High-performance liquid chromatography analysis of alkaloids in various parts of lotus extracts with ion mobility spectrometry and mass spectrometry dual detection.

Using high-performance liquid chromatography coupled with electrospray ionization-ion mobility spectrometry and mass spectrometry, we proposed a dual-detection method for the identification and profiling of alkaloids in various lotus parts including leaf, plumule, stem, seed epicarp, and receptacle. The eluent from high-performance liquid chromatography was split and conducted to electrospray ionization-ion mobility spectrometry and time-of-flight mass spectrometry separately to facilitate the compound identification. In total, 23 kinds of alkaloids were identified based on m/z, drift time, and retention time, including alkaloid isomers such as lirinidine, N-nornuciferine, and O-nornuciferine with identical m/z that are difficult to differentiate using mass spectrometry alone. Using this method, we investigated the changing dynamics of alkaloid accumulation in lotus leaves and lotus stems at different harvesting periods. The total alkaloid content showed an increasing trend with the growth and development of leave and stem. Overall, the developed dual detection method has the advantages of high peak capacity and high sensitivity compared with the conventional detection method and facilitates the identification of detected compounds.

Evaluation of Fourier deconvolution ion mobility spectrometer as high-performance gas chromatography detector for the analysis of plant extract flavors.

The Fourier deconvolution ion mobility spectrometer (FDIMS) offers multiplexing and improves the resolving power and signal-to-noise ratio. To evaluate the FDIMS as a detector for gas chromatography for the analysis of complex samples, we connected a drift tube ion mobility spectrometer to a commercial gas chromatograph and compared the performance including resolving power, sensitivity, and linear range using 2,6-di­tert-butylpyridine. Mixed standards were also injected into the tandem system to evaluate the performance under optimized conditions. A complex plant extract sample used as natural flavoring was investigated using the resulting system. The results show that the instrument implemented with the Fourier deconvolution multiplexing method demonstrated higher performance over the traditional signal averaging method including higher resolving power, better limit of detection, and wider linear range for a variety of compounds and natural plant extract flavorings.

Performance of wetland applied to ecological remediation of abandoned fish ponds: A case study in Gonghu Bay, Tai lake.

To restore the abandoned fish ponds to "near natural" state, the wetland restoration was carried out in Gonghu Bay lakeside, and its long-term performance of controlling external load was studied for 5 years. The findings showed that water quality and biodiversity had been improved dramatically after the preliminary transformation. The concentrations of permanganate index (CODMn), total nitrogen (TN), and total phosphorus (TP) obviously decreased from 12.91 mg L-1 to 4.32 mg L-1, from 3.46 mg L-1 to 1.42 mg L-1, and from 0.27 mg L-1 to 0.04 mg L-1, respectively. The proportion of Cyanophyta was effectively reduced from 31.82% to 18.89%, and favored the growth of diatoms (31.82%-37.78%) to be the dominant algae species. Aquatic plant species and coverage gradually increased from 16 to 56 and from 5% to 60%, respectively. An in-deep monitoring done for 5 years (2013-2017) showed that the wetland achieved a satisfactory removal efficiency of 58.95% for TN, 64.60% for TP, and up to 77.83% for chlorophyll-a. Besides, three pollution scenarios, such as stormwater runoff, algal bloom, and continuous water transfer, were selected to explore the tolerance of the wetland to the suddenly increased pollution loads. The results dedicated that even if the inlet load was up to 1.0 × 105 m3 d-1, the removal rate coefficients of wetland for chlorophyll-a, TP, and TN were 0.135-0.239 d-1, 0.041-0.112 d-1, and 0.030-0.109 d-1, respectively, which were equivalent to the well-running wetlands. This study confirmed that the wetland was not only a promising ecological remediation technique to contaminated abandoned fish ponds, but also could withstand high pollution load, which had the prospect of sustainable utilization.

The application of steel slag in a multistage pond constructed wetland to purify low-phosphorus polluted river water.

To investigate the effect of a constructed wetland (CW) with steel slag as the filler on water contaminated by low phosphorus levels, a multistage pond CW system was designed in this study. Low-phosphorus polluted river water was used as the research object. This study explored the effects of using steel slag as a CW filler on phosphorus removal and the total phosphorus (TP) purification effect of the wetland system. The results showed that the TP removal rates in the ecological pond, oxidation pond, surface flow wetlands and submerged plant pond were 5.17%, 8.02%, 21.56%, and 16.31%, respectively. Intermittent increases in phosphorus concentration were observed in the reactors and were caused by the decay of plant tissues, which released pollutants. Because steel slag was added to the filler, the TP concentrations in the effluent of the first- and second-level horizontal subsurface CWs increased by 151.13% and 16.29%, respectively, compared to the influent concentration. The 20th to 40th days of the test run was a period of rapid phosphorus release of the system. The use of steel slag has a potential risk of phosphorus release when applied in CWs used to purify low-phosphorus contaminated water bodies.

CTGF induces tenogenic differentiation and proliferation of adipose-derived stromal cells.

Intrasynovial tendons are paucicellular and hypovascular, resulting in a poor response to injury. Surgical repair of ruptured or lacerated tendons often lead to complications such as adhesions, repair site gapping, and repair site rupture. Adipose-derived stem cells (ASCs) have shown promise for enhancing tendon repair, as they have the capacity to differentiate into tendon fibroblasts and augment the healing response. Furthermore, connective tissue growth factor (CTGF) has been shown to promote tendon regeneration via the stimulation of endogenous tendon stem cells. Here, we evaluated the potential of CTGF to promote tenogenic differentiation of ASCs in vitro. Gene and protein expression, cell proliferation, and FAK and ERK1/2 signaling were assessed. CTGF increased tenogenic genes in mouse ASCs in a dose- and time-dependent manner. Western blot and immunostaining analyses demonstrated increases in tenogenic protein expression in CTGF-treated ASCs at all timepoints studied. CTGF increased ASC proliferation in a dose-dependent manner. CTGF induced phosphorylation of ERK1/2 within 5 min and FAK within 15 min; both signals persisted for 120 min. Blocking FAK and ERK1/2 pathways by selective inhibitors SCH772984 and PF573228, respectively, attenuated the CTGF-induced tenogenic differentiation and proliferation of ASCs. These results suggest that CTGF induces tenogenic differentiation of ASCs via the FAK and ERK1/2 pathway. Statement of clinical significance: Although prior research has led to advances in tendon operative techniques and rehabilitation methods, clinical outcomes after tendon repair remain variable, with high rates of repair site gapping or rupture. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.

Synchrotron radiation micro-CT as a novel tool to evaluate the effect of agomir-210 in a rat spinal cord injury model.

MicroRNA-210 (miR-210) was initially reported to be associated with hypoxia and plays a vital role in modulating angiogenesis. However, the potential effect and underlying mechanisms of miR-210 activity in rat spinal cord injury (SCI) have not yet been fully illuminated. In the present study, differential microRNA expression after SCI was determined by Microarray analysis. To explore the effect of miR-210 after SCI, we intrathecally injected agomir-210 with Alzet Osmotic Pumps to up-regulated the endogenous miR-210 expression. Then, synchrotron radiation micro-CT (SRµCT) imaging was used to investigate the effect of agomir-210 in rat SCI model. We found that the endogenous miR-210 expression could be up-regulated by intrathecal agomir-210 injection. The administration of agomir-210 significantly promoted angiogenesis, as evidenced by increased vessel number and volume detected by SRµCT, attenuated the lesion size and improved functional recovery after SCI. Additionally, agomir-210 attenuated cellular apoptosis and inflammation in the injured rat spinal cord. Expression levels of pro-apoptotic protein (Bax) and pro-inflammatory cytokines (TNF-α and IL-1ß) were significantly decreased after agomir-210 treatment, whereas expression levels of anti-apoptotic (Bcl-2) and anti-inflammatory (IL-10) proteins were up-regulated. In conclusion, our results indicated that SRµCT is a powerful imaging tool to evaluate the effects of angiogenesis after agomir-210 administration in rat SCI model. The up-regulation of endogenous miR-210 expression following agomir-210 administration promoted angiogenesis and anti-apoptotic protein expression, and attenuated inflammation. MiR-210 played a positive role in neurological functional recovery and could be a potential new therapeutic target for SCI.

Enhanced patella-patellar tendon healing using combined magnetic fields in a rabbit model.

BACKGROUND: A combined magnetic field (CMF) is a composite of a dynamic sinusoidal magnetic field and a magnetostatic field. Stimuli from CMFs has proved to be an effective tool for healing problem fractures and spinal fusion procedures. HYPOTHESIS: Combined magnetic field technology will enhance healing of bone-tendon junction repair via endochondral ossification for regeneration of the fibrocartilage zone. STUDY DESIGN: Controlled laboratory study. METHODS: Forty-eight mature rabbits were randomly divided into CMF-treated and placebo-treated (control) groups. A partial patellectomy model was created. The CMF-treated group was subjected to CMF stimulation from the third postoperative day for 30 minutes per day up to weeks 8 or 16. At each time point, tissue samples were harvested and evaluated biomechanically and histomorphologically. The area of newly formed bone and the thickness of fibrocartilage were measured in hematoxylin and eosin-stained sections and toluidine blue-stained sections, respectively, while the density of fibrocartilage cells and the amount of proteoglycans were calculated using safranin O-stained sections. A biomechanical analysis was carried out to ascertain tensile strength. RESULTS: Quantitative histological measurements showed that the newly formed bone and regenerated fibrocartilage zone in the CMF-treated group increased by a respective 99.2% and 41.9% compared with the control group at week 8 and a respective 97.8% and 22.8% at week 16. In the CMF-treated group at postoperative week 16, the amount of proteoglycans was 36.9% more than that of the control group, but the density of fibrocartilage cells was just 71.4% of the control group; there were no significant differences at week 8. Mechanical test results showed that energy to failure was not significantly different between the 2 groups at week 8. Yet, at week 16, load to failure, ultimate strength, and energy to failure in the CMF-treated group (311.0 ± 59.4 N, 8.46 ± 1.41 MPa, and 0.87 ± 0.17 J, respectively) were significantly higher than those in the control group (247.1 ± 65.6 N, 6.84 ± 1.12 MPa, and 0.52 ± 0.15 J, respectively). CONCLUSION: Biophysical stimulation with CMFs enhances healing after bone-tendon junction injuries in a rabbit model. CLINICAL RELEVANCE: These results demonstrate the feasibility of using CMFs for stimulating bone-tendon healing after repair.

Tetramethylpyrazine accelerates the function recovery of traumatic spinal cord in rat model by attenuating inflammation.

In the present study, we explored the effects of tetramethylpyrazine (TMP), an alkaloid extracted from the Chinese herbal medicine Ligusticum wallichii Franchat (chuanxiong), on a rat model of contusion spinal cord injury (SCI). The contusion SCI model was induced in rats by a modified Allen's weight-drop method with a severity of 5 g × 50 mm impacting on the T10 segment. In the TMP treatment group, rats were injected intraperitoneally (i.p.) with TMP (200mg/kg), every 24h for 5 days, starting half an hour after contusion SCI. The control group was treated with saline. Compared with the control group, the TMP group significantly ameliorated the recovery of hindlimb function of rats. TMP treatment significantly reduced the expression of macrophage migration inhibitory factor, nuclear factor κappa B, pro-inflammatory cytokine interleukin-18 and neutrophil infiltration. On the other hand, TMP enhanced the expression of inhibitor κappa B and anti-inflammation cytokine interleukin-10. In conclusion, our results demonstrate that TMP inhibits the development of inflammation and tissue injury associated with spinal cord contusion in rats which may improve the rats' hindlimb function.

Three-dimensional alteration of microvasculature in a rat model of traumatic spinal cord injury.

Acute spinal cord injury (SCI) always leads to severe destruction of the microvascular networks. To investigate the three-dimensional (3D) alterations of microvasculature following SCI, we utilized an established rat SCI model. Based on the hypothesis that the spinal cord would undergo reorganization and postinjury modification of the vascular networks after SCI, we reconstructed the normal and injured angioarchitecture using micro-CT images of silicone rubber microsphere-perfused specimens. Several morphometric parameters were used to study the 3D vascular alterations in the SCI rat model, including the casting-based vessel volume fraction, connectivity density, separation, thickness and thickness distribution. Our results indicated that the microvascular spatial conformations were significantly different between the normal and injured spinal cord segments. The morphometric changes showed an increase of the vessel volume fraction and separation and a decrease of vessel connectivity density during the vascular healing process after SCI. Our results may contribute to elucidation of the mechanisms of compensatory vascular reconstitution in traumatized spinal cord. The method used here has the potential to improve our understanding of changes in the spatial architecture of vascular networks after SCI compared to the conventional histomorphology techniques. In summary, we developed a new methodology to analyze neurovascular pathology based on 3D vascular network patterns and features in an experimental rat SCI model. This technique could be used as a complementary tool to investigate the efficacy and side effects of therapeutic drugs or rehabilitation regimens.