Osteoporosis and Stress Urinary Incontinence in Women: A National Health Insurance Database Study.

We aimed to determine the influence of osteoporosis and stress urinary incontinence in women. We hypothesized that women with osteoporosis had an increased risk of stress urinary incontinence. This retrospective study used data from the Taiwan Longitudinal Health Insurance database from 2005-2009. The study population was screened to identify women (age ≥ 40 years) newly diagnosed with osteoporosis (ICD-9-CM code = 733.0, 733.1). The osteoporosis cohort included 6125, and the non-osteoporosis cohort included 12,250 participants. The newly diagnosed stress urinary incontinence incidence was calculated to determine the influence of osteoporosis and stress urinary incontinence. We used the Cox proportional hazards model to predict the effects of stress urinary incontinence and the Kaplan-Meier analysis to estimate the cumulative incidence of stress urinary incontinence in women. Participants with osteoporosis experienced a 1.79 times higher risk than that of the non-osteoporosis group (95% CI = 1.28-2.51) for stress urinary incontinence, regardless of age. We did not observe a higher risk of stress urinary incontinence in participants with pathological fractures compared to those with simple osteoporosis. Our data emphasized that physicians and nurses should conduct urinary incontinence screening in women with osteoporosis to recommend proper treatment, medical help or to bring the disorder to light.

Optimal Initial Blood Pressure in Intensive Care Unit Patients with Non-Traumatic Intracranial Hemorrhage.

Blood pressure (BP) control is crucial for minimizing the risk of mortality and hematoma growth in patients with acute intracranial hemorrhage (ICH). We aimed to determine the optimal BP range associated with improved patient outcomes. From the Medical Information Mart for Intensive Care-III database, we identified 1493 patients (age, 18-99 years) admitted to the intensive care unit (ICU) with non-traumatic ICH. The 3-day and 14-day mortality of ICU admissions were compared at different BP ranges. Generalized additive models were used to assess the optimal range of initial mean arterial pressure, systolic blood pressure (SBP), and diastolic blood pressure, and these were identified to be 70-100, 120-150, and 60-100 mmHg, respectively. The 3-day or 14-day mortality showed U-shaped correlations with BP ranges. Our results show that an initial SBP between 120 and 150 mmHg is associated with minimal risk of mortality risk. This recommendation can assist physicians to achieve better outcomes for patients with ICH.

Interleukin-4 receptor-targeted liposomal doxorubicin as a model for enhancing cellular uptake and antitumor efficacy in murine colorectal cancer.

Our previous studies showed that colorectal tumor has high interleukin-4 receptor α (IL-4Rα) expression, whereas adjacent normal tissue has low or no IL-4Rα expression. We also observed that human atherosclerotic plaque-specific peptide-1 (AP1) can specifically target to IL-4Rα. In this study, we investigated the therapeutic efficacy and systemic toxicity of AP1-conjuagted liposomal doxorubicin. AP1 bound more strongly to and was more efficiently internalized into IL-4Rα-overexpressing CT26 cells than CT26 control cells. Selective cytotoxicity experiment revealed that AP1-conjugated liposomal doxorubicin preferentially killed IL-4Rα-overexpressing CT26 cells. AP1-conjugated liposomal doxorubicin administered intravenously into mice produced significant inhibition of tumor growth and showed decreased cardiotoxicity of doxorubicin. These results indicated that AP1-conjugated liposomal doxorubicin has a potent and selective anticancer potential against IL-4Rα-overexpressing colorectal cancer cells, thus providing a model for targeted anticancer therapy.

A new approach to reduce toxicities and to improve bioavailabilities of platinum-containing anti-cancer nanodrugs.

Platinum (Pt) drugs are the most potent and commonly used anti-cancer chemotherapeutics. Nanoformulation of Pt drugs has the potential to improve the delivery to tumors and reduce toxic side effects. A major challenge for translating nanodrugs to clinical settings is their rapid clearance by the reticuloendothelial system (RES), hence increasing toxicities on off-target organs and reducing efficacy. We are reporting that an FDA approved parenteral nutrition source, Intralipid 20%, can help this problem. A dichloro (1, 2-diaminocyclohexane) platinum (II)-loaded and hyaluronic acid polymer-coated nanoparticle (DACHPt/HANP) is used in this study. A single dose of Intralipid (2 g/kg, clinical dosage) is administrated [intravenously (i. v.), clinical route] one hour before i.v. injection of DACHPt/HANP. This treatment can significantly reduce the toxicities of DACHPt/HANP in liver, spleen, and, interestingly, kidney. Intralipid can decrease Pt accumulation in the liver, spleen, and kidney by 20.4%, 42.5%, and 31.2% at 24-hr post nanodrug administration, respectively. The bioavailability of DACHPt/HANP increases by 18.7% and 9.4% during the first 5 and 24 hr, respectively.

Hyperthermia-mediated local drug delivery by a bubble-generating liposomal system for tumor-specific chemotherapy.

As is widely suspected, lysolipid dissociation from liposomes contributes to the intravenous instability of ThermoDox (lysolipid liposomes), thereby impeding its antitumor efficacy. This work evaluates the feasibility of a thermoresponsive bubble-generating liposomal system without lysolipids for tumor-specific chemotherapy. The key component in this liposomal formulation is its encapsulated ammonium bicarbonate (ABC), which is used to actively load doxorubicin (DOX) into liposomes and trigger a drug release when heated locally. Incubating ABC liposomes with whole blood results in a significantly smaller decrease in the retention of encapsulated DOX than that by lysolipid liposomes, indicating superior plasma stability. Biodistribution analysis results indicate that the ABC formulation circulates longer than its lysolipid counterpart. Following the injection of ABC liposome suspension into mice with tumors heated locally, decomposition of the ABC encapsulated in liposomes facilitates the immediate thermal activation of CO2 bubble generation, subsequently increasing the intratumoral DOX accumulation. Consequently, the antitumor efficacy of the ABC liposomes is superior to that of their lysolipid counterparts. Results of this study demonstrate that this thermoresponsive bubble-generating liposomal system is a highly promising carrier for tumor-specific chemotherapy, especially for local drug delivery mediated at hyperthermic temperatures.

Pharmacokinetic analysis of 111 in-labeled liposomal Doxorubicin in murine glioblastoma after blood-brain barrier disruption by focused ultrasound.

The goal of this study was to evaluate the pharmacokinetics of targeted and untargeted (111)In-doxorubicin liposomes after these have been intravenously administrated to tumor-bearing mice in the presence of blood-brain barrier disruption (BBB-D) induced by focused ultrasound (FUS). An intracranial brain tumor model in NOD-scid mice using human brain glioblastoma multiforme (GBM) 8401 cells was developed in this study. (111)In-labeled human atherosclerotic plaque-specific peptide-1 (AP-1)-conjugated liposomes containing doxorubicin (Lipo-Dox; AP-1 Lipo-Dox) were used as a microSPECT probe for radioactivity measurements in the GBM-bearing mice. Compared to the control tumors treated with an injection of (111)In-AP-1 Lipo-Dox or (111)In-Lipo-Dox, the animals receiving the drugs followed by FUS exhibited enhanced accumulation of the drug in the brain tumors (p<0.05). Combining sonication with drugs significantly increased the tumor-to-normal brain doxorubicin ratio of the target tumors compared to the control tumors. The tumor-to-normal brain ratio was highest after the injection of (111)In-AP-1 Lipo-Dox with sonication. The (111)In-liposomes micro-SPECT/CT should be able to provide important information about the optimum therapeutic window for the chemotherapy of brain tumors using sonication.

Focused ultrasound and interleukin-4 receptor-targeted liposomal doxorubicin for enhanced targeted drug delivery and antitumor effect in glioblastoma multiforme.

The clinical application of chemotherapy to brain tumors has been severely limited because the blood-brain barrier (BBB) often prevents therapeutic levels from being achieved. Here we show that pulsed HIFU and human atherosclerotic plaque-specific peptide-1 (AP-1)-conjugated liposomes containing doxorubicin (AP-1 Lipo-Dox) act synergistically in an experimental brain tumor model. We developed an intracranial brain-tumor model in NOD-scid mice using human brain glioblastoma multiforme (GBM) 8401 cells. Pulsed HIFU was used to transcranially disrupt the BBB in these mouse brains by delivering ultrasound waves in the presence of microbubbles. Prior to each sonication, AP-1 Lipo-Dox or unconjugated Lipo-Dox was administered intravenously, and the concentration in the brains was quantified by fluorometer. Compared to control animals treated with injections of AP-1 Lipo-Dox or unconjugated Lipo-Dox, animals receiving the drug followed by pulsed HIFU exhibited enhanced accumulation of the drug in tumor cells. Drug injection with sonication increased the tumor-to-normal brain doxorubicin ratio of the target tumors by about twofold compared with the control tumors. Moreover, the tumor-to-normal brain ratio was highest after the injection of AP-1 Lipo-Dox with sonication. Combining sonication with AP-1 Lipo-Dox also significantly inhibited tumor growth compared with chemotherapy alone. There was a modest but significant increase in the median survival time in mice treated with AP-1 Lipo-Dox followed by pulsed HIFU, compared to those treated with AP-1 Lipo-Dox without sonication. The use of AP-1-conjugated liposomes carrying cytotoxic agents followed by pulsed HIFU represents a feasible approach for enhanced targeted drug delivery in brain tumor therapies.

Mechanisms of cellular uptake and intracellular trafficking with chitosan/DNA/poly(γ-glutamic acid) complexes as a gene delivery vector.

Chitosan (CS)-based complexes have been considered as a vector for DNA delivery; nonetheless, their transfection efficiency is relatively low. An approach by incorporating poly(γ-glutamic acid) (γ-PGA) in CS/DNA complexes was developed in our previous study to enhance their gene expression level; however, the detailed mechanisms remain to be understood. The study was designed to investigate the mechanisms in cellular uptake and intracellular trafficking of CS/DNA/γ-PGA complexes. The results of our molecular dynamic simulations suggest that after forming complexes with CS, γ-PGA displays a free γ-glutamic acid in its N-terminal end and thus may be recognized by γ-glutamyl transpeptidase in the cell membrane, resulting in a significant increase in their cellular uptake. In the endocytosis inhibition study, we found that the internalization of CS/DNA complexes took place via macropinocytosis and caveolae-mediated pathway; by incorporating γ-PGA in complexes, both uptake pathways were further enhanced but the caveolae-mediated pathway played a major role. TEM was used to gain directly understanding of the internalization mechanism of test complexes and confirmed our findings obtained in the inhibition experiments. After internalization, a less percentage of co-localization of CS/DNA/γ-PGA complexes with lysosomes was observed when compared with their CS/DNA counterparts. A greater cellular uptake together with a less entry into lysosomes might thus explain the promotion of transfection efficiency of CS/DNA/γ-PGA complexes. Knowledge of these mechanisms involving CS-based complexes containing γ-PGA is critical for the development of an efficient vector for DNA transfection.

Detection of Stage I nasopharyngeal carcinoma by serologic screening and clinical examination / 癌症

In a prospective study, 42 048 adults residing in Zhongshan City, Guangdong, China, were followed for 16 years, and 171 of them developed nasopharyngeal carcinoma (NPC). Although Epstein-Barr virus (EBV) antibody levels of the cohort fluctuated, the antibody levels of 93% of the patients with NPC were raised and maintained at high levels for up to 10 years prior to diagnosis. This suggests that the serologic window affords an opportunity to monitor tumor progression during the preclinical stage of NPC development, facilitating early NPC detection. We reviewed the clinical records of the 171 patients with NPC in the prospective study to assess the efficacy of early NPC detection by serologic screening and clinical examination. Of the 171 patients, 51 had Stage I tumor (44 were among the 73 patients detected by clinical examination and 7 were among the 98 patients presented to outpatient department). Initial serologic screening predicted 58 (95.1%) of the 61 patients detected within 2 years. The risk of the screened population (58/3093) raised 13 times relative to cohort (61/42 048) during this period. Clinical examination detected all the 58 predicted cases, and 35 (60.3%) of which were diagnosed with Stage I tumor. The serologic prediction rate fell to 33.6% (37/110) 2 to 16 years after screening. The proportion of cases detected by clinical examination fell to 40.5% (15/37). The proportion of Stage I tumors among the cases detected by clinical examination during both periods remained at about 60%. We concluded that early detection of NPC can be accomplished by repeated serologic screening to maintain high prediction rates and by promptly examining screened subjects to detect tumors before the symptoms develop.

Effects of the nanostructure of dendrimer/DNA complexes on their endocytosis and gene expression.

Cationic dendrimers constitute a potential nonviral vector for gene therapy due to their ability of forming electrostatic complexes with DNA (dendriplexes). However, the supramolecular structure of dendriplexes and its impact on the cellular uptake and gene transfection remain largely unknown. Using synchrotron small angle X-ray scattering, here we show that DNA in complexes with poly(amidoamine) (PAMAM) G4 dendrimers exhibited three distinct packaging states modulated by the degree of their protonation (dp). Our structure characterization suggests that the nanostructure of DNA in dendriplexes transformed from square-packed straightened chains (dp/0.1) to hexagonally-packed superhelices (dp/0.3) and eventually to a beads-on-string configuration (dp/0.6 and dp/0.9). The transfection efficiency in HT1080 cells significantly enhanced when the dp value was increased from 0.1 to 0.3. This enhancement was due to a higher positive surface charge of dendriplexes formed at higher dp, which facilitated adherence of test dendriplexes to the negatively charged cell membranes for the subsequent endocytosis. Although the surface charge of dendriplexes still increased accordingly, further increase of the dendrimer dp value to 0.9 reduced the transfection efficiency. This unexpected suppression of transfection may be attributed to the wrapping of DNA around dendrimers that frustrates the interaction between dendrimer and cholesterol in the membrane raft via the caveola-mediated endocytosis. These results can be used for the rational design of dendrimer-based gene delivery devices.

Effects of incorporation of poly(gamma-glutamic acid) in chitosan/DNA complex nanoparticles on cellular uptake and transfection efficiency.

Chitosan (CS)/DNA complex nanoparticles (NPs) have been considered as a vector for gene delivery. Although advantageous for DNA packing and protection, CS-based complexes may lead to difficulties in DNA release once arriving at the site of action. In this study, an approach through modifying their internal structure by incorporating a negatively charged poly(gamma-glutamic acid) (gamma-PGA) in CS/DNA complexes (CS/DNA/gamma-PGA NPs) is reported. The analysis of small angle X-ray scattering results revealed that DNA and gamma-PGA formed complexes with CS separately to yield two types of domains, leading to the formation of "compounded NPs". With this internal structure, the compounded NPs might disintegrate into a number of even smaller sub-particles after cellular internalization, thus improving the dissociation capacity of CS and DNA. Consequently, after incorporating gamma-PGA in CS/DNA complexes, a significant increase in their transfection efficiency was found. Interestingly, in addition to improving the release of DNA intracellularly, the incorporation of gamma-PGA in CS/DNA complexes significantly enhanced their cellular uptake. We further demonstrated that besides a non-specific charged-mediated binding to cell membranes, there were specific trypsin-cleavable proteins involved in the internalization of CS/DNA/gamma-PGA NPs. The aforementioned results indicated that gamma-PGA played multiple important roles in enhancing the cellular uptake and transfection efficiency of CS/DNA/gamma-PGA NPs.

The use of biodegradable polymeric nanoparticles in combination with a low-pressure gene gun for transdermal DNA delivery.

Gold particles have been used as a carrier for transdermal gene delivery, which may cause adverse side effects when accumulated. In this study, biodegradable nanoparticles, composed of chitosan (CS) and poly-gamma-glutamic acid (gamma-PGA), were prepared by an ionic-gelation method for transdermal DNA delivery (CS/gamma-PGA/DNA) using a low-pressure gene gun. The conventional CS/DNA without the incorporation of gamma-PGA was used as a control. Small-angle X-ray scattering (SAXS) was used to examine the internal structures of test nanoparticles, while identification of their constituents was conducted by Fourier transformed infrared (FT-IR) spectroscopy. The CS/gamma-PGA/DNA were spherical in shape with a relatively homogeneous size distribution. In contrast, CS/DNA had a heterogeneous size distribution with a donut, rod or pretzel shape. Both test nanoparticles were able to effectively retain the encapsulated DNA and protect it from nuclease degradation. As compared with CS/DNA, CS/gamma-PGA/DNA improved their penetration depth into the mouse skin and enhanced gene expression. These observations may be attributed to the fact that CS/gamma-PGA/DNA were more compact in their internal structures and had a greater density than their CS/DNA counterparts, thus having a larger momentum to penetrate into the skin barrier. The results revealed that CS/gamma-PGA/DNA may substitute gold particles as a DNA carrier for transdermal gene delivery.