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 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 explore the protective mechanism of Celosin Ⅰ on retinal ganglion cells by regulating reactive oxygen species (ROS) mediated mitochondrial apoptosis pathway.

Twenty-four New Zealand white rabbits were randomly divided into sham-operation, model, control and experimental groups, with 6 rabbits per group. The sham-operation group only had the bulbar conjunctiva cut and sutured, the other 3 groups were received with optic nerve clamp injury to contruct the optic nerve injury model. The sham-operation and model groups were given equal volumes of physiological saline by gavage, the control group was given a dose of 0.15 mg·kg^-1 mecobalamine solution by gavage, and the experimental group was given a dose of 30 mg·kg^-1 Celosin Ⅰ solution by gavage. Four groups were administered a daily gavage treatment for four consecutive weeks. The Tunnel method was used to detect apoptosis of retinal ganglion cells, and immunofluorescence was employed to detect the expression of ROS, superoxide dismutase (SOD) and caspase in retinal tissues. The expression levels of B-cell lymphoma-2 (Bcl-2), Bcl-2-like protein 1 (Bcl-xl), Bcl-2-associated X protein (Bax), and cytochrome C (CytC) proteins in retinal tissues were assessed using Western blotting.

After four weeks of intervention, the apoptosis rates of ganglion cells in the experimental, control, model and sham-operated groups were (7.65±1.20)%, (11.65±1.29)%, (17.66±0.93)% and (0.97±0.14)%; the expression levels of ROS in retinal tissues were 6.14±0.57, 8.25±0.96, 16.51±1.07 and 2.05±0.23; the expression levels of SOD were 2.45±0.33, 1.48±0.23, 0.74±0.13 and 3.19±0.30; the expression levels of caspase-3 were 8.55±1.11, 12.68±0.92, 18.74±1.97 and 3.08±0.20; the expression levels of caspase-9 were 3.69±1.20, 7.79±1.13, 12.81±1.72 and 2.31±0.30; the relative expression levels of Bcl-2 were 0.46±0.02, 0.64±0.07, 0.77±0.05 and 0.20±0.03; the relative expression levels of Bcl-xl were 0.46±0.05, 0.61±0.05, 0.78±0.03, and 0.20±0.03; the relative expression levels of Bax were 0.50±0.07, 0.69±0.18, 0.82±0.13 and 0.14±0.04; the relative expression levels of CytC were 0.43±0.15, 0.80±0.14, 1.08±0.19 and 0.26±0.07. The differences in the aforementioned indicators in the experimental group compared with control group were statistically significant (all P<0.05).

Celosin Ⅰ reduces ROS levels in retinal tissue of a model of optic nerve injury, inhibits the expression of mitochondrial apoptosis related proteins in retinal ganglion cells, and reduces retinal ganglion cell apoptosis.

To establish a method for the quantification of aripiprazole in male SD rats muscle, and apply it to the research of drug residue at injection site in rats after administration of long-acting aripiprazole injection (LAI), and to compare the pharmacodynamics in mice.

An ultra-high performance liquid chromatography tandem mass spectroscopy (UPLC-MS/MS) assay was used to determine the concentration of aripiprazole in rats muscle. Sample pre-treatment: Muscle tissue was homogenized in a 30% acetonitrile solution containing 5% acetic acid, and then precipitated with 0.1% formic acid in acetonitrile for detection. Chromatography column: Waters HSS T3^TM C₁₈ column (2.1 mm×50.0 mm, 1.8 μm). Mobile phase: 5% acetonitrile and 95% acetonitrile both containing 0.1% formic acid. Flow rate: 0.6 mL·min^-1. Injection volume: 5 μL. Mass spectrometry conditions: electrospray ion-source positive ions mode, multiple reaction monitor (MRM) scanning mode. After intramuscular administration of 25 mg·mL^-1 aripiprazole microspheres for injection (MS) or long-acting aripiprazole for injection (brand name: Abilify Maintena^®, AM), the drug residue in male SD rats was determined using an UPLC-MS/MS method. The hyperlocomotion model was established by acute administration of phencyclidine (PCP). And a comparative study was conducted on the efficacy of MS (40, 80, 240 mg·kg^-1) and AM (80 mg·kg^-1).

After 28 days of administration, the drug residue ratio of MS group was 6.27%-12.38%, while that of AM group was 23.31%. After 35 days of administration, the average drug residue ratios were 0.58%-2.94% and 10.28%-18.87%, respectively. Both the AM group and each dose (40, 80, 240 mg·kg^-1) of MS group could inhibit the spontaneous movement of mice induced by PCP.

The inhibitory effect of Aripiprazole Microsphere for injection is comparable to that of Abilify Maintena on the spontaneous movement of mice caused by PCP. And there is a certain dose-response relationship for MS, while the drug residue at the injection site of MS is much lower than that of Abilify Maintena.

To analysis the relationship between urinary albumin creatinine ratio (UACR) and heart rate variability (HRV) in patients with type 2 diabetes mellitus (T2DM).

Patients with T2DM were divided into control group (patients with T2DM only) and treatment group (patients with T2DM and proteinuria) using the cohort method. The treatment group was further divided into a microalbuminuria subgroup (UACR 30 - 300 mg·g^-1) and a macroalbuminuria subgroup (UACR≥300 mg·g^-1). Collected general clinical data, biochemical indicators and 24-hour holter electrocardiogram reports from patients, recorded HRV-related indicators [including pairs of normal N-N Intervals differ by more than 50 ms (PNN50), standard deviation of normal-to-normal R-R intervals (SDNN), etc] of each group of patients, and calculated body mass index (BMI) and estimated glomerular filtration rate (eGFR). Compared the differences in clinical data and HRV parameters among groups, analyzed the impact of UACR on HRV and their interrelationships, and used the receiver operating characteristic curve (ROC) to predict the optimal cutoff point for the occurrence of proteinuria in T2DM patients.

The treatment group enrolled 190 cases, and the control group enrolled 184 cases; the microalbuminuria subgroup enrolled 120 cases, and the macroalbuminuria subgroup enrolled 70 cases. The levels of UACR in the treatment and control groups were 113.99 and 12.76 mg·mmol^-1 Cr, the levels of UAER were 74.81 and 10.92 μg·min^-1, the levels of UA were (353.83±96.41) and (326.17±81.64) μmol·L^-1, the levels of PNN50 were 2.25 and 3.95, the levels of SDNN were 102.83±38.10 and 114.14±31.23, the levels of HRV triangular index were 20.80 and 25.55, respectively; the differences of above results were statistically significant between two groups (all P<0.05). In the macroalbuminuria and microalbuminuria subgroups, the levels of UACR were 1 088.17 and 64.64 mg·mmol^-1 Cr, the levels of UAER were 878.65 and 44.26 μg·min^-1, the levels of UA were (375.88±97.58) and (340.97±93.74) μmol·L^-1, the levels of PNN50 were 1.50 and 2.70, the levels of SDNN were 88.00 and 108.00, the levels of HRV triangular index were 19.49±7.77 and 24.48±8.84, respectively; the differences of above results were statistically significant between two subgroups (all P<0.05). Logistic regression analysis showed that the duration of diabetes mellitus, fasting blood glucose (FPG) and HRV triangle index were the influencing factors for the occurrence of proteinuria in patients with type 2 diabetes mellitus. ROC showed that HRV triangle index had the largest area under ROC curve, with 66.10% sensitivity and 63.00% specificity.

UACR in T2DM patients is closely related to the duration of DM, FPG and HRV triangle index, with the increase of UACR in T2DM patients, the risk of cardiovascular autonomic neuropathy increases.

To explore the regulation by cardamonin through targeting family 110 member A (FAM110A) regulate Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway on human breast cancer cells MCF-7 and effects of aerobic glycolysis and epithelial-mesenchymal transformation.

The breast cancer cells MCF-7 were divided into control group (cultured normally), experimental group (cells treated with 20 μmol·L^-1 cardamonin), oe-NC group (transfected with negative control plasmid oe-NC on the basis of experimental group), oe-FAM110A group (transfected with overexpressed plasmid oe-FAM110A on the basis of experimental group). Quantitative real-time polymerase chain reaction was used to detect the relative expression levels of FAM110A messenger RNA (mRNA) in each group of cells. The relative expression levels of FAM110A protein and the expression of E-cadherin and N-cadherin, which are related to epithelial mesenchymal transformation, were detected by immunofluorescence assay. Detection indexes of aerobic glycolysis in each group were detected by kit. The JAK2/STAT3 signaling pathway related protein phosphorylation level in each group was detected by Western blot.

The relative expression levels of FAM110A mRNA in control group and experimental group were 1.00±0.12 and 0.54±0.10, respectively, and the relative expression levels of FAM110A protein were 1.00±0.15 and 0.13±0.03, respectively; the mRNA and protein relative expression levels of FAM110A in experimental group were significantly different from those in control group (all P<0.05). The relative expression levels of E-cadherin protein in control group, experimental group, oe-NC group and oe-FAM110A group were 1.00±0.14, 2.98±0.51, 3.00±0.54 and 1.47±0.26, respectively; the relative expression levels of N-cadherin protein were 1.00±0.11, 0.41±0.08, 0.47±0.10 and 0.94±0.17; the relative lactic acid production were 1.00±0.19, 0.61±0.11, 0.59±0.09 and 0.98±0.16, respectively; the relative glucose absorption values were 1.00±0.16, 0.41±0.08, 0.42±0.08 and 0.88±0.15, respectively; the relative adenosine triphosphate production were 1.00±0.20, 0.29±0.05, 0.33±0.06 and 0.96±0.18, respectively; the phospho-JAK2/JAK2 levels were 1.00±0.17, 0.54±0.11, 0.58±0.10 and 0.91±0.15, respectively; the phospho-STAT3/STAT3 levels were 1.00±0.14, 0.37±0.06, 0.38±0.06 and 0.92±0.12, respectively; the above indexes in the experimental group were significantly different from those in the control group, and those in the oe-FAM110A group were significantly different from those in the oe-NC group (all P<0.05).

Cardamonin can inhibit aerobic glycolysis and epithelial-mesenchymal transition in breast cancer cells MCF-7, and the mechanism may be through down-regulating FAM110A and inhibiting JAK2/STAT3 signaling pathway.