To investigate the effect of microRNA-183-5p (miR-183-5p) targeting RNA-binding motif single-stranded-interacting protein 1 (RBMS1) on autophagy and migration of breast cancer cells and its mechanism.

Human breast cancer cells MCF-7 were divided into control group, NC inhibitor group (transfected with NC inhibitor) and miR-183-5p inhibitor group (transfected with miR-183-5p inhibitor), miR-183-5p inhibitor+si-NC group (transfected with miR-183-5p inhibitor and empty vector si-NC), miR-183-5p inhibitor+si-RBMS1 group (transfected with miR-183-5p inhibitor and RBMS1 knockdown plasmid si-RBMS1). The expression levels of autophagy and phosphatidylinositol-3-kinase (PI3K)/ protein kinase B (Akt)/ mammalian target of rapamycin (mTOR) pathway-related proteins in each group were detected by Western blot, and the mobility of cells in each group was detected by scratch test.

Control group, NC inhibitor group, miR-183-5p inhibitor group, miR-183-5p inhibitor+si-NC group and miR-183-5p inhibitor+si-RBMS1 group the relative expression levels of light chain 3 Ⅱ (LC3-Ⅱ)/light chain 3 Ⅰ (LC3-Ⅰ) protein were 0.29±0.03, 0.31±0.03, 0.86±0.10, 0.84±0.09 and 0.43±0.05, respectively; the relative expression levels of Beclin-1 protein were 0.18±0.02, 0.20±0.02, 0.74±0.08, 0.78±0.09 and 0.35±0.04, respectively; the relative expression levels of sequestosome-1 (P62) protein were 0.93±0.12, 0.89±0.10, 0.51±0.06, 0.54±0.06 and 0.86±0.10, respectively; the 48 h cell mobility was (62.87±7.26)%, (59.23±6.89)%, (24.13±3.49)%, (26.14±4.72)% and (40.11±5.71)%, respectively; the relative expression levels of phospho- (p-) PI3K/PI3K protein were 0.67±0.12, 0.64±0.10, 0.32±0.06, 0.35±0.06 and 0.74±0.09, respectively. The above indexes of miR-183-5p inhibitor+si-RBMS1 group were compared with those of miR-183-5p inhibitor+si-NC group, and those of miR-183-5p inhibitor group were compared with those of NC inhibitor group, and the differences were statistically significant (all P<0.05).

Inhibition of miR-183-5p expression can promote autophagy and inhibit cell migration in MCF-7 cells, while down-regulation of RBMS1 has the opposite effect, which is related to the PI3K/Akt/mTOR pathway.

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 develop a high performance liquid chromatographic method for determination of epinephrine in epinephrine hydrochloride injection.

The separation was carried out on a Waters XBridge C₁₈ column (150.0 mm×4.6 mm, 5 μm) with isocratic elution . The mobile phase consisted of 5.0 g·L^-1 potassium dihydrogen phosphate and 2.6 g·L^-1 octanesulfonic acid sodium solution (adjusting pH to 3.7 by phosphoric acid) -acetonitrile (85∶15) with the flow rate of 1.0 mL·min^-1, the column temperature of 40 ℃, the detection wavelength of 280 nm and the injection volume of 10 μL. The specificity, standard curve and lower limit of quantitation (LLOQ), system suitability, stability, precision and recovery were investigated.

The calibration curve of epinephrine was linear in the range of 2.09 - 83.66 μg·mL^-1. The standard curve of epinephrine was y=8.74×10³x-2.72×10³ (r=1.000 0). The LLOQ was 2.09 μg·mL^-1. The system applicability was proved to be good since the relative standard deviation (RSD) of the main peak area was 0.19% and the RSD of retention time was 0.18%. The RSD of the test solution at high, medium and low concentrations was less than 0.5%, and the average recovery rate was 105.09%-107.86%. The test solution was stable within 4 h.

The method validation proveded that the proposed method was stable, recovery, precise and specificity, which could be used for the measurement of epinephrine in epinephrine hydrochloride injection administration preparation.

To investigate the factors affecting the quality of warfarin anticoagulation therapy in elderly patients with nonvalvular atrial fibrillation (NVAF) and to assess the effect of the combined physician-pharmacist outpatient model on improving the quality of anticoagulation.

In this study, elderly NVAF patients treated with warfarin were divided into combined physician-pharmacist outpatient group and general outpatient group to assess the impact of different anticoagulation management strategies on patients. On this basis, patients were further classified into good anticoagulation quality group (TTR≥60%) and poor anticoagulation quality group (TTR<60%) based on the percentage of time (TTR) that their international normalised ratio (INR) was within the therapeutic target range, and the clinical prognosis and adverse events that occurred in both groups were analysed.

The mean TTR of patients in the combined physician-pharmacist outpatient group was (64.00±24.40)%, which was significantly higher than that of the general outpatient group, which was (42.10±30.20)% (P<0.05). Multifactorial logistic regression analysis showed that the number of comorbidities≥4 was an independent risk factor for poor anticoagulation quality (OR: 0.44, 95%CI: 0.23-0.87, P<0.05), whereas the combined physician-pharmacist outpatient clinic model significantly improved the anticoagulation quality of warfarin (OR: 3.50, 95%CI: 1.85-6.60, P<0.05). Although there were no significant differences between the two groups in the incidence of thromboembolic and bleeding events, the model showed potential advantages in the management of complex patients.

The combined physician-pharmacist outpatient model effectively improves the quality of anticoagulation therapy in elderly patients with NVAF by providing personalized anticoagulation treatment plans and medication management services. This model has a particularly positive impact on patients with multiple comorbidities, demonstrating its significant value in clinical practice.

To analyze the correlations between topoisomerase Ⅱ alpha (TOP2A) and prognosis and immune infiltration in lung cancer.

The expression difference of TOP2A in lung cancer patients was examined by tumor immune estimation resource (TIMER) database. The prognostic value of TOP2A in lung cancer was evaluated using the Kaplan-Meier Plotter and PrognoScan databases. Additionally, the correlation between immune infiltration related gene marker sets and TOP2A expression was analyzed by TIMER database. The gene-gene and protein-protein interactions were determined using GeneMANIA and STRING for network construction, respectively. The possible regulatory network of TOP2A was explored by miRWalk and DIANA-LncBase v2 databases.

The expression of TOP2A in lung adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC) was significantly upregulated and high expression of TOP2A was significantly associated with reduced overall survival, first progression survival, and post-progression survival in lung cancer patients. TOP2A expression was significantly correlated with the infiltration of immune cells in lung cancer, including monocytes, neutrophils, tumor-associated macrophages, helper T cells 1, regulatory T cells, and exhausted T cells. The majority of genes or proteins associated with TOP2A were involved in the regulation of gene transcription and cell cycle. An lncRNA (ENSG00000279978) was identified as being related to the progression of LUAD and LUSC.

TOP2A is an immune infiltration-related prognostic biomarker for lung cancer and may serve as a potential therapeutic target.

To establish a hollow fiber centrifugal ultrafiltration (HFCF-UF) coupled with ultra-high performance liquid chromatography (UPLC) method for the analysis of free methotrexate (MTX) concentration in human plasma and apply to clinical therapeutic drug monitoring.

Plasma samples 500 μL were prepared with HFCF-UF. A Waters UPLC BEH C₁₈ column (50.0 mm×2.1 mm, 1.7 μm) was used. The mobile phase consisted of methanol and 0.05 mol·L^-1 phosphate buffer (pH 6.2) (17∶83, v/v) at a flow rate of 0.2 mL·min^-1. The column temperature was maintained at 30 ℃. The detected wavelength was 302 nm. The specificity, linear relationship, lower limit of quantification (LLOQ), precision, recovery rate and stability of this method were investigated, and it was applied to the determination of clinical plasma samples.

The good linear relationship were obtained between concentration of free MTX in human plasma from 0.05-10.00 μmol·L^-1, y=0.297x+0.001 (r²=0.999). The LLOQ of MTX free plasma concentration analysis was 0.05 μmol·L^-1. The method recovery rates of free MTX were 95.77%-100.52%. The absolute recovery rates of free MTX was 84.09%-92.93%. The intra-day and inter-day relative standard deviation (RSD) were all less than 7.0 %. It was successfully used to analyze free MTX concentration in 52 plasma samples from patients with MTX chemotherapy. The average free plasma concentration of MTX in these 52 patients was 0.68 μmol·L^-1, with the maximum value of 9.32 μmol·L^-1 and the minimum value of 0.056 μmol·L^-1.

The developed method is simple, accurate and sensitive, which is suitable for the analysis of free MTX on clinical therapeutic drug monitoring.

To evaluate the bioequivalence of domestic bromhexine hydrochloride granules and imported bromhexine hydrochloride fine granules in Chinese healthy subjects under fasting and fed states.

A single-center, randomized, open-label, fasting and fed single-dose, two-preparation, two-sequence, and two-period crossover study design was used. Forty-eight Chinese healthy subjects were enrolled in fasting and fed trial, respectively. A random crossover, single-dose of bromhexine hydrochloride granules 0.4 g(containing 8 mg bromhexine) or bromhexine hydrochloride fine granules 0.4 g (containing 8 mg bromhexine) were orally given to subjects. Blood samples were collected at different time points, and plasma concentrations of bromhexine were measured by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Phoenix WinNonlin 8.3 software was used for data analysis.

In the fasting group, the main pharmacokinetic parameters of bromhexine in plasma after taking the test and reference preparations: C_max were (18.29±7.80) and (19.92±10.23) ng·mL^-1, AUC_0-t were (31.52±12.23) and (32.22±12.32) ng·h·mL^-1, AUC_0-∞ were (34.43±13.60) and (34.84±13.36) ng·h·mL^-1, respectively. In the fed group, the main pharmacokinetic parameters of bromhexine in plasma after taking the test and reference preparations: C_max were (12.55±6.27) and (12.64±6.51) ng·mL^-1, AUC_0-t were (58.86±24.38) and (60.60±26.44) ng·h·mL^-1, AUC_0-∞ were (68.27±30.76) and (94.01±113.13) ng·h·mL^-1, respectively. The 90% confidence intervals of the geometric mean ratio of the two preparations in fasting group: C_max was 88.79%-101.49%, AUC_0-t was 93.69%-102.15%, AUC_0-∞ was 94.59%-102.95%; in fed group: C_max was 93.08%-106.82%, AUC_0-t was 94.30%-102.80%, AUC_0-∞ was 92.97%-103.89%.

In this study, the test preparation bromhexine hydrochloride granules and the reference preparation bromhexine hydrochloride fine granules were bioequivalent in healthy Chinese subjects.