[Preparation and effect against cervical cancer invasion in vitro of harmine-loaded photosensitive liposomes].

Despite the development of HPV vaccines and screening programs, cervical cancer is still a serious threat to women's health. Early-stage cervical cancer is mainly treated by surgery. However, considering the serious complications after surgery, hyperthermia is recommended to enhance the effect of chemotherapy, retain the integrity of cervix, improve the treatment effect, which provides a therapeutic basis for the early treatment of cervical cancer. The photosensitive liposomes containing harmine and dye IR-780 were prepared by thin-film dispersion method and separated by Sephadex G-50 dextran gel column. The preparation conditions were optimized as the mass ratio of phospholipid to cholesterol membrane material being 8∶1 and that of drug to lipid being 1∶20. The results of HPLC showed that the encapsulation efficiency of harmine was 55.6%±0.18%. The prepared photosensitive liposomes were round and evenly distributed under transmission electron microscope, with the particle size of(125.2±0.62) nm determined by Marvin particle size analyzer and the Zeta potential of(-2.55±0.76) mV. Additionally, the photosensitive liposomes had the photothermal conversion efficiency, an important property of photothermal agent, of 27.1%±0.86%. The photosensitive liposomes stored at 4 ℃ showed stable encapsulation efficiency in the first 14 days without flocculation. The sulforhodamine B(SRB) assay was employed to determine the inhibitory effect of the liposomes on the proliferation of HeLa cells under near-infrared(NIR) irradiation or not, which showcased stronger inhibitory effect under NIR irradiation. The results of Transwell assay indicated that the prepared liposomes significantly inhibited the invasion and migration of HeLa cells in vitro. The findings of this study provide a basis for the treatment of cervical cancer with harmine.

[Effect of paidu baoshen pill in retarding the progression of chronic renal failure].

OBJECTIVE: To assess the impact of Paidu Baoshen Pill (PBP, modified Dahuang Zhechong Pill), in retarding the procession of chronic renal failure (CRF) of stage II-III. METHODS: The 283 patients of CRF stage II-III were randomly assigned to two groups, 151 patients in the treatment group treated with oral administration of PBP 3 g twice a day, and 132 patients in the control group with oxidative amylase aldehyde enveloped capsule 5-10 capsules thrice a day after meal. The course for both groups was 2 months, and the changes after 1 or 2 courses treatment in scoring of quality of life (QOL) and clinical symptoms, also in laboratory indexes including serum levels of creatinine (Cr), urea nitrogen (UN), and intrinsic creatinine clearance rate were observed. RESULTS: The total effective rate was 70. 86% (107/151 cases) in the treatment group and 44.70% (59/132 cases) in the control group, showing significant difference between them (X2 = 18.69, P < 0.01). Significant differences between groups were also shown in comparisons of scores of QOL and clinical symptoms after treatment. Inter-group comparison showed no difference in all the three indexes detected before treatment, but they did show statistical significance respectively after 1 and 2 courses of treatment (P < 0.05 and P < 0.01). CONCLUSION: PBP could effectively retard the progression of chronic renal failure and significantly improve the QOL of patients.

Coordinating Etk/Bmx activation and VEGF upregulation to promote cell survival and proliferation.

Etk/Bmx, a member of the Tec family of non-receptor tyrosine kinase, is characterized by an N-terminal PH domain and has recently been shown to be involved in the regulation of various cellular processes, including proliferation, differentiation, motility and apoptosis. Since VEGF and the activation of its signaling pathway have been implicated in modulating a variety of biological responses, we characterized the role of Etk-dependent signaling pathways involved in the upregulation of VEGF expression, and explored the functional implications of this enhancement in sustaining cell proliferation and survival. Using Northern and Western analyses, transient transfections, and pharmacological agents, we demonstrate that Etk activation alone is sufficient to transcriptionally induce VEGF expression, independent of the previously identified hypoxia response element (HRE), in both Pa-4 epithelial and TR-BBB endothelial cells under normoxia. In addition, Etk utilizes both MEK/ERK and PI3-K/Pak1 signaling pathways in concert to activate VEGF transcription. Functionally, Etk activation elicits a profound stimulatory effect on TR-BBB cell proliferation and formation of capillary-like networks in Matrigel containing reduced levels of growth factors. Finally, antisense oligonucleotides against either endogenous VEGF or Etk abrogate the proliferation of Etk-activated TR-BBB cells, and exogenous VEGF treatment stimulates endogenous Etk tyrosine phosphorylation in HUVECs. Taken together, these results indicate that VEGF is both an Etk downstream target gene and an Etk upstream activator, constituting a reciprocal Etk-VEGF autoregulatory loop. These findings, to our knowledge, are the first delineation of a network of positive feedforward signaling pathways that converge on the Etk-VEGF axis, causally associating Etk-mediation of VEGF induction with enhanced cellular processes in both epithelial and endothelial cells.

Etk/Bmx mediates expression of stress-induced adaptive genes VEGF, PAI-1, and iNOS via multiple signaling cascades in different cell systems.

We recently showed that Etk/Bmx, a member of the Tec family of nonreceptor protein tyrosine kinases, promotes tight junction formation during chronic hypoxic exposure and augments normoxic VEGF expression via a feedforward mechanism. Here we further characterized Etk's role in potentiating hypoxia-induced gene expression in salivary epithelial Pa-4 cells. Using transient transfection in conditionally activated Etk (DeltaEtk:ER) cells, we demonstrated that Etk enhances hypoxia-response element-dependent reporter activation in normoxia and hypoxia. This Etk-driven reporter activation is ameliorated by treatment with wortmannin or LFM-A13. Using lentivirus-mediated gene delivery and small interfering RNA, we provided direct evidence that hypoxia leads to transient Etk and Akt activation and hypoxia-mediated Akt activation is Etk dependent. Northern blot analyses confirmed that Etk activation led to induction of steady-state mRNA levels of endogenous VEGF and plasminogen activator inhibitor (PAI)-1, a hallmark of hypoxia-mediated gene regulation. We also demonstrated that Etk utilizes a phosphatidylinositol 3-kinase/Akt pathway to promote reporter activation driven by NF-kappaB, another oxygen-sensitive transcription factor, and to augment cytokine-induced inducible nitric oxide synthase expression in endothelial cells. To establish the clinical relevance of Etk-induced, hypoxia-mediated gene regulation, we examined Etk expression in keloid, which has elevated VEGF and PAI-1. We found that Etk is overexpressed in keloid (but not normal skin) tissues. The differential steady-state Etk protein levels were further confirmed in primary fibroblast cultures derived from these tissues, suggesting an Etk role in tissue fibrosis. Our results provide further understanding of Etk function within multiple signaling cascades to govern adaptive cytoprotection against extracellular stress in different cell systems, salivary epithelial cells, brain endothelial cells, and dermal fibroblasts.

Adsorption and activity of Candida rugosa lipase on polypropylene hollow fiber membrane modified with phospholipid analogous polymers.

Efforts have recently been made toward the study of interactions of phospholipid with various enzymes. It seems that phospholipids may be directly involved in regulating the enzyme activity. In this work, three phospholipid analogous polymers (PAPs), containing hydrophobic octyloxy, dodecyloxy, and octadecyloxy groups (abbreviated as 8-PAP, 12-PAP, and 18-PAP, respectively), were tethered on polypropylene hollow fiber microfiltration membrane (PPHFMM) to create a biocompatible interface for lipase immobilization. Lipase from Candida rugosa was immobilized on these PPHFMMs by adsorption. The adsorption capacity, activity, and thermal stability of enzyme on the PAP-modified PPHFMMs were compared with those of enzyme on the nascent ones. It was found that, as for the PAP-modified PPHFMMs, the adsorption capacities of lipase are lower than that of the nascent ones, while the activity retention of immobilized lipase increases from 57.5% to 74.1%, 77.5%, and 83.2% respectively for the 8-PAP-, 12-PAP-, and 18-PAP-modified PPHFMMs. In addition, the experimental results of thermal stability show that the residual activity of the immobilized lipase at 50 degrees C for 2 h is 62% for the 8-PAP-modified PPHFMM, 59% for the 12-PAP-modified PPHFMM, and 66% for the 18-PAP-modified PPHFMM, which are also higher than that of the nascent ones.