The Role for GABAergic Interneurons Subtypes in the Antidepressant Efficacy of Traditional Chinese Medicine Formula Lily Bulb and Rehmannia Decoction

Background Depression is associated with reduced γ-aminobutyric acid (GABA) and alterations of GABAergic interneurons in medial prefrontal cortex (mPFC), which can be normalized by traditional Chinese medicine formula lily bulb and Rehmannia decoction (LBRD) standard decoction. However, the impact of GABAergic interneurons subtypes including parvalbumin (PV), somatostatin (SST), and vasoactive intestinal peptide (VIP), in depressive episodes as well as in the LBRD standard decoction antidepressant effects remains to be elucidated. Methods We assessed the effects of LBRD standard decoction intervention on the expression of GABA associated miRNA/mRNA and GABAergic subtype-specific markers in chronic stress induced depression by western blot and qRT-PCR. The lncRNA/miRNA/GABA regulatory axis was verified by luciferase reporter assay, RNA immunoprecipitation, RNA pull-down assay and then detect its changes in LBRD standard decoction administration with use of immunofluorescence staining and RNA-fluorescence in situ hybridization (FISH). Results In the current study, we found that LBRD standard decoction exerted antidepressant effects based on the results of behavioral tests. Meanwhile, LBRD also revised the reduced GABA levels in depression by increasing the expression of lncRNA Neat1 and Malat1, as well as decreasing miRNA-144-3p and miRNA-15b-5p. Moreover, the expression of SST mRNA and protein in the mPFC tissues from depression group, was significantly lower than those in the control cases, while LBRD standard decoction treatment revised these changes. Whereas, neither chronic stress nor treatment can obviously change the level of PV and VIP mRNA and protein expression. In the SST-positive neurons of mPFC tissues, treatment with LBRD standard decoction resulted in the elevation of Gad-67, VGAT, GAT-3 and an reduction of miRNA-144-3p expression. Conclusion Taken together, these findings suggest that SST interneurons may be served as a preferentially vulnerable GABAergic neuronal subtype in depression and LBRD standard decoction antidepressant activities potentially relate to against the SST cells deficits by regulating the miRNA-144-3p mediated GABA synthesis, release.

Calibration and Verification Test of Lily Bulb Simulation Parameters Based on Discrete Element Method

In the simulation analysis of the lily harvesting process, the intrinsic parameters of the lily bulb and the contact parameters between the lily bulb and the lily mechanized harvesting equipment (Q235 steel) are deficient. Thus, the three-axis size, density, moisture content, Poisson’s ratio, elastic modulus, and other parameters of lily bulbs are measured in this paper with lily bulbs as the research object. Moreover, the discrete element model of the lily bulb was established using 3D scanning technology. The contact parameters between the lily bulb and Q235 steel were calibrated through bench test and simulation parameter test. The relative error between the measured value of the lily bulb accumulation angle and the simulated value is taken as response value to calibrate three parameters (collision recovery coefficient, static friction coefficient, and dynamic friction coefficient between lily bulbs). A regression model of the relative error of the stacking angle and three parameters is established, and the response surface is optimized. The results demonstrate that collision recovery coefficient, static friction coefficient, and dynamic friction coefficient between lily bulb and Q235 steel are 0.301, 0.423, and 0.063, respectively; these coefficients between lily bulbs are 0.455, 0.425, and 0.158, respectively. Additionally, a better combination of parameters is adopted to perform the simulation stacking test. The measured stacking angle is 32.31°, which is 0.34% in error with the stacking angle obtained by the physical stacking test. The test results suggest that the discrete element model and contact parameters of the lily bulb can be used in the discrete element simulation test. Furthermore, these research results could provide references for simulation tests, such as mechanized harvesting and post-harvest processing, of lily.

Integrated analysis of the chemical-material basis and molecular mechanisms for the classic herbal formula of Lily Bulb and Rehmannia Decoction in alleviating depression

Background Lily Bulb and Rehmannia Decoction (LBRD), is a traditional Chinese formula that has been shown to be safe and effective against depression; however, its material basis and pharmacological mechanisms remain unknown. Methods Here, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) and high-performance liquid chromatography (HPLC) were used to identify the chemical spectrum and qualitatively identify the major active ingredients in the LBRD standard decoction, respectively. Subsequently, we assessed the behavior, neuronal function and morphology, neurotransmitter levels, hypothalamic–pituitary–adrenal (HPA)-axis associated hormones, inflammatory cytokine levels, and miRNA/mRNA expression alterations in an in vitro/vivo depression model treated by the LBRD standard decoction. Finally, miRNA/mRNA regulatory networks were created through bioinformatics analysis, followed by functional experiments to verify its role in LBRD standard decoction treatment. Results A total of 32 prototype compounds were identified in the LBRD standard decoction, and the average quality of verbascoside in the fresh lily bulb decoction, fresh raw Rehmannia juice, and the LBRD standard decoction were 0.001264%, 0.002767%, and 0.009046% (w/w), respectively. Administration of the LBRD standard decoction ameliorated chronic unpredictable mild stress (CUMS)-induced depression-like phenotypes and protected PC12 cells against chronic corticosterone (CORT)-induced injury. The levels of neurotransmitter, cytokine, stress hormones and neuronal morphology were disrupted in the depression model, while LBRD standard decoction could work on these alterations. After LBRD standard decoction administration, four differentially expressed miRNAs, rno-miR-144-3p, rno-miR-495, rno-miR-34c-5p, and rno-miR-24-3p, and six differentially expressed mRNAs, Calml4, Ntrk2, VGAT, Gad1, Nr1d1, and Bdnf overlapped in the in vivo/vitro depression model. Among them, miR-144-3p directly mediated GABA synthesis and release by targeting Gad1 and VGAT, and miR-495 negatively regulated BDNF expression. The LBRD standard decoction can reverse the above miRNA/mRNA network-mediated GABA and BDNF expression in the in vivo/vitro depression model. Conclusion Collectively, the multi-components of the LBRD standard decoction altered a series of miRNAs in depression through mediating GABAergic synapse, circadian rhythm, and neurotrophic signaling pathway etc., thereby abolishing inhibitory/excitatory neurotransmitter deficits, recovering the pro-/anti-inflammatory cytokine levels and regulating the HPA-axis hormone secretion to achieve balance of the physiological function of the whole body.

Design and Operating Parameter Optimization of a Harvester for Efficient Lily Bulb Cultivation

Highlights An urgent need exists to mechanize lily harvesting to meet growing demand. A new lily bulb harvester suited to the agronomy of lily planting sites in China was designed. Operation parameters were optimized and validated via field tests. Bulb loss and stem damage rates of 2.31% and 1.82%, respectively, were achieved. Abstract . At present, lily bulbs are harvested primarily by manual excavation in most countries, whereas harvesting machinery is used for other tuber and rhizome crops. However, this approach results in several problems, such as high manual excavation costs, low efficiency, and incomplete separation of soil and lily bulbs. To address these problems, we propose a harvesting machine designed specifically for lily harvesting. Our design process involved measuring and analyzing the physical and mechanical properties of soil and lily bulbs during the harvest period, designing key mechanical components through theoretical calculations, and performing a single-factor simulation of the harvesting process to determine the test value range of each factor. We considered the bulb loss and stem damage rates as evaluation indicators and the machine forward speed, elevator linear speed, and elevator inclination as test factors. We conducted a three-factor, three-level rotation combined test, and the working parameter values were optimized according to the regression analysis results. The test results show that for a forward speed, linear elevator speed, and elevator inclination of 1.4 m/s, 1.8 m/s, and 25°, respectively, the bulb loss and stem damage rates were limited to 2.31% and 1.82%, respectively, thus maximizing the quality and yield of the harvest. Field experiment results showed that the optimized results were consistent with the experimental results and met the requirements of lily harvesting. Keywords: Harvester, Rhizome harvesting, Lily cultivation, Lily bulb properties, Soil parameters, Simulation analysis.