Pharmacokinetic study of tanshinol and ligustrazine in rat plasma after intravenous administration of tanshinol and Danshen Chuanxiongqin Injection.

To investigate the effect of ligustrazine on the pharmacokinetic profile of tanshinol after intravenous administration in rats, a sensitive liquid chromatography tandem mass spectrometry method was developed and validated for quantitative determination of tanshinol and ligustrazine in rat plasma. After prepared by protein precipitation, the analytes were separated on a Waters Acquity HSS T3 column (100 × 2.1 mm, 1.8µm) and eluted by 0.1% formic acid in water and acetonitrile at a flow rate of 0.4 ml/min. The precursor-product ion transitions were m/z 197.0 → 135.0 for tanshinol, m/z 417.1 → 255.1 for liquiritin (internal standard) in negative ion mode and m/z 137.1 → 55.0 for ligustrazine in positive ion mode. To avoid the interference of tanshinol metabolite transformation, the stability of analytes in samples collected after administration was assessed. The validated method was successfully applied to a pharmacokinetic study after intravenous administration of single tanshinol and Danshen Chuanxiongqin Injection. After Danshen Chuanxiongqin injection administration, the values of elimination half-time, area under the concentration-time curve and Co were 0.36 ± 0.13 h, 1.29 ± 0.37 µg/ml h and 10.51 ± 2.58 µg/ml for male rats, respectively. In the single tanshinol group, the corresponding values were 0.56 ± 0.24 h, 1.85 ± 0.44 µg/ml h and 14.11 ± 2.26 µg/ml for male rats-30-40% higher than those for the Danshen Chuanxiongqin Injection group. There was a significant different between male and female rats. This study provided information on the influence of ligustrazine on the pharmacokinetic characteristics of tanshinol after intravenous administration of Danshen Chuanxiongqin Injection in rats, which will be helpful for its clinical application.

The protective effect of magnesium lithospermate B against glucose-induced intracellular oxidative damage.

OBJECTIVES: To investigate the effects of magnesium lithospermate B (LAB) on intracellular reactive oxygen species (ROS) production induced by high dose of glucose or H(2)O(2), we explored the influences of LAB on the expression of heme oxygenase-1 (HO-1) and nuclear factor E2-related factor-2 (Nrf2) in HEK293T cells after treatment with high dose of glucose. MATERIALS AND METHODS: The total nuclear proteins in HEK293T cells were extracted with Cytoplasmic Protein Extraction Kit. The ROS level was determined by flow cytometry. The mRNA and protein expression of HO-1 and Nrf2 were determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot. RESULTS: LAB reduced the ROS production in HEK293T cells cultured under oxidative stress. High dose of glucose enhanced the expression of HO-1 mRNA and HO-1 protein in a time-dependent manner. LAB enhanced the expression of HO-1 mRNA and HO-1 protein in a dose-dependent manner treated with high dose of glucose. The amount of Nrf2 translocation was enhanced after cells were pretreated with 50µmol/L or 100µmol/L LAB. Silencing of Nrf2 gene eliminated the enhanced expression of HO-1 protein induced by high dose of glucose plus LAB. CONCLUSIONS: LAB plays an important role against glucose-induced intracellular oxidative damage. The enhanced expression of HO-1 mRNA and HO-1 protein caused by LAB is regulated via Nrf2 signal pathway.

Effect of bronchoalveolar lavage on the clinical efficacy, inflammatory factors, and immune function in the treatment of refractory pneumonia in children.

BACKGROUND: Refractory pneumonia is a special type of pneumonia in children. This study aimed to analyze the effect of bronchoalveolar lavage (BAL) on the clinical efficacy, inflammatory factors, and immune function in the treatment of pediatric refractory pneumonia. METHODS: A total of 196 children with refractory pneumonia admitted to our hospital from January 2017 to January 2020 were enrolled and allocated to a study group (n=99) and a control group (n=97). The study group was treated with BAL treatment plus conventional treatment, and the control group was treated with conventional treatment. The clinical efficacy, time of fever regression, time of cough relief, and length of hospital stay were compared between groups. Changes in inflammatory factors, immune function, pulmonary ventilation function, and complications were analyzed. The levels of inflammatory factors in BAL fluid were compared. RESULTS: The times of fever remission, cough relief, and hospital stay of the study group was shorter than those of the control group, and the total clinical effective rate of the study group was higher. At any time after treatment, the levels of interleukin-6 (IL-6), C-reactive protein (CRP), and tumor necrosis factor-alpha (TNF-α) in the study group were lower than the control group. After 3-day of treatment, IL-6, CRP and TNF-α in BAL fluid in the study group were significantly decreased compared with before treatment. Immunoglobulin A (IgA) and immunoglobulin G (IgG) levels in the study group were higher than those in the control group at any time after treatment, and immunoglobulin M (IgM) levels were lower than in the control group. The levels of oxygenation index (OI), lung dynamic compliance (Cdyn), and work of breathing (WOB) in the study group were higher than those in the control group at any time after treatment. CONCLUSIONS: BAL treatment can effectively relieve the inflammatory response, improve immune function and lung ventilation function in children with refractory pneumonia. The clinical effect is remarkable and worthy of promotion.

Effect of docetaxel on expression of eIF3a in human lung cancer A549 cell line.

OBJECTIVE: To explore the dose-dependent and time-dependent effect of docetaxel on the expression of mammalian eukaryotic initiation factor 3 subunit A (eIF3a) in lung cancer cell line. METHODS: The human lung cancer cell line A549 was treated with gradient concentrations of docetaxel for different time. Real-time PCR and Western blot were used to detect mRNA and protein expression levels of eIF3a and alpha-tubulin, respectively. RESULTS: Docetaxel did not affect alpha-tubulin expression at either mRNA level or protein level. When A549 cells were treated with high concentration of docetaxel (30 µg/L), the expression level of eIF3a mRNA tended to increase in a time-dependent manner. Protein expression level of alpha-tubulin was not associated with eIF3a expression significantly in cells treated by docetaxel. CONCLUSION: Docetaxel could slightly increase the expression of eIF3a mRNA, and eIF3a does not regulate the expression of alpha-tubulin in A549 cells treated by docetaxel.

TNF-α and MTHFR Polymorphisms Associated with Cerebral Palsy in Chinese Infants.

This study aims to examine whether the presence of polymorphisms in TNF-α (rs361525 and rs1799724) and MTHFR (rs1476413 and rs9651118) genes is associated with the pathogenesis of cerebral palsy (CP). A total of 105 CP patients and 114 age-, gender-, and ethnicity-matched healthy controls were genotyped for the selected polymorphisms, using TaqMan allelic discrimination assay. Odds ratios (OR) and 95 % confidence intervals (CI) were determined to measure the strength of associations of TNF-α and MTHFR polymorphisms with CP. The proportion of subjects with the gestational age more than 37 weeks or asphyxia was much larger in cases compared with controls (gestational age 63.8 vs. 34.2 %; asphyxia 25.7 vs. 7.9 %). The genotype frequencies of TNF-α rs1799724 were similar between groups (P > 0.05), yet the allele distributions were significantly different (P < 0.05). Both the allele and genotype distributions of MTHFR rs9651118 polymorphism varied significantly between the groups (P < 0.05). Subgroup analysis based on gestational age indicated a significant association between rs361525 and rs9651118 and CP with or without premature. TNF-α protein concentrations were significantly increased among patients with rs361525 GG genotype compared with controls. Also, a significant increase in the risk of CP was observed to be associated with the interactions of TNF-α rs1799724 and MTHFR rs9651118 (OR 2.75, 95 % CI 1.23-6.13). These data suggest that polymorphisms in TNF-α and MTHFR genes might be involved in the pathogenesis of CP in Chinese infants.