To investigate the effects of long non-coding RNA (lncRNA) MAGI1-IT1 on the proliferation, invasion, migration and epithelial mesenchymal transformation (EMT) of breast cancer cells by regulating the miR-485-5p/actin gamma 1 (ACTG1) axis.

HCC1937 cells were cultured in vitro and randomly divided into control, si-MAGI1-IT1, miR-485-5p mimics, negative cotransfection and positive cotransfection groups. Control group was cultured normally without transfection; si-MAGI1-IT1 group was transfected with si-MAGI1-IT1; miR-485-5p mimics group was transfected with miR-485-5p mimics; negative cotransfection group was transfected with si-NC and NC-miR-485-5p; positive cotransfection group was given si-MAGI1-IT1 and miR-485-5p inhibitor combined transfection. Cell proliferation was detected by cell counting kit-8 method, cell migration was detected by cell scratch assay, cell invasion was detected by Transwell assay, and expression levels of lncRNA MAGI1-IT1, miR-485-5p and ACTG1 were detected by real-time fluorescence quantitative polymerase chain reaction method. The expression of proliferation and EMT-related proteins were detected by immunohistochemical staining.

The cell activities (optical density) of the control, si-MAGI1-IT1, miR-485-5p mimics, negative cotransfection and positive cotransfection groups were 0.89±0.12, 0.36±0.05, 0.32±0.03, 0.89±0.10 and 0.85±0.11; the migration rates were (91.82±11.13)%, (38.40±7.64)%, (33.17±6.03)%, (92.43±10.87)% and (86.10±10.76)%; the number of invasions was 296.50±23.48, 93.00±17.36, 76.50±15.01, 302.50±25.32 and 269.00±24.92; the relative expression levels of lncRNA MAGI1-IT1 were 0.98±0.11, 0.29±0.06, 0.96±0.10, 1.00±0.12 and 0.30±0.07; the relative expression levels of miR-485-5p were 0.99±0.13, 2.12±0.16, 2.20±0.18, 0.98±0.15 and 1.04±0.16; the relative expression levels of ACTG1 were 1.01±0.12, 0.34±0.05, 0.31±0.07, 1.02±0.14 and 0.97±0.11; the positive expression levels of proliferating cell nuclear antigen (optical density) were 1.65±0.16, 0.53±0.12, 0.45±0.08, 1.64±0.14 and 1.60±0.17; the positive expression levels of Cyclin D1 (optical density) were 1.53±0.15, 0.46±0.11, 0.40±0.06, 1.52±0.14 and 1.48±0.13; the positive expression levels of Vimentin (optical density) were 1.49±0.14, 0.45±0.07, 0.39±0.06, 1.50±0.12 and 1.45±0.13, respectively. The above indexes in the si-MAGI1-IT1 group were compared with the control group, the above indexes in the miR-485-5p mimics group (except lncRNA MAGI1-IT1) were compared with the control group, the above indexes in the positive cotransfection group (except lncRNA MAGI1-IT1) were compared with the si-MAGI1-IT1 group, the differences were statistically significant (all P<0.05).

Knockdown of lncRNA MAGI1-IT1 can reduce the expression of ACTG1 by up-regulating miR-485-5p, thereby inhibiting the proliferation, migration, invasion and EMT of breast cancer cells.

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 investigate the protective effects of Angelicae and Astragalus ultra-filtration (AAU) on H₉C₂ myocardial cell injury induced by X-ray radiation based on NOD-like receptor thermal protein domain-associated protein 3 (NLRP3) inflammasome.

The H₉C₂ myocardial cells were divided into blank group, model group and experimental -L, -M, -H groups. The H₉C₂ myocardial cells injury model of model group and experimental -L, -M, -H groups were constructed with X-ray 6 Gy irradiation. After successful modeling, the blank and model groups were treated with 0.9% NaCl; the experimental -L, -M, -H groups were treated with 2%, 5%, 10% AAU drug-containing serum 0.5 mL, respectively. After 24 h of treatment, EdU staining was used to observe cell survival rate, Western blot was used to detect the expression levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) and Caspase-1 protein.

The cell viability rates of the experimental -M, -H groups, model group and blank group were (89.91±1.16)%, (91.66±1.93)%, (78.61±1.30)% and (98.10±0.20)%; the relative expression levels of NLRP3 protein were 0.79±0.05, 0.74±0.04, 0.94±0.02 and 0.53±0.12; the relative expression levels of ASC protein were 0.40±0.04, 0.41±0.07, 0.48±0.01 and 0.22±0.02; the relative expression levels of Caspase-1 protein were 0.25±0.03, 0.23±0.01, 0.58±0.04 and 0.18±0.01; the relative expression levels of IL-18 protein were 0.67±0.03, 0.62±0.02, 0.91±0.07 and 0.61±0.03; the relative expression levels of IL-1β protein were 0.41±0.06, 0.38±0.02, 0.78±0.02 and 0.34±0.09, respectively. Compared with the experimental -M, -H groups, the differences of above indicators in the model group were statistically significant (all P<0.05).

AAU may protect H₉C₂ myocardial cells from injury induced by X-ray radiation by regulating NLRP3 inflammasome.

To explore the effects and mechanism of zoledronic acid on bone metabolism in osteoporosis rats under hypoxic environment.

The rat model of osteoporosis in hypoxia environment was established by bilateral ovary extraction combined with low-pressure oxygen chamber feeding, and was randomly divided into model group, control group and experimental -L, -M, -H groups, with 10 rats per group; another 10 normal rats were selected as the normal group. Experimental -L, -M, -H groups were given 2.5, 5.0 and 10.0 mg·kg^-1 zoledronic acid by subcutaneous injection, once a week. Control group was given 0.105 mg·kg^-1 estradiol valerate by intragastric administration, once a day. Normal and model groups were injected with equivalent 0.9% NaCl by subcutaneous injection. Six groups were treated for 4 weeks. The serum levels of tartrate-resistant acid phosphatase (TRAP), alkaline phosphatase (ALP) and bone gla protein (BGP) were detected by enzyme-related immunosorbent assay. The bone mineral density (BMD) was measured by dual-energy X-ray. The expression levels of mitogen extracellular signal-regulated kinase (MEK)/extracellular regulated protein kinases (ERK) in femur were detected by Western blot.

The TRAP levels of experimental-M, experimental -H, control, model and normal groups were (78.06±1.16), (69.39±1.44), (70.19±1.24), (94.39±1.04) and (64.95±1.19) U·L^-1; the ALP levels were (61.72±3.10), (45.40±0.82), (43.53±1.52), (88.69±2.29) and (42.01±1.56) U·L^-1; the BGP levels were (14.82±0.28), (12.29±0.28), (12.92±0.29), (19.96±0.46) and (11.79±0.59) ng·mL^-1; the BMD levels were (0.28±0.02), (0.35±0.04), (0.38±0.03), (0.16±0.02) and (0.41±0.06) g·cm^-2; the relative expression levels of p-MEK protein were 0.64±0.06, 0.84±0.05, 0.78±0.01, 0.15±0.06 and 0.88±0.04; the relative expression levels of p-ERK protein were 0.70±0.03, 0.95±0.02, 0.84±0.07, 0.29±0.03 and 0.97±0.06, respectively. Compared with the model group, the above indexes in the experimental-M, -H groups and control group were statistically significant (all P<0.05).

Zoledronic acid can improve bone mineral density and bone metabolism in hypoxic osteoporosis rats, the mechanism may be related to the regulation of MEK/ERK pathway.

To develop a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of crizotinib in human plasma and apply this method to determine its concentration in patients with non-small cell lung cancer after administrating.

Osimertinib was employed as the internal standard. The plasma samples were performed through protein precipitation with acetonitrile and chromatographed on Agilent Eclipse XDB C₁₈ (2.1 mm×100.0 mm, 3.5 μm), mobile phase was consisted of acetonitrile containing 0.1% formic acid - 10 mmol·L^-1 ammonium acetate containing 0.1% formic acid, flow rate was 0.25 mL·min^-1, the column temperature was 30 ℃. The whole analytical time was 3 min. The detection was performed by positive ion electrospray ionization in multiple reaction monitoring mode, the quantitative analysis ion pairs were m/z 450.1→259.2 and m/z 500.2→72.1. The specificity, standard curve and lower limit of quantitation, precision and recovery, stability, matrix effect and residual effect of the method were investigated. After full validation, the method was applied to the determination of crizotinib in human plasma.

Good linearity was observed over the concentration range of 10.0-2 500.0 μg·L^-1, the calibration curve was y=4.54×10^-3x+3.22×10^-2 (r=0.992 7). The accuracies were between 97.66%-105.55%. The relative standard deviation of inter-day and intra-day precision were less than 8.49%. The average recoveries and matrix effects were between 80.10%- 86.05% and 95.22%-110.66%.

The LC-MS/MS method is developed which meets the requirements of biological sample analysis and it can quickly, accurately monitor the concentration of crizotinib.

To establish in vitro and in vivo correlations from in vitro dissolution data and in vivo data from animals.

In vitro method: an aqueous gelatin solution was first prepared with alkaline gelatin, paliperidone palmitate was added into the solution and mixed well, gelatin hydrogel was obtained by refrigerating in the refrigerator for 1-2 h. The hydrogel was put into paddle dissolution apparatus for dissolution experiments, the concentration of the drug was detected by high-performance liquid chromatography and the degree of release was calculated. In vivo method: Timing was started after paliperidone palmitate was injected into Beagle dogs via intramuscular injection, blood was collected at 0 h before and 6 h-36 d after drug administration, respectively, the blood concentration was detected by liquid chromatography-tandem mass spectrometry (LC/MS/MS) technique and the blood concentration-time curve was plotted. Finally, the obtained in vitro data were fitted by Weibull function, the in vivo data were transformed by Nelson-Wagner method and area under the curve method. Graphs were constructed to evaluate the correlation between in vivo and in vitro drug release.

The in vitro release was approaching 100% by day 13, while the in vivo release was approaching 100% by day 30-36. Both Nelson-Wagner method and area under the curve method yielded that in vivo and in vitro were correlated.

It is feasible to establish in vitro correlations from in vivo dissolution data and in vivo data from animals.

To observe the clinical efficacy and safety of citric acid solution anticoagulant hemodialysis combined with tacrolimus tablets in the treatment of patients with nephrotic syndrome complicated with acute kidney injury.

The patients with nephrotic syndrome complicated with acute kidney injury were randomly divided into control group and treatment group. The control group was given low molecular weight heparin anticoagulation (initial dose 2 000-3 000 U, additional dose 500-1 000 U·h^-1) hemodialysis + oral tacrolimus capsule, oral tacrolimus capsules at an initial dose of 0.05 mg·kg^-1·d^-1, twice a day. The treatment group was given 180 mL·h^-1 4% sodium citrate anticoagulant hemodialysis + oral tacrolimus capsules, with an initial dose of 0.05 mg·kg^-1·d^-1 twice a day. Two groups were treated for 4 weeks. The clinical efficacy, renal function, coagulation function and safety were compared between the two groups.

Treatment group was enrolled 85 cases, 2 cases dropped out, and 83 cases were finally included in the statistical analysis. Control group was enrolled 84 cases, 3 cases dropped out, and 81 cases were finally included in the statistical analysis. After treatment, the total effective rates of treatment group and control group were 96.38% (80 cases/83 cases) and 92.59% (75 cases /81 cases), without statistical significance (P>0.05). After treatment, the albumin levels of treatment group and control group were (40.19±7.01) and (35.72±6.12) g·L^-1, the serum creatinine levels were (82.39±12.66) and (73.65±11.27) μmoL·L^-1, the 24 h urinary proteins were (1.41±0.26) and (1.75±0.31) g, the prothrombin time was (15.32±1.61) and (18.71±1.74) s, the activated partial thrombin time was (46.29±3.08) and (50.23±3.12) s, the thrombin time was (18.01±1.73) and (21.04±1.85) s, the urine output was (1 358.79±397.22) and (1 283.02±378.97) mL·d^-1, and the differences of above indexes were statistically significant between two groups (all P<0.05). The adverse drug reactions of treatment group were metabolic alkalosis, metabolic acidosis, hypocalcemia and citric acid accumulation, while those in the control group were bleeding, metabolic alkalosis and metabolic acidosis. The incidences of total adverse drug reactions in treatment and control groups were 16.87% and 14.81%, without statistical significance (P>0.05).

Compared with lower molecular weight heparin calcium anticoagulant hemodialysis combined with tacrolimus tablets, sodium citrate anticoagulant hemodialysis combined with tacrolimus tablets can more effectively improve renal function and coagulation function in patients with nephrotic syndrome and acute kidney injury, and will not increase the incidence of adverse drug reactions.

To observe the effects of different dosage forms of Yupingfeng powder on lipopolysaccharide induced pyroptosis of mouse lung macrophages.

The RAW264.7 cells were randomly divided into normal, model, decoction piece and granule groups. Except for normal group, the other 3 groups were used lipopolysaccharide concentration to conduct a cellular inflammation model. No drug will be given to the normal group, and normal cultivation; the model group was treated with lipopolysaccharide; the decoction piece group was treated with lipopolysaccharide and decoction piece containing serum; the granule group was treated with lipopolysaccharide and granule containing serum. The macrophage inflammatory factor was detected by enzyme-linked immunosorbent assay. The expression levels of cell pyroptosis pathway protein were detected by Western blot.

The tumor necrosis factor -α (TNF-α) contents in cell supernatant of normal, model, decoction piece and granule groups were (48.19±12.00), (154.48±34.84), (125.80±29.66) and (125.73±28.88) pg·mL^-1; the interleukin (IL) -18 levels were (21.36±4.36), (78.33±14.41), (49.77±9.50) and (49.27±9.40) pg·mL^-1; the Caspase-1 levels were (39.72±10.58), (149.49±20.92), (93.25±16.73) and (91.93±16.45) pg·mL^-1; the IL-1β levels were (17.14±3.41), (74.43±16.04), (45.01±9.40) and (43.34±8.96) pg·mL^-1; the relative expression levels of TNF-α mRNA were 1.00±0.05, 2.17±0.07, 1.50±0.20 and 1.35±0.08; the relative expression levels of nuclear factor kappa-B mRNA were 1.00±0.01, 2.02±0.01, 1.52±0.01 and 1.50±0.03; the relative expression levels of IL-18 mRNA were 1.00±0.03, 1.48±0.04, 1.18±0.04 and 1.13±0.10; the relative expression levels of Caspase-1 mRNA were 1.00±0.04, 1.90±0.03, 1.59±0.04 and 1.49±0.10; the relative expression levels of IL-1β mRNA were 1.00±0.10, 1.74±0.08, 1.26±0.06 and 1.23±0.13, respectively. Compared with the model group, there were statistically significant differences of the above indicators in the granule group and the decoction piece group (P<0.05, P<0.01), but there were no statistically significant differences in the above indicators between the decoction piece group and the granule group (all P>0.05).

Both formulations of Yupingfeng powder can reduce the expression level of proteins on the cell pyroptosis pathway and inhibit inflammatory response.

To observe the clinical efficacy and safety of salmon calcitonin nasal spray combined with calcium carbonate D₃ tablets in the treatment of disuse osteoporosis patients after ankle fracture surgery.

The disuse osteoporosis patients with postoperative ankle fractures were randomly divided into control group and treatment group using a random number table method. The control group was treated with calcium carbonate D₃ tablets 600 mg per time, twice daily, while the treatment group received salmon calcitonin nasal spray 200 U per time, once daily, in addition to the treatment in the control group. Both groups were treated for 6 months. The clinical efficacy, bone metabolic markers (osteocalcin, alkaline phosphatase), changes in bone mineral density (BMD) and postoperative pain level (VAS) were compared between the two groups, the safety evaluation was also conducted.

A total of 3 cases dropped out during the trial. Ultimately, the treatment group and the control group included 63 and 62 patients, respectively. After treatment, the overall effective rates for the treatment group and the control group were 93.65% (59 cases/63 cases) and 75.81% (47 cases/62 cases), respectively, with statistically significant difference (P<0.01). After 3 months of treatment, the BMD of the control and treatment groups were (0.87±0.09) and (0.93±0.08) g·cm^-2, respectively; serum osteocalcin levels were (20.10±3.20) and (22.80±3.50) ng·mL^-1, respectively; alkaline phosphatase levels were (74.50±12.30) and (80.10±13.20) U·L^-1, respectively; after 6 months of treatment, the BMD values for the control and treatment groups were (0.90±0.08) and (1.01±0.07) g·cm^-2, respectively; serum osteocalcin levels were (21.00±3.40) and (24.60±3.40) ng·mL^-1, respectively; alkaline phosphatase levels were (76.00±12.50) and (84.90±13.80) U·L^-1, respectively; statistically significant differences were observed for the above indicators between the experimental and control groups (P<0.05, P<0.001). After 3 months of treatment, the visual analog scale (VAS) scores of the control and treatment groups were (5.30±1.30) and (4.50±1.20) points, respectively; after 6 months of treatment, the VAS scores were (4.20±1.10) and (2.90±1.00) points, respectively; the differences between the treatment and control groups were statistically significant (all P<0.001). The main adverse drug reactions in the treatment group were nasal discomfort and nausea, while the control group were gastrointestinal discomfort. The total incidence of adverse drug reactions in the treatment and control groups were 7.94% (5 cases/63 cases) and 4.84% (3 cases/62 cases), respectively, without statistically significant difference between the two groups (P>0.05).

Salmon calcitonin nasal spray combined with calcium carbonate D₃ tablets can significantly increases BMD in disuse osteoporosis patients after ankle fracture surgery, reduce the risk of disuse osteoporosis, with good safety.