To investigate the mechanism of action of bezafibrate (BEZ) targeting nuclear factor of activated T cells 5 (NFAT5) through microribonucleic acid (miR)-486-5p on the kidney tissues of mice with diabetic nephropathy.

Db/m mice were used as control group. Db/db mice were used to construct a diabetic nephropathy model, which was divided into 4 groups according to the random number table method: Model group, experimental group (75 mg·kg^-1·d^-1 BEZ), miR-486-5p antagomir group (75 mg·kg^-1·d^-1 BEZ+10 mg·kg^-1 miR-486-5p antagomir), sh-NFAT5 group (75 mg·kg^-1·d^-1 BEZ+10 mg·kg^-1 miR-486-5p antagomir+recombinant adenovirus containing NFAT5 knockdown plasmid). The kit was used to detect the levels of renal function and oxidative stress in mice; and the expression levels of glucose metabolism and renal fibrosis-related proteins in renal tissues of mice in each group were detected by Western blot.

The levels of malondialdehyde (MDA) in the control group, model group, experimental group, miR-486-5p antagomir group, and sh-NFAT5 group were (2.78±0.56), (4.96±0.97), (3.02±0.54), (4.51±0.79) and (3.24±0.58) nmol·mg^-1, respectively; the levels of aldose reductase (AR) were 1.00±0.12, 3.61±0.43, 1.82±0.29, 3.09±0.58 and 2.25±0.47, respectively; the levels of pyruvate dehydrogenase (PDH) were 1.00±0.11, 0.53±0.10, 0.91±0.15, 0.64±0.12 and 0.82±0.14, respectively; the levels of pyruvate dehydrogenase E1 subunit α1 (PDHA1) were 1.00±0.14, 0.36±0.07, 0.83±0.16, 0.51±0.11 and 0.74±0.13, respectively; the levels of pyruvate dehydrogenase kinase (PDK) were 1.00±0.10, 2.59±0.52, 1.34±0.31, 2.11±0.43 and 1.56±0.39, respectively; the levels of transforming growth factor-β1 (TGF-β1) were 1.00±0.11, 3.15±0.52, 1.43±0.38, 2.72±0.43 and 1.89±0.57, respectively; the levels of fibronectin (FN) were 1.00±0.09, 2.84±0.57, 1.46±0.41, 2.59±0.52 and 1.73±0.49, respectively; the levels of collagen type I (Collagen I) were 1.00±0.13, 4.28±0.71, 2.03±0.46, 3.64±0.68 and 2.51±0.53, respectively. The above indicators of the control group were compared with the model group, the above indicators of the model group were compared with the experimental group, the above indicators of the experimental group were compared with the miR-486-5p antagomir group, and the above indicators of the miR-486-5p antagomir group were compared with the sh-NFAT5 group. The differences were statistically significant (P<0.05, P<0.01, P<0.001).

BEZ can effectively protect the kidney tissue of mice with diabetic nephropathy, and the mechanism may be realized through the miR-486-5p/NFAT5 signaling pathway.

To explore the mechanism of tacrolimus mediating the protective effect on chronic glomerulonephritis (CGN) rats through regulating the expression of microRNA-145-5p (miR-145-5p) and modulating the C-X-C motif chemokine ligand 16/Ras homolog family member A/Rho-associated coiled-coil-containing protein kinase (CXCL16/RhoA/ROCK) pathway.

SD rats were randomly divided into the control group (administered an equal volume of 0.9% NaCl), model group (CGN model + equal volume of 0.9% NaCl), experimental group (CGN model + 3 mg·kg^-1·d^-1 tacrolimus) and combined group (experimental group + tail vein injection of 140 nmol·L^-1 miR-145-5p antagomir). Enzyme-linked immunosorbent assay was used to measure 24-hour urinary protein (PRO), blood urea nitrogen (BUN), and serum creatinine (SCr) levels. Real-time quantitative polymerase chain reaction was used to detect the relative expression level of miR-145-5p in rat kidney tissue; Western blotting was used to determine protein expression levels.

The 24-hour PRO levels in the control, model and experimental groups were (7.44±2.16), (85.29±17.35) and (49.81±9.52) mg, respectively; the BUN levels were (5.82±1.03), (9.65±1.78) and (6.97±1.14) mmol·L^-1, respectively; the SCr levels were (46.31±7.69), (93.57±16.94) and (58.72±8.36) μmol·L^-1, respectively. The relative expression levels of miR-145-5p in the control, model, experimental and combined groups were 1.00±0.14, 0.56±0.07, 0.83±0.12 and 0.68±0.09, respectively; the TNF-α levels were (116.42±14.53), (237.56±26.18), (164.79±18.35) and (205.12±21.64) pg·mL^-1, respectively; the IL-1β levels were (15.76±1.84), (52.41±6.27), (21.35±2.92) and (43.63±4.58) pg·mL^-1, respectively; the IL-6 levels were (26.28±3.47), (71.53±8.64), (38.92±5.06) and (59.78±6.21) pg·mL^-1, respectively; the CXCL16 expression levels were 1.00±0.12, 1.62±0.17, 1.23±0.14 and 1.45±0.15, respectively. Statistical analysis revealed significant differences between the model group and the control group, between the experimental group and the model group, and between the combined group and the experimental group for these indicators (P<0.05, P<0.01, P<0.001).

Tacrolimus can upregulate miR-145-5p, activate the CXCL16/RhoA/ROCK pathway, reduce inflammatory factors, and improve kidney damage in CGN rats.

To investigate the mechanism of soyisoflavones (SIF) alleviate the rats with polycystic ovary syndrome (PCOS) by regulating the miR-29a-3p/insulin-like growth factor 1 (IGF1) molecular axis.

The female SD rats were divided into normal, model and experimental groups, with 12 rats in each group. The model and experimental groups were induced with gavage letrozole to construct the PCOS rat model. After successful modeling, the normal and model groups were given 100 mg·kg^-1 cellulose sodium carboxymethyl by gavage, and the experimental group was given 100 mg·kg^-1 SIF by gavage. Three groups were treated for 21 d with once a day. The apoptosis rate of ovarian tissues was detected by TdT-mediated dUTP nick-end labeling, the serum sex hormone levels were detected by enzyme-linked immunosorbnent assay, the expression levels of miR-29a-3p and IGF1 were detected by real-time fluorescence quantitative plymerase chain reaction, and the positive expression level of IGF1 was detected by immunohistochemistry in ovarian tissues. Ovarian granulosa cells from PCOS model rats were divided into blank group (normal culture), experimental group (100 μg·L^-1 SIF), miR-29a-3p group (after transfection with miR-29a-3p inhibitor, 100 μg·L^-1 SIF culture was given) and IGF1 group (after transfection with miR-29a-3p inhibitor and si-IGF1, 100 μg·L^-1 SIF culture was given). Apoptosis of ovarian granulosa cells was measured by mitochondrial membrane potential analysis, and the expression level of apoptosis-related proteins was detected by Western blotting.

In the animal experiments, the apoptosis rates of experimental, model and normal groups were (16.14±4.09)%, (23.75±5.68)% and (10.83±3.54)%; the luteinizing hormone levels were (26.81±5.99), (41.55±9.84) and (17.63±3.19) mU·mL^-1; the follicle stimulating hormone levels were (51.09±8.72), (41.66±9.87) and (67.91±18.58) pg·mL^-1; the relative expression levels of miR-29a-3p were 0.89±0.08, 0.47±0.06 and 1.00±0.11; the relative expression levels of IGF1 mRNA were 1.42±0.39, 2.36±0.54 and 1.00±0.07; the positive expression levels of IGF1 were 1.37±0.31, 1.98±0.41 and 1.00±0.12, respectively. The differences of above indexes in the experimental and normal groups were statistically different from those in the model group (all P<0.05). In the cell experiments, the mitochondrial membrane potentials of the experimental, miR-29a-3p, IGF1 and blank groups were 1.28±0.13, 0.66±0.05, 1.47±0.19 and 0.54±0.06; the relative expressions levels of the B-cell lymphoma-2-associated X protein were 0.73±0.06, 0.97±0.11, 0.62±0.07 and 1.00±0.08; the relative expression levels of B-cell lymphoma-2 were 1.74±0.23, 1.14±0.18, 2.11±0.37 and 1.00±0.05, respectively. The differences of above indexes in the experimental and IGF1 groups were statistically different from those in the miR-29a-3p group (all P<0.05).

SIF may delay the progression of PCOS rats by regulating miR-29a-3p/IGF1 pathway to mediate the apoptotic process in ovarian cells.

To investigate the possible mechanism of action of sinomenine (Sin) in the treatment of collagen-induced arthritis (CIA) rats by regulating macrophage polarization.

Wistar rats were randomly divided into control group (normal diet and care), model group (CIA model was established with bovine type Ⅱ collagen at the tail), experimental-L, -M, -H groups (all on the basis of the model group, 40, 80 and 160 mg·kg^-1 Sin by gavage respectively), inh-NC group and inh-miR-885-5p group[on the basis of the experimental-H group, the two groups were injected with a negative control of microRNA-885-5p (miR-885-5p) and miR-885-5p inhibitor in the tail vein, respectively)], for four weeks, with 10 rats in each group. The arthritis index (AI) and plantar swelling of rats in each group were statistically analyzed. The pathological conditions of the synovial tissue of the ankle joint were observed by hematoxylin-eosin staining. Real-time fluorescent quantitative polymerase chain reaction was performed to examine the relative expression level of miR-885-5p in rat synovial tissue. Western blot was carried out to detect the expression of related proteins in the synovial tissue of rats. Enzyme-linked immunosorbent assay was used to detect the level of serum inflammatory factors. Terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) was used to detect the apoptosis of rat synovial cells.

The AI scores of the control group, model group and experimental-H group were 0, (7.20±1.38) and (5.50±1.10) scores, respectively; the paw swelling were 35.32±7.05, 72.46±14.47 and 51.65±10.31, respectively; the relative expression levels of miR-885-5p were 1.00±0.18, 0.52±0.10 and 0.71±0.14, respectively. The relative expression levels of suppressor of cytokine signaling 1 (SOCS1) protein in the control, model group, experimental-H group, inh-NC group and inh-miR-885-5p group were 1.00±0.18, 0.51±0.10, 0.75±0.15, 0.71±0.14 and 0.55±0.11, respectively; the levels of interleukin (IL)-6 were (35.29±7.05), (215.82±43.15), (121.33±24.25), (115.62±23.08) and (207.39±41.36) pg·mL^-1, respectively; the apoptosis rates were (31.47±6.29)%, (12.19±2.43)%, (22.03±4.41)%, (20.16±4.03)% and (13.37±2.66)%, respectively. The above indexes in the experimental-L, -M, -H groups were dose-dependent, and the differences in the above indexes between the experimental-H group and the control group were statistically significant compared with the model group (P<0.05, P<0.01, P<0.001), and the differences in the above indexes between the inh-miR-885-5p group and the inh-NC group were also all statistically significant (all P<0.05).

Sin can regulate macrophage polarization, and improve synovial inflammation in CIA rats, which may be related to the up-regulation of miR-885-5p expression and activation of SOCS1/signal transducer and activator of transcription 3 (STAT3) signaling pathway.

To evaluate the bioequivalence of rotigotine patches after a single dose in healthy Chinese subjects.

Using randomized, open, fasting, single-dose, two-preparation, four-cycle, two-sequence exact repeat crossover design, 24 healthy subjects applied one of the test preparation or reference preparation (4.5 mg·10 cm^-2) to the abdominal skin. To determine the drug concentration in human plasma by liquid chromatography-tandem mass spectrometry. Pharmacokinetic software was used to calculate the relevant pharmacokinetic parameters, and the bioequivalence evaluation was carried out.

The C_max of the test preparation and the reference preparation of rotigotine patches were (275.04±98.93) and (277.11±96.78) pg·mL^-1, the AUC_0-t were (5 011.91±2 015.32) and (4 934.28±2 095.76) h·pg·mL^-1, the AUC_0-∞ were (5 092.68±2 033.23) and (5 160.61±2 052.04) h·pg·mL^-1, respectively. C_max, AUC_0-t and AUC_0-∞ geometric mean (90% confidence interval) were all in the equivalent interval of 80.00%-125.00%.

The test preparation and reference preparation of rotigotine patches are bioequivalent in healthy Chinese subjects.

To establish a method for the determination of carfilzomib in rat plasma by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), and to investigate the pharmacokinetic characteristics of carfilzomib in rat plasma.

The rat plasma samples were processed by a protein precipitation approach using methanol∶acetonitrile (1∶1, v/v). Chromatographic column: Thermo Scientific^TM Hypersil GOLD^TM C₁₈ Column (2.1 mm×50.0 mm, 3 μm); mobile phase: 2 mmol·L^-1 ammonium acetate-0.02% formic acid water (A)-methanol (B) in gradient elution mode; flow rate: 0.45 mL·min^-1; injection volume: 5 μL. Electrospray ionization source in positive ion mode and multiple reaction monitoring were adopted. The specificity, standard curve and lower limit of quantification, residue effect, precision and extraction recovery rate, dilution reliability, matrix effect and stability of this method were examined. After a single intravenous injection of 4 mg·kg^-1 carfilzomib into 6 healthy male SD rats, blood was collected from the inner corner of the eye, and the concentration of carfilzomib in the plasma was detected. The pharmacokinetic parameters were calculated using DAS 3.0.

Carfilzomib exhibited good specificity and linearity within the range of 0.5 to 1 250.0 ng·mL^-1, with a standard curve of y=2.40×10^-2x+6.81×10^-2 (r=0.996 4), the quantitative limit of detection was 0.5 ng·mL^-1, the sample residues were 7.64% and 1.00% of the quantitative limit of detection and the internal standard, respectively, the intra-day and inter-day precision were ≤7.51% and ≤8.16%, respectively, the intra-day and inter-day accuracy were -4.54% to 5.14% and -4.41% to 8.26%, respectively, the extraction recovery were 92.49% to 108.33%, and the methodological evaluation results of sample dilution effect, matrix effect and stability were all in accordance with the requirements for the validation of biological sample analysis methods. The main pharmacokinetic parameters of carfilzomib were: C_max was (1.10×10⁶±5.45×10⁵) ng·L^-1, t_1/2 was (23.10±7.38) min, AUC_0-t was (1.85×10⁷±9.85×10⁶) ng·L^-1·min.

The approach formulated in this study is operationally facile, highly discriminatory, sensitive, precise, dependable, and is appropriate for the determination and analysis of carfilzomib in rat plasma specimens.

To devolope a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of furmonertinib in human plsama and apply this method to analyze the blood concentration of non-small cell lung cancer (NSCLC) patients after taking medicine.

Furmonertinib -d3 was employed as the internal standard. The analyte and internal standard were extracted from plasma by protein precipitation with acetonitrile and chromatographed on Agilent Poroshell 120 SB-C₁₈(2.1 mm×100.0 mm, 3.5 μm); mobile phase consisted of acetonitrile-10 mmoL^-1 ammonium acetate(both of containing 0.1% formic acid); the whole analytical time was 3 min. The quantitative analysis were ionized with an electrospray ionization (ESI) source and operated in positive ion, multiple reaction monitoring (MRM) mode, the specificity, linear range, precision and accuracy, extraction recovery rate, matrix effect, dilution validation, and stability were all investigated.

The standard curves were demonstrated to be liner in the range of 0.50-100.00 μg·L^-1 with y=3.95×10^-2x-4.16×10^-4 (r=0.995 4). The accuracy was 95.8%-107.3%. The coefficient of variation (CV) of inter-day (n=6) and intra-day (n=3) for four different concentration levels were less than 15%. The average recoveries and matrix effects were between 83.1%-88.2% and 94.6%-105.8%.

The established LC-MS/MS method for determining the concentration of vormetinib meets the requirements of biological sample analysis and can be used for the concentration determination of vormetinib in clinical practice.

To establish a quantitative analysis method for serum vitamin D (VD) based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) and evaluate its clinical application value in pediatric nutritional support.

Serum samples were processed with acetonitrile, using 25-hydroxyvitamin D₂-d₆ (25-OH-VD₂-d₆) and 25-hydroxyvitamin D₃-d₆ (25-OH-VD₃-d₆) as internal standards. The chromatographic column was Kinetex C18 (50.0 mm × 3.0 mm, 2.6 μm). The mobile phase consisted of 0.1% formic acid aqueous solution and 0.1% formic acid methanol solution, with gradient elution at a flow rate of 0.5 mL·min^-1 and an injection volume of 10 μL. The column temperature was set to 30 ℃. Electrospray ionization was used in positive ion mode, with multiple reaction monitoring. The LC-MS/MS method was evaluated for specificity, calibration curves, lower limits of quantification, precision, recovery rate, stability, and matrix effects. The method was applied to detect VD levels in the serum of infants from the external group (oral 10 μg·kg^-1 VD drops, qd) and the parenteral group [parenteral nutrition solution with injectable fat-soluble vitamin (Ⅰ) administered intravenously, it was equivalent to 10 μg·mL^-1 VD, qd].

Both 25-OH-VD₂ and 25-OH-VD₃ showed good linearity in the range of 5-100 ng·mL^-1, with the 25-OH-VD₂ standard curve of y=1.00x+2.34×10^-2 (R=0.999 8), and a lower limit of quantification (LLOQ) of 0.5 ng·mL^-1; 25-OH-VD₃ standard curve of y=1.00x+5.6×10^-3 (R=0.999 8), and LLOQ of 1 ng·mL^-1. The recovery rate of 25-OH-VD₂ quality control samples was 95.59%-105.38%, with intra- and inter-day precision ≤ 4.75% and ≤ 4.68%, respectively. The recovery rate of 25-OH-VD₃ was 98.38%-113.63%, with intra- and inter-batch precision ≤5.64% and ≤7.83%, respectively. Stability of 25-OH-VD₂ and 25-OH-VD₃ samples at different concentrations was good under various conditions, with matrix effects ranging from 85%-110% and 88%-112%, respectively. After enteral or parenteral VD supplementation, the serum levels of 25-OH-VD₂ and 25-OH-VD₃ in both groups of preterm infants significantly increased, with the parenteral group showing significantly higher concentrations of 25-OH-VD₂ and 25-OH-VD₃ compared to the enteral group (P<0.05). The daily weight gain rate of preterm infants in the parenteral group was also significantly higher than those in the enteral group (all P<0.05).

The quantitative analysis method of serum VD based on LC-MS/MS has strong specificity and high sensitivity, which is suitable for the monitoring of VD level in premature infants.

To evaluate the efficacy, safety, and economy of trastuzumab emtansine (T-DM1) in the treatment of human epidermal growth factor receptor 2(HER2)-positive metastatic breast cancer based on the rapid health technology assessment methodology, and to provide a reference for the rational use of the drug in the clinic.

A systematic search of Chinese and English databases such as PubMed, Embase, the Cochrane Library, CNKI, Wanfang, SinoMed, etc. was performed to screen the literature according to the inclusion and exclusion criteria, extract the data, evaluate the quality of the literature, and qualitatively analyze the results.

T-DM1 had comparable or even superior efficacy in terms of progression-free survival (PFS) and overall survival (OS) compared with conventional anti-HER2 treatment regimens as well as other chemotherapeutic regimens. Some regimens, such as docetaxel+trastuzumab+patuximab, and pyrrolitinib+capecitabine, might be superior to T-DM1 in terms of PFS and OS. Adverse effects of T-DM1 were mainly liver function abnormalities and thrombocytopenia, and overall the safety profile was better, with a lower probability of serious adverse effects. In terms of economy, T-DM1 was more favorable than T-Dxd, and further evaluation was still needed compared with other regimens.

T-DM1 has favorable efficacy and safety, with economics to be further evaluated.

To explore the design and application of digital intelligence quality control technology in drug clinical trials, aiming to improve the quality and efficiency of these trials.

Based on the characteristics and requirements of quality control in drug clinical trials, a digital intelligence quality control model was designed and applied throughout the entire process of drug clinical trials. The system’s performance and security were evaluated.

The digital intelligence quality management system can significantly enhance the accuracy and completeness of clinical trial data, improve the efficiency of drug clinical trials, and strengthen real-time monitoring capabilities to ensure compliance and safety of the trials.

The digital intelligence quality management system can optimize resource allocation, shorten trial duration, and reduce overall costs. It provides new perspectives and methodologies for quality control in drug clinical trials, laying a reliable foundation for the digital transformation of the biopharmaceutical industry.