To explore effect and mechanism of microRNA 142a-3p (miR-142a-3p) on adriamycin-induced nephropathy (AN).

BALB/c mice were randomly divided into normal group and model group. The model group was injected with 10.5 mg·kg^-1 doxorubicin at a time through the tail vein, and the normal group was injected with equal volume 0.9% NaCl by the same method. After 28 days of feeding, both sides of the kidney tissues were taken for experiment. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-142a-3p in the renal tissues. The expression levels of autophagy related protein 16-like protein 1 (Atg16l1) and inflammatory NOD-like receptor heat protein domain associated protein 3 (NLRP3), interleukin-1β (IL-1β) and interleukin-18 (IL-18) in the renal tissues were detected by Western blotting. Human embryonic kidney 293T cells were divided into Atg16l1-WT+NC transfection group (transfected pmirGLO-Atg16l1-WT and NC mimics) and Atg16l1-WT+miRNA-142a-3p group (transfected pmirGLO-Atg16l1-WT and miR-142a-3p mimics). Dual luciferase assay was used to detect the targeted regulation of miR-142a-3p on Atg16l1.

The expression levels of miR-142a-3p in kidney tissues of normal group and model group were 1.20±0.27 and 2.02±0.14; the expression levels of Atg16l1 were 0.81±0.13 and 0.57±0.12, NLRP3 were 0.49±0.11 and 0.82±0.14; IL-1β were 0.55±0.11 and 0.84±0.08; IL-18 levels were 0.41±0.15 and 0.74±0.09, respectively, and the above indexes in the model group were significantly different from those in the normal group (all P<0.05). The relative luciferase activity in Atg16l1-WT+NC transfection group and Atg16l1-WT+miRNA-142a-3p transfection group were 1.26±0.07 and 0.92±0.10, respectively, with statistical significance (all P<0.05).

MiR-142a-3p down-regulates the expression of target protein Atg16l1 in renal tissue of adriamycin nephropathy mice, which activates NLRP3 pathway of inflammatory bodies and promotes the progressive development of adriamycin-induced nephropathy.

To observe the effect of Tetrahydropalmatine on the epidermal growth factor receptor / phosphatidylinositol 3-kinase/protein kinase B (EGFR/PI3K/AKT) signaling pathway in endometriosis dysmenorrhoea SD rats.

The endometriosis dysmenorrhoea model was replicated by auto-transplantation in 60 female SD rats of SPF grade. After successful modelling, the rats were randomly divided into model group, control group and experimental -L, -M, -H groups according to body mass, with 12 rats in each group; and another 12 normal rats were taken as the blank group. The blank and model groups were given 10 mL·kg^-1 0.9% NaCl by gavage, and the control group was given 0.25 mg·kg^-1 progesterone suspension by gavage. The experimental-L, experimental-M, experimental-H groups were given 10, 20 and 40 mg·kg^-1 Tetrahydropalmatine suspension by gavage, respectively. Six groups of rats were administered once daily for 4 weeks. After the last administration, 2 U of oxytocin was given to induce contractions, and the rats were observed for torsional responses. The serum levels of tumor necrosis factor -α (TNF-α), epidermal growth factor (EGF), and EGFR were measured by enzyme-linked immunosorbent assay in each group of rats, and the expression levels of EGFR, PI3K and AKT proteins in ectopic endometrial tissues were detected by Western blotting.

The number of twists in the experimental -M, -H groups and control group, model group, blank group were 9.25±1.42, 8.33±1.88, 11.17±2.41, 28.67±2.15 and 0; the inhibition rates were 67.74%, 70.95%, 61.05%, 0 and 0; serum TNF-α levels were (281.96±13.06), (278.75±10.39), (282.12±14.47), (303.59±9.09) and (274.26±15.33) ng·L^-1; serum EGF levels were (313.82±13.23), (308.26±10.90), (311.39±19.52), (336.04±17.60) and (301.67±27.17) pg·mL^-1; serum EGFR levels were (315.71±7.56), (311.43±11.04), (313.75±7.26), (333.56±14.22) and (307.86±17.94) ng·L^-1; the relative expression levels of EGFR protein were 1.60±0.39, 1.26±0.30, 1.40±0.40, 2.08±0.41 and 1.00±0.22; the relative expression levels of PI3K protein were 1.60±0.42, 1.21±0.18, 1.27±0.24, 2.01±0.41 and 1.00±0.19; the relative expression levels of AKT protein were 1.27±0.18, 1.11±0.10, 1.27±0.16, 1.64±0.28 and 1.00±0.07, respectively. Statistically significant differences were found between the above indicators in the experimental -M, -H groups compared to the model group (P<0.01, P<0.05).

Tetrahydropalmatine may play a therapeutic role in endometriosis dysmenorrhoea by interfering with the EGFR/PI3K/AKT signaling pathway.

To investigate the antitumor activity and mechanism of a pyrazole [1,5-a]pyrimidine derivative (BPPA) on human tongue squamous cell carcinoma Cal33 cells.

Cal33 cells were divided into blank group (normal culture), experimental-L, -M, -H groups (1, 2.5 and 5 μmol·L^-1BPPA) and combined group [2.5 μmol·L^-1BPPA and 5 mmol·L^-1 N-acety-L-cysteine (NAC)]. Cell viability was determined by thiazole blue assay. Cell apoptosis was measured using flow cytometry, while apoptosis-related protein expression levels were determined by Western blot analysis. Changes in reactive oxygen species and mitochondrial membrane potential were evaluated using immunofluorescence assay.

The survival rates of Cal33 cells in blank, experimental-L,-M,-H and combined groups were (99.56±0.91)%, (58.31±2.31)%, (47.93±1.67)%, (29.35±4.11)% and (70.27±1.21)%, respectively; cell apoptotic rates were (8.07±1.01)%, (44.04±0.94)%, (49.85±1.75)%, (66.79±0.83)% and (24.33±1.04)%, respectively; the relative fluorescence intensities of reactive oxygen species (ROS) were 1.23±0.21, 3.37±0.35, 15.53±1.46, 20.28±1.24 and 5.59±0.52, respectively; the mitochondrial membrane potential red fluorescence/green fluorescence values were 4.69±0.11, 4.24±0.12, 0.86±0.16, 0.46±0.05 and 2.99±0.11, respectively. Compared with blank group, the above indexes in the experimental-L, -M, -H groups were statistically significant (P<0.01, P<0.001). Compared with experimental-M group, the above indexes in combined group were statistically significant (P<0.01, P<0.001). The relative levels of pro-apoptotic B-cell lymphoma-2 (Bcl-2)-Associated X protein (Bax) in blank, experimental-L,-M,-H groups were 1.00±0.02, 1.94±0.03, 3.73±0.06 and 4.64±0.08, respectively; the relative levels of anti-apoptotic protein Bcl-2 were 1.00±0.02, 0.75±0.04, 0.62±0.02 and 0.46±0.03, respectively. Compared with blank group, the above indexes in the experimental-L, -M, -H groups were statistically significant (P<0.01, P<0.001).

BPPA exhibits anti-head and neck cancer activity by ROS mediating the decrease in mitochondrial membrane potential, thereby inducing apoptosis.

To investigate the effects of baicalein on apoptosis and immune escape of laryngeal carcinoma cells and its mechanism.

Human laryngeal carcinoma TU686 cells were randomly divided into control group(routine culture), baicalein group (50 μmol·L^-1 baicalein), Vector group (transfected with Vector+50 μmol·L^-1 baicalein) and CD47 group [transfected with differentiation cluster 47(CD47) +50 μmol·L^-1 baicalein]. Cell proliferation and apoptosis were detected by 5-acetylidene-2′-deoxyuridine (Edu) method and Annexin V-FITC/PI double staining method. The expression of proliferative and apoptotic proteins were detected by Western blot. Human peripheral lymphocytes were co-cultured with cells in each group to detect the killing rate of TU686 cells. The phagocytosis rate of TU686 cells was observed.

The Edu positive cell rates in control group, baicalein group, Vector group and CD47 group were (36.24±2.74)%, (13.33±1.93)%, (14.22±1.48)% and (27.80±2.74) %, respectively; the apoptosis rates were (3.94±0.40)%, (28.40±1.76)%, (26.60±1.41)% and (12.33±0.84)%, respectively; the relative expressions of CD47 protein were 1.00±0.11, 0.42±0.07, 0.44±0.06 and 1.21±0.12, respectively; the relative expression levels of CyclinD1 protein were 0.86±0.08, 0.37±0.05, 0.36±0.04 and 0.74±0.08, respectively; the relative expression levels of cleaved cysteine aspartate proteinase-7(Cl-Caspase-7) protein were 0.35±0.04, 0.99±0.12, 1.00±0.11 and 0.50±0.06, respectively; the killing rates of human peripheral lymphocytes to TU686 cells (1∶1 ratio) were (10.61±1.19)%, (32.02±2.27)%, (31.86±2.06)% and (14.39±1.07)%, respectively; the phagocytosis rates were (6.35±0.71)%, (11.99±1.64)%, (12.03±0.76)% and (7.06±0.57)%, respectively. The above indicators, baicalein group compared with the control group, Vector group compared with CD47 group, the differences were statistically significant (P<0.05, P<0.01, P<0.001).

Baicalein may induce the apoptosis of laryngeal carcinoma cells and inhibit the immune escape of laryngeal carcinoma cells by inhibiting the expression of CD47, thus slowing down the progression of laryngeal carcinoma.

To investigate the efficacy of the hypoxia-inducible factor-1α (HIF-1α) inhibitor PX-478 in improving radiation-induced apoptosis in cardiomyocytes and to elucidate its molecular mechanisms.

H9c2 cells were divided into blank group, model group, experimental-L group and experimental-H group. The experimental-L, -H groups were cultured with PX-478 at concentrations of 5 and 10 mol·L^-1, respectively, while the other groups were cultured in normal medium. After 24 h of culture, all groups except the blank group were irradiated with 6 Gy X-rays to establish a model of radiation-induced cardiac injury and were subsequently cultured for another 24 h. Cell viability was assessed using the cell counting kit-8 assay; relative protein expression levels were determined by Western blotting; and apoptosis was detected using Annexin V/PI flow cytometry.

The relative expression levels of HIF-1α protein in the blank group, model group, experimental-L group and experimental-H group were 0.89±0.01, 1.05±0.01, 0.64±0.02 and 0.52±0.02; the relative expression levels of aquaporin-1 (AQP1) protein were 0.79±0.02, 0.94±0.02, 0.74±0.01 and 0.59±0.01; AQP4 protein levels were 0.89±0.01, 1.03±0.01, 0.68±0.01 and 0.50±0.01; B-cell lymphoma-2 (Bcl-2) protein levels were 0.78±0.02, 0.58±0.02, 1.00±0.03 and 0.82±0.03; Bcl-2-associated X protein (Bax) levels were 0.66±0.01, 0.96±0.01, 0.77±0.01 and 0.27±0.03; Caspase-3 levels were 0.83±0.01, 1.10±0.01, 0.71±0.02 and 0.30±0.01; the apoptosis rates were (4.05±0.60)%, (17.82±0.63)%, (9.35±0.41)% and (9.07±0.57)%, respectively. The above indicators in the blank, experimental-L, experimental-H groups showed statistically significant differences compared to the model group (all P<0.05).

PX-478 can improve radiation-induced cardiomyocyte edema and reduce apoptosis by inhibiting the abnormal activation of the HIF-1α/AQPs axis in H9c2 cells after irradiation, thereby regulating fluid metabolism.

To explore the effect of honey-processed Hedysari Radix on the metabolism of short-chain fatty acids (SCFAs) in the cecum of rats with spleen qi deficiency.

A total of 48 SPF male SD rats were randomly divided into blank group, model group, control group and experimental group, with 8 rats in each group. The blank group was fed normally, drank water and ate freely for 15 days, and the other groups used the three-factor composite modeling method of bitter cold diarrhea, overwork and hunger and satiety to construct a spleen qi deficiency model rats for 15 days. After the successful replication of the model, the rats in the control group were given 62.5 mg·kg^-1 bifidobacterium Lactobacillus triple viable tablets suspension, the experimental group was given 12.6 g·kg^-1 honey-processed Hedysari Radix decoction, and the blank group was given the same dose of distilled water, and the model group continued to model for 15 days. Gas chromatography-mass spectrometry (GC-MS) was used to determine the contents of SCFAs in the cecal contents of rats in each group.

The contents of acetic acid, isobutyric acid, butyric acid, isovaleric acid and caproic acid in the cecal contents of rats in the blank group were (3 706.24±401.84), (99.34±39.11), (2 567.95±529.44), (86.50±54.11) and (393.14±103.33) μg·g^-1, respectively. Compared with the blank group, the contents of acetic acid, isobutyric acid, butyric acid, isovaleric acid and caproic acid in the cecum contents of spleen-qi deficiency rats in the model group were significantly reduced, which were (3 049.48±590.62), (67.27±12.99), (2 058.00±417.46), (43.10±11.12) and (27.98±14.40) μg·g^-1, respectively (P<0.05, P<0.01). Compared with the model group, the contents of isovaleric acid and caproic acid in the cecum contents of rats in the experimental group were significantly increased, which were (90.59±26.03) and (60.05±19.89) μg·g^-1, respectively (all P<0.01).

Honey-processed Hedysari Radix can play a role in strengthening the spleen and replenishing qi by improving the metabolism level of cecal contents, regulating the metabolism levels of SCFAs in cecal contents, protecting the intestinal mucosal barrier, and reducing intestinal inflammation.

To analyze the effect of insulin degludec/insulin aspart injection combined with metformin extended-release tablets on insulin resistance in patients with type 2 diabetes mellitus (T2DM).

T2DM patients were divided into control group and treatment group using a cohort method. The control group received oral metformin extended-release tablets (0.5 g, tid) and subcutaneous injections of insulin glargine injection (10 U, qd), while the treatment group received oral metformin extended-release tablets (0.5 g, tid) and subcutaneous injections of insulin degludec/insulin aspart injection (0.1-0.2 U·kg^-1·d^-1, evenly divided before breakfast and dinner). Both groups were treated for 12 weeks. The efficacy, blood glucose control [fasting blood glucose, 2-hour postprandial blood glucose, glycated hemoglobin A1c(HbA1c)], insulin function [fasting insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR) and homeostasis model assessment-β (HOMA-β)]were compared, and safety was evaluated.

A total of 49 cases were enrolled in the treatment group and 51 cases in the control group. The overall effective rates in the treatment and control groups were 93.88% (46 cases/49 cases) and 80.39% (41 cases/51 cases), respectively, showing a statistically significant difference (P<0.05). After treatment, the fasting blood glucose levels in the treatment and control groups were (5.83±0.79) and (6.53±0.81) mmol·L^-1; the 2-hour postprandial blood glucose levels were (7.73±0.86) and (8.41±0.97) mmol·L^-1; HbA1c levels were (6.34±0.88)% and (6.80±0.92)%; FINS levels were (6.02±1.13) and (7.13±1.04) mU·L^-1; HOMA-IR values were 1.56±0.20 and 2.07±0.23; HOMA-β values were 51.67±3.18 and 47.06±3.31, respectively, with all differences being statistically significant (all P<0.05). The adverse drug reactions in the treatment group mainly included hypoglycemia, nausea, dizziness, and diarrhea, while the adverse drug reactions in the control group mainly included hypoglycemia and nausea. The incidence of adverse drug reactions was 10.20% (5 cases/49 cases) in the treatment group and 11.76% (6 cases /51 cases) in the control group, with no statistically significant difference (P<0.05).

The combination of insulin degludec/insulin aspart injection and metformin extended-release tablets can improve insulin resistance and regulate blood glucose levels in T2DM patients, with good safety profiles.