To investigate the effect and of hydrogen sulfide (H₂S) on the osteogenic differentiation of osteoblast precursor cells (MC3T3-E1) by regulating the polarization of macrophages (RAW264.7).

RAW264.7 cells were divided into four groups (n=10 in each group), including control group (untreated), M1 induction group [added with 100 ng·mL^-1 lipopolysaccharide (LPS)], M2 induction group (added with 20 ng·mL^-1 interleukin-4+20 ng·mL^-1 interleukin-10, IL-4+IL-10), GYY4137 intervention group (H₂S donor GYY4137 100 μmol·L^-1). Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to detect the mRNA and protein expression levels of M1 macrophage related factors including tumor necrosis factor-α (TNF-α), interleukin-1 β (IL-1β) and inducible nitric oxide synthase (i-NOS) and M2 macrophage related factors including interleukin-10 (IL-10), arginase-1 (Arg-1) and mannose receptor (CD206), and to verify the effect of polarization induction and the intervention of GYY4137; cell counting kit-8 (CCK-8) was used to detect the activity of RAW264.7 cells treated with GYY4137 at different concentrations for 24 h and 72 h (6 wells in each group). The expression levels of Arg-1 and CD206 in control group and GYY4137 group (100 μmol·L^-1) were detected by flow cytometry (n=10 in each group). MC3T3-E1 cells were divided into four groups (n=10), which were blank group (without osteogenic induction medium), osteogenic control group (with osteogenic induction medium), co-culture group (co-cultured with RAW264.7 cells Transwell) and GYY4137 pretreatment co-culture group (co-cultured with 100 μmol·L^-1 GYY4137 pretreated RAW264.7 cells). The mRNA expression levels of osteogenic related genes including Runt-related transcription factor 2 (Runx-2), osteopontin (OPN), bone morphogenetic protein-2 (BMP-2) and bone sialoprotein (BSP) mRNA were detected by RT-qPCR; alkaline phosphatase (ALP) staining and alizarin red staining were used to detect osteogenic differentiation.

The relative expression levels of TNF-α mRNA in control group, M1 induction group and GYY4137 intervention group were 1.56±0.22, 16.49±3.56 and 0.57±0.11, respectively; the relative expression levels of IL-1β mRNA were 1.06±0.24, 3.37±0.76 and 1.43±0.34, respectively. The above indexes in M1 induced group were significantly higher than those in control group, and the above indexes in GYY4137 intervention group were significantly lower than those in M1 induced group (all P<0.001). The relative expression levels of Arg-1 mRNA in GYY4137 group and control group were 1.38±0.36 and 0.91±0.12, respectively; the relative expression levels of CD206 mRNA were 17.43±2.84 and 1.14±0.21, respectively; the relative expression levels of CD206 protein were 0.89±0.25 and 0.20±0.04, respectively; the relative expression levels of i-NOS protein were 0.72±0.19 and 0.25±0.07, respectively; the relative expression levels of Arg-1 were 28.65±3.21 and 12.31±2.15, respectively; the relative expression levels of CD206 were 31.24±2.89 and 10.8±1.98, respectively. The above indexes in GYY4137 intervention group were significantly higher than those in control group, and the differences were statistically significant (P<0.05, P<0.001). The relative expression levels of Runx-2 mRNA in GYY4137 pretreatment and co-culture groups were 3.21±0.45 and 1.56±0.22, respectively; the relative expression levels of OPN mRNA were 2.89±0.38 and 1.43±0.19, respectively; the relative expression levels of BMP-2 mRNA were 3.56±0.52 and 1.67±0.25, respectively; the relative expression levels of BSP mRNA were 2.98±0.41 and 1.51±0.20, respectively. The above indexes in GYY4137 pretreatment co-culture group were significantly higher than those in co-culture group, and the differences were statistically significant (all P<0.05). The OD values of ALP in GYY4137 pretreatment co-culture group and osteogenic control group were (0.89±0.12) and (0.56±0.08) μm², respectively; the areas of alizarin red stained calcium nodules were (2 865.32±215.67) and (1 568.45±189.32) μm², respectively; the above indexes of GYY4137 pretreatment co-culture group were significantly higher than those of osteogenic control group, and the differences were statistically significant (all P<0.05).

100 μmol·L^-1 GYY4137 can enhance the osteogenic differentiation of MC3T3-E1 cells by promoting the polarization of RAW264.7 cells to M2 macrophages, up regulating the expression of osteogenic related genes.

To investigate the effects of baicalein combined with 5-fluorouracil (5-FU) on the proliferation, apoptosis and migration of H446 cells, and to explore the sensitization mechanism.

Lung cancer H466 cells were allocated into blank, control, experimental and combination groups. The blank group was treated with 10 μmol·L^-1 dimethyl sulfoxide, the control group with 20 μmol·L^-1 baicalein, the experimental group with 20 μmol·L^-1 5-FU, and the combination group with 10 μmol·L^-1 baicalein plus 10 μmol·L^-1 5-FU. Cell viability was determined using the cell counting kit-8 assay. Apoptosis and reactive oxygen species (ROS) levels were analyzed by flow cytometry. Cell migration was evaluated with Transwell assays, and alterations in mitogen-activated protein kinase (MAPK) pathway protein expression were examined by Western blotting.

After 24 h of incubation, cell viability in blank, control, experimental and combination groups were (114.34±2.88)%, (79.87±3.48)%, (57.66±1.82)% and (40.31±2.73)%, respectively; the corresponding apoptosis rates were (14.47±1.36)%, (33.94±4.33)%, (49.20±3.82)% and (49.17±2.13)%; and cell migration rates were (8.43±1.94)%, (11.25±1.55)%, (26.74±3.31)% and (68.83±4.32)%. Compared with control group, the above indexes of experimental group and combined group were significantly increased, and combined group were significantly higher than experimental group (all P<0.01). After adding ROS scavenger NAC to combination group, the apoptosis rates of blank group, control group, experimental group, combination group and combination group+NAC were (7.82±0.94)%, (12.31±2.13)%, (33.48±4.92)%, (52.42±3.17)% and (30.39±2.72)%, respectively; the cell migration rates were (4.36±0.54)%, (4.92±0.71)%, (31.43±3.14)%, (52.64±5.26)% and (27.61±3.52)%, respectively. Compared with combined group, the above indexes of combined group+NAC were significantly decreased (all P<0.01). The relative expression levels of p-JNK protein in blank, experimental, control, combination and combination group+NAC were 0.78±0.05, 0.70±0.04, 0.73±0.03, 0.98±0.02 and 0.73±0.05, respectively; the relative expression levels of p-p38 were 0.56±0.02, 0.51±0.03, 0.60±0.04, 0.76±0.06 and 0.54±0.03, respectively; the relative expression levels of p-ERK were 0.34±0.03, 0.30±0.04, 0.28±0.04, 0.18±0.03 and 0.33±0.05, respectively. The relative expression levels of p-JNK and p-p38 protein in combination group were significantly higher than those in control group, and the relative expression levels of p-ERK protein were significantly lower (all P<0.01); the relative expression levels of p-JNK and p-p38 low protein in combined group+NAC were significantly decreased, while the relative expression levels of p-ERK protein were significantly increased (all P<0.01).

The combination of baicalein and 5-FU can enhance the inhibitory effect on proliferation, induce apoptosis, and suppress migration in H446 cells by upregulating ROS levels, activating the JNK and p38 pathways, and inhibiting the ERK pathway.

To explore the effects of ropivacaine on the cyclic guanosine monophosphate synthase interferon gene stimulating factor (cGAS/STING) pathway in glial cells of septic encephalopathy (SAE) model mice, and its related mechanisms of action.

Mice were randomly divided into blank group, model group, experimental group and control group, with 15 mice in each group. Except for the blank group and control group, the other two groups were used to establish sepsis mouse models by intraperitoneal injection of lipopolysaccharide. The experimental group and control group were intraperitoneally injected with 0.3 mg·kg^-1·d^-1 ropivacaine, while the blank group and model group were intraperitoneally injected with an equal volume of 0.9% NaCl. The four groups of mice were administered once a day for 7 consecutive days. The Morris water maze was used to detect the cognitive function of the mouse brain, real-time fluorescent quantitative polymerase chain reaction was used to detect the mRNA levels of brain inflammatory factors, Western blot was used to detect the expression levels of brain cGAS and STING proteins, and immunofluorescence staining was used to detect the expression of apoptosis-related proteins.

The exploration time proportions of the blank group, model group, experimental group and control group were (0.48±0.10)%, (0.23±0.05)%, (0.37±0.04)% and (0.46±0.07)%, respectively; the platform crossing times were (4.67±0.48), (1.03±0.11), (2.50±0.65) and (4.99±0.52) times, respectively; the escape latency time was (11.92±0.98), (12.49±1.22), (11.17±1.43) and (12.21±1.52) min, respectively; the relative expression levels of interleukin-1β (IL-1β) mRNA were 1.00±0.10, 3.80±0.28, 1.87±0.13 and 1.06±0.29, respectively; the relative expression levels of IL-18 mRNA were 1.00±0.12, 1.61±0.27, 1.06±0.08 and 0.97±0.05, respectively; the relative expression levels of tumor necrosis factor (TNF)-α mRNA were 1.00±0.13, 2.02±0.18, 1.05±0.09 and 1.13±0.11, respectively; the relative expression levels of IL-6 mRNA were 1.00±0.08, 1.56±0.11, 1.05±0.09 and 0.93±0.06, respectively; the relative expression levels of cGAS protein were 1.00±0.08, 1.92±0.19, 1.42±0.11 and 1.03±0.10, respectively; the relative expression levels of STING protein were 1.00±0.12, 2.01±0.22, 1.18±0.09 and 1.08±0.16, respectively; the relative fluorescence intensities of cGAS protein were 1.00±0.18, 1.88±0.15, 1.57±0.09 and 1.03±0.11, respectively; the relative fluorescence intensities of STING were 1.00±0.16, 1.79±0.26, 1.14±0.06 and 1.20±0.29, respectively; the relative fluorescence intensities of caspase-3 protein were 1.00±0.12, 1.51±0.08, 1.23±0.10, and 1.11±0.15, respectively. The above indexes of the experimental group and the control group were statistically different from the model group (all P<0.05).

Ropivacaine can alleviate apoptosis of glial cells in the hippocampus of SAE mice by inhibiting the cGAS/STING pathway, and has a protective effect on the brain of SAE mice.

To explore the antidepressant effect and its mechanism of effective components of Lilium brownii based on brain-derived neurotrophic factor/protein kinase B/mammalian target of rapamycin (BDNF/AKT/mTOR) pathway.

In cellular experiments, the experiments were divided into control group (cell culture solution), model group (corticosterone 400 μmol·L^-1), lily saponin low dose group (corticosterone 400 μmol·L^-1+0.2 mg·L^-1 liliy saponin), lily saponin high dose group (corticosterone 400 μmol·L^-1+0.6 mg·L^-1 liliy saponin), lily polysaccharide low dose group (corticosterone 400 μmol·L^-1+0.2 mg·L^-1 liliy polysaccharide), and lily polysaccharide high dose group (corticosterone 400 μmol·L^-1+0.6 mg·L^-1 liliy polysaccharide). Except for the blank group, the remaining groups were modeled with corticosterone to establish human neuroblastoma cells (SH-SY5Y) injury, and cell survival was detected. In the animal experiments, 48 male ICR mice were randomly divided into 6 groups: model group (drinking water with the same amount), low dose of lily saponin (50 mg·kg^-1), high dose of lily saponin (100 mg·kg^-1), low dose of lily polysaccharide (50 mg·kg^-1), high dose of lily polysaccharide (100 mg·kg^-1) and positive drug control group (fluoxetine,10 mg·kg^-1). Each group was given acute stress modeling in mice, and changes in behavioral indexes were observed. Serum corticosterone levels were measured by enzyme-linked immunosorbent assay. The relative expression levels of proteins such as postsynaptic densifier 95 (PSD95) and synapsin in the prefrontal cortex of mice were detected by protein immunoblot analysis.

In the cellular experiments, the cell survival rates of each subgroup were (71.46±1.92)%, (95.97±1.11)%, (105.51±1.80)%, (107.48±0.27)% and (107.66±3.61)%, respectively. The differences between the administered groups and the model group were all statistically significant (all P<0.01). In the animal experiments, the cumulative immobilization time of mice with suspended tails in model group, low and high dose of lily saponin groups, low and high dose of lily polysaccharide group and positive drug group was (122.75±4.77), (90.75±2.09), (66.13±3.23), (79.75±2.84), (54.38±4.40), and (42.00±6.34) s, respectively; the levels of corticosterone in the serum of mice were (236.94±6.17), (213.88±3.95), (165.52±2.19), (203.40±3.27), (170.18±2.18) and (104.96±3.87) ng·mL^-1, respectively; the relative expression levels of brain-derived neurotrophic factor (BDNF) were 0.26±0.03, 0.74±0.25, 0.86±0.03, 0.97±0.11, 0.92±0.13 and 1.05±0.23, respectively; the relative expression levels of phosphorylated protein kinase B/protein kinase B (p-AKT/AKT) were 0.26±0.03, 0.60±0.19, 0.75±0.09, 0.82±0.05, 0.94±0.23 and 0.58±0.07, respectively; the relative expression levels of phosphorylated mammalian target of rapamycin/mammalian target of rapamycin protein (p-mTOR/mTOR) were 0.46±0.04, 0.52±0.19, 0.60±0.18, 1.01±0.01, 0.98±0.06 and 0.93±0.10, respectively; the relative expression levels of postsynaptic density protein-95 (PSD95) were 0.36±0.05, 0.50±0.08, 0.60±0.09, 0.73±0.09, 0.84±0.18 and 1.00±0.12, respectively; the relative expression levels of synapsin protein were 0.13±0.04, 1.03±0.02, 1.16±0.06, 1.02±0.00, 1.00±0.04 and 0.65±0.15, respectively. Except the p-mTOR/mTOR and PSD95 in high dose of lily saponin, the differences of the above indexes between the model group and the high dose groups were all statistically significant (P<0.05, P<0.01).

Lily saponins and polysaccharides may improve synaptic plasticity in prefrontal cortex by modulating the BDNF/AKT/mTOR signaling pathway, which improves depressive-like behavior and suppresses serum corticosterone levels in behavioral despair mice. Among them, lily polysaccharides were slightly better than lily saponins in improving synaptic plasticity.

To investigate the effects of fucoxanthin on endothelial cell function in coronary heart disease (CHD) and its mechanism.

The cell experiment section divided the human coronary artery endothelial cells (HCAEC) into blank group, ox-LDL group (40 μg·mL^-1), fucoxanthin group (ox-LDL+25 μmol·L^-1 fucoxanthin) and miR-483-3p inhibitor group (transfected with miR-483-3p inhibitor+ox-LDL+25 μmol·L^-1 fucoxanthin). The expression levels of miR-483-3p and pyroptosis related proteins NLR family pyrin domain containing 3 (NLRP3) were detected by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting (WB), respectively. The animal experiment section divided the rats into control group, model group (constructed CHD model), experimental group (modeling +30 mg·kg^-1 fucoxanthin) and inhibitor group (modeling+transfection of miR-483-3p inhibitor+30 mg·kg^-1 fucoxanthin). The relative expression levels of miR-483-3p, endothelium-related factors and pyroptosis related proteins NLRP3 were detected by RT-qPCR, enzyme-linked immunosorbent assay (ELISA) and Western blot, respectively.

In cell experiment, the relative expression levels of miRNA-483-3p in cells of blank group, ox-LDL group, fucoxanthin group and miR-483-3p inhibitor group were 1.00±0.11, 0.39±0.07, 0.74±0.06 and 0.22±0.03, respectively; NLRP3 protein relative expression levels were 0.39±0.02, 1.03±0.07, 0.52±0.04 and 0.85±0.10, respectively. ox-LDL group was compared with blank group, fucoxanthin group was compared with ox-LDL group, miR-483-3p inhibitor group was compared with fucoxanthin group, the above indicators all showed statistically significant differences. (all P<0.001). In animal experiments, the relative expression levels of miR-483-3p in control group, model group, experimental group and inhibitor group were 1.00±0.12, 0.51±0.06, 0.70±0.08 and 0.31±0.04, respectively; nitric oxide (NO) levels were (46.93±3.18), (20.21±2.43), (36.13±4.87) and (24.43±1.60) μmol·L^-1, respectively; the NLRP3 protein relative expression levels were 0.29±0.04, 0.97±0.09, 0.56±0.09 and 0.86±0.08, model group was compared with control group, experimental group was compared with model group, inhibitor group was compared with experimental group, the above indicators all showed statistically significant differences. (all P<0.001).

Fucoxanthin can up-regulate miR-483-3p to improve endothelial cell pyroptosis and relieve CHD inflammatory response.

To investigate the effects of total alkaloids of Aconitum carmichaelii on myocardial function in rats and its underlying mechanisms.

SPF-grade Wistar rats were randomly assigned into five groups (n=10 each): blank group (0.9% NaCl), control group (isoproterenol 10 mg·kg^-1), experimental-L group (2.72 g·kg^-1 total alkaloids of A. carmichaelii), experimental-M group (3.68 g·kg^-1), and experimental-H group (4.98 g·kg^-1). All groups were gavaged once daily for 5 consecutive days. Thirty minutes after the last injection, T-wave amplitude on electrocardiogram was recorded. An isolated heart perfusion model was established to evaluate cardiac function and myocardial contractility. Myocardial cell activity and malondialdehyde content were measured, and real-time quantitative PCR was used to detect mRNA expression levels of peroxisome proliferator-activated receptor gamma coactivator-1 alpha (Pgc-1α), B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax).

The T-wave amplitudes for the experimental -L, -M, -H groups, blank group and control group were (0.17±0.07), (0.29±0.05), (0.37±0.04), (0.30±0.07) and (0.36±0.04) mV, respectively; the myocardial cell activities were (0.46±0.05), (0.44±0.08), (0.35±0.07), (0.45±0.06) and (0.33±0.05) Abs, respectively; the while malondialdehyde contents were (2.56±0.28), (2.55±0.26), (2.86±0.28), (2.49±0.34) and (2.83±0.25) mmol·mL^-1, respectively; the relative mRNA expression levels of Pgc-1α were 0.49±0.05, 0.41±0.07, 0.27±0.05, 0.48±0.06 and 0.35±0.05, respectively; the relative mRNA expression levels of Bax were 1.10±0.08, 1.25±0.09, 1.36±0.07, 1.09±0.09 and 1.33±0.07, respectively; the relative mRNA expression levels of Bcl-2 were 0.58±0.06, 0.51±0.05, 0.44±0.04, 0.56±0.08 and 0.45±0.03, respectively. Compared with the experimental -M, -H groups and blank group, the above indexes of the experimental -L group were statistically significant (all P<0.05).

Lower doses of Aconitum carmichaelii total alkaloids positively influence ECG T-wave amplitude, cardiac function, and myocardial contractility, improve myocardial cell activity, and reduce oxidative stress, whereas high doses exert cardiotoxic effects.

To explore the effect and potential mechanism of hyperuricemia (HUA) on acetaminophen (APAP)-induced liver injury.

Male mice were randomly divided into four groups: group A (control), group B [establishment of a hyperuricemia (HUA) model], group C (administered by oral gavage at a dose of 300 mg·kg^-1 APAP), and group D (establishment of a HUA model combined with administered by oral gavage at a dose of 300 mg·kg^-1 APAP). The HUA model was established by intraperitoneal injection of potassium oxonate (350 mg·kg^-1) and gavage of xanthine (450 mg·kg^-1) for 14 consecutive days. On day 13, mice in group C and group D received APAP via gavage. All mice were sacrificed 24 h later. Serum levels of uric acid (UA), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were determined by biochemical analyzer. Liver histopathology was evaluated by hematoxylin-eosin (H&E) staining. Levels of malondialdehyde (MDA), reactive oxygen species (ROS), glutathione (GSH), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in liver tissues were measured using commercial kits. Immunofluorescence was used to detect the relative expression of nuclear factor erythroid 2-related factor 2 (Nrf2).

The serum uric acid levels in the group A, group B, group C and group D were (186.20±15.61), (521.50±56.81), (325.20±31.74) and (682.00±49.51) μmol·L^-1, respectively; the liver coefficients were (4.51±0.14)%, (4.77±0.23)%, (4.98±0.21)%, and (5.51±0.30)%, respectively; the serum ALT levels were (55.80±15.86), (78.09±10.92), (27 688.81±7 166.01), and (34 571.89±7 793.74) U·L^-1, respectively; the AST levels were (159.90±29.72), (143.70±26.46), (10 880.00±4 291.01), and (19 824.84±7 874.64) U·L^-1, respectively; the hepatic MDA contents were (5.42±0.38), (7.77±0.32), (6.07±0.31), and (9.08±0.36) mol·g^-1·prot^-1, respectively; the ROS contents were (28.09±3.52), (39.67±2.70), (32.22±4.71), and (45.67±2.71) μg·g^-1·prot^-1, respectively; the GSH levels were (2.47±0.27), (1.72±0.16), (2.24±0.16), and (1.37±0.11) μg·g^-1·prot^-1, respectively; the IL-1β contents were (22.16±0.88), (31.74±1.82), (25.35±1.07), and (38.82±1.68) μg·g^-1·prot^-1, respectively; the TNF-α contents were (114.90±10.60), (180.20±3.72), (142.20±4.69), and (228.10±17.33) μg·g^-1·prot^-1, respectively; the relative protein expression of Nrf2 were 218.30±5.16, 63.60±29.02, 100.80±23.60, and 9.31±8.83, respectively. The indices in groups B and C were significantly different from those in group A, and the indices in group D were significantly different from those in group C, with all differences showing statistical significance (P<0.05).

In the hyperuricemia (HUA) mouse model, the liver damage induced by acetaminophen (APAP) is aggravated, and this effect may be related to the downregulation of NRF2 expression.

To establish a rat model of renal fibrosis induced by unilateral ureteral obstruction (UUO), and investigate the mechanism of the Yishen Qufeng Formula (YQF) improves renal fibrosis through regulating the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) axis to mediate ferroptosis.

Eight-week-old SD rats were randomly divided into the sham-operation group, model group, positive control group (dapagliflozin at 20 mg·kg^-1·d^-1), and experimental -L,-M,-H groups (Yishen Qufeng Formula at 15.21, 22.82 and 30.42 g·kg^-1·d^-1). Except for the sham operation group, rats in the other groups were subjected to left ureteral ligation to establish a renal interstitial fibrosis (RIF) model. After successful modeling, the positive control group and the -L, -M, and -H dose experimental groups were given the corresponding drugs by gavage, while the sham operation group and model group were given an equal volume of 0.9% NaCl by gavage. All rats in the 6 groups were administered once a day for 14 consecutive days. Serum creatinine, urea nitrogen and urine protein levels were detected using a fully automatic integrated biochemical analyzer. The content of transforming growth factor-β1 (TGF-β1) was measured by enzyme-linked immunosorbent assay (ELISA). The changes in the relative expression levels of fibrosis-related, oxidative stress-related and ferroptosis-related proteins in renal tissues were determined by Western blotting.

The levels of serum creatinine in the experimental -M, -H groups, sham operation group, model group and positive control group were (50.24±7.81), (49.15±8.87), (41.96±5.25), (59.44±11.93) and (52.14±8.38)μmol·L^-1, respectively; the levels of urea nitrogen were (4.58±0.56), (4.45±0.55), (4.32±0.60), (6.00±1.03) and (6.55±1.02)mmol·L^-1, respectively; the levels of urine protein were (640.59±205.70), (538.74±237.98), (526.51±192.62), (982.91±349.13) and (964.26±456.84)mg·d^-1, respectively; the relative protein expression levels of glutathione peroxidase 4 (GPX4) were 1.62±0.15, 3.01±0.41, 2.25±0.27, 1.01±0.51 and 2.15±0.63, respectively; the relative protein expression levels of cystine/glutamate antiporter (xCT) were 0.62±0.10, 1.02±0.06, 1.65±0.13, 0.52±0.20 and 1.18±0.13, respectively; the relative protein expression levels of Nrf2 were 0.66±0.12, 1.04±0.16, 1.20±0.10, 0.54±0.03 and 1.19±0.14, respectively; the relative protein expression levels of HO-1 were 1.13±0.15, 1.21±0.18, 2.21±0.17, 0.64±0.44 and 1.57±0.26, respectively; the relative protein expression levels of α-smooth muscle actin (α-SMA) were 1.63±0.47, 1.13±0.57, 0.49±0.91, 2.88±0.97 and 1.05±0.60, respectively. There were statistically significant differences in the above indicators between the high-dose experimental group and the model group (all P<0.05).

Yishen Qufeng Formula may inhibit ferroptosis by activating the Nrf2/HO-1 signaling pathway, thereby effectively alleviating renal interstitial fibrosis in UUO rats. This provides important theoretical evidence for the mechanism by which the Yishen Qufeng Formula improves renal fibrosis through regulation of ferroptosis.

To investigate the effect of Gardenia jasminoides Ellis (Zhi-zi) in alleviating the hepatotoxicity induced by its constituent, genipin, which is called the "self-detoxification" effect, and to explore its mechanism through targeted metabolomics and integrated network pharmacology.

Male mice of the Kunming breed were randomly divided into three groups (n=8) including blank group, control group and experimental group. Blank group was administered a blank solvent 0.5% carboxymethyl cellulose (CMC-Na) by gavage; control group was administered genipin by gavage at a dose of 100 mg·kg^-1·d^-1, and experimental group was administered 100 mg·kg^-1·d^-1 genipin and 520 mg·kg^-1·d^-1 Zhi-zi aqueous extract powder by gavage. After 28 days of continuous administration, serum biochemical indicators were measured using an automated biochemical analyzer, liver tissue pathology was examined using hematoxylin and eosin (HE) staining, and differential metabolites were quantified and analyzed using ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for targeted metabolomics analysis. The targets associated with active compounds in Zhi-zi, genipin, and liver damage were predicted separately. Finally, the results of metabolomics and network pharmacology were integrated to screen the potential targets related to the self-detoxification effect of Zhi-zi.

The alanine transaminase (ALT) levels in experimental group, control group and blank group were (35.75±8.50), (53.00±14.43) and (37.38±3.11) U·L^-1, respectively; the aspartate transaminase (AST) levels were (123.38±31.40), (146.50±19.73) and (115.75±17.44) U·L^-1, respectively. Compared with control group, experimental group showed significant decrease in ALT levels, with statistically significant difference (P<0.05). Compared with control group, the AST levels in experimental group showed no statistically significant difference (P>0.05). Targeted metabolomics identified 8 characteristic endogenous metabolites including fumaric acid, isocitric acid, maleic acid, ornithine, aminocaproic acid, isoLCA, pipecolic acid and pyrrole-2-carboxylic acid, and the metabolic pathways induced by these 8 differentially expressed metabolites were primarily associated with arginine biosynthesis and the tricarboxylic acidcycle (TCA) metabolic pathways.

Zhi-zi can improve the metabolic disorders of endogenous metabolites in the liver caused by genipin by affecting the arginine biosynthesis and the TCA metabolic pathway, and improve inflammation and oxidative stress by acting on the corresponding targets to alleviate the hepatic injuries caused by genipin and exerting the self-detoxification effect.

To establish and validate a high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method for simultaneously determining the serum concentration of lamotrigine(LTG), levetiracetam(LEV), oxcarbazepine(OXC) and monohydroxycarbazepine(MHD), which is applied for clinical blood concentration detection.

A simple one-step precipitation method was used for the serum sample pretreatment, and stable isotopes of two antiepileptic drugs (OXC-D4, LEV-D6) as the internal standard. The mobile phases were aqueous solution of 0.1% formic acid and 5 mM ammonium acetate(A) and methyl alcohol(B), and the gradient elution was performed at the speed of 0.6 mL·min^-1 for 6 min on Phenomenex C₁₈(100.0 mm×4.6 mm, 3.0 μm). The column temperature was 40 ℃. Positive ion scan analysis was carried out in multi-reaction monitoring mode, using electrospray ion source. The specificity, standard curve and lower limit of quantification (LLOQ), accuracy and precision, extraction recovery, matrix effect and stability were evaluated. The patients treated with LRG, LEV or OXC in hospital were included as the research object. Blood samples were collected after the blood concentration reached a steady state and the serum concentration was detected by the method developed in this study.

The method demonstrated good specificity. For LTG, LEV, OXC and MHD, outstanding linearity was observed in the ranges of (0.20-25.00), (0.39-50.00), (0.04-5.00), (0.31-40.00) μg·mL^-1, respectively. The linear equation of LTG was y=0.99x+0.02 (r=0.999 2), LEV was y=0.55x+0.01 (r=0.999 4), OXC was y=4.35x+0.04 (r=0.998 9), and MHD was y=0.56x+0.01 (r=0.998 7). The LLOQs were 0.20, 0.39, 0.04, 0.31 μg·mL^-1, respectively. Acceptable accuracy as well as intra-batch and inter-batch precision were achieved; relative error (RE) and relative standard deviation (RSD) were all <15.00%. The extraction recoveries reached the standard (88.56%~112.18%, RSD<15.00%), without significant matrix effects and carryover effects. The serum samples of LTG, LEV, OXC, and MHD exhibited stability under the following conditions: storage at -20 ℃ for 14 days, three freeze-thaw cycles (-20 ℃ to room temperature), room temperature exposure for 3 hours, and post-treatment storage in the autosampler (4 ℃) for 24 hours, with all RSDs ≤10.72%. The analysis of 145 clinical samples showed that the linear range could meet the clinical needs.

This study successfully established a HPLC-MS/MS method to simultaneously quantify the serum concentration of LTG, LEV, OXC and MHD. The method is simple, economical accurate and efficient, which has promotional value.