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.     

Suppression of puerarin on polymethylmethacrylate-induced lesion of peri-implant by inhibiting NF-κB activation in vitro and in vivo

Puerarin (PR), a natural isoflavone isolated from Chinese traditional plant pueraria lobata, has attracted considerable attention due to its important biological and pharmacological activities. However, its effects on lesion of peri-implant and related mechanism of action are still not clear, which require further investigation. In this study, we evaluated the effects of PR on polymethylmethacrylate (PMMA)-induced lesion of peri-implant in vitro and in vivo, and explored its possible mechanism of action. Our results indicated that PR could inhibit PMMA-induced osteoclastogenesis in RAW264.7 cells with a dose-dependent manner in vitro and effectively down-regulate mRNA and protein expressions of matrix metalloprotein 9 (MMP-9), tumor necrosis factor (TNF)-α, interleukin (IL)-6, and receptor activator of nuclear factor (NF)-κB (RANK), primarily via the suppression of NF-κB signaling. Furthermore, we found that PMMA induction could directly cause the phosphorylation of IκB and significantly promote the nuclear translocation of p65 in RAW264.7 cells. In other words, PR was able to dose-dependently attenuate the PMMA-induced nuclear translocation of p65 in RAW264.7 cells. In vivo, PR was observed to attenuate PMMA-induced osteoclastogenesis, osteolysis, mRNA expressions of receptor activator of nuclear factor (NF)-κB ligand (RANKL) and RANK, as well as protein levels of MMP-9, TNF-α, IL-6, and p65 in a murine calvarial osteolysis model. These findings suggested that PR might be a potential therapeutic drug to lesion of peri-implant, and provided new insights for understanding its possible mechanism.     

Botulinum toxin blocks mast cells and prevents rosacea like inflammation

Background

Rosacea is a chronic inflammatory skin condition whose etiology has been linked to mast cells and the antimicrobial peptide cathelicidin LL-37. Individuals with refractory disease have demonstrated clinical benefit with periodic injections of onabotulinum toxin, but the mechanism of action is unknown.

Single stage immediate breast reconstruction with acellular dermal matrix and implant: Defining the risks and outcomes of post-mastectomy radiotherapy

IntroductionThe objective of this study is to evaluate outcomes and complications in patients with single-stage ADM-implant based immediate breast reconstruction with and without radiotherapy (RT), highlighting the effects of RT on the reconstruction.Materials and methodsThis prospective study recruited 91 consecutive patients who underwent skin-sparing, nipple-sparing or wise-pattern skin reduction mastectomy with direct-to-implant breast reconstruction with ADMs using sub-pectoral or pre-pectoral approach at the two breast units. Early and late complications like seroma, delayed wound healing, wound breakdown, infection, capsular contracture, implant loss and revision surgery were evaluated in the RT and non-RT groups.ResultsIn the total cohort of 91 patients, 29 received adjuvant RT and 62 did not need RT. In the RT group, 3–7% of them had early complications like seroma, wound infections and delayed healing. 20.7% had post-RT capsular contractures which either required revision surgery with autologous flap (6.9%) or capsulotomy with exchange of implant (6.9%). In the non-RT group, 7–9% cases had seroma & wound infections, 3.06% had delayed wound healing and 7.25% had capsular contracture. 13.04% required revision surgery due to infection, implant loss or failure to achieve expectations. The total loss of implants in the cohort was 7.14% (RT group 6.9% and non-RT group 7.25%). The need for PMRT could have been predicted pre-operatively in the RT group in 55.17% cases based on the extent of disease, multifocality, tumour grade and positive LN status on imaging.ConclusionADM based reconstruction in patients anticipated to receive adjuvant RT is always debatable. Though there is no significant difference in the revision surgeries in our study of the 2 groups, the rate of capsular contracture as expected, was higher in the RT group. Hence, pre-operative discussion on the need for RT highlighting the risks and complications will help patients make a better-informed choice.     

液相色谱-电喷雾串联质谱分析人全血中环孢素的基质效应评估及其机理探究

目的： 评估液相色谱-电喷雾串联质谱（LC-ESI-MS/MS）分析人全血中环孢素的基质效应，并探究其发生机理。方法： 全血裂解物的蛋白沉淀上清液用作全血基质。分别用乙腈水溶液和全血基质配制等浓度环孢素A（CyA）、环孢素C（CyC）的质控样品。分别在添加H⁺、NH₄⁺和Na⁺的流动相中，用LC-ESI-MS/MS的MRM模式检测质控样品中CyA，CyC的[M+H]⁺、[M+NH₄]⁺和[M+Na]⁺（M=CyA，CyC）峰面积，以其计算基质效应因子；用MS Scan模式检查质控样品中CyA、CyC色谱流出峰的质谱离子组成情况，用于判断基质效应原因。结果： 在添加H⁺、NH₄⁺和Na⁺的流动相中，全血基质中CyA、CyC基质效应因子分别为-76.6%~-54.0%、-86.0%~-55.3%和-42.8%~-34.1%。在加H⁺、加NH₄⁺流动相中，标准液加全血基质的CyA、CyC色谱峰的质谱中出现丰度很高的[M+Na]⁺和[M+K]⁺，在加Na⁺流动相中它们出现丰度很高的[M+K]⁺，而对应的定量离子[M+H]⁺、[M+NH₄]⁺或[M+Na]⁺的丰度显著降低。结论： 全血基质对环孢素的LC-ESI-MS/MS分析产生明显的基质效应，其原因与基质中Na⁺、K⁺同添加在流动相中用于辅助电离的H⁺、NH₄⁺或Na⁺竞争与环孢素加合，导致定量目标加合离子丰度显著降低有关。     

中药酊剂临床外用技术规范(草案)

基于对中药酊剂外用技术的数据挖掘,结合临床实际应用研究,经外治学会专家多次论证,形成中药酊剂临床外用技术规范(草案),包括临床适用范围、操作步骤以及外用酊剂的方法、剂量、频率、时间、注意事项、不良反应及护理要点。以期规范中药外用酊剂的临床应用,提高其疗效并减少不良反应。     

Exosomes secreted by prostate cancer cells under hypoxia promote matrix metalloproteinases activity at pre-metastatic niches

Approximately, 30 000 men die from prostate cancer (PCa) every year in the United States, mainly due to the metastasis. Thus, the key events associated with PCa metastasis are under rigorous investigation, with recent studies showing that preparation of pre-metastatic niches (PMN) in distant organs is an important step. However, the molecular basis for PMN preparation is still unclear. Hypoxia in primary tumors promotes aggressiveness; however, its precise role in metastasis is not clear. We recently reported that exosomes secreted by PCa cells under hypoxia promote stemness and invasiveness in naïve PCa cells; however, whether these extracellular vesicles also influence PMN remains unknown. In the present study, we isolated exosomes from human PCa PC3 cells under normoxic (21% O₂, exosomes secreted under normoxic condition [Exo^Normoxic]) and hypoxic (1% O₂, exosomes secreted under hypoxic condition [Exo^Hypoxic]) conditions, and characterized their effect (10 µg exosomes, intraperitoneal (IP) treatment every 48 hours for 4 weeks) on key biomarkers associated with PMN in nude mice. Whole animal fluorescence imaging showed that Exo^Hypoxic treatment promotes matrix metalloproteinases (MMPs) activity in several putative metastatic sites. Histological studies confirmed that Exo^Hypoxic treatment enhanced the level of MMP2, MMP9, and extracellular matrix proteins (fibronectin and collagen) as well as increased the number of CD11b+ cells at selective PMN sites. Furthermore, proteomic profiling of exosomes by liquid chromatography/mass spectrometry identified cargo proteins in Exo^Normoxic and Exo^Hypoxic as well as distinct canonical pathways targeted by them. These results suggest that exosomes secreted by PCa cells under hypoxia plausibly remodel distant PMN, and thus, could be a potential target to control metastatic PCa.     

The key role of macrophage depolarization in the treatment of COPD with ergosterol both in vitro and in vivo

Macrophages are the most abundant immune cells in the lung, which play an important role in COPD. The anti-inflammatory and anti-oxidation of ergosterol are well documented. However, the effect of ergosterol on macrophage polarization has not been studied. The objective of this work was to investigate the effect of ergosterol on macrophage polarization in CSE-induced RAW264.7 cells and Sprague-Dawley (SD) rats COPD model. Our results demonstrate that CSE-induced macrophages tend to the M1 polarization via increasing ROS, IL-6 and TNF-α, as well as increasing MMP-9 to destroy the lung construction in both RAW264.7 cells and SD rats. However, treatment of RAW264.7 cells and SD rats with ergosterol inhibited CSE-induced inflammatory by decreasing ROS, IL-6 and TNF-α, and increasing IL-10 and TGF-β, shuffling the dynamic polarization of macrophages from M1 to M2 both in vitro and in vivo. Ergosterol also decreased the expression of M1 marker CD40, while increased that of M2 marker CD163. Moreover, ergosterol improved the lung characters in rats by decreasing MMP-9. Furthermore, ergosterol elevated HDAC3 activation and suppressed P300/CBP and PCAF activation as well as acetyl NF-κB/p65 and IKKβ, demonstrating that HDAC3 deacetylation was involved in the effect of ergosterol on macrophage polarization. These results also provide a proof in immunoregulation of ergosterol for therapeutic effects of cultured C. sinensis on COPD patients.