Proteases and their inhibitors as prognostic factors for high-grade serous ovarian cancer

Ovarian carcinoma is one of the most lethal malignancies, but only very few prognostic biomarkers are known. The degradome, comprising proteases, protease non-proteolytic homologues and inhibitors, have been involved in the prognosis of many cancer types, including ovarian carcinoma. The prognostic significance of the whole degradome family has not been specifically studied in high-grade serous ovarian cancer. A targeted DNA microarray known as the CLIP-CHIP microarray was used to identify potential prognostic factors in ten high-grade serous ovarian cancer women who had early recurrence (<1.6 years) or late/no recurrence after first line surgery and chemotherapy. In women with early recurrence, we identified seven upregulated genes (TMPRSS4, MASP1/3, SPC18, PSMB1, IGFBP2, CFI – encoding Complement Factor I – and MMP9) and one down-regulated gene (ADAM-10). Using immunohistochemistry, we evaluated the prognostic effect of these 8 candidate genes in an independent cohort of 112 high-grade serous ovarian cancer women. Outcomes were progression, defined according to CA-125 criteria, and death. Multivariate Cox proportional hazard regression models were done to estimate the associations between each protein and each outcome. High ADAM-10 expression (intensity of 2–3) was associated with a lower risk of progression (adjusted hazard ratio (HR): 0.51; 95% confidence interval (CI): 0.29-0.87). High complement factor I expression (intensity 2–3) was associated with a higher risk of progression (adjusted HR: 2.30, 95% CI: 1.17–4.53) and death (adjusted HR: 3.42; 95% CI: 1.72–6.79). Overall, we identified the prognostic value of two proteases, ADAM-10 and complement factor I, for high-grade serous ovarian cancer which could have clinical significance.     

Implementing Substance Use Screening in Family Planning

Although substance use has negative health effects on women, especially during the reproductive years, family planning practices in which nurse practitioners are key care providers, generally do not adequately screen and intervene for alcohol and drugs. Screening, brief intervention, and referral to treatment (SBIRT) can limit the effects of substance use on women and families. This study explored barriers and facilitators to SBIRT through qualitative analysis of focus groups with family planning providers. Results suggest family planning providers are favorable to implementation. Barriers include training and organizational support; facilitators include use of electronic health records and setting patient expectations.     

Distributive differences of P2Xs between the forelimb and hind limb of adjuvant arthritis rats and intervention by Notopterygh rhizoma et radix

Context: Notopterygium incisum Ting ex H. T. Chang (Umbelliferae) (NI) specializes in treatment of upper limb rheumatoid arthritis (RA), but the exact mechanism is unclear. P2Xs are useful targets for inflammatory pain therapy. It led us to hypothesize that NI may preferentially act on particular P2Xs and these receptors may be unevenly distributed in the upper/lower limb.

Objective: To investigate P2Xs distribution in the upper/lower limb and NI's targets in upper limb RA.

Materials and methods: The SD rats were randomized into 11 groups of 10 animals each. Eight experimental groups were established by the injection of 0.1?mL FCA into the plantar surface of rat paw. Three control groups suffered the same volume of saline. The articular cavities were then taken on the seventh day to detect P2Xs expression. NI (3?g/kg) and prednisone (10?mg/kg) were respectively given by oral gavage once daily for 14?d. The swelling degree and P2Xs were evaluated individually.

Results: In normal rats, the expressions of P2X₃ and P2X₆ in forelimb were markedly higher than that of in hind limb (P?1, P2X₃, P2X₄, P2X₅ and P2X₇ were increased significantly (P?3. In NI treatment rats, swelling degree of the 7th/14th day in forelimb was 68.24%/38.89%, whereas that of in hind limb was 88.72%/79.92%. P2X₃ mRNA and protein expression was significantly reduced as contrasted with the control group (P?Conclusions: P2X₃ receptor was predominantly expressed in the forelimb RA rat. NI relieved the FCA-induced RA by inhibiting upper limb’s P2X₃ receptor.     

川芎-天麻配伍对川芎中2种成分在大鼠脑内药动学的影响

目的探讨川芎⁃天麻配伍对川芎中川芎嗪、阿魏酸在大鼠脑内药动学的影响。方法18只大鼠随机分成川芎组、川芎⁃天麻(1∶0.25)组、川芎⁃天麻(1∶1)组,每组6只,建立血瘀型偏头痛模型。灌胃给药后,于大鼠脑内插入微透析探针,收集不同时间点脑透析液,UPLC⁃MS/MS法检测川芎嗪、阿魏酸含有量,绘制血药浓度⁃时间曲线,计算药动学参数。结果与单味药组比较,配伍组2种成分T1/2[川芎⁃天麻(1∶1)组阿魏酸除外]、MRT0~∞、Cmax、AUC0~∞升高(P<0.05,P<0.01);川芎⁃天麻(1∶0.25)组两者T1/2、MRT0~∞、AUC0~∞高于川芎⁃天麻(1∶1)组(P<0.05,P<0.01),川芎嗪Cmax降低(P<0.05)。结论天麻可提高川芎中川芎嗪、阿魏酸脑组织吸收程度,延长作用时间,减缓消除速率,增加蓄积,其作用强度与天麻剂量有关。     

Small‐Angle X‐Ray Scattering Measurements on Amphiphilic Polymer Conetworks Swollen in Orthogonal Solvents

Amphiphilic polymer conetworks (APCNs), which combine two different polymer nanophases, have a broad range of applications that involve their unique potential to separately swell one of these nanophases in a selective solvent. Little is known about the structural changes of such APCNs upon swelling in dependence on the topology. Here, conetworks composed of poly(2‐ethylhexyl acrylate) crosslinked by poly(2‐methyl‐2‐oxazoline) (PMOx) are investigated with small‐angle X‐ray scattering in dry and swollen state using the orthogonal solvents water and toluene. The data clearly show that the structural changes induced by swelling are strongly dependent on the topology of the APCNs. While water leads to fusion of PMOx phases resulting in larger structures than found in the dry APCN, toluene is only swelling the hydrophobic phases without structural changes.     

Indirect activation of pregnane X receptor in the induction of hepatic CYP3A11 by high-dose rifampicin in mice

Rifampicin (RIF), a typical ligand of human pregnane X receptor (PXR), powerfully induces the expression of cytochrome P450 3A4 (CYP3A4) in humans. Although it is thought that RIF is not a ligand of rodent PXR, treatment with high-dose RIF (e.g. more than 20?mg/kg) increases the expression of CYP3A in the mouse liver. In this study, we investigated whether the induction of CYP3A by high-dose RIF in the mouse liver is mediated via indirect activation of mouse PXR (mPXR). The results showed that high-dose RIF increased the expression of CYP3A11 and other PXR-target genes in the liver of wild-type mice but not PXR-knockout mice. However, the results of reporter gene and ligand-dependent assembly assays showed that RIF does not activate mPXR in a ligand-dependent manner. In addition, high-dose RIF stimulated nuclear accumulation of mPXR in the mouse liver, and geldanamycin and okadaic acid attenuated the induction of Cyp3a11 and other PXR-target genes in primary hepatocytes, suggesting that high-dose RIF triggers nuclear translocation of mPXR. In conclusion, the present study suggests that high-dose RIF stimulates nuclear translocation of mPXR in the liver of mice by indirect activation, resulting in the transactivation of Cyp3a11 and other PXR-target genes.     

Characterization of CYP2C Induction in Cryopreserved Human Hepatocytes and Its Application in the Prediction of the Clinical Consequences of the Induction

CYP2C enzymes play key roles in drug metabolism, and clinical drug-drug interactions caused by CYP2C induction have been reported. The aim of this study was to establish a method to predict the potency of CYP2C inductions considering the mechanism. We first investigated the relations of CYP2C induction with CYP3A4 or CYP2B6 induction in human hepatocytes after 48-h exposure with 19 inducers. The fold-induction values of CYP2C8 and CYP2C9 were well correlated with those of CYP3A4, whereas the inducers were separated into 2 groups showing different correlations with CYP2B6 induction for CYP2C8 and CYP2C9 induction. In the regression models established, the fold-induction values of CYP2C8 and CYP2C9 were well expressed as the functions of those of CYP3A4 and CYP2B6, while no such obvious correlation was observed for CYP2C19 induction. These results suggest that CYP2Cs are not simply coinduced with CYP3A4 and that CYP2C8 and CYP2C9 inductions are regulated by both pregnane X receptor and constitutive androstane receptor with different contributions. Finally, simple correlations were proposed using the experimental E_max values obtained and plasma concentrations of CYP2C9 substrates from the literature, and positive correlations were observed. These data provide methods to estimate the clinical impact of CYP2C9 induction from in vitro data.     

去甲斑蝥素抑制黑色素瘤M14细胞增殖和诱导细胞凋亡的研究

Objective To investigate the effects of norcantharidin on the proliferation and apoptosis related proteins Bax and Bcl-2 of human melanoma M14 cells. Methods Melanoma M14 cell line cultured in vitro was treated with concentrations of 50, 100 and 500 μg / L norcantharidin respectively, and untreated cells were used as control. Cell proliferation was detected by CCK-8 method after treatment for 24 h and 48 h. Morphology of cells was observed under inverted microscope. DAPI nuclear staining was used to observe the morphological changes of apoptosis under fluorescence microscope. The expressions of Bax and Bcl-2 were detected by Western blot assay. Results CCK-8 results showed that concentrations of 100 and 500 μg/L norcantharidin inhibited the proliferation of melanoma M14 cells after treatment for 24 h and 48 h (P＜0.05), and the higher the concentration, the more significant the inhibitory effect. The cells showed irregular contours, nucleus shrinkage, small volume, and poor cell adhesion of apoptosis changes under inverted microscope and fluorescence microscope. Western blot analysis showed that the relative expression of Bcl-2 decreased gradually with the increased concentration of norcantharidin (P＜0.05), and the relative expression of Bax increased gradually (P＜0.05).Conclusion Norcantharidin can inhibit the proliferation of melanoma M14 cells, and its mechanism may be related to the promotion of apoptosis.