To investigate the effects of etomidate on the expression of mammalian sirolimus like target protein (mTOR)/silencing information regulatory factor 1 (SIRT1) gene to inhibit the proliferation, invasion and migration of bladder cancer BIU-87 cells.

The BIU-87 cells were divided into normal group, positive control group and experimental -L, -M, -H groups. The normal group was given normal culture; the positive control group was treated with MEM medium containing paclitaxel at a final concentration of 15 mg·mL^-1; the experimental -L, -M, -H groups were treated with MEM medium containing etomidate at final concentrations of 112.50, 225.00 and 450.00 mg·L^-1, respectively. After 24 hours of intervention, the migration ability of cells was detected using the cell scratching method, the invasion ability of cells was detected using the Transwell method, the levels of glucose and lactate in cells were detected using enzyme-linked immunosorbent assay, and the expression levels of mTOR, SIRT1 and matrix metalloproteinase (MMP) proteins were detected using Western blotting.

The number of invasive cells in the experimental -M, -H groups, positive control group and normal group was 248.56±5.67, 117.37±16.72, 76.54±8.23 and 376.40±53.41; the migration distances were (22.30±2.89), (13.67±5.46), (8.63±2.07) and (38.24±5.24) μm; the glucose levels were (38.97±2.43), (29.75±3.58), (21.52±4.20) and (64.26±7.26) μmol·L^-1; the lactate levels were (37.36±3.29), (20.32±5.67), (14.76±2.37) and (51.52±3.10) μmol·L^-1; the phosphorylated mTOR/mTOR ratios were 0.26±0.05, 0.20±0.04, 0.15±0.03 and 0.47±0.07; the relative expression levels of SIRT1 protein were 0.23±0.01, 0.18±0.02, 0.09±0.02 and 0.72±0.13; the relative expression levels of MMP-9 protein were 0.19±0.05, 0.14±0.01, 0.10±0.01 and 0.28±0.04, respectively. Compared with positive control group and normal group, the differences of above indexes in experimental -M, -H groups were statistically significant (all P<0.05).

Etomidate can effectively inhibit the proliferation, invasion and migration of BIU-87 cells by suppressing the expression of mTOR/SIRT1.

To explore the therapeutic effect of hydroxysafflor yellow A (HSYA) on acute exacerbation of chronic obstructive pulmonary disease (AECOPD) mice and its mechanism of action.

Mice were randomly divided into control group, model group and experimental -L, -M, -H groups, with 20 mice per group. The control group was raised normally, and the other groups were exposed to cigarette smoke (CS) and non-typeable Haemophilus influenzae (NTHi) infection to establish an AECOPD mouse model. Starting from the 12th week of CS exposure, experimental -L, -M, -H groups were administered 25, 50 and 100 mg·kg^-1·d^-1 HSYA by gavage, and control and model groups were given 0.9% NaCl by gavage. Five groups were administered once a day for 2 weeks. The functional residual capacity (FRC) of the mice was measured using the Buxco lung function test system. The percentage of neutrophils in bronchoalveolar lavage fluid (BALF) was measured using flow cytometry. The expression levels of phosphatidylinositol-3-kinase (PI3K) and protein kinase B (Akt) in lung tissues were detected by Western blot.

The FRC of experimental -M, -H groups, model group and control group were (0.31±0.01), (0.27±0.01), (0.37±0.02) and (0.25±0.01) L; the neutrophil percentages were (37.96±2.88)%, (28.41±1.64)%, (60.27±2.12)% and (11.33±0.31)%; the phosphorylated PI3K/PI3K ratios were 1.59±0.08, 1.30±0.06, 2.28±0.08 and 1.00±0.06; the phosphorylated Akt/Akt ratios were 1.33±0.11, 1.15±0.03, 1.76±0.11 and 1.00±0.06, respectively. Compared with model group, the differences of above indexes in experimental -M, -H groups were statistically significant (all P<0.05).

HSYA may reduce neutrophil activity by inhibiting the PI3K/Akt pathway, thereby inhibiting inflammatory responses and alleviating lung injury.

To explore the mechanism of prasugrel (PRAS) in alleviating oxidative stress of myocardial ischemia reperfusion (IR) by inducing pyroptosis of rat cardiomyocytes based on nuclear factor erythroid derived 2-like 2 (Nrf2)/heme oxygenase-1 (HO-1)/NAD(P)H quinone oxidoreductase 1 (NQO1) signaling pathway.

SD rats were randomly divided into sham operation group (only the heart was exposed, no IR model was constructed, and the same amount of normal saline was injected into the stomach), model group (IR rat model was constructed, and the stomach was given an equal amount of 0.9% NaCl), the experimental-L group (model group rats were intragastric with 5 mg·kg^-1 PRAS), the experimental-M group (model group rats were intragastric with 10 mg·kg^-1 PRAS), and experimental-H group (model group rats were intragastric with 15 mg·kg^-1 PRAS). The serum levels of myocardial injury markers and oxidative stress factors were detected by enzyme-linked immunosorbent assay. The relative expression levels of pyroptosis-related proteins and Nrf2/HO-1/NQO1 signaling pathway-related proteins in myocardial tissue were detected by Western blot.

The contents of cardiac troponin Ⅰ (cTnⅠ) in sham operation group, model group, experimental-L group, experimental-M group and experimental-H group were (264.52±39.04), (558.40±101.68), (477.65±64.33), (429.71±75.94) and (313.84±52.26) pg·mL^-1, respectively; the creatine kinase isoenzyme (CK-MB) levels were (0.68±0.11), (2.76±0.45), (1.92±0.27), (1.53±0.23) and (1.11±0.19) U·L^-1, respectively; the levels of malondialdehyde (MDA) were (4.05±0.68), (10.12±1.74), (8.44±1.26), (6.79±1.05) and (5.47±1.02) nmol·L^-1, respectively; the relative expression levels of nucleotide binding oligomerization domain-like receptor protein 3 (NLRP3) were 1.00±0.14, 2.16±0.32, 1.72±0.25, 1.54±0.28 and 1.28±0.21, respectively; the relative expression levels of Nrf2 were 1.00±0.15, 0.32±0.04, 0.39±0.05, 0.61±0.10 and 0.80±0.13, respectively. The above indexes of model group were compared with those of sham-operation group, and those of experimental-L, experimental-M and experimental-H groups were compared with those of model group (P<0.05, P<0.01, P<0.001).

PRAS may resist oxidative stress and reduce pyroptosis and apoptosis of myocardial cells by activating Nrf2/HO-1/NQO1 signaling pathway, thereby alleviating IR injury and protecting cardiac function.

To study the effects of carbocisteine on airway inflammation and airway remodeling in mice with Mycoplasma pneumoniae pneumonia and its mechanism.

Mice were randomly divided into control group, model group, experimental group, negative transfection group and positive transfection group, with 12 mice in each group. The control group were inoculated with 0.9% NaCl by nasal drip, the other groups were inoculated with Mycoplasma pneumoniae suspension to establish Mycoplasma pneumoniae pneumonia model. After successful modeling, the experimental group was given 10 mg·kg^-1 carbocisteine by intragastric administration; the negative transfection group was given 10 mg·kg^-1 carbocisteine by gavage + 1.0×10⁹ PFU of adenovirus containing tumor necrosis factor-α-induced protein 8-like 2 (TIPE2) blank control plasmid in the tail vein, the positive transfection group was given 10 mg·kg^-1 carbocisteine by intragastrial + 1.0×10⁹ PFU containing sh-TIPE2 plasmid was injected into the tail vein, the control and model groups were given 0.9% NaCl by intragastric administration. Five groups were treated for 2 weeks with once a day. The levels of inflammatory factors in lung tissue were detected by enzyme-linked immunosorbent assay. Airway remodeling index and p38 mitogen activated protein kinase (p38MAPK) /activated protein-1 (AP-1) signaling pathway related protein expression were detected by Western blot.

The levels of tumor necrosis factor-α in control, model, experimental, negative transfection and positive transfection groups were (9.18±1.92), (134.32±24.56), (42.20±8.61), (39.95±7.77) and (74.03±13.59) pg·mL^-1; the levels of interleukin-6 were (5.80±1.21), (65.12±11.81), (22.94±4.47), (26.11±5.08) and (42.67±7.97) pg·mL^-1; the relative protein expression levels of matrix metallopeptidase 9 were 1.00±0.16, 3.64±0.62, 1.45±0.22, 1.39±0.20 and 2.16±0.38; the relative protein expression levels of tissue inhibitor of metalloproteinases 1 were 1.00±0.14, 0.52±0.11, 0.89±0.16, 0.84±0.15 and 0.65±0.13; the phospho-p38MAPK/p38MAPK ratios were 1.00±0.16, 2.74±0.48, 1.57±0.29, 1.54±0.27 and 2.31±0.36; the relative protein expression levels of cellular proto-oncogene Jun were 1.00±0.14, 2.25±0.38, 1.34±0.21, 1.37±0.22 and 1.98±0.32; the relative protein expression levels of cellular proto-oncogene Fos were 1.00±0.18, 3.07±0.56, 1.86±0.32, 1.91±0.35 and 2.51±0.44, respectively. The above indexes in the model group were compared with the experimental and control groups, the above indexes in the positive transfection group were compared with the negative transfection group, and the differences were statistically significant (all P<0.05).

Carbocisteine inhibits airway inflammation and airway remodeling in mice with Mycoplasma pneumoniae pneumonia by inhibiting the p38MAPK/AP-1 signaling pathway.

To establish an animal model of polycystic ovary syndrome (PCOS) characterized by kidney deficiency and blood stasis, and to evaluate its efficacy.

SD rats were randomly divided into normal group, PCOS group, model group and formula counter-evidence group, with 10 rats in each group. The normal group was given 1 mg·kg^-1 1% sodium carboxymethyl cellulose daily by intragastric administration; PCOS group was given 1 mg·kg^-1 letrozole daily by intragastric administration; on the basis of PCOS group, the model group was injected subcutaneously with 0.6 mg·kg^-1 epinephrine hydrochloride solution daily for 2 weeks from day 21. On the basis of the model group, the formula counter-evidence group was given 12.51 g·kg^-1 Zhu’s Tiaojing decoction daily for 2 weeks from the 21st day. After 35 days of intervention, the indexes of ovarian index, testosterone, vitamin D and whole blood low shear rate were compared.

The ovarian indexes of normal, PCOS, model and formula counter-evidence groups were (6.39±0.58)×10^-4, (5.32±0.71)×10^-4, (5.05±0.29)×10^-4 and (8.17±0.65)×10^-4; testosterone concentrations were (7.10±0.97), (9.22±0.43), (9.06±1.21) and (7.73±0.76) nmol·L^-1; vitamin D levels were (1 850.98±227.95), (986.10±152.50), (1 220.67±64.35) and (1 598.20±294.75) pg·mL^-1; whole blood low shear rates were (13.00±1.79), (13.37±0.54), (16.38±1.51) and (11.43±2.77) mPa·s, respectively. The above indexes in the model group were statistically significant compared with the formula counter-evidence group and the normal group (all P<0.05).

Letrozole combined with epinephrine hydrochloride can stably replicate the PCOS rat model with kidney deficiency and blood stasis syndrome.

To observe the protective effect of resveratrol on kidney of diabetic rats through lipid metabolism pathway.

The SD rats were randomly divided into control group, model group and experimental -L, -M, -H groups. The control group was fed with normal feed, while the other four groups were all based on high-fat and high-sugar feed feeding + intraperitoneal injection of streptozotocin to establish diabetic rat models. After successful modeling, the control group and the model group were given the same amount of normal saline, while the experimental-L, -M, -H groups were given 5, 25, 50 mg·kg^-1 concentration of resveratrol once a day, and were given continuous gavage for 10 weeks. Kidney function levels [cystatin C (Cys-C), serum creatinine (SCr), blood urea nitrogen (BUN)]were determined by enzyme-linked immunosorbent assay. The expressions of peroxissome proliferator-activated receptor γ (PPARγ), fatty acid- binding protein (FABP1), peroxisomal acyl-coenzyme A oxidase 1 (ACOX) 2 and ACOX1 were detected by Western blot.

The Cys-C levels of experimental -M, -H groups, control group, model group were (1.85±0.15), (1.39±0.11), (0.78±0.11) and (2.42±0.32) mg·L^-1; SCr levels were (55.74±7.02), (48.15±6.97), (32.41±4.22) and (68.15±8.48) μmol·L^-1; BUN levels were (10.37±1.02), (9.41±0.87), (7.12±1.12) and (14.48±2.45) mmol·L^-1; the relative expression levels of PPARγ protein were 0.74±0.08, 0.92±0.10, 1.22±0.15 and 0.39±0.05; the relative expression levels of FABP1 protein were 0.64±0.08, 0.81±0.07, 0.98±0.09 and 0.30±0.05; the relative expression levels of ACOX2 protein were 0.59±0.06, 0.72±0.07, 0.85±0.13 and 0.27±0.05; the relative expression levels of ACOX1 protein were 0.58±0.06, 0.67±0.05, 0.78±0.06 and 0.24±0.03. The differences of above indexes were statistically significant between the experimental-M, -H groups and control group and the model group (all P<0.05).

Resveratrol may play a protective role on kidney of diabetic rats by regulating lipid metabolism pathway.

To investigate the effects of Tonglagat-5, a Mongolian drug, on bone microstructure and bone metabolism in osteoporotic mice, and to explore the possible mechanisms of action based on phosphatase and tensin homologue-induced kinase 1 (PINK1)/Parkin-mediated mitophagy.

Osteoporotic mouse model was established by ovariectomy (OVX). Osteoporotic model mice were randomly divided into model group (equal volume 0.9% NaCl by gavage), control group (estradiol valerate 1 mg·kg^-1 by gavage), experimental group (0.05 g·kg^-1 Tonglagat-5 solution by gavage) and combined group (0.05 g·kg^-1 Tonglagat-5 solution by gavage + 50 mg·kg^-1 Mdivi-1 by intraperitoneal injection), with 10 mice per group; ten mice were randomly selected as the sham-operation group. After 10 consecutive weeks of treatment, the bone mineral density (BMD) was measured by Micor-CT assay, the bone metabolism marker assay [C-terminal telopeptide of type Ⅰ collagen (CTx-Ⅰ), alkaline phosphatase (ALP) and bone Gla-protein (BGP)] were detected by kit, and the protein expression levels of translocase of outer mitochondrial membrane 20 (Tomm20), p62, PINK1 and Parkin were performed by Western blot.

The BMD in the sham-operation, model, control, experimental and combined groups were (0.23±0.02), (0.09±0.01), (0.22±0.03), (0.23±0.01) and (0.11±0.02) g·mm^-3; the levels of CTx-Ⅰ were (14.58±1.22), (26.62±2.04), (16.44±2.31), (12.31±1.98) and (22.76±2.52) ng·mL^-1; the levels of ALP were (4.87±1.12), (1.41±1.10), (3.03±1.07), (4.44±1.15) and (2.05±1.41) U·L^-1; the levels of BGP were (9.35±1.06), (5.28±0.84), (7.76±0.35), (8.34±0.87) and (6.12±0.81) ng·mL^-1; the relative expression levels of Tomm20 protein were 0.93±0.08, 0.10±0.01, 0.37±0.03, 0.84±0.06 and 0.28±0.03; the relative expression levels of p62 protein were 0.15±0.02, 0.93±0.09, 0.65±0.06, 0.37±0.04 and 0.86±0.06; the relative expression levels of PINK1 protein were 0.87±0.09, 0.14±0.02, 0.32±0.03, 0.69±0.06 and 0.26±0.03; the relative expression levels of Parkin protein were 0.79±0.08, 0.12±0.02, 0.26±0.04, 0.58±0.05 and 0.23±0.03, respectively. The above indexes in model group were compared those in sham-operation, control and experimental groups, the differences were statistically significant (all P<0.05).

Tonglagat-5 is able to improve bone microstructure and bone metabolism and reduce bone loss in OVX mice, and the mechanism may be related to the activation of PINK1/Parkin pathway-mediated mitophagy to exert osteoprotective effects.

To investigate the protective effect of icariin on recurrent spontaneous abortion (RSA) mice and its potential mechanism.

Female CBA/J mice were randomly divided into control group (mated with male BALB/c mice), model group (modeling mating with male DBA/2 mice), experimental-L group (after modeling, 25.00 mg·kg^-1 icariin was given by intragastric administration), experimental-H group (50.00 mg·kg^-1 icariin was given by intragastric administration after modeling), positive group (2.6 mg·kg^-1 desdrogesterone was given by intragastream after modeling), uterine tissue was collected after 14 days of treatment for follow-up experiments. Embryo absorption rate was observed and recorded. The levels of interleukin (IL) -4 and interferon γ (IFN-γ) were detected by enzyme linked immunosorbent assay (ELISA). The mRNA expressions of Foxp3 and Toll-like receptor 4 (TLR4) were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). Western blot assay was used to detect the expression of TLR4 and phosphorylated nuclear transcription factor-κB (p-NF-κB) protein.

The absorption rates of mouse embryos in control group, model group, experimental-L group, experimental-H group and positive group were 7.37% (7 cases/95 cases), 37.33% (28 cases/75 cases), 20.93% (18 cases/86 cases), 13.79% (12 cases/87 cases) and 12.22% (11 cases/90 cases), respectively; IL-4 expression levels were (33.84±5.71), (10.71±2.34), (16.86±1.13), (26.91±3.04) and (19.56±1.79) pg·mL^-1, respectively; the expression levels of IFN-γ were (25.97±1.40), (63.38±5.39), (47.23±6.78) (34.80±4.08) and (29.61±3.25) pg·mL^-1, respectively; the relative expression levels of Foxp3 mRNA were 1.00±0.07, 0.57±0.05, 0.72±0.06, 0.90±0.05 and 0.66±0.06, respectively; the relative expression levels of TLR4 mRNA were 1.00±0.10, 2.32±0.17, 1.78±0.14, 1.40±0.08 and 2.05±0.26, respectively; the relative expression levels of TLR4 protein were 0.31±0.05, 0.90±0.11, 0.68±0.05, 0.55±0.05 and 0.79±0.10, respectively; the relative expression levels of p-NF-κB protein were 0.27±0.02, 0.82±0.13, 0.66±0.04, 0.42±0.05 and 0.75±0.09, respectively. The above indicators in the model group compared with the control group, the experimental-L and experimental-H groups were compared with the model group, and the differences were statistically significant (all P<0.05).

Icariin may play a protective role in improving RSA immune response by inhibiting TLR4/NF-κB pathway.

To explore the effects of essential oil of Ligusticum chuanxiong (LCEO) on primary dysmenorrhea (PD) and preliminarily its mechanism of action.

Forty-eight female non-pregnant KM mice were randomly divided into blank group, model group, positive control group and experimental -L, -M, -H groups. Except for the blank group, the remaining groups were gavaged with estradiol valerate to establish the PD mouse model. Starting from the 7th day, the blank group and the model group were gavaged with equal volumes of distilled water in the morning, the positive control group was administered 120 mg·kg^-1 of ibuprofen solution by gastric lavage, while the low, medium, and high dose experimental groups were gavaged with 22, 44 and 88 mg·kg^-1 of LCEO, respectively, for 5 consecutive days. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of oxytocin (OT) and prostaglandin F_2α (PGF_2α) in mice uterine. Using the oxytocin-induced rat isolated uterine contraction model, the study explored the mechanism of LCEO in relaxing uterine smooth muscle through preincubation with different blocking agents (Indomethacin, phentolamine mesylate, propranolol hydrochloride, atropine sulfate, diphenhydramine hydrochloride, ranitidine hydrochloride and nifedipine).

The OT contents in the blank group, model group, positive control group and experimental -L, -M, -H groups were (3.57±0.81), (5.54±0.60), (4.63±0.75), (4.15±0.76), (4.04±1.19) and (3.67±0.66) pg·mg^-1; the PGF_2α contents were (1.22±0.38), (2.53±0.45), (1.84±0.52), (1.30±0.31), (1.21±0.35) and (0.48±0.15) pg·mg^-1, respectively. The above indexes in the model group were compared with the experimental -L, -M, -H and blank, positive control groups, there were statistically significant differences in all comparisons (P<0.01, P<0.05). After preincubation with indomethacin, phentolamine mesylate, and atropine sulfate, the relaxant effect on uterine smooth muscle of LCEO was significantly weakened. After preincubation with propranolol hydrochloride, diphenhydramine hydrochloride, ranitidine hydrochloride and nifedipine, the relaxant effect on uterine smooth muscle of LCEO did not show any significant changes.

LCEO has a significant effect on primary dysmenorrhea, and its mechanism may be related to inhibiting cyclooxygenase and blocking receptors α and M.

To investigate the effect of Acorus calamus L. on wound healing of radioactive skin injury in rats based on the mitogen-activated protein kinase (MAPK)/nuclear factor κB (NF-κB) signaling pathway.

A rat model of radioactive skin injury was induced by the radionuclide irradiation, the successful model rats were randomly divided into the model group, the control group, and the experimental -H, -M, -L groups with 12 rats in each group, and another 12 normal rats were taken as the normal group. Experimental-H, -M, -L goups was applied with Acorus calamus L. 600, 400, 200 g·kg^-1, respectively. Control group was applied with triethanolamine cream 400 g·kg^-1 on rat wounds; normal and model groups were treated with 0.9% NaCl. Six groups were treated once a day for 45 days. The healing rate of wounds was compared among the six groups. The levels of interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α in serum and hydroxyproline (HYP) in wound tissue were measured by enzyme linked immunosorbent assay. The protein levels of IL-1β, IL-6, TNF-α, mitogen-activated protein kinase (p38) and nuclear factor κB p65 subunit (p65) were detected by Western blot.

The wound healing rates of experimental-H, -M, -L groups, control group and model group were (70.24±3.17)%, (62.41±4.52)%, (49.89±4.61)%, (50.35±2.44)% and (29.42±4.23)%, respectively. The levels of IL-1β in experimental-H, -M, -L groups, control group, model group and normal group were (41.22±2.98), (52.35±1.69), (61.32±2.74), (45.26±3.12), (79.24±5.64) and (28.85±2.11) pg·L^-1; the levels of IL-6 were (25.68±1.65), (32.55±2.64), (51.23±2.34), (32.32±1.56), (61.12±2.35) and (18.88±1.12) pg·L^-1; the levels of TNF-α were (62.38±5.66), (73.56±4.52), (88.63±5.68), (68.33±4.32), (112.48±6.52) and (55.21±3.89) pg·L^-1; the levels of HYPs of (3.77±0.41), (3.51±0.26), (2.98±0.39), (3.62±0.37), (2.21±0.48) and (4.22±0.55) μg·mg^-1; the relative expression levels of IL-1β were 0.31±0.04, 0.43±0.07, 0.60±0.03, 0.51±0.04, 0.69±0.06 and 0.11±0.07; the relative expression levels of IL-6 were 0.50±0.03, 0.60±0.04, 0.69±0.04, 0.62±0.07, 0.83±0.08 and 0.28±0.06; the relative expression levels of TNF-α were 0.49±0.06, 0.61±0.03, 0.73±0.06, 0.59±0.04, 0.83±0.05 and 0.28±0.04; the phosphorylated p38/p38 ratios were 0.46±0.06, 0.56±0.04, 0.67±0.03, 0.56±0.05, 0.84±0.04 and 0.34±0.08; the phosphorylated p65/p65 ratios were 0.37±0.06, 0.54±0.03, 0.63±0.04, 0.47±0.05, 0.79±0.04, and 0.32±0.06, respectively. Compared with model group, the above indexes in the experimental-H, -M groups were statistically significant (all P<0.05).

Acorus calamus L. can promote the healing of radioactive skin injury in rats, inhibit the inflammation level of the wound tissue, and promote collagen deposition, and its mechanism may be related to the regulation of MAPK/NF-κB pathway.