Recovery from prolonged disorders of consciousness: A dual-center prospective cohort study in China.

BACKGROUND: Recent innovations in intensive care have improved the prognosis of patients with severe brain injuries and brought more patients with disorders of consciousness (DoC). Data are lacking regarding the long-term outcomes of those patients in China. It is necessary to study the long-term outcomes of patients with prolonged DoC in light of many factors likely to influence crucial decisions about their care and their life. AIM: To present the preliminary results of a DoC cohort. METHODS: This was a two-center prospective cohort study of inpatients with vegetative state (VS)/unresponsive wakefulness syndrome (UWS). The study outcomes were the recovery from VS/UWS to minimally conscious state (MCS) and the long-term status of patients with prolonged DoC considered in VS/UWS or MCS for up to 6 years. The patients were evaluated using the Glasgow coma scale, coma recovery scale-revised, and Glasgow outcome scale. The endpoint of follow-up was recovery of full consciousness or death. The changes in the primary clinical outcome improvement in clinical diagnosis were evaluated at 12 mo compared with baseline. RESULTS: The study population included 93 patients (62 VS/UWS and 31 MCS). The post-injury interval range was 28-634 d. Median follow-up was 20 mo (interquartile range, 12-37 mo). At the endpoint, 33 transitioned to an emergence from MCS or full consciousness, eight had a locked-in syndrome, and there were 35 patients remaining in a VS/UWS and 11 in an MCS. Seven (including one locked-in syndrome) patients (7.5%) died within 12 mo of injury. Compared with the unresponsive group (n = 52) at 12 mo, the responsive group (n = 41) had a higher proportion of males (87.8% vs 63.5%, P = 0.008), shorter time from injury (median, 40.0 d vs 65.5 d, P = 0.006), higher frequency of vascular etiology (68.3% vs 38.5%, P = 0.007), higher Glasgow coma scale score at admission (median, 9 vs 6, P < 0.001), higher coma recovery scale-revised score at admission (median, 9 vs 2.5, P < 0.001), at 1 mo (median, 14 vs 5, P < 0.001), and at 3 mo (median, 20 vs 6, P < 0.001), lower frequency of VS/UWS (36.6% vs 90.0%, P < 0.001), and more favorable Glasgow outcome scale outcome (P < 0.001). CONCLUSION: Patients with severe DoC, despite having strong predictors of poor prognosis, might recover consciousness after a prolonged time of rehabilitation. An accurate initial diagnosis of patients with DoC is critical for predicting outcome and a long-term regular follow-up is also important.

Epidemiological analysis of malaria surveillance in Jinan City from 2012 to 2016 / 中国血吸虫病防治杂志

Objective To analyze the epidemiological characteristics of malaria reported in the malaria surveillance system in Jinan City from 2012 to 2016,so as to provide the evidence for improving the ability of diagnosis,treatment,prevention and control of malaria.Methods The data of all reported malaria cases in the malaria surveillance system were retrospectively ana-lyzed.Results From 2012 to 2016,91 malaria cases were found in Jinan City,of which one patient died.All the cases were imported and 95.60%(87 cases)of them came from Africa(note:most of the people were Chinese residents who had been in Af-rica for the export of labor service and came back China).Falciparum malaria accounted for the most(82.42%,75 cases).All the cases were adult males,and were mainly migrant workers.The median time from onset to being confirmedly diagnosed was 5 days,and the median time from seeing a doctor to being diagnosed was 1 day.The medical institutions where the patients first visited were mainly municipal medical institutions(42 cases,46.15%).The misdiagnosis rate was 100% in village clinics and township health centers(8/8 and 2/2,respectively). The misdiagnosis rate was lowest in the municipal medical institutions (3/42,7.14%).There were 41 malaria patients(45.05%)with complications.Conclusions The situation of overseas imported malaria in Jinan City is becoming more and more serious.It is necessary to further strengthen the related professional training for doctors and strengthen the multi-sectoral cooperation for health education,etc.in order to find the cases in time and conduct the active standardized treatment,so as to prevent the second generation cases.

[Epidemiological analysis of malaria surveillance in Jinan City from 2012 to 2016].

OBJECTIVE: To analyze the epidemiological characteristics of malaria reported in the malaria surveillance system in Jinan City from 2012 to 2016, so as to provide the evidence for improving the ability of diagnosis, treatment, prevention and control of malaria. METHODS: The data of all reported malaria cases in the malaria surveillance system were retrospectively analyzed. RESULTS: From 2012 to 2016, 91 malaria cases were found in Jinan City, of which one patient died. All the cases were imported and 95.60% (87 cases) of them came from Africa (note：most of the people were Chinese residents who had been in Africa for the export of labor service and came back China). Falciparum malaria accounted for the most (82.42%, 75 cases). All the cases were adult males, and were mainly migrant workers. The median time from onset to being confirmedly diagnosed was 5 days, and the median time from seeing a doctor to being diagnosed was 1 day. The medical institutions where the patients first visited were mainly municipal medical institutions (42 cases, 46.15%). The misdiagnosis rate was 100% in village clinics and township health centers (8/8 and 2/2, respectively). The misdiagnosis rate was lowest in the municipal medical institutions (3/42, 7.14%). There were 41 malaria patients (45.05%) with complications. CONCLUSIONS: The situation of overseas imported malaria in Jinan City is becoming more and more serious. It is necessary to further strengthen the related professional training for doctors and strengthen the multi-sectoral cooperation for health education, etc. in order to find the cases in time and conduct the active standardized treatment, so as to prevent the second generation cases.

Evaluation of Granulated Lactose as a Carrier for Dry Powder Inhaler Formulations 2: Effect of Drugs and Drug Loading.

Previously, granulated lactose carriers were shown to improve uniformity and aerosolization of a low-dose model drug. In the present study, the blending uniformity and aerosol dispersion performance were assessed for 2 model drugs salbutamol sulfate (SS) and rifampicin (RIF), blended at high loadings (10% or 30% drug) with granulated lactose carriers. The model drug powders differed in particle size distribution, morphology, density, and surface energies. Content uniformity of RIF blends was better than that of SS. Aerosolization studies showed that all blend formulations had acceptable emitted fractions (>70%). The SS blends showed low induction-port deposition (6%-10%) compared to RIF (5%-30%). This difference was greater at high flow rates. At 90 L/min, the low induction port deposition of SS blends allowed high fine particle fraction (FPF) of 73%-81%, whereas the FPF of the RIF blends was around 43%-45% with higher induction port deposition. However, SS blends exhibited strong flow rate-dependent performance. Increasing the flow rate from 30 L/min to 90 L/min increased SS FPF from approximately 20% to 80%. Conversely, RIF blends were flow rate and drug loading independent. It was concluded that the aerosolization of high drug-loaded dry powder inhaler formulations using granulated lactose, particularly flow rate dependency, varies with active pharmaceutical ingredient properties.

Formulation of a novel fixed dose combination of salmeterol xinafoate and mometasone furoate for inhaled drug delivery.

Co-administration of an inhaled corticosteroid and long acting beta agonist for chronic obstructive pulmonary disease has reduced mortality compared to either drug alone. This combination reduces exacerbations, hospitalization, emergency department visits and health care costs. A novel fixed-dose combination of the long acting beta-2 agonist salmeterol xinafoate (SX) and the corticosteroid mometasone furoate (MF) were prepared in a composite particle formulation as brittle matrix powder (BMP) and investigated for suitability as an inhaled combination product. In this study, BMP fixed dose combinations of SX and MF with or without stabilizing excipients (lactose, mannitol, glycine and trehalose) were prepared and characterized with respect to their thermal properties, morphology, aerodynamic performance and physical stability. BMP combination formulations of SX and MF exhibited improved aerodynamic properties when delivered by dry powder inhalation as compared to the micronized blends of the same substances. Aerodynamic evaluation was carried out by next generation pharmaceutical impactor (NGI) with a marketed DPI device. Results demonstrated that co-deposition occurred when SX and MF were formulated together as composite particles in a BMP, while physical blends resulted in inconsistent deposition and dose uniformity. As a result of the bottom-up particle engineering approach, combination BMP formulations allow for dual API composite formulations to be dispersed as aerosolized particles. Aerosolized BMP combination formulations resulted in delivered dose uniformity and co-deposition of each API. Further, an excipient-free formulation, BMP SXMF, delivered approximately 50% of the loaded dose in the respirable range and demonstrated stability at ambient conditions for 6months. Single dose 24-h pharmacokinetic studies in rats demonstrated that lung tissue deposition and blood circulation (AUC0-24h) of two APIs were higher for the BMP combination group exhibiting a significantly higher lung concentration of drugs than for the crystalline physical blend. While high system drug levels are generally undesirable in lung targeted therapies, high blood levels in this rodent study could be indicative of increased pulmonary tissue exposure using BMP formulations.

Bilateral Medial Medullary Infarction with Nondominant Vertebral Artery Occlusion.

Bilateral medial medullary infarction (MMI) is a rare stroke subtype. Here, we report a case with bilateral MMI caused by nondominant vertebral artery occlusion confirmed by brain digital subtraction angiography and magnetic resonance imaging basi-parallel-anatomical-scanning. We highlight that anterior spinal arteries could originate from a unilateral vertebral artery (VA). Radiologists and neurologists should pay attention to the nondominant VA as bilateral MMI may be induced by occlusion of nondominant VA that supplies the bilateral anteromedial territories of the medulla.

[Discussion of epidemic trend and control strategies of malaria in Jinan City from 1989 to 2013].

OBJECTIVE: To explore the epidemic trend of malaria in Jinan City , so as to provide the evidence for improving the prevention and control of malaria. METHODS: The surveillance and annual report data of malaria were collected and analyzed epidemiologically in Jinan City from 1989 to 2013. RESULTS: The prevalence of malaria was low in Jinan City from 1989 to 2013. Totally 79 cases of malaria were reported, and 14 cases (7.82%) were locally infected and 165 cases (92.18%) were imported. CONCLUSION: Most malaria cases were imported since the disease was basically eliminated in Jinan City. The overseas workers from high prevalence areas of malaria should be well managed.

Connecting the P300 to the diagnosis and prognosis of unconscious patients.

The residual consciousness of unconscious patients can be detected by studying the P300, a wave among event-related potentials. Previous studies have applied tones, the subject's name and other names as stimuli. However, the results were not satisfactory. In this study, we changed the constituent order of subjects' two-character names to create derived names. The subject's derived names, together with tones and their own names, were used as auditory stimuli in event-related potential experiments. Healthy controls and unconscious patients were included in this study and made to listen to these auditory stimuli. In the two paradigms, a sine tone followed by the subject's own name and the subject's derived name followed by the subject's own name were used as standard and deviant stimuli, respectively. The results showed that all healthy controls had the P300 using both paradigms, and that the P300 in the second paradigm had a longer latency and two peaks. All minimally conscious state patients had the P300 in the first paradigm and the majority of them had the P300 in the second paradigm. Most vegetative state patients had no P300. Patients who showed the P300 in the two paradigms had more residual consciousness, and patients with the two-peak P300 had a higher probability of awakening within a short time. Our experimental findings suggest that the P300 event-related potential could reflect the conscious state of unconscious patients.

[Relationship between CCL20/CCR6/Th17 axis and vascular invasion and metastasis in patients with primary hepatocellular carcinoma].

OBJECTIVE: To investigate the role of CCL20/CCR6/Th17 axis in vascular invasion and metastasis of primary hepatocellular carcinoma (HCC). METHODS: Expression levels of CCL20 mRNA in the normal human liver cell line L-02, and human hepatocellular carcinoma cell lines Hep3B, Huh7 and HepG2 were quantified by using SYBR green real time PCR. CCL20 secretions from these cell lines were quantified by using ELISA. The chemotactic effect of HCC cell line Hep3B on human peripheral blood mononuclear cells was determined by using transwell chemotaxis assay. Pre-therapy serum levels of IL-1α, IL-1ß, IL-6, IL-8, IL-10, IL-17, IL-23, IFN-γ, TNF-α and CCL20 in 93 patients with HCC were measured by using 9-plex array and ELISA. All the patients were chronic hepatitis B virus associated HCC, and 51 cases were those with vascular invasion and metastasis (metastasis group) and 42 cases were not (non-metastasis group). CCL20 and CCR6 mRNA expressions in the HCC and tumor-adjacent tissues were determined by using SYBR Green real time PCR in 41 patients, among them, 20 cases were from the group of patients with metastasis and 21 cases were from the group of patients without metastasis. The CCL20 expression was further determined by immunohistochemistry. RESULTS: The HCC cell lines expressed and secreted higher amount of CCL20, which effectively recruited CCR6(+) T cells. Pre-therapy serum levels of CCL20 in 93 HCC patients were (38.2 ± 28.4)pg/ml, significantly increased than those with benign hepatic hemangiomas [(7.8 ± 17.8)pg/ml, P < 0.01]. In addition, the serum levels of CCL20 were positively correlated with the tumor diameters in HCC patients (r = 0.32, P = 0.0018). CCL20 was dominantly expressed in the cytoplasm in HCC cells, and it was also expressed by some infiltrating immune cells. The mRNA expression levels of CCL20 of the tumor tissues were significantly higher than that in the tumor-adjacent tissues (P < 0.05). Multivariate logistic regression analysis showed that serum levels of IL-17 and CCL20 were independent risk factors of metastasis in HCC patients (P < 0.05 for both). CCL20 mRNA showed no statistically significant differences between patients with metastasis and without metastasis in both tumor tissues and tumor-adjacent tissues (P > 0.05 for both). But the patients with metastasis showed significantly higher expressions of CCR6 both in their tumor [5.75 (1.79, 19.13)]and tumor-adjacent tissues [7.99 (4.49, 19.54)] than those with non-metastasis [1.69 (0.76, 2.87) and 3.58 (1.84, 4.32), P < 0.05 for both]. CONCLUSION: CCL20/CCR6/Th17 axis may promote vascular invasion and metastasis hepatocellular carcinoma.

Improved Biofilm Antimicrobial Activity of Polyethylene Glycol Conjugated Tobramycin Compared to Tobramycin in Pseudomonas aeruginosa Biofilms.

The objective of this study was to develop a functionally enhanced antibiotic that would improve the therapeutic activity against bacterial biofilms. Tobramycin was chemically conjugated with polyethylene glycol (PEG) via site-specific conjugation to form PEGylated-tobramycin (Tob-PEG). The antibacterial efficacy of Tob-PEG, as compared to tobramycin, was assessed on the planktonic phase and biofilms phase of Pseudomonas aeruginosa. The minimum inhibitory concentration (MIC80) of Tob-PEG was higher (13.9 µmol/L) than that of tobramycin (1.4 µmol/L) in the planktonic phases. In contrast, the Tob-PEG was approximately 3.2-fold more effective in eliminating bacterial biofilms than tobramycin. Specifically, Tob-PEG had a MIC80 lower than those exhibited by tobramycin (27.8 µmol/L vs 89.8 µmol/L). Both confocal laser scanning microscopy and scanning electron microscopy further confirmed these data. Thus, modification of antimicrobials by PEGylation appears to be a promising approach for overcoming the bacterial resistance in the established biofilms of Pseudomonas aeruginosa.

Effect of platelet-derived growth factor-B on renal cell carcinoma growth and progression.

BACKGROUND: Platelet-derived growth factor-B (PDGF-B) expression promotes the proliferation of mural cells surrounding the blood vessels during angiogenesis. The effect of PDGF-B involved in angiogenesis on tumor growth and progression in clear cell renal cell carcinoma (ccRCC) is unknown. METHODS: We examined the expression of PDGF-B and its receptor PDGFR-ß in 174 patients with ccRCC by microarray analysis. Cancer-specific survival was estimated using the Kaplan-Meier method. PDGF-B-transfected and mock-transfected ACHN cells were implanted into mice to induce tumor formation and tumor growth, respectively, and progression in mice models was assessed using immunohistochemistry and histomorphology. The role of PDGF-B during angiogenesis in vitro was evaluated by flow cytometry analysis, cell migration, and tube formation assay. RESULTS: High expression of PDGF-B was associated with significantly decreased risk of cancer-specific mortality (P ≤ 0.001). The data indicated significant inhibition of tumor growth (P ≤ 0.05) and a reduction in proliferating tumor cells (P = 0.019) in vivo. PDGF-B also inhibits tumor metastasis and invasion events in tumor-bearing mice models. In vitro studies revealed that the tube formation capability of vascular smooth muscle cells (VSMCs), which are believed to be the precursors to pericytes in vivo, significantly induced by PDGF-B. The PDGF-B overexpression also results in a tendency to reside in S and G2/M phases of the cell cycle (P = 0.001) and increasing migration capability of VSMCs (P ≤ 0.001). CONCLUSION: Our results demonstrated that PDGF-B, which increased VSMCs proliferation and migration capability during angiogenesis, limited tumor growth and progression in ccRCC. Therefore, PDGF-B may be a novel and promising prognostic marker.

Swellable ciprofloxacin-loaded nano-in-micro hydrogel particles for local lung drug delivery.

Incorporation of drug-loaded nanoparticles into swellable and respirable microparticles is a promising strategy to avoid rapid clearance from the lung and achieve sustained drug release. In this investigation, a copolymer of polyethylene glycol grafted onto phthaloyl chitosan (PEG-g-PHCs) was synthesized and then self-assembled with ciprofloxacin to form drug-loaded nanoparticles. The nanoparticles and free drug were encapsulated into respirable and swellable alginate micro hydrogel particles and assessed as a novel system for sustained pulmonary drug delivery. Particle size, morphology, dynamic swelling profile, and in vitro drug release were investigated. Results showed that drug-loaded nanoparticles with size of 218 nm were entrapped into 3.9-µm micro hydrogel particles. The dry nano-in-micro hydrogel particles exhibited a rapid initial swelling within 2 min and showed sustained drug release. Preliminary in vivo pharmacokinetic studies were performed with formulations delivered to rats by intratracheal insufflation. Ciprofloxacin concentrations in plasma and in lung tissue and lavage were measured up to 7 h. The swellable particles showed lower ciprofloxacin levels in plasma than the controlled group (a mixture of lactose with micronized ciprofloxacin), while swellable particles achieved higher concentrations in lung tissue and lavage, indicating the swellable particles could be used for controlling drug release and prolonging lung drug concentrations.

Evaluation of granulated lactose as a carrier for DPI formulations 1: effect of granule size.

The objective of this study was to investigate the effect of large granulated lactose carrier particle systems on aerosol performance of dry powder inhaler formulations. Granulated lactose carriers with average sizes ranging from 200 to 1,000 µm were prepared and subsequently fractionated into separate narrow size powders. The fractionated granulated lactose (GL) samples were characterized in terms of size, specific surface area, surface roughness, morphology, density, flowability, and solid-state. The in vitro aerosolization performance was performed on the different size fractions of GL samples from a commercial inhaler device (Aerolizer®) with a model formulation (2% w/w salbutamol sulfate). The cascade impaction parameters employed were 60 or 90 L/min with standard (aperture size, 0.6 mm) or modified piercing holes (aperture size, 1.2 mm) of the inhaler loaded capsules. It was shown that the largest size fraction formulation (850-1000 µm) had a slight improvement in the fine particle fraction (FPF) compared to immediately preceding size fractions, explained by a smaller adhesive force between drug and carrier. Compared to commercial piercing holes, enlarged piercing holes generated a slight decreasing trend of FPF as the lactose powder sizes increased from 200-250 µm to 600-850 µm, perhaps due to the reduced detachment force by flow forces. The size, surface roughness, density, and flowability of lactose carrier as well as device design all contributed to the aerosol dispersion performance of granulated lactose-based adhesive mixtures. It was concluded that poorer or enhanced redispersion performance is not an inherent property to the significantly large size of granulated lactose carriers as previously contended.

Mitochondrial DNA haplogroup confers genetic susceptibility to nasopharyngeal carcinoma in Chaoshanese from Guangdong, China.

Recent studies have shown association of mtDNA background with cancer development. We analyzed mitochondrial DNA (mtDNA) control region variation of 201 patients with nasopharyngeal carcinoma (NPC) and of 201 normal controls from Chaoshan Han Chinese to discern mtDNA haplogroup effect on the disease onset. Binary logistic regression analysis with adjustment for gender and age revealed that the haplogroup R9 (P = 0.011, OR = 1.91, 95% CI = 1.16-3.16), particularly its sub-haplogroup F1 (P = 0.015, OR = 2.43, 95% CI = 1.18-5.00), were associated significantly with increased NPC risk. These haplogroups were further confirmed to confer high NPC risk in males and/or individuals ≥ 40 years of age, but not in females or in subjects <40 years old. Our results indicated that mtDNA background confers genetic susceptibility to NPC in Chaoshan Han Chinese, and R9, particularly its sub-haplogroup F1, is a risk factor for NPC.

Hydrogels for controlled pulmonary delivery.

A significant number of research articles have focused on pulmonary delivery as an alternative administration route owing to no first-pass metabolism, low protease activity, thin epithelium barrier and large surface area in the lung system. Controlled release in the pulmonary delivery system further reduces loading dose, frequency of dosing and systemic side effects, and also increases duration of action and patient compliance. Compared with other microparticles used in controlled-release pulmonary administration, hydrogels (3D polymeric matrix networks) have recently been investigated due to their swelling and mucoadhesive properties that could help bypass pulmonary delivery barriers. This review introduces controlled-release drug delivery to the lung, followed by a summary of currently available approaches for controlled-release pulmonary drug delivery. Lastly, the origin, advantages, detailed applications and concerns of hydrogels in pulmonary delivery are discussed.

Dry powder insufflation of crystalline and amorphous voriconazole formulations produced by thin film freezing to mice.

Attention has begun to focus on the pulmonary delivery of antifungal agents for invasive fungal infections as inhalation of the fungal spores is often the initial step in the pathogenesis of many of these infections, including invasive pulmonary aspergillosis (IPA). IPA in immunocompromised patients has high mortality rates despite current systemic (oral or intravenous) therapies. In this study, particulate voriconazole (VRC) formulations were designed with suitable properties for inhalation using thin film freezing (TFF), a particle engineering process capable of producing low-density porous aggregate particles. Nanostructured amorphous morphology of VRC was less favorable in vitro and in vivo than microstructured crystalline morphology, despite being a poorly water-soluble compound. Using a Handihaler dry powder inhaler (DPI), microstructured crystalline TFF-VRC and nanostructured amorphous TFF-VRC-PVP K25 (1:3) had fine particle fractions of 37.8% and 32.4% and mass median aerodynamic diameters of 4.2 and 5.2 µm, respectively. Single dose 24-h pharmacokinetic studies were conducted in ICR mice. AUC(0-24h) in the lung tissue and plasma was 452.6 µg h/g wet lung weight and 38.4 µg h/mL, respectively, following a 10mg/kg insufflated dose of TFF-VRC directly into the lungs of the mice, while AUC(0-24 h) in the lung tissue and plasma was 232.1 µg h/g wet lung weight and 18.6 µg h/mL, respectively, following a 10mg/kg insufflated dose of TFF-VRC-PVP K25 (1:3). High concentrations of VRC in lung tissue coupled with clinically relevant plasma concentrations suggest that pulmonary delivery of microstructured crystalline VRC could potentially be a beneficial strategy for administration of VRC to patients with invasive pulmonary fungal infections.

PEGylated Poly(amidoamine) dendrimer-based dual-targeting carrier for treating brain tumors.

A dual-targeting drug carrier (PAMAM-PEG-WGA-Tf) based on the PEGylated fourth generation (G = 4.0) PAMAM dendrimer with transferrin (Tf) and wheat germ agglutinin (WGA) on the periphery and doxorubicin (DOX) loaded in the interior was synthesized and its BBB penetration and tumor targeting properties were explored. DLS and TEM measurements revealed the size of PAMAM-PEG-WGA-Tf was in the range of 14-20 nm. It reduced the cytotoxicity of DOX to the normal cells greatly, while efficiently inhibited the growth rate of the C6 glioma cells. The assay of transport across the BBB showed that PAMAM-PEG-WGA-Tf delivered 13.5% of DOX in a period of 2 h, demonstrating an enhanced transport ratio as compared to the ratio of 8% for PAMAM-PEG-WGA, 7% for PAMAM-PEG-Tf and 5% for free DOX in the same period of time. The accumulation of DOX in the tumor site was increased due to the targeting effects of both Tf and WGA, leading to the complete breakage of the avascular C6 glioma spheroids in vitro.

Effects of stealth liposomal daunorubicin plus tamoxifen on the breast cancer and cancer stem cells.

PURPOSE: The cancer stem cells play an important role in the invasion, metastasis and relapse of cancers as they are resistant to regular chemotherapy. In the present study, stealth liposomal daunorubicin plus tamoxifen was developed for eradicating breast cancer cells together with cancer stem cells. METHODS: Inhibitory effects were performed on the bulk human breast cancer cells (MCF-7), the sorted MCF-7 cancer stem-like cells (side population, SP), and the sorted MCF-7 cancer cells (NSP), respectively. Antitumor activity and TUNEL analysis were evaluated on the MCF-7 xenografts in nude mice. RESULTS: The encapsulation efficiencies of daunorubicin and tamoxifen were 95% and 90%, respectively. The mean particle size of the stealth liposomes was about 100 nm. Breast cancer stem cells were identified by the specific markers CD44+/CD24-, and isolated from bulk MCF-7 cells. When applying stealth liposomal daunorubicin plus tamoxifen, the inhibitory effects on both the breast cancer cells and the cancer stem cells were significantly increased in vitro, respectively. In the MCF-7 xenografts in mice, stealth liposomal daunorubicin plus tamoxifen showed the most favorable antitumor activity due to the passive targeting the tumor tissue and the synergistic effects in eliminating breast cancer cells and cancer stem cells. CONCLUSION: Stealth liposomal daunorubicin plus tamoxifen could have the potentials in eliminating both breast cancer cells and cancer stem cells.

Enhanced efficacy of functionalized epirubicin liposomes in treating brain glioma-bearing rats.

PURPOSE: The restriction of drug transporting across the blood-brain barrier (BBB) and the limit of drug penetrating into the tumor tissue remain the major obstacles for brain tumor chemotherapy. In the present study, we developed a functionalized liposomal nanoconstruct, epirubicin liposomes modified with tamoxifen (TAM) and transferrin (TF), for transporting drug across the BBB and afterwards targeting the brain glioma. METHODS: Evaluations were performed on the murine C6 glioma cells, the C6 glioma spheroids, the BBB model in vitro and the brain glioma-bearing rats. RESULTS: When compared with controls, epirubicin liposomes modified with TAM and TF showed the strongest inhibitory effect to C6 glioma cells or glioma spheroids in vitro, significant transport ability across the BBB model in vitro, an evident effect of targeting the brain tumor cells in vitro, and an extended median survival time in the brain glioma-bearing rats. CONCLUSION: Epirubicin liposomes modified with TAM and TF significantly improve the therapeutic efficacy of brain glioma in vitro and in animals, hence providing a new strategy for brain tumor chemotherapy.

Dual-targeting daunorubicin liposomes improve the therapeutic efficacy of brain glioma in animals.

Chemotherapy for brain glioma has been of limited value due to the inability of transport of drug across the blood-brain barrier (BBB) and poor penetration of drug into the tumor. For overcoming these hurdles, the dual-targeting daunorubicin liposomes were developed by conjugating with p-aminophenyl-alpha-D-manno-pyranoside (MAN) and transferrin (TF) for transporting drug across the BBB and then targeting brain glioma. The dual-targeting effects were evaluated on the BBB model in vitro, C6 glioma cells in vitro, avascular C6 glioma tumor spheroids in vitro, and C6 glioma-bearing rats in vivo, respectively. After applying dual-targeting daunorubicin liposomes, the transport ratio across the BBB model was significantly increased up to 24.9%. The most significant uptake by C6 glioma was evidenced by flow cytometry and confocal microscope. The C6 glioma spheroid volume ratio was significantly lowered to 54.7%. The inhibitory rate to C6 glioma cells after crossing the BBB was significantly enhanced up to 64.0%. The median survival time of tumor bearing rats after administering dual-targeting daunorubicin liposomes (22 days) was significantly longer than that after giving free daunorubicin (17 days, P=0.001) or other controls. In conclusion, the dual-targeting daunorubicin liposomes are able to improve the therapeutic efficacy of brain glioma in vitro and in animals.