A method for simultaneous isolation and clean-up pyrethroid insecticides (methothrin, fenpropathrin,cyhalothrin, permethrin, cypermethrin, fenvalerate and deltamethrin)with a solid phase extractionmethod (Sep-Pak C18 cartridge) from biological samples (human plasma and urine) is presented.The systematic separation of the insecticides was excellently performed by wide-bore capillary gaschromatography with flame ionization detection and temperature programming. The recoveries werebetween 81% and 93% for plasma, and 90% and 102% for urine. Mixing the samples with 70%methanol and eluting with chloroform demonstrated excellent adsorption on C18 and the best recoveries,respectively.

Based on the Pharmaceutical Comprehensive Knowledge and Skills Outline in the year of 2003, 2007, 2011 and 2015,the changes and the trend in the chapters and contents of Pharmaceutical Comprehensive Knowledge and Skills were analyzed and discussed,the meaning of versions was elaborated,and the shortcomings were discussed as well. The catalog of the chapters and contents conformed to the economic and social development in China,which is moving toward the direction of clinical pharmacy service.

Objective:To explore the annexin A2 (ANXA2) expression and distribution in dorsal root ganglion (DRG) after chronic compression of DRG (CCD) in rat models.Methods:One hundred and two adult male Wistar rats were randomly divided into control group ( n=24), CCD model group A (7 d after modeling, n=30), CCD model group B (14 d after modeling, n=24), and CCD model group D (28 d after modeling, n=24). Rats in the later 3 groups were established CCD models with the help of "U" rod screw. Mechanical withdrawal threshold (MWT) and thermal radiation paw withdrawal latency (TWL) were measured by mechanical pain stimulator and thermal pain stimulator. The ANXA2 protein expression in the DRG was detected by Western blotting and immunofluorescent staining. The distributions of ANXA2 and class III β-tubulin (TUBB3) positive cells in DRG were detected by immunofluorescence double staining. Results:As compared with those in the control group, MWT and TWL in the CCD model group A and CCD model group B were significantly decreased ( P<0.05). Western blotting showed that ANXA2 protein expression in the DRG of CCD model group A was statistically increased as compared with that in the control group ( P<0.05). Immunofluorescent staining showed that the immunoreactivity of ANXA2 in DRG of CCD model group A was enhanced as compared with that in control group. Immunofluorescence double staining showed that ANXA2 was mainly expressed in the cell membrane of neurons in the DRG of CCD model group A. Conclusion:The mechanical and thermal pain thresholds are decreased, while the ANXA2 protein expression at the pressure side of DRG is up-regulated and the immunoreactivity is increased in CCD models; ANXA2 may be involved in the occurrence and development of pathological neuralgia after CCD.

Pituitary tumors are usually benign but can occasionally exhibit hormonal and proliferative behaviors. Dysregulation of the G1/S restriction point largely contributes to the over-proliferation of pituitary tumor cells. F-box protein S-phase kinase-interacting protein-2 (SKP2) reportedly targets and inhibits the expression of p27(Kip1), a well-known negative regulator of G1 cell cycle progression. In this study, SKP2 expression was found to be upregulated while p27(Kip1) expression was determined to be downregulated in rat and human pituitary tumor cells. Furthermore, SKP2 knockdown induced upregulation of p27(Kip1) and cell growth inhibition in rat and human pituitary tumor cells, while SKP2overexpression elicited opposite effects on p27(Kip1) expression and cell growth. The expression of microRNA-186 (miR-186) was reported to be reduced in pituitary tumors. Online tools predicted SKP2 to be a direct downstream target of miR-186, which was further confirmed by luciferase reporter gene assays. Moreover, miR-186 could modulate the cell proliferation and p27(Kip1)-mediated cell cycle alternation of rat and human pituitary tumor cells through SKP2. As further confirmation of these findings, miR-186 and p27(Kip1) expression were downregulated, while SKP2 expression was upregulated in human pituitary tumor tissue samples; thus, SKP2 expression negatively correlated with miR-186 and p27(Kip1) expression. In contrast, miR-186 expression positively associated with p27(Kip1) expression. Taken together, we discovered a novel mechanism by which miR-186/SKP2 axis modulates pituitary tumor cell proliferation through p27(Kip1)-mediated cell cycle alternation.
Animals ; Cell Cycle ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p27 ; Genes, Reporter ; Humans ; Luciferases ; Pituitary Neoplasms ; Rats ; Up-Regulation

Pulmonary hypertension (PH) is an extremely malignant pulmonary vascular disease of unknown etiology. ADAR1 is an RNA editing enzyme that converts adenosine in RNA to inosine, thereby affecting RNA expression. However, the role of ADAR1 in PH development remains unclear. In the present study, we investigated the biological role and molecular mechanism of ADAR1 in PH pulmonary vascular remodeling. Overexpression of ADAR1 aggravated PH progression and promoted the proliferation of pulmonary artery smooth muscle cells (PASMCs). Conversely, inhibition of ADAR1 produced opposite effects. High-throughput whole transcriptome sequencing showed that ADAR1 was an important regulator of circRNAs in PH. CircCDK17 level was significantly lowered in the serum of PH patients. The effects of ADAR1 on cell cycle progression and proliferation were mediated by circCDK17. ADAR1 affects the stability of circCDK17 by mediating A-to-I modification at the A5 and A293 sites of circCDK17 to prevent it from m1A modification. We demonstrate for the first time that ADAR1 contributes to the PH development, at least partially, through m1A modification of circCDK17 and the subsequent PASMCs proliferation. Our study provides a novel therapeutic strategy for treatment of PH and the evidence for circCDK17 as a potential novel marker for the diagnosis of this disease.