Determination of the components of danyikangtai powder into the plasma and its pharmacodynamic study.

Danyikangtai powder has a definite therapeutic effect on pancreatitis. However, the internal mechanism is unclear. The purpose of this experiment is to quickly identify the blood components of danyikangtai powder and evaluate its efficacy. 25 blood components were identified by comparing the components with the same mass spectrometry information from in vivo and in vitro samples. The AR42J cells of the pancreatitis model were treated with drug-containing plasma, and the drug efficacy was evaluated by investigating the amylase release rate. This study provides a scientific reference for its pharmacological research and rational clinical application.

[New Method of Measuring Arterial Oxygen Saturation].

The traditional method of measuring arterial oxygen saturation is that R value, the ratio of alternating component of the logarithmic photoplethysmography, is firstly computed and then the linear regression model is established by experiment. The R value computation is a dimension reduction process based on Lambert-beer law, which aims at eliminating the influence of optical path and minimizing the impact of individual differences. When taking scattering into consideration, the dimension reduction process loses information, introduces the system error and limits the precision of measurement. In order to reduce the measurement error resulting from the scattering effects, this paper presents a new method that the peak and valley values of dual-wavelength logarithmic photoplethysmography waves are used as the independent variables to develop a linear regression model to predict the arterial oxygen saturation. During the experiment, the in-vivo measurements were carried out on 23 healthy volunteer and 133 samples of photoplethysmography waves and the reference value of oxygen saturation were recorded. To compare the predictive performance between the new method and the R value method, 90 samples were randomly selected as modeling sets and the remaining 43 samples were used as prediction sets. Random selection of modeling sets and prediction are executed 10 times. The average related coefficients of the prediction sets of the new method and the R value method are 0.890 6 and 0.846 8, and the average root mean square errors are 0.889 6% and 1.037 3% respectively. Results indicate that the performance of the new method is better than the one of the R value method, and the predictivemodel based on 4 parameters can improve the stability and accuracy of measurement. And the new method has guiding significance to the measurement of human body's blood physiological information based on limited wavelength spectrum data.

A new multi-template molecularly imprinted polymer for separation and purification of dioscin, protodioscin, and diosgenin from purple yam.

Saponin is an essential natural compound in purple yams with high nutritional and medicinal value. In this work, a multitemplate molecule-imprinted polymer (MMIP) was synthesized with dioscin, protodioscin, and diosgenin templates. The MMIPs were characterized with scanning electron microscopy, thermogravimetric analysis, Brunauer-Emmett-Teller (BET) adsorption, and Fourier transform infrared spectroscopy. The efficacy of the MMIPs was assessed with static, dynamic, selective adsorption, desorption, and reusability experiments. The three saponins were selectively extracted and determined by MMIP-high-performance liquid chromatography. The polymer morphology was regular and spherical. The amount of the MMIP adsorbed was 74.825 mg/g, and the imprinting factor was 2.1. The MMIP adsorbed the three saponins from purple yam extract, with recovery rates of 95.5-103.43 % and desorption rates of 85 %-98 %. In addition, the MMIPs were reused at least six times. These results demonstrated that the MMIPs efficiently and selectively extracted dioscin, protodioscin, and diosgenin from food matrices at high rates.

A high-permeability method for extracting purple yam saponins based on ultrasonic-assisted natural deep eutectic solvent.

The extraction of active ingredients from traditional Chinese medicine has received considerable attentions. In this study, 16 kinds of natural deep eutectic solvent (NADES) with ultrasonic were selected to extract saponins from purple yam root and the extraction mechanism was investigated. The results showed that chloride/acrylic acid (1:2; n/n) had the highest extraction yield for saponins. The optimal extraction process parameters were 24% water content, 20 mL/g liquid-solid ratio, and ultrasonic extraction for 85 min (81 °C, 600 W). The extraction rate (ER) of purple yam saponins was 0.935%, close to the fitted result of 96.5 mg/g. Molecular dynamics simulations and FT-IR results showed that the NADES may extract the saponin constituents from purple yam through hydrogen bonding. Compared with traditional extraction methods and molecularly imprinted polymer methods, NADES has a higher ER and lower cost (1.53 $/g), which provides a reference for subsequent industrial quantitative production.

[Inhibitory effect of imperatorin and isoimperatorin on activity of cytochrome P450 enzyme in human and rat liver microsomes].

Imperatorin (IM) and isoimperatorin (ISOIM) are major active components of common herbal medicines from Umbelliferae plants, and widely used in clinic. This article studies the inhibitory effect of IM and ISOIM on the activity of cytochrome P450 (CYP) enzyme, and assesses their potential drug-drug interaction. IM and ISOIM were incubated separately with human or rat liver microsomes for 30 min, with phenacetin, bupropion, tolbutamide, S-mephenytoin, dextromethorphan and midazolam as probe substrates. Metabolites of the CYP probe substrates were determined by LC-MS/MS, and IC50 values were calculated to assess the inhibitory effect of the two drugs on human CYP1A2, 2B6, 2C9, 2C19, 2D6 and 3A4 enzymes, as well as on rat CYP1A2, 2B6, 2D2 and 3A1/2, and grade their inhibitory intensity. In human liver microsomes, IM and ISOIM showed different inhibitory effects on all of the six CYP isoenzymes. They were strong inhibitors for 1A2 and 2B6. The IC50 values were 0.05 and 0.20 micromol x L(-1) for 1A2, and 0.18 and 1.07 micromol x L(-1) for 2B6, respectively. They also showed moderate inhibitory effect on 2C19, and weak effect on 2C9, 2D6 and 3A4. In rat liver microsomes, IM and ISOIM were identified as moderate inhibitors for 1A2, with IC50 values of 1.95 and 2.98 micromol x L(-1). They were moderate and weak inhibitors for 2B6, with IC50 values of 6.22 and 21.71 micromol x L(-1), respectively. They also had weaker inhibitory effect on 2D2 and 3A1/2. The results indicated that IM and ISOIM had extensive inhibitory effects on human CYP enzymes. They are strong inhibitors of CYP1 A2 and 2B6 enzymes. However, it is worth noting the interaction arising from the inhibitory effect of CYP enzymes in clinic.

Reduced inhibitory and excitatory input onto parvalbumin interneurons mediated by perineuronal net might contribute to cognitive impairments in a mouse model of sepsis-associated encephalopathy.

Sepsis-associated encephalopathy (SAE) is commonly defined as diffuse brain dysfunction and can manifest as delirium to coma. Accumulating evidence has suggested that perineuronal net (PNN) plays an important role in the modulation of the synaptic plasticity of central nervous system. We here investigated the role of PNN in SAE induced by lipopolysaccharide (LPS) injection. Behavioral tests were performed by open field, Y-maze, and fear conditioning tests at the indicated time points. The densities of vesicular γ-aminobutyric acid transporter, vesicular glutamate transporter 1, PNN, and parvalbumin (PV) in the hippocampus were evaluated by immunofluorescence. Matrix metalloproteinases-9 (MMP-9) expression and its activity were detected by Western blot and gel zymography, respectively. Local field potential was recorded by in vivo electrophysiology. LPS-treated mice displayed significant cognitive impairments, coincided with activated MMP-9, decreased PNN and PV densities, reduced inhibitory and excitatory input onto PV interneurons enwrapped by PNN, and decreased gamma oscillations in hippocampal CA1. Notably, MMP-9 inhibitor SB-3CT treatment rescued most of these abnormalities. Taken together, our study demonstrates that active MMP-9 mediated PNN remodeling, leading to reduced inhibitory and excitatory input onto PV interneurons and abnormal gamma oscillations in hippocampal CA1, which consequently contributed to cognitive impairments after LPS injection.

LC-MS guided isolation of phenolic glycosides from Viburnum luzonicum Rolfe leaves and their α­amylase and α-glucosidase inhibitory activities.

Viburnum luzonicum Rolfe is widely used in China as folk medicine. The bioactivity evaluation indicated that the n-BuOH fraction of V. luzonicum leaves (VLLB) could significantly inhibit α­amylase and α-glucosidase. In order to clarify its active constituents, the phytochemical analysis on VLLB was first performed using HPLC-QTOF-MS/MS, and three new phenolic compounds, viburosides A-C (1-3), along with seven known analogues (4-10) were isolated through preparative HPLC. The undescribed compounds were determined by extensive spectroscopic analyses (1H and 13C NMR, HSQC, HMBC, HRESIMS, and ORD) and enzymatic hydrolysis. In the in vitro enzyme assays, compounds 1-8 showed potent α­amylase and α-glucosidase inhibitory activities. The enzymatic kinetics and molecular docking of the strongest inhibitors 2 and 3 against the corresponding target enzyme were also performed.

[Chemical constituents of Rauvolfia verticillata].

The study on the Rauvolfia verticillata (Lour.) Baill., which belongs to Apocynaceae, was carried out to look for its chemical constituents and pharmacological activity. The isolation and purification were performed by chromatography on silica gel, Sephadex LH-20 and ODS (octadecyl silane) open column. The structures of obtained compounds were elucidated on the basis of physicochemical properties and spectral analysis. Three indole alkaloids and one acridone alkaloid were isolated from chloroform layer extract and identified as ajmalicine B (1), sandwicine (2), raunescine (3) and 7-hydroxynoracronycine (4) separately. Ajmalicine B (1) is a new compound belonging to indole alkaloid. Compound 4 as an acridone alkaloid was a new type compound isolated from Rauvolfia genus for the first time. We also did some biological activity research on the new type compound (4) to explore other pharmacological activities in addition to antihypertensive activity.

Two new phenolic allopyranosides and their analogues from the stems of Viburnum luzonicum Rolfe guided by LC-MS.

Two new phenolic allopyranosides, named viburluzosides A and B (1, 2), together with eight known phenolic glycosides (3 - 10) were discovered from the stems of Viburnum luzonicum Rolfe under the guidance of LC-MS analyses coupled with bioactivity evaluation. They were purified through various chromatography methods and identified by extensive spectroscopic analyses (1H and 13C NMR, HSQC, HMBC, and HRESIMS) and chemical methods. The in vitro evaluation on α-glucosidase and aldose reductase (AR) inhibitory activities of isolated compounds were conducted. Compounds 1 - 4 and 6 - 9 exhibited α-glucosidase inhibitory activities with IC50 values of 5.35 - 21.34 µM and AR inhibitory activities with IC50 values of 6.21 - 40.06 µM. Moreover, the inhibitory kinetics analyses of compounds 1 and 2 were also performed.

FOXG1 as a Potential Therapeutic Target for Alzheimer's Disease with a Particular Focus on Cell Cycle Regulation.

BACKGROUND: Several recent findings have revealed that targeting of cell cycle reentry and (or) progression may provide an opportunity for the therapeutic intervention of Alzheimer's disease (AD). FOXG1 has been shown to play important roles in pattern formation, cell proliferation, and cell specification. Thus far, the roles of FoxG1 and its involvement in AD are largely unknown. OBJECTIVE: Our study aimed to explore the intervention effect of FOXG1 on AD pathology and its potential mechanism with a particular focus on cell cycle regulation. METHODS: We investigated the association of Foxg1 gene variants with AD-like behavioral deficits, p21 expression, neuronal apoptosis, and amyloid-ß (Aß) aggregate formation; we further determined whether targeting FOXG1-regulated cell cycle has therapeutic potential in AD. RESULTS: Paralleling AD-like behavioral abnormalities, neuronal apoptosis, and Aß deposits, a significant reduction in the expression of FOXG1 was observed in APP/PS1 mice at 6 months of age. Using the APP/PS1;Foxg1fl/fl-CreAAV mouse line, we found that FOXG1 potentially antagonized cell cycle reentry by negatively regulating the levels of p21-activated kinase (PAK3). By reducing p21cip1-mediated arrest at the G2 stage and regulating cyclin A1- and cyclin B-dependent progression patterns of the cell cycle, FOXG1 blocked neuronal apoptosis and Aß deposition. CONCLUSION: These results indicate that FOXG1 contributes to the regulation of the neuronal cell cycle, thereby affecting brain abnormalities in AD. An elevation of the FOXG1 level, either pharmacologically or through other means, could present a therapeutic strategy for AD.

In vitro and in vivo evaluation of levofloxacin sustained-release capsules.

OBJECTIVE: To reduce the frequency of administration and improve patient compliance, novel levofloxacin sustained-release capsules with suitable in vitro release profiles and good bioavailability were developed. MATERIALS AND METHODS: A fluidized bed was used to prepare levofloxacin pellets by spraying the drug solution onto blank pellets. Then the pellets were coated with either Surelease water dispersion or Eudragit® NE30D water dispersion to achieve sustained-release characteristics. The mixed pellets containing 15% of the uncoated pellets and 85% of the coated pellets were filled into the hard gelatin capsules. In vitro release test was performed with the capsules. A single-dose pharmacokinetic study of the capsules was carried out in beagle dogs. RESULTS: Although Eudragit® NE30D-coated pellets and Surelease-coated pellets showed similar sustained-release profiles in vitro, their in vivo pharmacokinetic characteristics exhibited significant difference. Unsuccessful in vivo-in vitro correlation was shown in Eudragit® NE30D-coated pellets with a relative bioavailability of only 41.5%, whereas Surelease-coated pellets achieved best sustained-release feature both in vitro and in vivo with a relative bioavailability of 103.0%. CONCLUSION: Statistical analysis indicated that the capsules containing Surelease-coated pellets had a satisfactory sustained-release behavior and a desired pharmacokinetic property.

Histone deacetylase 3 in hippocampus contributes to memory impairment after chronic constriction injury of sciatic nerve in mice.

ABSTRACT: Chronic neuropathic pain is frequently accompanied by memory impairment, yet the underlying mechanisms remain unclear. Here, we showed that mice displayed memory impairment starting at 14 days and lasting for at least 21 days after chronic constriction injury (CCI) of unilateral sciatic nerve in mice. Systemic administration of the pan histone deacetylase (HDAC) inhibitor sodium butyrate attenuated this memory impairment. More specifically, we found that hippocampus HDAC3 was involved in this process because the levels of its mRNA and protein increased significantly in the hippocampus at 14 and 21 days after CCI, but not sham surgery. Systemic administration of the selective HDAC3 antagonist RGFP966 attenuated CCI-induced memory impairment, improved hippocampal long-term potentiation impairment, and rescued reductions of dendritic spine density and synaptic plasticity-associated protein in the hippocampus. In addition, HDAC3 overexpression in the hippocampus led to memory impairment without affecting basal nociceptive responses in naive mice. Our findings suggest that HDAC3 contributes to memory impairment after CCI by impairing synaptic plasticity in hippocampus. Histone deacetylase 3 might serve as a potential molecular target for therapeutic treatment of memory impairment under neuropathic pain conditions.

[Research progress of Wif1 in development of nervous system].

Wnt inhibitor factor-1 (WIF-1) is an extracellular antagonist of Wnts secreted proteins, first characterized as an expressed sequence tag in the human retina. WIF-1 belongs to the secreted Frizzled-related protein (sFRP) class, which can directly bind to Wnt proteins, prevent Wnts from binding to their receptors in vertebrates. Wif1 is expressed in the nervous system of mouse, Xenopus, zebrafish and human. It has been shown that WIF-1 affects the formation of somites in Xenopus embryos and inhibits rod production in retinal histogenesis by binding to Wnt4 in mice. Histological information of Wif1 expression during the development of the central nervous system has been reported in mouse, Xenopus and zebrafish and the strong embryonic expression suggests Wif1 may play an essential role in the spatial and temporal regulation of Wnt signals in development of central nervous system. The Wnt pathway plays a key role in the patterning of the nervous system. However, insights into the function of Wif1 in the development of the central nervous system are rather limited. Selecting suitable stage and target according to the expression pattern may contribute to understanding the function of Wif1.

[Study on pharmacokinetics of PVP coated beta-elemene liposome in rats].

OBJECTIVE: To study the pharmacokinetics of PVP costed beta-elemente liposmes in rats. METHODS: Gas chromatography was established to determine the concentration of beta-elemene in plasma of rats after administered through i.g. RESULTS: The pharmacokinetics parameters was: T(1/2) = 95.07 +/- 20.46 min, AUC = 348.72 +/- 32.49 microg x min/ml, Cmax = 4.39 +/- 0.33 microg/ml, Tmax = 60 min. CONCLUSION: The bioavailability of PVP coated beta-elemene liposomes is 140.2 +/- 7.5% compared with conventional liposomes.

Sensory input is required for callosal axon targeting in the somatosensory cortex.

BACKGROUND: Sensory input is generally thought to be necessary for refining and consolidating neuronal connections during brain development. We here report that cortical callosal axons in somatosensory cortex require sensory input for their target selection in contralateral cortex. RESULTS: Eliminating sensory input to either hemisphere by unilateral transection of infraorbital nerve (ION) prevents target selection of callosal axons in contralateral cortex. Strikingly, blocking sensory input bilaterally, by simultaneously transecting both IONs, results in rescued callosal projection. In contrast, non-simultaneous bilateral ION transection has the same effect as unilateral transection. Similar results are obtained by lesion of whisker hair follicles. c-Fos-positive neurons in brain slices treated with KCl is decreased more in contralateral cortex with unilateral removal of sensory input, but decreased similarly in both cortices in mice with simultaneous bilateral removal of sensory input. Frequency of sEPSC of cortical neurons is also reduced in contralateral cortex with the unilateral removal of sensory input, but equally reduced on both sides with the bilateral removal of sensory input, suggesting that unbalanced bilateral sensory input might lead to mismatched neuronal activity between the two cortices and contribute to the formation of callosal projection. CONCLUSION: Our data demonstrate a critical role of balanced bilateral somatosensory input in the formation of callosal connections, and thus reveal a new role of sensory input in wiring brain circuits.