To investigate the cardioprotective mechanism of Ilexin A in ameliorating hyperlipidemia through the regulation of microRNA-133a-3p (miR-133a-3p) and uncoupling protein 2 (UCP2).

Rats were randomly divided into control group (gavage with an equal volume of distilled water as the experimental group), model group (gavage with an equal volume of distilled water as the experimental group), experimental group (gavage with 40 mg·kg^-1 Ilexin A), miR-133a-3p inhibitor group (gavage with 40 mg·kg^-1 Ilexin A + tail vein injection of 1.95×10¹² vg·mL^-1 adenovirus containing miR-133a-3p inhibitor), and OE-UCP2 group (gavage with 40 mg·kg^-1 Ilexin A + tail vein injection of 1.95×10¹² vg·mL^-1 adenovirus containing both miR-133a-3p inhibitor and OE-UCP2). Blood lipid levels, including low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C), were measured by enzyme-linked immunosorbent assay. Cardiac function parameters, such as the mitral inflow peak early diastolic velocity to late diastolic velocities (E/A) and left ventricular ejection fraction (LVEF), were assessed via echocardiography. The relative expression levels of mRNA were analyzed by real-time quantitative polymerase chain reaction (RT-qPCR), while protein levels were detected by Western blot.

In the model and experimental groups, LDL-C levels were (0.92±0.12) and (0.76±0.09) mmol·L^-1; HDL-C levels were (0.82±0.10) and (1.14±0.13) mmol·L^-1; E/A ratios were 0.89±0.11 and 1.17±0.14; LVEF values were (41.31±5.07)% and (59.92±7.86)%; the relative expression levels of miR-133a-3p were 0.33±0.06 and 1.22±0.18; uncoupling protein 2 (UCP2) mRNA levels were 0.57±0.08 and 0.79±0.09; cleaved caspase-3 protein levels were 1.00±0.15 and 0.71±0.09; B-cell lymphoma-2 (Bcl-2) protein levels were 1.00±0.17 and 1.94±0.27, respectively. The differences between the model and experimental groups for these parameters were statistically significant (all P<0.05).

Ilexin A ameliorates hyperlipidemia by regulating miR-133a-3p and UCP2. Its mechanism may involve downregulating Bcl-2 and activating cleaved caspase-3 to induce apoptosis, thereby exerting cardioprotective effects and improving cardiac function.

To explore the effect and mechanism of piperlongumine on hepatic steatosis induced by sleep deprivation.

Mice were randomly divided into control group (normal feeding), model group (sleep deprivation treatment), low-, middle- and high dose groups (2.5, 5.0 and 10.0 mg·kg^-1 piperlongumine was given on the basis of sleep deprivation), and the mice were given continuous gavage for 14 days. Liver index was detected after 14 days of continuous gavage. Western blot assay was used to detect protein expression in hepatic tissue, lipid levels in hepatic tissue were detected by kit, and reactive oxygen species (ROS) levels in hepatic tissue were detected by dihydroethidium (DHE).

The liver index of control group, model group, and high-dose group were (2.99±0.40)%, (4.32±0.17)% and (3.30±0.37)%, respectively; triglyceride (TG) levels were (0.16±0.011), (0.29±0.02) and (0.17±0.02) mmol·mgprot^-1, respectively; fatty acid synthase (FAS) protein levels were 0.31±0.03, 0.69±0.08 and 0.35±0.03, respectively; hypoxia-inducing factor-1 α (HIF-1α) protein levels were 0.38±0.04, 0.94±0.08 and 0.44±0.04, respectively; the protein levels of adaptor p66Shc (p66Shc) were 0.21±0.03, 0.82±0.07 and 0.37±0.04, respectively; ROS levels were 1.00±0.09, 4.72±0.44 and 1.30±0.07, respectively. The above indexes in the model group were significantly different from those in the control group, and the above indexes in the high-dose groups were significantly different from those in the model group (all P<0.05).

Piperlongumine can improve liver steatosis induced by sleep deprivation in mice, which may be related to the regulation of HIF-1α/p66Shc signaling pathway.

To study the effect of hydroxylsafflower yellow A (HSYA) on epiphyseal injury of tibial in rats through neurotrophic factor-3 (NT-3).

Fifty male SD rats were randomly divided into control group, model group, HSYA group, NC group and sh-NT-3 group, with 10 rats in each group. Except the control group, the injury model of tibial epiphyseal was established in the other 4 groups by high speed dental abrasive drill. The HSYA group was given 10 mg·kg^-1 HSYA by intragastric administration once a day for 6 weeks. NC group and sh-NT-3 group were respectively injected with 1 mg·kg^-1 sh-NC and sh-NT-3 through the tail vein, once every two days for 6 weeks. The control group and model group were given the same amount of normal saline intragastric and caudal intravenous injection. Quantitative real-time polymerase chain reaction was used to detect the expression level of NT-3 mRNA. The expression levels of bone repair markers and Notch receptor 1 (Notch1) signaling pathway related proteins were detected by Western blot.

The mRNA levels of NT-3 in control group, model group and HSYA group were 1.00±0.17, 3.31±0.59 and 5.16±0.84, respectively. The relative expression of runt related transcription factor 2 protein in control group, model group, HSYA group, NC group and sh-NT-3 group were 1.00±0.17, 3.67±0.63, 7.36±1.35, 7.31±1.42, 5.74±1.32, respectively; the relative expressions of osteoblast-specific transcription factor protein were 1.00±0.18, 5.76±1.01, 8.49±1.42, 8.56±1.54, 6.48±1.16, respectively; the relative expressions of Notch1 protein were 1.00±0.18, 1.44±0.23, 1.79±0.34, 1.83±0.35, 1.52±0.27, respectively. The above indexes in the model group were compared with the control group, the HSYA group was compared with the model group, and the sh-NT-3 group was compared with the NC group, and the above indexes were statistically significant (all P<0.05).

HSYA improves tibial epiphyseal injury in rats by promoting NT-3 expression and activating Notch1 signaling pathway.

To study the effect and mechanism of galangin in improving depression-like behavior in vascular dementia (VD) mice through microRNA-134 (miRNA-134).

Seventy C57BL/6J mice were randomly divided into sham group (0.9% NaCl), model group (0.9% NaCl), low-dose group (25 mg·kg^-1 galangin), medium-dose group (50 mg·kg^-1 galangin), high-dose group (100 mg·kg^-1 galangin), and mimic NC group (100 mg·kg^-1 galangin and mimic NC 15 nmol), miRNA-134 mimic group (100 mg·kg^-1 galangin and miRNA-134 mimic 15 nmol). The degree of depression was detected by tail suspension test and forced swimming test; the level of corticosterone was detected by enzyme-linked immunosorbent assay; the level of miRNA-134 was detected by quantitative real time polymerase chain reaction; the apoptosis rate of brain tissue cells was detected by TdT-mediated dUTP nick-end labeling; and the level of synaptic plasticity related protein was detected by Western blot.

The suspension time of the sham group, the model group and the high-dose group were (105.32±15.22), (185.32±24.62) and (112.65±15.08) s, respectively; the swimming time were (86.52±13.25), (152.32±22.22) and (96.22±16.35) s respectively; the levels of corticosterone were (32.65±5.02), (86.62±14.32) and (48.52±5.55) ng·L^-1, respectively; the levels of miRNA-134 were 1.00±0.12, 2.32±0.35 and 1.21±0.18, respectively. The apoptosis rates of the sham group, model group, high group, mimic NC group and miRNA-134 mimic group were (5.32±0.62)%, (62.33±8.65)%, (14.32±1.78)%, (15.85±2.22)% and (43.21±7.05)%, respectively; synaptophysin protein levels were 1.00±0.18, 0.32±0.05, 0.88±0.12, 0.82±0.11 and 0.30±0.05, respectively; the postsynaptic density protein-95 protein levels were 1.00±0.14, 0.61±0.07, 0.72±0.09, 0.70±0.08 and 0.45±0.06, respectively. The above indexes in the model group were compared with those in the sham group, those in the high-dose group were compared with those in the model group, and those in the miRNA-134 mimic group were compared with those in the mimic NC group, and the differences were statistically significant (all P<0.05).

Galangin may improve the depression-like behavior of VD mice by inhibiting the expression of miRNA-134, improving synaptic plasticity and inhibiting cell apoptosis.

To study the pharmacokinetic profile of oseltamivir phosphate dry syrup in Chinese healthy subjects and to evaluate the bioequivalence and safety of the test formulation (T) and the reference formulation (R) under fasting and fed conditions.

In a randomized, open, two-agent, two-sequence, two-cycle, double-crossover trial design, 36 healthy subjects were enrolled in the fasting and fed groups, respectively, who received a single oral dose of 2.5 g of oseltamivir phosphate dry syrup of either the T or R. The blood concentrations of oseltamivir were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and the pharmacokinetic parameters were calculated by using the WinNonlin (8.2) software and evaluated for bioequivalence and safety.

Under fasting conditions, the C_max of oseltamivir of the T and R were (64.40±17.50) and (64.10±20.90) ng·mL^-1, AUC_0-t were (142.76±43.64) and (151.60±35.10) h·ng·mL^-1, AUC_0-∞ were (147.67±37.60) and (153.05±35.30) h·ng·mL^-1, respectively. Under fed conditions, the C_max of oseltamivir of the T and R were (47.70±18.30) and (46.90±17.20) ng·mL^-1, AUC_0-t were (170.52±26.28) and (168.70±24.51) h·ng·mL^-1, AUC_0-∞ were (173.08±26.50) and (171.30±24.59) h·ng·mL^-1, respectively. The 90% confidence intervals for the geometric mean ratios of C_max, AUC_0-t and AUC_0-∞ for the T and R fell in the range of 80.00% to 125.00% for both the fasting and fed groups. Adverse drug reaction rates were 27.78% and 44.44% in the fasting and fed groups, respectively.

Two kinds of oseltamivir phosphate dry syrups were bioequivalent and had favorable safety profiles in Chinese healthy subjects in both fasted and fed states.

To establish a high performance liquid chromatography-mass spectrometry (HPLC-MS/MS) method for the determination of nicorandil concentrations in human plasma and for pharmacokinetic studies in healthy subjects.

Human plasma samples were pretreated with acetonitrile extraction. Chromatographic cdumn: Acquity HPLC BEH C₁₈ (1.7 μm, 2.1 mm×50.0 mm), mobile phase: methanol (0.1% formic acid)-5 mmoL· L^-1 ammonium acetate (with 0.1% formic acid). Flow rate: 0.2 mL·min^-1, column temperature: 40 ℃, injection volume: 5 μL. Gradient elution, electrospray ion source, multiple reaction monitoring, positive ion scanning, nicorandil-d4 isotope as internal standard. The specificity of the method, the standard curve and the lower limit of quantification, the carryover effect and dilution reliability, the precision and recovery, the matrix effect, and the stability of the method were investigated.

The specificity of the established method was good, nicorandil had a good linear relationship between 0.2-200.0 ng·mL^-1, the standard curve was y=1.08×10^-1x+2.66×10^-2 (r=0.999 7), the lower limit of quantification was 0.2 ng·mL^-1, there was no residual effect, the dilution results were reliable, and the intra-day and inter-day precision of quality control samples were 0.64%-8.70% and 0.91%-8.70%, respectively. The intra-day and inter-day accuracy were 100.00%-115.00% and 98.00%-115.00%, respectively. The extraction recoveries of nicorandil at low, medium and high quality control concentrations were 92.78%-95.35%, with no obvious matrix effect and good stability.

This method is highly specific, accurate, stable, reproducible, and simple to operate, and can be used to determine the concentration of drug in plasma of nicorandil.

To establish an ultra-high performance liquid chromatography tandem high resolution mass spectrometry (UHPLC-HRMS) method for the determination of serum demthylzeylasteral concentration in diabetic nephropathy patients, which treated by tripterygium wilfordii polyglycosides tablets (TGTs), and to apply this method in the exploratory study of clinical pharmacodynamic marker.

Serum samples and ¹⁸O-demethylzeylasteral which was used as the internal standard were processed by solid phase extraction method and took the supernatant for the determination. Chromatographic separation of the analytes was accomplished using a BEH C₁₈ column (2.1 mm×50.0 mm, 1.7 μm) under gradient elution, where the mobile phase consisted of a mixture of 0.1% formic acid aqueous solution and acetonitrile, as well as 0.1% formic acid aqueous solution. The flow rate was set at 0.3 mL·min^-1 and the column temperature at 30 ℃. The mass spectrometer was equipped with an electrospray ionization source with positive multiple reaction monitoring mode. The specificity, standard curve and lower limit of quantification, precision and recovery rate, matrix effect and stability were investigated. This method was applied to determine the serum concentration of demethylzeylasteral from 15 patients with oral TGTs. Receiver operating characteristic (ROC) curve and area under the ROC curve (AUC) were utilized to explore the correlation between serum concentration of demethylzeylasteral and efficacy in treating diabetic nephropathy.

The calibration curve of demethylezylasteral was linear over the range of 1-200 ng·mL^-1, and the lower limit of quantification was 1 ng·mL^-1. The precision and accuracy relative standard deviation of intra-day and inter-day was less than 10% and 15%, respectively. The average absolute recovery was more than 89%, no matrix effect has been affected and all stability tests met the acceptance criteria. The AUC of the efficacy of TGTs predicted by serum demethylzeylasteral quantitation was 0.66 (95% confidence interval was 0.53-0.78), the cut-off value was 61.73 ng·mL^-1, the sensitivity was 83.3% and the specificity was 46.7% (P<0.05).

The UHPLC-HRMS method established in this study has the advantages of simple operation, high sensitivity, accurate and reliable results, which can be suitable for determining serum concentration of demethylzsylasteral and studying clinical drug efficacy markers of TGTs.

Acute lung injury (ALI)/acute respiratory distress syndrome is a life-threatening acute diffuse inflammatory lung injury characterised by increased pulmonary vascular permeability, inflammatory cell infiltration, pulmonary oedema, diffuse alveolar damage and reduced respiratory output. Its pathogenesis is complex, and oxidative stress is a key mechanism leading to ALI exacerbated by alveolar epithelial cell dysfunction. The Kelch-like ECH associated protein 1 /nuclear factor -E2-related factor 2 (Keap1/Nrf2) signalling pathway is an important signalling pathway of traditional Chinese medicine in the treatment of ALI by regulating oxidative stress. This paper analyses and summarizes the relevant Chinese medicine formulas and active ingredients for the treatment of ALI, and finds that a large number of antioxidant active ingredients can regulate oxidative stress through the Keap1/Nrf2 signaling pathway and play a role in improving ALI, which also provides a basis for the prevention and treatment of ALI disease and further drug development.