Elucidation of the mechanism of the Yinhua Miyanling Tablet against urinary tract infection based on a combined strategy of network pharmacology, multi-omics and molecular biology.

ETHNOPHARMACOLOGICAL RELEVANCE: Yinhua Miyanling Tablet (YMT), a traditional Chinese medicine consisting of 10 herbs, has been widely used clinically to treat urinary tract infections (UTIs), however, its therapeutic mechanism is not fully understood. AIM OF THE STUDY: To investigate the mechanism of YMT in treating UTIs through network pharmacology, multi-omics and experimental validation. MATERIALS AND METHODS: Clinically, blood and urine samples from YMT-treated UTI patients were collected for transcriptomic and metabolomic analyses. Computationally, compounds that are related to YMT were obtained from the databases, relevant targets were identified, and UTI-related targets were analyzed to determine the core signaling pathways. Subsequently, an integrated approach combining multi-omics and network pharmacology assisted in identifying the key pathways underlying therapeutic effects of YMT on UTI. Finally, a mouse model of UTI was established using uropathogenic Escherichia coli (UPEC), and the therapeutic mechanism of YMT on UTI was validated by ELISA, qRT-PCR and Western blotting. RESULTS: After taking YMT, patients showed reduced levels of urinary bacteria, white blood cells, and serum inflammatory factors (CRP, IL-6 and TNF-α). Multi-omics analysis combined with network pharmacology demonstrated that YMT significantly inhibited the TLR/MAPK/NFκB signaling pathway. In vivo experiments confirmed that YMT attenuated UPEC-induced pathological changes in bladder structural, reduced the expression of bladder proteins (TLR4, MyD88, p-p38 MAPK and p-p65 NFκB), increased protein expression of IκB-α, and attenuated the release of inflammatory factors (TNF-α, IL-6 and IL-1ß) in mice. CONCLUSION: YMT is effective in treating UTI by down-regulating the TLR4/p38MAPK/p65NFκB pathway, thereby providing a scientific basis for its clinical application.

Harnessing a Novel Machine-Learning-Assisted Evolutionary Algorithm to Co-optimize Three Characteristics of an Electrospun Oil Sorbent.

The optimization of materials is challenging as it often involves simultaneous manipulation of an assembly of condition parameters, which generates an enormous combinational space. Thus, optimization models and algorithms are widely adopted to accelerate material design and optimization. However, most optimization strategies can poorly handle multiple parameters simultaneously with limited prior knowledge. Herein, we describe a novel systematic optimization strategy, namely, machine-learning-assisted differential evolution, which combines machine learning and the evolutionary algorithm together, for zero-prior-data, rapid, and simultaneous optimization of multiple objectives. The strategy enables the evolutionary algorithm to "learn" so as to accelerate the optimization process, and also to identify quantitative interactions between the condition parameters and functional characteristics of the material. The performance of the strategy is verified by in silico simulations, as well as an application on simultaneously optimizing three characteristics, namely, water contact angle, oil absorption capacity, and mechanical strength, of an electrospun polystyrene/polyacrylonitrile (PS/PAN) material as a potential sorbent for a marine oil spill. With only 50 tests, the optimal fabrication parameters were successfully located from a combinatorial space of 50 000 possibilities. The presented platform technique offers a universal enabling technology to identify the optimal conditions rapidly from a daunting parameter space to synthesize materials with multiple desired functionalities.

Chemical Profiling of Xueshuan Xinmaining Tablet by HPLC and UPLC-ESI-Q-TOF/MS.

Xueshuan Xinmaining Tablet (XXT) is a widely used traditional Chinese medicine for the treatment of stroke, chest pain, coronary heart disease, and angina pectoris caused by blood stasis. Having a multiple-component preparation, it is still far from meeting the requirements of modernization and standardization because its detailed chemical basis and action mechanism have not been clarified. In this work, the different batches of XXT samples were analyzed by HPLC and the typical sample was analyzed by UPLC-ESI-Q-TOF/MS to understand its chemical profiling. As a result, 77 chromatographic peaks were detected, among which 63 constituents were identified or tentatively characterized based on the comparison of retention time and UV spectra with authentic compounds as well as by summarized MS fragmentation rules and matching of empirical molecular formula with those of published components. This is the first systematic report on the chemical profiling of the commercial XXT products, which provides the sufficiently chemical evidence for the global quality evaluation of XXT products.

Feedback System Control Optimized Electrospinning for Fabrication of an Excellent Superhydrophobic Surface.

Superhydrophobic surface, as a promising micro/nano material, has tremendous applications in biological and artificial investigations. The electrohydrodynamics (EHD) technique is a versatile and effective method for fabricating micro- to nanoscale fibers and particles from a variety of materials. A combination of critical parameters, such as mass fraction, ratio of N, N-Dimethylformamide (DMF) to Tetrahydrofuran (THF), inner diameter of needle, feed rate, receiving distance, applied voltage as well as temperature, during electrospinning process, to determine the morphology of the electrospun membranes, which in turn determines the superhydrophobic property of the membrane. In this study, we applied a recently developed feedback system control (FSC) scheme for rapid identification of the optimal combination of these controllable parameters to fabricate superhydrophobic surface by one-step electrospinning method without any further modification. Within five rounds of experiments by testing totally forty-six data points, FSC scheme successfully identified an optimal parameter combination that generated electrospun membranes with a static water contact angle of 160 degrees or larger. Scanning electron microscope (SEM) imaging indicates that the FSC optimized surface attains unique morphology. The optimized setup introduced here therefore serves as a one-step, straightforward, and economic approach to fabricate superhydrophobic surface with electrospinning approach.

Protective Effect of Yinhua Miyanling Tablet on Lipopolysaccharide-Induced Inflammation through Suppression of NLRP3/Caspase-1 Inflammasome in Human Peripheral Blood Mononuclear Cells.

Yinhua Miyanling Tablet (YMT), the Chinese formula, has long been administrated in clinical practice for the treatment of acute pyelonephritis and acute urocystitis. In the current study, we aimed to investigate the anti-inflammatory effect of YMT in vitro and to evaluate the association between anti-inflammation and innate immune response. Human peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll density gradient centrifugation and then were stimulated by Lipopolysaccharide (LPS). The differential gene expression of inflammation-related genes after drug administration was assessed using PCR array, and the protein levels of differential genes were measured by ELISA and Western blot. The result showed that YMT significantly inhibited the expression of NLRP3, Caspase-1, and the downstream cytokine IL-1ß and suppressed the production of inflammatory mediators TNF-α, IL-6, IL-10, and MCP-1 in a dose-dependent manner compared to the LPS group (P < 0.01). The finding indicated that YMT exhibited anti-inflammatory effect in vitro by suppressing the NLRP3/Caspase-1 inflammasome, and that may have therapeutic potential for the treatment of inflammatory diseases.

The Activation of Nrf2 and Its Downstream Regulated Genes Mediates the Antioxidative Activities of Xueshuan Xinmaining Tablet in Human Umbilical Vein Endothelial Cells.

Epidemiological studies have verified the critical role that antioxidative stress plays in protecting vascular endothelial cells. The aims of the present study were to investigate the antioxidative activities and differential regulation of nuclear erythroid-related factor 2- (Nrf2-) mediated gene expression by Xueshuan Xinmaining Tablet (XXT), a traditional Chinese medicine with the effect of treating cardiovascular diseases. The antioxidative activities of XXT were investigated using quantitative real-time PCR (qPCR), a PCR array, and western blotting. Our results indicated that XXT exhibited potent antioxidative activities by suppressing the levels of hydrogen peroxide- (H2O2-) induced reactive oxygen species (ROS) in human umbilical vein endothelial cells (HUVECs). We were also conscious of strong Nrf2-mediated antioxidant induction. XXT enhanced the expressions of Keap1, Nrf2, and Nrf2-mediated genes, such as glutamate-cysteine ligase modifier subunit (GCLM), NAD(P)H: quinine oxidoreductase 1 (NQO1), heme oxygenase 1 (HMOX1), and glutathione peroxidase (GPX) in HUVECs. In summary, XXT strongly activated Nrf2 and its downstream regulated genes, which may contribute to the antioxidative and vascular endothelial cell protective activities of XXT.

[Leak subtraction analysis of Axon patch clamp system].

AIM AND METHODS: Using blind whole-cell recording techniques on rat hippocampal slices, the function and mechanisms of several leak subtraction methods of Axon patch clamp system (Axopatch amplifier and pClamp software) were analyzed. That how to select and use scaled P/N leak subtraction, patch clamp amplifier leak subtraction and Clampfit leak subtraction were emphasized in our present study. RESULTS: The noise induced by scaled P/N leak subtraction of Clampex soft ware was smaller than that of P/N leak subtraction. Axon patch clamp amplifier leak subtraction could subtract the leak current produced by single depolarizing pulse but not the leak current produced by a series of different step depolarizing pulses. Due to its assumption that leak current would be produced if only potential difference arises across membrane, Clampfit leak subtraction was not suitable to subtracting the steady-state leak current while recording voltage-gated channel currents. CONCLUSION: P/N- and scaled P/N leak subtraction, but not Clampfit leak subtraction, cad be used to subtract steady-state leak current while recording voltage-gated channel currents.

[Difference in action sites between mecamylamine and hexamethonium on nicotinic receptors of sympathetic neurons].

To compare the difference in action sites between mecamylamine (MEC) and hexamethonium (HEX) on nicotinic receptors of sympathetic neurons, we investigated the effects of MEC and HEX on the nicotine-induced currents in cultured superior cervical ganglion neurons by whole-cell patch clamp technique. The IC(50) of MEC and HEX for antagonizing the effect of 0.08 mmol/L nicotine was 0.0012 and 0.0095 mmol/L, respectively. Both MEC and HEX accelerated the desensitization of nicotinic receptors. Furthermore, by comparing their effects at holding potentials 30, 70 and 110 mV, it was indicated that their suppressing effect on the nicotine-induced currents was voltage-dependent. However, different from that of HEX, the inhibitory effect of MEC increased with administering the mixture of MEC and nicotine at intervals of 3 min, indicating a use-dependent effect of MEC. It is concluded that the action site of MEC on nicotinic receptors of sympathetic neurons is different from that of HEX.