material. The increased activity might be ascribed to the improvement of total phenolic content, hydrolyzation of glucosides and esters of phenolics and metabolism of saccharides according to ultraviolet and nuclear paramagnetic resonance spectroscopy. This paper suggested that fermenting Stemonae Radix with M. circinelloides T2-12 is effective to increase antioxidant and anti-tyrosinase effects and Stemonae Radix fermented by M. circinelloides T2-12 might be newly alternative of natural antioxidant and tyrosinase inhibitor. The present study is the first to report that pure strain fermentation processing is effective in improving the antioxidant and anti-tyrosinase activities of Stemonae Radix.

Rynchopeterine F, unique heterodimer of phenethanol-phenylacetyl linking with lactic acid from the medicinal insect Blaps rynchopetera.

A new heterodimer, rynchopeterine F (1), a new natural product, rynchopeterine G (2), and eleven known phenolics were isolated from Blap rynchopetera Fairmaire, a kind of medicinal insect utilized by the Yi and Bai Nationality in Yunnan Province of China. Their structures were established on the basis of extensive spectroscopic analyses (1D and 2D NMR, HR-MS) along with calculated electronic circular dichroism method. Rynchopeterine F was a unusual heterodimer of a 3,4-dihudroxy phenylethanol unit fused to a 3,4-dihudroxy phenylacetyl group through two ester bonds with lactic acid, and rynchopeterine G was a 3,4-dihudroxy phenylethanyl monoester succinate. Attributed to the adjacent dihydroxyl grops, compounds 1 and 2 exhibited significant anti-radical activity with an IC50 value of 3.52 and 7.83 µg/mL for DPPH radical-scavenging, similar with that of the positive controls, vitamin C, 6.92 µg/mL and rutin, 8.28 µg/mL.

Improving the acetylcholinesterase inhibitory effect of Illigera aromatica by fermentation with Clonostachys rogersoniana.

Illigera aromatica was fermented by Clonostachys rogersoniana. The acetylcholinesterase (AChE) inhibitory effects of unfermented and fermented I. aromatica revealed that C. rogersoniana-fermented I. aromatica (CFIA) induced significantly more AChE inhibitory activity (IC50: 35.4 ± 2.1 µg/mL). The biotransformation of actinodaphnine (1) into (4R,6aS)-4-hydroxyactinodaphnine (2) was found during the fermentation, which played an important role in the improvement of the AChE inhibitory activity of I. aromatica. Subsequently, the fermentation conditions-including the solid-liquid ratio, fermentation temperature, and fermentation time-were optimized. I. aromatica immersed in 100-200% water and fermented with C. rogersoniana at ambient temperature for 30 days was conducive to the biotransformation of actinodaphnine (1) and improved the AChE inhibitory activity of I. aromatica. The present study provides a novel approach for improving the pharmacological effect of I. aromatica and suggests that CFIA may be used as an alternative AChE inhibitor.

[Pollution Load Characteristics of Runoff from Urban Roofs of Different Materials].

It is of great significance to analyze the runoff pollution load characteristics of different roof materials to improve the estimation accuracy of urban non-point source pollution loads. Yangzhou City was selected as the study area. There, three types of roofs were chosen for rainfall-runoff monitoring, including a Chinese style tile roof, cement tile roof, and concrete flat roof. The pollutant concentrations, scour law, and first flush effect of the three types of roofs were compared. The results show that the event mean concentration (EMC) of total nitrogen (TN), total phosphorus (TP), permanganate index, and total suspended solids (TSS) in the runoff of Chinese style tile roofs are around 4-9 times that in the runoff of cement tile roofs. The rainfall intensity exhibits stronger effect on the change in pollutant concentrations of runoff from the Chinese style tile roof than that from the cement tile roof. The Pearson correlation coefficients (r) of rainfall intensity against TP and TSS in time series were 0.853 and 0.822, respectively. The first flush intensities of the three types of roof materials were in the order cement tile roof > concrete flat roof > Chinese style tile roof. It was found that 60.0% of the roof runoff pollution load could be reduced by intercepting 31.5%, 58.0%, and 60.4% of the initial runoff for the Chinese style tile roof, the cement tile roof, and the concrete flat roof, respectively. The actual emissions of TN, TP, and TSS, and the permanganate index in rainstorm events would be significantly underestimated when roof materials are not distinguished. This would have negative effects on the pollution control of urban non-point sources. It is demonstrated that the fine distinction of roof materials is able to improve the estimation accuracy of urban non-point source loads.

A new menthane-type monoterpenoid from fermented Illigera aromatica with Clonostachys rogersoniana 828H2.

A new menthane-type monoterpenoid, illigerate E (1), as well as two known ones, (1R*,3R*,4S*,6R*)-6,8-dihydroxymenthol (2) and cis-4-hydroxy-5-(1-hydroxy-1-methylethyl)-2-methyl-2-cyclohexene-1-one (3), were isolated from fermented Illigera aromatica with Clonostachys rogersoniana 828H2. Their structures were identified by HRESIMS and 1D/2D NMR spectra. Their inhibitory effects of NO production in RAW 264.7 macrophages were estimated.

Application of a proton quantitative nuclear magnetic resonance spectroscopy method for the determination of actinodaphnine in Illigera aromatica and Illigera henryi.

Illigera aromatica S. Z. Huang et S. L. Mo and Illigera henryi W. W. Sm., belonging to the genus Illigera (Hernandiaceae), are used as herbal medicines for promoting blood circulation and treating tuberculosis. Actinodaphnine, the major bioactive alkaloid, plays an important role in the quality controls of the herbs. In the present study, a rapid, simple, accurate, and precise proton quantitative nuclear magnetic resonance (1H-qNMR) method was developed to determine the content of actinodaphnine in I. aromatica and I. henryi. DMSO-d6 enabled satisfactory separation of the signals to be integrated in 1H NMR spectrum. 1,4-Dinitrobenzene was selected as an internal standard. The limits of determination and quantitation were 0.005 and 0.038 mg/mL, respectively. This work implied that 1H-qNMR represents a feasible alternative to HPLC-based methods for quantitation of actinodaphnine in I. aromatica and I. henryi and is suitable for the quality control of I. aromatica and I. henryi.

Simultaneous determination of alkaloids dicentrine and sinomenine in Stephania epigeae by 1H NMR spectroscopy.

Stephania epigaea Lo is an important herbal medicine used as antiphlogistic and analgesic drugs. Its major components are dicentrine (1) and sinomenine (2). In the present study, a rapid, accurate, and precise method for simultaneous quantitation of dicentrine (1) and sinomenine (2) in S. epigeae using 1H NMR spectra was developed. The deuterated solvent of DMSO-d6 enabled satisfactory separation of the signals to be integrated in 1H NMR spectrum and dimethyl terephthalate was selected as an internal standard. The feature signals of δ 7.57 and 5.70 were selected for quantifying the dicentrine (1) and sinomenine (2), respectively. Validation of the quantitative method was performed in terms of specificity, accuracy, precision, and stability. This work implied that quantitative 1H NMR represents a feasible alternative to high-performance liquid chromatography-based methods for quantitation of dicentrine (1) and sinomenine (2) in S. epigeae and is suitable for the quality control of S. epigeae.

A novel sesquiterpene derivative with a seven-membered B ring from Illigera aromatica.

A novel sesquiterpene derivative with a seven-membered B ring, illigerine (1), along with four known compounds, 1-epi-chiliophyllin (2), 3,4-dihydroxyphenethyl alcohol (3), coniferyl alcohol (4) and phenylpropionic acid (5), were isolated from Illigera aromatica S. Z. Huang et S. L. Mo. Their structures were identified by 1D/2D NMR, HRESIMS and electronic circular dichroism spectra and the cytotoxic activity and inhibitory effect of NO production in LPS-stimulated RAW264.7 were also evaluated. This is the first report of sesquiterpene isolated from the genus Illigera.

Cytotoxicity of the Defensive Secretion from the Medicinal Insect Blaps rynchopetera.

Blaps rynchopetera Fairmaire has long been used as a folk medicine by the Yi and Bai ethnic groups in China to treat fever, cough, gastritis, boils, and tumors. In the present study, the cytotoxicity of the defensive secretion (TDS) of B. rynchopetera against AGS Caco-2, HepG2 U251 and Bel-7402 was tested, and the results revealed that TDS had potent cytotoxicity against testing cells with IC50 values of 45.8, 17.4, 53.6, 98.4 and 23.4 µg/mL, respectively. Gas chromatography-mass spectrometry (GC-MS) analysis was employed to clarify the cytotoxic constituents in TDS of B. rynchopetera and five volatile compounds, including 2-ethyl-2,5-cyclohexadiene-1,4-dione (3, 31.00%), 1-tridecene (5, 28.02%), 2-methyl-2,5-cyclohexadiene-1,4-dione (2, 22.86%), hydroquinone (4, 1.33%), and p-benzoquinone (1, 1.01%), were identified. Chemical constituent investigation on TDS further supported the presence of 5 above compounds. A cytotoxic assay indicated that compounds 1, 2, 3 and 4 exhibited significant cytotoxicity against the testing cell lines, implying that benzoquinones and hydroquinone played important roles in the cytotoxicity of TDS of B. rynchopetera. TDS is a cytotoxic natural material and further studies investigating mechanisms and inhibitory activities on other cell lines is warranted.

Five New Phenolic Compounds with Antioxidant Activities from the Medicinal Insect Blaps rynchopetera.

Five new phenolic compounds rynchopeterines A-E (1-5), in addition to thirteen known phenolics, were isolated from Blaps rynchopetera Fairmaire, a kind of medicinal insect utilized by the Yi Nationality in Yunnan Province of China. Their structures were established on the basis of extensive spectroscopic analyses (1D and 2D NMR, HR-MS, IR) along with calculated electronic circular dichroism method. Rynchopeterines A-E (1-4) exhibited significant antioxidant activities with IC50 values of 7.67-12.3 µg/mL measured by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. Besides, rynchopeterines B (2) and C (3) showed mild cytotoxicity against tumor cell Caco-2 and A549.

Improving the acetylcholinesterase inhibitory effect of Illigera henryi by solid-state fermentation with Clonostachys rogersoniana.

Illigera henryi, an endemic traditional Chinese medicine, contains abundant aporphine alkaloids that possess various bioactivities. In the present study, tubers of I. henryi were fermented by several fungi, and the acetylcholinesterase (AChE) inhibitory activities of non-fermented and fermented I. henryi were measured. The results showed that the fermentation of I. henryi with Clonostachys rogersoniana 828H2 is effective for improving the AChE inhibitory activity. A key biotransformation was found during the C. rogersoniana fermentation for clarifying the improvement of the AChE inhibitory activity of I. henryi: (S)-actinodaphnine (1) was converted to a new 4-hydroxyaporphine alkaloid (4R,6aS)-4-hydroxyactinodaphnine (2) that possessed a stronger AChE inhibitory activity, with an IC50 value of 17.66±0.06 µM. This paper is the first to report that the pure strain fermentation processing of I. henryi and indicated C. rogersoniana fermentation might be a potential processing method for I. henryi.

A new phenylpropanoid glucoside and a chain compound from the roots of Allium tuberosum.

A new phenylpropanoid glucoside tuberosinine D (1) and a chain compound (Z)-11R,12S,13S-trihydroxy-9-octadecenoate (2) were isolated from the roots of Allium tuberosum. The absolute configuration of 1 was established by comparing of experimental and calculated electronic circular dichroism. The absolute configuration of 2 was determined using the modified Mosher's method for the first time.

Monoterpene esters and aporphine alkaloids from Illigera aromatica with inhibitory effects against cholinesterase and NO production in LPS-stimulated RAW264.7 macrophages.

Three new monoterpene phenylpropionic acid esters, illigerates A-C (1-3), and one new aporphine alkaloid, illigeranine (4), as well as four known ones, actinodaphnine (5), nordicentrine (6), 8-hydroxy carvacrol (7), and 3-hydroxy-α,4-dimethyl styrene (8), were isolated from the tubers of Illigera aromatica. The structures of 1-4 were identified by HRESIMS, 1D and 2D NMR, and electronic circular dichroism spectra. Compound 1 potently inhibited NO production in LPS-stimulated RAW264.7 cells with an IC50 value of 18.71 ± 0.85 µM; compound 1, 3, and 4 showed moderate butyrylcholinesterase inhibitory activities with the IC50 values of 46.86 ± 0.65, 53.51 ± 0.71, and 31.62 ± 1.15 µM, respectively. Compound 4 showed weak AChE inhibitory activity with an IC50 value of 81.69 ± 2.07 µM, and compounds 5 and 6 possessed moderate AChE inhibitory activities with the IC50 values of 47.74 ± 1.66 and 40.28 ± 2.73 µM, respectively. This paper provides a chemical structure and bioactive foundation for using I. aromatica as an herbal medicine.

Production of a new tetracyclic triterpene sulfate metabolite sambacide by solid-state cultivated Fusarium sambucinum B10.2 using potato as substrate.

The aim of this work is to explore integracide analogues from secondary metabolites of microorganisms. A new tetracyclic triterpene sulfate was produced by solid-state fermentation (SSF) with Fusarium sambucinum B10.2. The tetracyclic triterpene sulfate was identified as (3S,5R,10S,11S,12S,13R,17R,20R)-4,4-dimethylergosta-8,14,24-triene-3,11,12-triol-12-acetate, 3-sulfate on the basis of HRESIMS, NMR and electronic circular dichroism (ECD) spectra and named sambacide (1). The antibacterial and antifungal assays of sambacide (1) showed significant antibacterial activities against Staphylococcus aureus and Escherichia coli. The fermentation conditions including culture media, fermentation temperature and time, were optimized. And potato was selected as the fermentation substrate, 28°C was used as the fermentation temperature, and 20-days fermentation time was determined for F. sambucinum-SSF to produce sambacide (1) with a high yield of 19.04±0.82g/kg. This paper provides an efficient approach to produce the antibacterial and antifungal agent sambacide (1) in a very high yield.

A new flavone C-glycoside and a new bibenzyl from Bulbophyllum retusiusculum.

A new flavone C-glycoside, apigenin 6-C-α-arabinofuranosyl 8-C-α-arabinopyranoside (1) and a new bibenzyl, bulbotetusine (2), were isolated from the tubers of Bulbophyllum retusiusculum. Their structures were established on the basis of extensive spectroscopic analyses. The absolute configuration of 2 was determined by the comparison of experimental and calculated electronic circular dichroism. Compounds 1 and 2 showed no obvious cytotoxic activity against any five human tumour cell lines with IC50 values >40 µM.

Two new peroxy fatty acids with antibacterial activity from Ophioglossum thermale Kom.

Two new peroxy fatty acids, thermalic acids A (1) and B (2), together with eight known compounds, (3ß)-methyl-3-hydroxy-urs-11-en-28 oate (3), luteolin (4), quercetin (5), 3-methoxyquercetin (6), ophioglonol (7), ophioglonol 4'-O-α-D-glucopyranoside (8), pedunculosumoside B (9), syringol (10), were isolated from the herba of Ophioglossum thermale Kom. The structures of 1 and 2 were identified by HRESIMS, EIMS, 1D and 2D NMR, and electronic circular dichroism (ECD) spectra. Both two acids exhibited potential antibacterial activities against Staphylococcus aureus, Bacillus subtilis, and Escherichia coli. This is the first report of peroxy fatty acids isolated from herbaceous plants of Ophioglossaceae.

Innovative Approach to the Accumulation of Rubrosterone by Fermentation of Asparagus filicinus with Fusarium oxysporum.

Rubrosterone, possessing various remarkable bioactivities, is an insect-molting C19-steroid. However, only very small amounts are available for biological tests due to its limited content from plant sources. Fungi of genus Fusarium have been reported to have the ability to convert C27-steroids into C19-steroids. In this study, Asparagus filicinus, containing a high content of 20-hydroxyecdysone, was utilized to accumulate rubrosterone through solid fermentation by Fusarium oxysporum. The results showed that F. oxysporum had the ability to facilitate the complete biotransformation of 20-hydroxyecdysone to rubrosterone by solid-state fermentation. The present method could be an innovative and efficient approach to accumulate rubrosterone with an outstanding conversion ratio.

Proaporphine and aporphine alkaloids with acetylcholinesterase inhibitory activity from Stephania epigaea.

An unusual proaporphine alkaloid bearing an isopropanenitrile group at isoquinoline nitrogen, named epiganine A (1) and a new aporphine alkaloid, epiganine B (2), together with eight known alkaloids, pronuciferine (3), dehydrodicentrine (4), romerine (5), romeline (6), N-methylcalycinine (7), phanostenine (8), dicentrine (9), and N-methyllaurotetanine (10), were isolated from the roots of Stephania epigaea. The absolute configuration of 1 was determined by calculating electronic circular dichroism (ECD) and comparing with experimental data. Compounds 2 and 4 showed strong acetylcholinesterase (AChE) inhibitory effects with the IC50 values of 4.36 and 2.98µM, respectively. Compounds 5-9 also exhibited potent AChE inhibitory activities.