To investigate the effect of paeonol (PAE) on epithelial-mesenchymal transition (EMT) and endoplasmic reticulum stress-induced apoptosis in ovarian cancer cells by regulating the microRNA-128-3p (miR-128-3p)/ never in mitosis gene A-related kinase 2 (NEK2) signaling axis.

SKOV3 cells were divided into the following groups: SKOV3 group (normal culture), PAE group (4 mmol·L^-1 PAE treatment), inhibitor-NC group (transfected with miR-128-3p inhibitor negative control), miR-128-3p inhibitor group (transfected with miR-128-3p inhibitor), si-NC group (transfected with si-NEK2 negative control), si-NEK2 group (transfected with si-NEK2), PAE+inhibitor-NC group (based on the PAE group, transfected with miR-128-3p inhibitor negative control), PAE+miR-128-3p inhibitor group (based on the PAE group, transfected with miR-128-3p inhibitor), PAE+miR-128-3p inhibitor+si-NC group (based on the PAE group, co-transfected with miR-128-3p inhibitor and si-NEK2 negative control) and PAE+miR-128-3p inhibitor+si-NEK2 group (based on the PAE group, co-transfected with miR-128-3p inhibitor and si-NEK2). Quantitative real-time polymerase chain reaction (PCR) was used to detect the relative expression levels of mRNA; immunofluorescence was used to detect the positive expression of NEK2 protein; Western blot was used to detect the relative expression levels of related proteins.

The relative expression levels of miR-128-3p in SKOV3 group, PAE group, PAE+inhibitor-NC group and PAE+miR-128-3p inhibitor group were 1.00±0.12, 2.34±0.27, 2.30±0.29 and 1.19±0.21, respectively; the relative expression levels of NEK2 mRNA were 1.00±0.13, 0.58±0.09, 0.61±0.12 and 0.93±0.18, respectively; the positive expression of NEK2 was 1.00±0.11, 0.29±0.05, 0.31±0.06 and 0.94±0.17, respectively; the relative expression levels of E-cadherin mRNA in SKOV3 group, PAE group, PAE+inhibitor-NC group, PAE+miR-128-3p inhibitor group, PAE+miR-128-3p inhibitor+si-NC group and PAE+miR-128-3p inhibitor+si-NEK2 group were 1.00±0.12, 2.25±0.29, 2.27±0.34, 1.30±0.21, 1.32±0.20 and 2.03±0.35, respectively; the relative expression levels of N-cadherin mRNA were 1.00±0.14, 0.60±0.09, 0.63±0.12, 0.92±0.17, 0.94±0.18 and 0.67±0.13, respectively; the relative expression levels of Vimentin mRNA were 1.00±0.13, 0.62±0.09, 0.66±0.11, 0.90±0.17, 0.92±0.18 and 0.71±0.13, respectively; the relative expression levels of activating transcription factor 4 (ATF4) protein were 1.00±0.11, 3.22±0.57, 3.31±0.60, 1.51±0.24, 1.49±0.21 and 2.97±0.44, respectively; the relative expression levels of CCAAT/ enhancer binding protein (C/EBP) homologous protein (CHOP) were 1.00±0.14, 3.85±0.47, 3.62±0.41, 1.31±0.17, 1.28±0.17 and 3.26±0.57, respectively; the relative expression levels of cleaved-caspase-3 were 1.00±0.13, 4.52±0.67, 4.61±0.71, 1.67±0.26, 1.69±0.31 and 4.11±0.76, respectively. The differences in the above indicators were statistically significant when comparing PAE group with SKOV3 group, PAE+miR-128-3p inhibitor group with PAE+inhibitor-NC group and PAE+miR-128-3p inhibitor+si-NEK2 group with PAE+miR-128-3p inhibitor+si-NC group (all P<0.05).

PAE suppresses EMT and promotes endoplasmic reticulum stress-induced apoptosis in ovarian cancer cells, potentially by upregulating miR-128-3p to inhibit NEK2 expression.

To observe the clinical efficacy and safety of nafurafine tablets in patients with pruritus due to uremia in China.

Patients diagnosed with pruritus due to uremia in the Hemodialysis Center of the Second People’s Hospital of Neijiang City and West China Fourth Hospital of Sichuan University were divided into treatment group and control group by treatment region. Control group was given conventional treatment, while treatment group was treated with nafurafine tablets (2.5 μg, orally every night, once a day, for a course of 3 months) on the basis of control group. The clinical efficacy, 5-D pruritus scale score, calcium and phosphorus metabolism indicators, blood urea nitrogen (BUN), parathyroid hormone level (PTH), β 2-microglobulin (β2-MG), substance P (SP), interleukin-6 (IL-6), interleukin-31 (IL-31) and quality of life evaluation of the two groups were compared. And conducted a safety evaluation.

A total of 120 patients were included, including 60 cases in control group and 60 cases in treatment group. After 3 months of treatment, the total effective rates of treatment group and control group were 95.00% (57 cases /60 cases) and 81.67% (49 cases /60 cases), respectively. The treatment group was significantly higher than control group (P<0.05). After 3 months of treatment, the 5-D pruritus scale scores of treatment and control group were (5.81±3.29) and (8.12±3.22) points, respectively; the scores of the dermatology life quality index (DLQI) scale were (24.56±2.87) and (21.23±2.45) points, respectively; the blood calcium levels were (2.43±0.43) and (2.65±0.37) mmol, respectively; the blood phosphorus levels were (1.34±0.26) and (1.50±0.31) mmol, respectively; the BUN levels were (17.76±3.87) and (19.86±4.25) μmol·L^-1, respectively; the PTH levels were (322.45±76.54) and (356.12±87.65) pg·mL^-1, respectively; the levels of β2-MG were (14.56±4.87) and (18.12±5.45) mg·L^-1, respectively; the SP levels were (82.45±18.76) and (93.06±20.45) pg·mL^-1, respectively; the levels of IL-6 were (7.56±2.47) and (10.12±2.45) pg·mL^-1, respectively; the levels of IL-31 were (52.45±10.87) and (58.11±12.45) pg·mL^-1, respectively. Compared with control group, the above indicators in treatment group showed statistically significant differences (all P<0.05). The main adverse drug reactions in treatment group were nausea in 3 cases, insomnia in 3 cases, dizziness in 1 case and constipation in 2 cases, while those in control group were nausea in 2 cases, insomnia in 1 case, dizziness in 2 cases and constipation in 2 cases. The total incidence of adverse drug reactions in treatment and control group were 15.00% and 11.66%, respectively, and there was no statistically significant difference (P>0.05).

Nafurafine tablets can significantly improve the symptoms of skin pruritus, calcium and phosphorus metabolism disorders, renal function and inflammatory response in Chinese patients with uremia, enhance the quality of life, and has good safety.

To develop a liquid chromatography-tandem mass spectrometry method for the detection of lenvatinib in quantitative dried blood spots.

Lenvatinom-[d₄] was used as the internal standard. The dried blood spot samples were extracted with acetonitrile/water (70/30) and separated on a ProChrom C₁₈ reversed-phase column (2.1 mm×100.0 mm, 3.0 μm). The mobile phase consisted of 10 mmol·L^-1 ammonium acetate solution and methanol with gradient elution. The flow rate was 0.3 mL·min^-1. The ionization was carried out under the electrospray ionization positive mode. Multiple reaction monitoring was used to detect the transitions of lenvatinib at m/z 427.1→370.0 and 338.0, as well as lenvatinib-[d₄] at m/z 431.1→370.0. The specificity, standard curve, carryover, precision and accuracy, extraction recovery, matrix effect and stability of the method were investigated.

Endogenous components in the dried blood spot matrix of blank human whole blood did not affect the determination. The optimized pretreatment employs a ostep extraction with acetonitrile/water (70/30). Lenvatinib in dried blood spots demonstrated good linearity in the range of 10-250 ng·mL^-1. The standard curve equations from three analytical batches were y=3.81×10^-2x+2.76×10^-2(r=0.999 7), y=2.0×10^-2x-4.87×10^-3(r=0.999 9), y=2.01×10^-2x-3.68×10^-3(r=0.999 7), respectively. The lower limit of quantification (LLOQ) was 10 ng·mL^-1. The intra- and inter-batch relative errors were within ±15%, and relative standard deviation (RSD) were below 15%. There was no significant matrix effect, and lenvatinib dried blood spots sample remained stable for at least 7 days at a high temperature of 45 ℃.

The method is convenient, efficient and accurate and it is suitable for the requirements of The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use M10 guideline "Bioanalytical Method Validation and Study Sample Analysis". It can be used for the determination of lenvatinib concentrations in dried blood spot samples.

To observe the clinical efficacy and safety of insulin degludec and liraglutide combination with metformin tablets in the treatment of type 2 diabetes mellitus (T2DM) patients with poorly controlled insulin and oral hypoglycemic drugs.

The T2DM patients with poor control of insulin and oral hypoglycemic drugs were randomly divided into control group and treatment group according to the queue method. Both groups of patients received metformin tablets for basic treatment. On this basis, the control group was given premixed insulin aspartic 30 injection multiple times a day for treatment. The initial injection dose was 0.2-0.4 U·kg^-1·d^-1, twice a day, adjusted according to the blood glucose level, once every 3-5 days, with an increase or decrease of 1-4 U each time. The treatment group was treated with insulin degludec and liraglutide injection. The initial injection was 10 to 16 dose units, administered subcutaneously once a day. The dose was adjusted every 1 to 2 weeks based on the blood glucose level, with an increase or decrease of 2 to 4 dose units each time. Both groups of patients were treated for 3 months. The clinical efficacy, blood glucose level, lipid metabolism level, pancreatic islet function and safety of the two groups of patients were compared.

A total of 40 cases in the control group and 40 cases in the treatment group were enrolled. After treatment, the total effective rates of the treatment group and the control group were 97.50% (39 cases/40 cases) and 80.00% (32 cases/40 cases), the difference was statistically significant (P<0.05). After 3 months of treatment, the glycosylated hemoglobin levels of the treatment group and the control group were (6.14±1.03)% and (7.20±1.08)%, respectively; the fasting blood glucose levels were (6.25±1.34) and (7.82±1.26) mmol·L^-1, respectively; the 2 h postprandial blood glucose levels were (6.37±0.64) and (7.15±0.67) mmol·L^-1, respectively; the triglyceride levels were (1.87±0.62) and (2.64±0.79) mmol·L^-1, respectively; the serum total cholesterol levels were (4.53±0.62) and (5.18±0.76) mmol·L^-1, respectively; and the fasting insulin levels were (7.16±0.71) and (8.56±0.94) μmol·L^-1, respectively; islet β cell function index levels were (58.36±5.87)% and (50.72±6.14)%, respectively; while insulin resistance index levels were 1.52±0.44 and 2.09±0.47, respectively, the differences were statistically significant between the two groups (all P<0.05). The main adverse drug reactions in the treatment group were gastrointestinal reactions, dizziness, increased heart rate and hypoglycemia, while those in the control group were mainly gastrointestinal reactions, dizziness and hypoglycemia. The total incidences of adverse drug reactions in the treatment group and the control group were 15.00% and 12.50%, respectively, and there was no statistically significant difference (P>0.05).

Insulin degludec and liraglutide injection has shown significant clinical efficacy in the treatment of T2DM with poor control of insulin and oral hypoglycemic drugs. It can effectively control patients’ blood glucose and lipid metabolism levels, improve pancreatic function, and has good safety.

To analyze the drug resistance phenotypes and genotypes of 2 307 Enterococcus faecium strains collected from 2011 to 2024.

The antimicrobial susceptibility testing of the strains was performed using the broth microdilution method, followed by genomic DNA extraction from the strains and polymerase chain reaction (PCR) amplification for screening of resistance genes.

From 2011 to 2012, the detection rate of vancomycin-resistant Enterococcus faecium was 1.67%, and the detection rate of vanM gene-positive vancomycin-susceptible strains was 0.34%. In contrast, from 2023 to 2024, the detection rate of vancomycin-resistant Enterococcus faecium increased to 5.11%, while the detection rate of vanM gene-positive vancomycin-susceptible strains reached as high as 4.75%. Additionally, 22 Enterococcus faecium strains co-harboring both vanA and vanM resistance genes were identified during the period from 2021 to 2024.

The detection rate of the vanM resistance gene in Enterococcus faecium has increased year by year, and its horizontal transfer may have contributed to the increased prevalence of Enterococcus faecium co-harboring both vanA and vanM resistance genes in recent years.

Based on the nuclear factor E2-related factor 2 (Nrf2)/nuclear factor kappa B inhibitor alpha (IκB-α)/nuclear factor-kappa B (NF-κB) signaling axis, this study explored the protective effects of quercetin on lipopolysaccharide (LPS)-induced acute kidney injury (AKI) in rats.

A total of 60 SD rats were randomly divided into control group, model group, dexamethasone (DEX) group, quercetin high dose (100 mg·kg^-1) experimental group and quercetin low (10 mg·kg^-1) dose experimental group, with 12 rats in each group. Except for the control group, all other groups received intraperitoneal injection of LPS (10 mg·kg^-1) to prepare an AKI rat model. Enzyme-linked immunosorbent assay was used to detect serum levels of cystatin C (CysC), kidney injury molecule-1 (KIM-1) and inflammatory factors; renal tissue pathological changes were observed, and levels of myeloperoxidase (MPO), catalase (CAT) and 4-hydroxynonenal (4-HNE) in renal tissues were measured by colorimetry; immunohistochemistry was used to detect protein relative expression levels of Nrf2, IκB-α and NF-κB.

The levels of CysC in control group, model group, quercetin high dose group, low dose group and DEX group were (93.18±8.29), (317.58±12.26), (132.73±10.52), (182.91±10.63) and (153.29±10.14) ng·mL^-1, respectively; levels of KIM-1 were (12.39±1.79), (52.19±4.26), (26.83±3.89), (35.71±4.03) and (29.07±4.12) pg·mL^-1, respectively; MPO were (72.52±8.23), (177.95±9.14), (116.37±8.93), (136.52±9.37) and (121.81±9.02) pg·mL^-1, respectively; CAT contents were (60.73±8.12), (17.92±8.32), (38.41±8.73), (29.72±7.95) and (36.68±9.26) U·mL^-1, respectively; 4-HNE were (59.82±7.38), (179.63±9.41), (97.38±8.83), (116.91±9.17) and (98.56±8.79) nmol·L^-1, respectively. The levels of tumor necrosis factor-α (TNF-α) were (8.96±2.69), (58.27±5.13), (19.14±4.82), (39.56±5.37) and (16.38±4.11) pg·mL^-1, respectively; IL-1β were (15.62±3.58), (52.09±5.76), (29.47±4.61), (36.17±5.23) and (25.76±4.29) pg·mL^-1, respectively; IL-6 were (10.46±2.89), (71.14±6.53), (21.93±5.57), (33.28±5.96) and (19.12±5.61) pg·mL^-1, respectively. The intergrated optical density (IOD) of Nrf2 protein were 92.56±6.13, 45.09±6.97, 73.22±6.02, 63.52±6.73 and 68.29±6.51; IκB-α protein IOD values were 85.62±5.75, 42.83±6.29, 70.12±6.51, 61.45±6.87 and 78.77±6.32; the IOD values of NF-κB p65 protein were 43.28±4.92, 87.59±6.35, 55.79±5.32, 68.83±6.13 and 49.18±5.08, respectively. There were significant differences in the above indexes between the control group and the model group, between the model group and the quercetin high and low dose groups, and the DEX group (all P<0.05).

Quercetin may alleviate inflammation and oxidative stress damage by regulating the expression of Nrf2/IκB-α/NF-κB signaling axis, thereby playing a protective role against LPS-induced acute kidney injury.

To explore the effects of midazolam on the proliferation, invasion and apoptosis of colorectal cancer cells by regulating the liver kinase B1 (LKB1)/AMP-activated protein kinase (AMPK) signaling pathway.

Cell counting kit-8 method was used to detect the effects of different concentrations (5, 10, 20 and 40 μg·mL^-1) midazolam on the proliferation of HCT116 cells. 20 μg·mL^-1 midazolam was selected for the subsequent experiments. HCT116 cells were grouped into blank group (no treatment), midazolam group (20 μg·mL^-1 midazolam), si-NC group (transfected with si-NC plasmid), si-LKB1 group (transfected with si-LKB1 plasmid), midazolam+si-NC group (transfected with si-NC plasmid+20 μg·mL^-1 midazolam) and midazolam+si-LKB1 group (transfected with si-LKB1 plasmid+20 μg·mL^-1 midazolam). Flow cytometry, cell countig kit-8 (CCK-8), EdU and Transwell experiments were used to detect changes in cell apoptosis, proliferation and invasion. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to measure the LKB1 and AMPK mRNAs. Western blot was used to measure the proliferating cell nuclear antigen (PCNA), cysteinyl aspartate-specific protease 3 (caspase-3), matrix metalloproteinase 9 (MMP-9), liver kinase B1 (LKB1) and AMP-activated protein kinase (AMPK) proteins.

The EdU positive rates of the blank group, midazolam group, si-NC group, si-LKB1 group, midazolam + si-NC group and midazolam + si-LKB1 group were (62.52±6.38)%, (35.26±3.64)%, (61.95±6.24)%, (88.63±8.94)%, (36.05±3.71)% and (55.62±5.67)%, respectively; the proliferation rates were (90.85±9.04)%, (48.85±4.92)%, (91.04±8.92)%, (129.32±13.05)%, (49.05±4.98)% and (79.86±8.11)%, respectively; the apoptosis rates were (5.05±0.53)%, (20.22±2.15)%, (4.86±0.49)%, (3.11±0.35)%, (19.86±2.08)% and (8.77±0.91)%, respectively; the invasion numbers were 198.64±20.45, 122.53±12.37, 195.32±20.34, 286.61±29.33, 123.64±12.46 and 176.64±18.06, respectively; the relative expression levels of LKB1 mRNA were 0.93±0.10, 2.86±0.31, 1.08±0.12, 0.57±0.07, 2.78±0.29 and 1.55±0.17, respectively; AMPK mRNA were 1.06±0.11, 2.25±0.24, 0.98±0.10, 0.42±0.05, 2.19±0.23 and 1.35±0.15, respectively; the relative protein expression levels of PCNA were 0.79±0.08, 0.42±0.05, 0.81±0.09, 1.22±0.14, 0.46±0.05 and 0.68±0.07, respectively; Caspase-3 were 0.92±0.10, 1.44±0.16, 1.03±0.11, 0.51±0.06, 1.48±0.16 and 1.07±0.11, respectively; MMP-9 were 0.98±0.10, 0.52±0.06, 1.01±0.12, 1.53±0.17, 0.58±0.06 and 0.82±0.09, respectively; LKB1 were 0.31±0.03, 0.76±0.08, 0.34±0.04, 0.15±0.02, 0.72±0.08 and 0.45±0.05, respectively; AMPK were 0.44±0.05, 0.85±0.09, 0.42±0.05, 0.23±0.03, 0.80±0.09 and 0.57±0.06, respectively. For all these indicators, compared between the midazolam group and the blank group, the si-LKB1 group and the si-NC group, the midazolam + si-LKB1 group and the midazolam + si-NC group, and the si-LKB1 group, all showed statistically significant differences (all P<0.05).

Midazolam activates the LKB1/AMPK signaling pathway, inhibits the proliferation and invasion of colorectal cancer cells, and induces their apoptosis.