To analyze the species distribution and antimicrobial resistance profiles of Acinetobacter baumannii complex (ABC) isolates from a Grade A tertiary hospital in Beijing (2017-2024), providing evidence for clinical microbiological identification and antimicrobial therapy.

ABC isolates were collected from the hospital’s antimicrobial resistance surveillance network. Species identification was performed by mass sectrometry and susceptibility testing was analyzed using WHONET 5.6. Statistical comparisons of resistance rates were performed with SPSS 26.0 (chi-square test).

Among 2 752 non-repetitive ABC isolates,Acinetobacter baumannii (ABA) accounted for 91.75%, followed by Acinetobacter pittii (APT, 6.90%) and Acinetobacter nosocomialis (ANO, 1.31%). Sputum and urine were the predominant specimen types, with ABA and APT frequently isolated from bloodstream infections. The resistance profile varied statistically significantly defferences across the bacterial species for all 12 common antimicrobials (all P<0.05). Among these, ABA demonstrated the highest resistance rates, ANO showed an intermediate profile, and APT was the most susceptible. Specifically, ABA exhibited resistance rates exceeding 80% to β-lactam antibiotics, including piperacillin/tazobactam, ceftazidime, imipenem and meropenem. In contrast, ANO showed resistance rates of 30%-50%, and APT only 10%-30% to the same agents. Resistance profiles between ABA and ANO differed statistically significantly except for trimethoprim-sulfamethoxazole and minocycline (all P>0.05). The single Acinetobacter calcoaceticus isolate was only resistant to doxycycline and trimethoprim-sulfamethoxazole, while remaining susceptible to all other β-lactam and fluoroquinolone agents tested.

ABC species exhibit distinct clinical distributions and resistance patterns, emphasizing the need for accurate species identification and ongoing resistance surveillance to guide appropriate empirical therapy.

To explore the clinical efficacy and safety of different doses of buspirone hydrochloride tablets combined with venlafaxine hydrochloride sustained-release tablets in the treatment of elderly patients with generalized anxiety disorder (GAD) accompanied by depression.

Elderly patients with GAD accompanied by depression were taken as the research subjects. The control group was given oral treatment with venlafaxine hydrochloride sustained-release tablets (75 mg·d^-1, once daily) and buspirone hydrochloride tablets (the initial dose was 5 mg each time, and it was increased by 5 to 10 mg every 2 to 3 days according to the patient’s tolerance, and then increased to 10 mg each time, twice daily after 2 weeks). The treatment group was treated with venlafaxine hydrochloride sustained-release tablets on the basis of the control group. Different doses of buspirone hydrochloride tablets were given orally for treatment (the initial dose was 5 mg each time, and it was increased by 5 to 10 mg every 2 to 3 days according to the patient’s tolerance, and then increased to 20 mg each time, twice a day after 2 weeks). Both groups were treated continuously for 8 weeks. The clinical efficacy, Hamilton anxiety scale (HAMA) scores, anxiety and depression scores, sleep conditions, levels of neurotransmitters for mood regulation, neurological function levels and endocrine metabolic levels of the two groups of patients were compared, and the safety was evaluated.

A total of 100 cases were screened, and 95 cases were enrolled, including 49 cases in the control group and 46 cases in the treatment group. In the control group, 3 cases dropped out due to loss to follow-up, and eventually 46 cases were included in the statistical analysis. There were no patients with dropout in the treatment group. Eventually, 46 cases were included in the statistical analysis. After 8 weeks of treatment, the HAMA scores of the treatment group and the control group were (17.93±2.85) and (19.51±2.96) points, respectively; the Hamilton depression scale scores were (18.85±3.11) and (20.37±3.08) points, respectively; the scores of the Beck despair scale were (4.64±1.44) and (5.37±1.53) points, respectively; the scores of the Chinese version of the self-rating scale for constrained neurosis were (25.63±4.17) and (27.73±4.31) points, respectively; the scores of melatonin were (38.83±5.78) and (36.24±5.27) pg·mL^-1, respectively; the cortisol levels were (109.36±11.92) and (115.41±12.17) ng·mL^-1, respectively; and the Pittsburgh sleep quality index scores were (8.28±2.35) and (9.48±2.46) points, respectively; the values of substance P were (112.17±10.66) and (117.61±11.37) ng·L^-1, respectively; and those of γ -aminobutyric acid were (5.65±0.65) and (5.31±0.67) μmol·L^-1, respectively; the glutamic acid were (25.02±3.22) and (26.75±3.28) mg·L^-1, respectively; the brain-derived neurotrophic factor were (14.33±1.56) and (13.54±1.46) ng·mL^-1, respectively; the glial cell-derived neurotrophic factor were (556.41±62.12) and (525.57±64.72) pg·mL^-1, respectively; and the glial fibrillary acidic protein were (2.51±0.28) and (2.67±0.35) ng·mL^-1, respectively; the free thyroxine were (13.33±1.96) and (12.41±1.85) pmol·L^-1, respectively; and the 25-hydroxyvitamin D₃ was (33.46±4.39) and (31.57±3.14) ng·mL^-1, respectively; the prolactin levels were (22.56±3.94) and (24.59±3.78) ng·mL^-1, respectively. The differences in the above indicators of the treatment group were statistically significant with control group (all P<0.05). Two groups of patients experienced adverse drug reactions such as dizziness, headache, nausea and vomiting, constipation and dry mouth. The total effective rates of the treatment group and the control group were 82.61% and 73.91%, respectively, and the total incidences of adverse drug reactions were 15.22% and 13.04%, respectively. There were no statistically significant differences between the two group (all P>0.05).

The application of 20 mg buconspirone tablets combined with venlafaxine tablets in the treatment of elderly patients with GAD and depression can improve sleep conditions and clinical symptoms, regulate neurological function and endocrine metabolism, and has certain safety.

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 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 observe the clinical efficacy and safety of continuous infusion of extremely low-dose dexmedetomidine hydrochloride injection combined with remifentanil hydrochloride for injection and propofol emulsion injection in elderly patients undergoing total hip arthroplasty (THA).

Elderly patients undergoing THA were divided into treatment group and control group according to the administration method. The control group was induced with intravenous injection of sufentanil citrate injection at a dose of 0.3 μg·kg^-1, propofol emulsion injection at a dose of 1.0-2.5 mg·kg^-1, and rocuronium bromide injection at a dose of 0.6-0.9 mg·kg^-1. Anesthesia was maintained with intravenous infusion of propofol emulsion injection at a dose of 4-6 mg·kg^-1·h^-1 and remifentanil hydrochloride for injection at a dose of 0.1-0.2 μg·kg^-1·h^-1. The treatment group received dexmedetomidine hydrochloride injection 0.2-0.5 μg·kg^-1 before anesthesia induction, followed by ultra-low dose dexmedetomidine hydrochloride injection 0.1-0.2 μg·kg¹·h^-1 intravenous infusion from after anesthesia induction to 30 min before the end of the operation, with other treatments the same as the control group. The vital signs of the two groups were compared after entering the room (T₀), immediately after tracheal intubation (T₁), at the time of upper tourniquet (T₂), when necrotizing bone and tissue were removed (T₃), when prosthesis was implanted (T₄), when tourniquet was released (T₅) and immediately after extubation (T₆) and the modified alert sedation/score (MOAA/S) was improved; the dosage of anesthetics, the occurrence of postoperative delirium, preoperative and postoperative neuronal injury markers, and neurotransmitter markers were compared between the two groups, and safety was evaluated.

A total of 120 patients were enrolled, including 63 cases in the treatment group and 57 cases in the control group. At T₁, the heart rate (HR) of treatment group and control group were (79.68±7.34) and (76.79±8.40) times·min^-1, respectively; at T₂, they were (81.43±7.37) and (78.68±7.49) times·min^-1, respectively; at T₃, they were (83.83±7.53) and (80.35±7.14) times·min^-1, respectively; at T₄, they were (82.63±6.90) and (79.77±7.39) times·min^-1, respectively; at T₅, they were (81.27±7.31) and (78.16±7.51) times·min^-1, respectively; at T₆, they were (86.16±7.32) and (83.74±7.31) times·min^-1, respectively. The heart rate of treatment group at each time were all significantly higher than those of control group (all P<0.05). At T₁, the mean arterial pressure (MAP) of treatment group and control group were (80.29±9.89) and (75.32±9.73) times·min^-1, respectively; at T₂, they were (82.10±10.00) and (77.18±9.82) times·min^-1, respectively; at T₃, they were (83.16±9.95) and (79.46±10.06) times·min^-1, respectively; at T₄, they were (82.83±9.58) and (77.19±9.92) times·min^-1, respectively; at T₅, they were (81.30±9.59) and (76.09±9.82) times·min^-1, respectively; at T₆, they were (84.14±9.87) and (82.12±9.97) times·min^-1, respectively. The MAP of treatment group at each time were all significantly higher than those of control group (all P<0.05). At T₁, the MOAA/S of treatment group and control group were (2.05±0.28) and (2.23±0.46) score, respectively; at T₂, the scores were (1.92±0.33) and (2.09±0.51) score, respectively; at T₃, the scores were (1.95±0.42) and (2.09±0.34) score, respectively; at T₄, the scores were (1.92±0.33) and (2.11±0.36) score, respectively; at T₅, the scores were (1.92±0.33) and (2.07±0.37) score, respectively; at T₆, the scores were (3.30±0.73) and (3.77±0.80) score, respectively. The MOAA/S scores of treatment group at each time were all significantly lower than those of control group (all P<0.05). The intraoperative propofol dosage in treatment group and control group were (381.81±46.48) and (407.25±49.36) mg, respectively; the intraoperative remifentanil dosage were (14.43±1.82) and (15.14±2.00) μg, respectively; the incidence of postoperative delirium at 72 h were 15.87% and 33.33%, respectively; the postoperative 24 h central nervous system-specific protein (S100β) levels were (0.59±0.11) and (0.66±0.14) μg·mL^-1, respectively; the postoperative 24 h glial fibrillary acidic protein (GFAP) levels were (0.87±0.29) and (1.02±0.22) ng·mL^-1, respectively; the postoperative 8 h 5-hydroxytryptamine (5-HT) levels were (168.91±18.25) and (159.35±17.54) μg·L^-1, respectively, with statistically significant differences (all P<0.05). The total incidence of adverse drug reactions were 12.70% in treatment group and 22.81% in control group, respectively, with no statistically significant difference (P>0.05).

Continuous infusion of extremely low-dose dexmedetomidine hydrochloride injection combined with remifentanil hydrochloride for injection and propofol emulsion injection in elderly patients undergoing THA can effectively improve intraoperative vital signs and postoperative recovery quality, enhance analgesia and sedation, reduce the dosage of remifentanil hydrochloride for injection and propofol emulsion injection, reduce the incidence of postoperative delirium, regulate the levels of neuronal injury markers and neurotransmitter markers, and has good safety.

To investigate the efficacy and safety of anlotinib combined with neoadjuvant chemotherapy (NAC) in elderly patients with triple-negative breast cancer (TNBC).

Elderly patients with TNBC who underwent NAC followed by modified radical mastectomy were enrolled and divided into control group and treatment group based on their treatment regimens. Control group received the TEC regimen as preoperative NAC, while treatment group received additional oral administration of anlotinib hydrochloride capsules (12 mg once daily, in a 2-weeks-on/1-week-off cycle) for 4 cycles. Carbohydrate antigen 153 (CA153), carcinoembryonic antigen (CEA), growth differentiation factor-3 (GdF3), carbohydrate antigen 19-9, Ki-67 positive rate, cyclooxygenase-2 (COX2), vascular endothelial growth factor (VGEF), bone sialoprotein, prolactin, insulin-like growth factor-1, clinical efficacy and long-term survival outcomes were compared between the two groups, and the safety was conducted.

A total of 93 patients were enrolled and divided into control group (n=48) and treatment group (n=45). After treatment, the objective response rates (ORR) were 43.75% (21 cases/48 cases) in control group and 64.44% (29 cases /45 cases) in treatment group, respectively; while the disease control rates (DCR) were 70.83% (34 cases /48 cases) and 88.89% (40 cases /45 cases), respectively; CA 153 levels were (24.59±3.57) and (22.21±3.86) μ·mL^-1, respectively; GdF3 levels were (117.53±18.29) and (102.24±15.47) ng·L^-1, respectively; CEA levels were (4.19±0.85) and (3.72±0.64) ng·L^-1, respectively; the Ki-67 positivity rates were 37.50% and 17.78%, respectively; COX-2 levels were (88.73±21.64) and (74.38±19.51) ng·L^-1, respectively; VEGF levels were (38.23±7.15) and (34.74±6.32) ng·L^-1, respectively; the progression-free survival (PFS) rates were 33.33% (16 cases/48 cases) and 46.67% (21 cases/45 cases), respectively. Statistically significant differences were observed in all the above indicators between the two groups (all P<0.05, P<0.01, P<0.001). The adverse drug reactions during chemotherapy in treatment group and control group included anemia, decreased appetite, neutropenia, leukopenia, thrombocytopenia, headache, hypertension, nausea and vomiting, fatigue, elevated thyroid hormones, diarrhea, constipation, infection, alopecia, impaired liver and kidney function, and hand-foot syndrome. The total incidence of adverse drug reactions was 64.58% (31 cases/48 cases) in control group and 71.11% (32 cases /45 cases) in treatment group, with no statistically significant difference between the groups (P>0.05). The overall survival (OS) rate was 60.42% (29 cases /48 cases) in control group and 68.89% (31 cases /45 cases) in treatment group, and the difference was not statistically significant (P>0.05).

Anlotinib combined with NAC regimen for elderly TNBC patients shows good efficacy, can improve Ki-67 positive rate and serum COX-2 and vascular endothelial growth factor levels, improve long-term survival, and has high 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.

To investigate the effect of total flavonoids from Patrinia on neuroprotection in rats with ischemic stroke (IS) by Rat sarcoma virus oncogene homolog (Ras) homolog gene family member (Rho) A/Rho-associated kinase (ROCK) signaling pathway.

The IS rat model was established by the suture method, and was randomly divided into B group (IS), C group (total senna flavonoids low dose), D group (total senna flavonoids high dose), E group (positive drug), F group (high dose of total senna flavonoids+U-46619F) and A group (the sham operation), where group A and group B were given intragastric and intraperitoneal injections of normal saline; group C was given 100 mg·kg^-1 total senna flavonoids by intragastric administration; group D was given 200 mg·kg^-1 total senna flavonoids by intragastric administration; group E was given 1.0 mg·kg^-1 nimodipine by intraperitoneal injection; group F was given intragastric administration of 200 mg·kg^-1 total senna flavonoids and 30 μg·kg^-1 U-46619 by intraperitoneal injection; each group was administered once a day for two consecutive weeks. After the intervention, the modified neurological severity (mNSS) score was used to evaluate the nerve injury; the triphenyltetrazolium chloride staining (TTC) was used to detect the ratio of cerebral infarction area; the brain tissue was separated, and hematoxylin-eosin was used for pathological changes; the expression of nerve growth factor (NGF) was detected by immunohistochemistry; the apoptosis changes of striatal dopaminergic neurons were detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling method (TUNEL); the relative expression levels of Bcl-2 related X protein (Bax), RhoA, ROCK and B-cell lymphoma factor 2 (Bcl-2) proteins were detected by immunoblotting.

The nerve injury scores for groups A, B, D, E, and F were (0.15±0), (11.24±1.26), (4.06±0.42), (3.52±0.38) and (7.18±0.73) points, respectively; the ratio of infarct area were (0±0)%, (27.86±2.81)%, (11.57±1.27)%, (12.07±1.24)% and (18.07±1.88)%; the relative expression levels of NGF were 34.11±3.51, 15.86±1.61, 32.07±3.28, 32.11±3.32 and 22.87±2.31; the apoptosis of dopaminergic neurons were (6.57±0.68)%, (27.81±2.82)%, (12.75±1.31)%, (12.52±1.27)% and (20.47±2.11)%; the aforementioned indicators of group B were compared with those of group A; those of groups C and D were compared with group B; and those of group F were compared with group D, all showing statistically significant differences (all P<0.05).

The total flavonoids of Cynanchum paniculatum have neuroprotective effects on IS rats, and the mechanism may be related to the activation of the RhoA/ROCK signaling pathway.

To explore the main active components, potential targets and mechanisms of Yinpu Jiedu tablets in the treatment of diabetic renal inflammation by using network pharmacological methods and molecular docking techniques.

The active ingredients of Yinpu jiede tables and therapeutic targets for diabetic kidney inflammation were retrieved from databases including TCMSP, PubChem, SwissTargetPrediction and GeneCards. A protein-protein interaction (PPI) network was constructed using Venny, STRING, Metascape, Bioinformatics and Cytoscape, followed by gene enrichment analysis. Molecular docking was performed using AutoDock, and the results were visualized with PyMOL. Furthermore, a lipopolysaccharide (LPS)-induced cellular model of diabetic kidney inflammation was established. The experiment included control group, model group (4 μmol·L^-1 LPS) and experimantol group (30 μg·mL^-1 Yinpu jiede tables). Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was used to measure the messenger ribonucleic acid (mRNA) relative expression levels of AKT1 serine/threonine kinase 1(Akt1), tumor protein p53(Tp53), tumor necrosis factor-α (Tnf-α) and interleukin-6 (IL-6) to validate the network pharmacology predictions.

A total of 30 active ingredients and 435 potential targets of Yinpu jiede tables were identified. The core active ingredients included quercetin, luteolin, acacetin, morin and pelargonin. Intersection analysis between disease-related targets and drug-targets yielded 71 key genes, from which 4 core targets, inducling AKT1, TNF-α, IL-6 and TP53. Molecular docking results indicated strong binding affinity between the core active ingredients and the core targets. gene ontolog (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses suggested that Yinpu jiede tables exerts its therapeutic effects on diabetic kidney inflammation primarily through pathways such as pathways in cancer, the JAK-STAT signaling pathway, the AMPK signaling pathway and the Wnt signaling pathway. Cellular experiments confirmed that LPS induction significantly upregulated the mRNA expression of Akt1, Tp53, TNF-α, and IL-6 in podocytes, while treatment with Yinpu jiede tables effectively reversed this trend, restoring their expression to near-normal levels, thereby validating its regulatory role on the predicted targets and inflammatory pathways.

Yinpu jiede tables may act on signal pathways such as JAK-STAT, AMPK, and Wnt through core targets such as AKT1, TNF, IL-6, and TP53, and exert therapeutic effects on diabetic kidney inflammation.

To explore the effect of Gallic acid on liver tissue damage in autoimmune hepatitis (AIH) mice by adjusting adenosine monophosphate-activated protein kinase (AMPK) / acetyl-coa carboxylase (ACC) signaling pathway.

An AIH mouse model was established by injecting 15 mg·kg^-1 concanavalin A through the tail vein. The mice were randomly divided into model group, low-dose experimental group (50 mg·kg^-1), high-dose experimental group (100 mg·kg^-1), AMPK inhibitor group (5 μmol·L^-1), gallic acid + AMPK inhibitor group and normal mice group with the same volume of phosphate-buffered solution as control group. After the intervention, the automatic biochemical analyzer was used to analyze glutamic pyruvic transaminase (GPT), the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in liver homogenate were detected by enzyme-linked immunosorbent assay (ELISA), and the levels of interleukin (IL)-10, IL-6 and tumor necrosis factor (TNF-α) in serum supernatant were also measured. The histopathological changes were detected by the hematoxylin-eosin (HE) staining method; the apoptosis status was examined by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method. The relative expression levels of phosphorylated (p-) AMPK/AMPK, Bcl-2 related X protein (Bax), B-cell lymphoma/leukemia-2 protein (Bcl-2) and ACC protein were detected by Western blotting.

The glutamic-oxalacetic transaminase (GOT) levels in control group, model group, low-dose experimental group, high-dose experimental group, AMPK inhibitor group and high-dose experimental+AMPK inhibitor group were (82.05±8.31), (210.20±21.14), (135.26±14.64), (91.02±9.21), (308.15±30.85) and (129.84±13.08) U·L^-1, respectively; the GPT levels were (50.23±5.11), (324.85±33.25), (152.34±15.48), (88.64±8.95), (482.62±50.02) and (148.85±15.02) U·L^-1, respectively; the TNF-α levels were (41.06±4.37), (149.85±15.28), (88.39±8.97), (51.06±5.21), (267.85±27.18) and (154.62±15.68) pg·mL^-1, respectively; the IL-6 levels were (48.42±4.92), (134.52±13.58), (75.42±7.61), (57.85±5.81), (215.84±21.98) and (78.62±7.98) pg·mL^-1, respectively; the IL-10 levels were (486.52±49.02), (188.72±19.08), (318.54±32.08), (445.67±45.11), (114.21±11.94) and (324.15±32.67) pg·mL^-1, respectively; the SOD levels were (48.26±4.99), (19.52±2.11), (32.25±3.44), (44.21±4.57), (11.37±1.21) and (31.24±3.22) U·mg^-1, respectively; the MDA levels were (0.28±0.03), (0.84±0.09), (0.56±0.06), (0.31±0.04), (1.31±0.15) and (0.64±0.07) nmol·mg^-1, respectively; the tissue pathological scores were 0, 2.64±0.27, 2.27±0.24, 1.22±0.14, 3.54±0.36 and 2.19±0.23, respectively; the apoptosis rates were (5.24±0.54)%, (23.62±2.44)%, (16.42±1.72)%, (8.52±0.89)%, (34.85±3.61)% and (17.06±1.75)%, respectively. The above indexes of model group were compared with those of control group; low-dose and high-dose experimental group were compared with those of model group and high-dose experimental+AMPK inhibitor group were compared with those of high-dose experimental group, there were statistically significant differences (all P<0.05).

Gallic acid can inhibit the inflammatory response and oxidative stress level in AIH mice, reduce cell apoptosis, improve liver function, alleviate liver tissue pathological damage, down-regulate Bax and ACC expression, up-regulate p-AMPK/AMPK and Bcl-2 expression; the AMPK inhibitor reverses the protective effect of high-dose gallic acid on AIH mice. Gallic acid activates the AMPK/ACC signaling pathway to alleviate liver tissue damage in AIH mice.

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 molecular mechanism of losartan in alleviating renal injury in sepsis.

In animal experiment, mice were randomly divided into the sham group, the model group (modeled by cecal ligation and perforation), losartan group (losartan treatment at 15 mg·kg^-1 after modeling) and antagonist group [treated with the G protein-coupled receptor 84 (GPR84) antagonist GLPG1205 at 30 mg·kg^-1 after modeling], each group contained 10 mice. Enzyme-linked immunosorbent assay (ELISA) experiments were used to detect serum renal injury indicators, and immunohistochemistry was used to detect the protein expressions of GPR84 and cluster of differentiation 86 (CD86) in renal tissues. The expression of cytokines in renal tissues was detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). In cell experiment, THP-1 cells were randomly divided into control group, induce group (induced to M1-type macrophages), medicine group (inducing +10 μg·mL^-1 losartan) and medicine+6-OAU group (induction +10 μg·mL^-1 losartan +1 μmol·L^-1 GPR84 antagonist 6-OAU), the relative expression levels of GPR84 protein was detected by Western blot assay, the relative expression level of CD86 was detected by immunofluorescence assay, and the expression of cytokines in cells was detected by RT-qPCR.

In animal experiments, the levels of blood urea nitrogen (BUN) in mice of the sham group, model group, losartan group and antagonist group were (48.55±4.54), (111.28±5.83), (71.52±4.79) and (79.10±8.12) mmol·L^-1, respectively; serum creatinine (SCr) were (13.33±1.58),(64.12±6.91), (30.43±5.80) and (40.09±4.49) μmol·L^-1, respectively; the average optical density values of GPR84 were 0.05±0.01, 0.08±0.01, 0.05±0.01 and 0.05±0.01, respectively; the average optical density values of CD86 were 0.03±0, 0.08±0.01, 0.05±0.01, 0.06±0.01, respectively; the relative expression levels of GPR84 mRNA were 1.00±0.13, 2.70±0.23, 1.78±0.15 and 1.12±0.09, respectively; interleukin (IL)-1β mRNA were 1.00±0.14, 1.78±0.24, 1.41±0.13 and 1.50±0.19, respectively; tumor necrosis factor-alpha (TNF-α) mRNA were 1.00±0.10, 1.85±0.12, 1.39±0.10 and 1.56±0.12, respectively; the relative expression levels of inducible nitric oxide synthase (iNOS) mRNA were 1.00±0.14, 3.33±0.23, 1.93±0.16 and 2.30±0.24, respectively. Compared between the model group and the sham group; compared the losartan group or the contagonist group with the model group and compared between the losartan group with antagonist group, the differences of the above-mentioned indicators were statistically significant (P<0.05, P<0.01, P<0.001). In cell experiments, the relative expression levels of GPR84 protein in the control group, induce group, medicine group and medicine+6-OAU group were 0.58±0.06, 0.94±0.11, 0.63±0.07 and 1.45±0.09, respectively; the fluorescence intensities of CD86 were (1 072.36±105.92), (3 209.59±206.38), (1 903.95±172.73) and (2 471.43±273.18) a.u.; the relative expression levels of iNOS mRNA were 1.00±0.11, 3.06±0.23, 1.98±0.27 and 2.85±0.24, respectively, the above-mentioned indicators compared between the control group and the induce group, compared between the medicine group and the induce group, and compared between the medicine+6-OAU group and the medicine group, the differences were all statistically significant (all P<0.001).

Losartan can alleviate renal injury in sepsis, which is related to the inhibition of GPR84-mediated M1 polarization of macrophages.

To explore the effects of geniposide on insulin resistance and oxidative stress damage in polycystic ovary syndrome (PCOS) rats by regulating the nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase-1(HO-1) signaling pathway.

A PCOS model was induced by gavage of letrozole for 21d and rats were randomly assigned into blank group (not modeled +normal saline), model group (modeled +normal saline), low-dose geniposide group (modeled+400 mg·kg^-1 geniposide by gavage), high-dose geniposide group (modeled+800 mg·kg^-1 geniposide by gavage), positive control group (modeled+0.34 mg·kg^-1 ethinylestradiol and cyproterone by gavage) and high-dose geniposide+ML385 group (modeled+800 mg·kg^-1 geniposide by gavage+30 mg·kg^-1 ML385 by by intraperitoneal injection), with 10 rats in each group. The levels of fasting blood glucose (FBG) and fasting insulin (FINS) were measured by blood glucose meter and enzyme-linked immunosorbent assay (ELISA) and then homeostatic model assessment of insulin resistance (HOMA-IR) was calculated. The levels of serum sex hormones and inflammatory were detected by ELISA and ovarian tissue oxidative stress index levels were detected by test kit, hematoxylin-eosin (HE) staining was used to measure the number of cystic follicles in rat ovarian tissue. TdT-media duTP nict end labeling (TUNEL) method was used to measure apoptosis of rat ovarian granulosa cells. Moreover, Western blot (WB) was used to detect apoptosis and Nrf2/HO-1 signaling pathway related protein expression in ovarian tissues.

The FBG levels in the low-dose geniposide group, high-dose geniposide group, positive control group, high-dose geniposide+ML385 group and control group, as well as the model group were (6.87±0.42), (5.21±0.28), (5.03±0.31), (8.32±0.51), (4.90±0.35) and (8.56±0.54) mmol·L^-1, respectively; FINS were (16.13±1.62), (10.46±1.37), (10.21±1.51), (20.67±1.82), (9.82±1.14) and (21.45±2.05) mIU·L^-1, respectively; HOMA-IR were 4.93±0.48、2.42±0.53、2.28±0.36、7.64±0.72、2.14±0.31 and 8.16±0.67, respectively; follicle-stimulating hormone (FSH) levels were (5.14±0.56), (7.49±0.62), (7.62±0.71), (2.95±0.54), (7.83±0.76) and (2.71±0.43) IU·L^-1, respectively; the superoxide dismutase (SOD) levels were (42.91±3.22), (56.45±4.16), (58.02±3.98), (32.16±2.85), (59.73±4.35) and (30.84±2.74) U·mg prot^-1, respectively. The relative protein expression levels of Nrf2 were 0.67±0.08, 1.10±0.12, 1.12±0.13, 0.26±0.07, 1.13±0.14 and 0.24±0.06, respectively; HO-1 were 0.61±0.07, 1.06±0.08, 1.08±0.10, 0.18±0.05, 1.09±0.11 and 0.16±0.04, respectively. The malondialdehyde (MDA) levels were (2.47±0.33), (1.39±0.21), (1.30±0.19), (3.38±0.42), (1.21±0.26) and (3.62±0.47) nmol·mg prot^-1, respectively. The number of ovarian antral follicles was 9.60±0.74, 4.30±0.57, 4.10±0.71, 14.20±0.98, 3.90±0.65 and 15.10±1.13, respectively. The ovarian granulosa cell apoptosis index was (16.27±1.16)%, (7.48±0.91)%, (7.19±0.76)%, (23.86±1.23)%, (6.94±0.85)% and (25.31±1.42)%, respectively. Compared blank group with the control group; compared model group with low-dose geniposide group, high-dose geniposide group and positive control group; compared low-dose geniposide group with high-dose geniposide group and compred high-dose geniposide group with high-dose geniposide+ML385 group, the differences of above indexes were all statistically significant (all P<0.05).

Geniposide can alleviate insulin resistance and oxidative stress damage in PCOS rats, possibly by activating the Nrf2/HO-1 signaling pathway.

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 explore the expression of thrombospondin-1 (THBS1) in pan-cancer and its relevance to tumor microenvironment (TME).

The discrepancies in the expression and prognostic significance of THBS1 were examined in 33 different types of tumors using TCGA and GTEx data. The correlation between THBS1 and the TME index, various factors like immune cell infiltration, tumor mutagenesis burden (TMB) and microsatellite instability(MSI) was evaluated using computational methods. The potential mechanism was also dissected by constructing interaction networks of THBS1 with other genes by GEPIA; this was validated using multiplex immunohistochemistry(mIHC) in pancreatic cancer (PAAD) tissues.

Pan-cancer analysis revealed THBS1 mRNA was differentially expressed in 17 common cancer types (all P<0.05) and differential expression of the THBS1 protein was observed in 9 cancer types (all P<0.001). Survival analysis indicated that higher THBS1 expression was significantly associated with poorer patient prognosis in various malignant tumors (P<0.05). THBS1 expression was statistically significantly correlated with cancer-associated fibroblast (CAFs) infiltration across all cancer types (P<0.05), and positively associated with the expression of multiple immune checkpoint genes, including CD274(P<0.001)、HAVCR2(P<0.001)、PDCD1LG2(P<0.001)、LAG3(P<0.001)、PDCD1(P<0.001)、CTLA4(P<0.001) and TIGIT(P<0.001). These findings suggested that THBS1 may contribute to reduced responsiveness to immunotherapy by shaping an immunosuppressive tumor microenvironment. Co-expression and enrichment analyses demonstrated that the integrin genes strongly correlated with THBS1 in pancreatic adenocarcinoma (PAAD) were ITGAV (r=0.60, P<0.001)、ITGB3 (r=0.54, P<0.001) and ITGB1 (r=0.69, P<0.001). These findings were validated in pancreatic cancer tissues: immunofluorescence co-localization analysis demonstrated a strong positive correlation between THBS1 expression and integrin ITGAV (r=0.75, P<0.001).

This study identifies THBS1 as a crucial pan-cancer prognostic biomarker and a key regulator of the TME, providing a compelling rationale for its potential as a therapeutic target in oncology.

To investigate the inhibitory effect of different doses of metformin on colorectal cancer (CRC) and its mechanism involving the regulation of bile acid metabolism and the cyclooxygenase-2 (COX-2)/prostaglandin E2 (PGE2) pathway.

A total of 60 BALB/c mice were divided into control, model, low-dose metformin, medium-dose metformin and high-dose metformin groups. A CRC model was induced using azoxymethane (AOM)/dextran sodium sulfate (DSS). Tumor number and diameter were measured with a caliper; fecal bile acid levels were detected by liquid chromatography-mass spectrometry (LC-MS); and protein expression of COX-2 and PGE2 in colon tissue was determined by immunohistochemistry.

Tumor numbers in the control, model, low-, medium- and high-dose groups were 0, 11.58±7.13, 7.83±3.38, 3.33±2.06 and 2.83±1.40, respectively; tumor diameters were 0, (2.91±0.32), (2.43±0.31), (2.11±0.37) and (1.98±0.38) mm, respectively; total bile acid concentrations were (20 989.00±306.69), (43 256.50±187.08), (32 265.26±325.08), (23 035.72±136.33) and (22 351.51±145.36) ng·g^-1, respectively; deoxycholic acid (DCA) concentrations were (3 160.14±113.31), (16 684.58±4.39), (11 544.47±7.86), (5 472.60±25.03) and (5 201.36±79.95) ng·g^-1, respectively; lithocholic acid (LCA) concentrations were (2 140.43±76.23), (14 452.6±47.82), (9 085.87±11.14), (3 145.09±54.20) and (2 955.51±38.02) ng·g^-1, respectively; cholic acid (CA) concentrations were (6 917.71±51.12), (2 051.41±88.47), (3 181.64±23.79), (5 564.01±35.45) and (5 866.10±63.80) ng·g^-1, respectively; chenodeoxycholic acid (CDCA) concentrations were (5 720.33±53.07), (1 135.43±48.38), (1 530.56±37.57), (4 060.14±37.09) and (4 542.83±27.39) ng·g^-1, respectively; taurocholic acid (TCA) concentrations were (497.24±59.29), (1 560.15±26.03), (958.13±28.00), (705.21±3.66) and (669.24±33.00) ng·g^-1, respectively; taurochenodeoxycholic acid (TCDCA) concentrations were (1 027.96±84.28), (2 918.60±122.90), (2 518.23±70.50), (1 411.36±49.15) and (1 389.87±5.80) ng·g^-1, respectively; COX-2 staining scores in the model, low-, medium- and high-dose groups were (5.75±0.97), (4.50±1.78), (2.75±1.66) and (2.67±0.89) points, respectively; PGE2 staining scores were (5.75±0.97), (4.50±1.93), (2.67±1.67) and (2.50±0.90) points, respectively; compared with the control group, the aforementioned indicators in the model group showed statistically significant differences; compared with the model group, the indicators in the low-, medium- and high-dose groups showed statistically significant differences (all P<0.05). The medium- and high-dose groups were significantly more effective than the low-dose group in reducing tumor number and diameter and regulating bile acid metabolism (P<0.05). The inhibition of COX-2/PGE2 expression was negatively correlated with the dose of metformin.

Metformin exerts anti-CRC effects through dose-dependent regulation of bile acid metabolism (reducing DCA, LCA, TCA, TCDCA and restoring CA and CDCA levels) and inhibition of the COX-2/PGE2 signaling pathway. The medium- and high-dose groups (250-500 mg·kg^-1·d^-1) show more significant effects.

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.

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 investigate and analyze the current state of professional training for clinical research coordinators (CRCs), explore the differences in CRC cultivation between clinical trial institutions and site management organizations (SMOs), and provide references for optimizing the CRC training system.

A total of 613 active CRCs was conducted using convenience sampling method and quota sampling method. The questionnaire includes basic information, induction teaching methods, training frequency, training content, training satisfaction and training needs, etc. Data were collected by the ‘questionnaire star’, statistically analyzed using SAS 9.3 software, and differences were compared using the chi-square test.

There were significant differences between institutions and SMO companies in the education, professional background and working years of CRC (All P<0.05). Institutional CRC generally has a higher degree and more nursing background, while SMO company CRC contains a certain proportion of people with non-medical background. In terms of training, institutional CRC prefers moderate frequency training (6 to 12 times per year), while SMO CRC prefers more frequent high frequency training (more than 12 times per year). SMO company has a high utilization rate of online training, and institutional CRC has some deficiencies in communication skills training. Overall, the CRC is relatively satisfied with the existing training system.

Institutions and SMO companies should develop reasonable training plans according to their own characteristics and reduce unnecessary training burden to improve the work efficiency and clinical trial quality of CRC. It is recommended to adopt more offline training forms, and to conduct targeted training according to the individual needs of CRC.