Alterations of Cardiac KATP Channels and Autophagy Contribute in the Late Cardioprotective Phase of Exercise Preconditioning.

The cardiac effects of exercise preconditioning (EP) are well established; however, the mechanisms involving cardiac ATP-sensitive potassium channel (KATP channel) subunits and autophagy are yet to be fully established. The present work aims to investigate the alterations of cardiac KATP channel subunits Kir6.2, SUR2A, and autophagy-related LC3 during the late cardioprotective phase of EP against exhaustive exercise-induced myocardial injury. Rats run on treadmill for four running time intervals, each with 10 minutes running and rest. Exhaustive exercise was performed 24 h after EP. Cardiac biomarkers, cTnI and NT-proBNP, along with the histological stain, were served as indicators of myocardial injury. Cardiac KATP channel subunits Kir6.2 and SUR2A were analyzed in this study, and autophagy was evaluated by LC3. The results revealed that EP reduced the exhaustive exercise-induced high level of serum cTnI and myocardial ischemia/hypoxia; however, it did not reveal any changes in the serum NT-proBNP level or cardiac BNP. Cardiac SUR2A mRNA significantly upregulated during the exhaustive exercise. The high levels of Kir6.2, SUR2A, LC3IIpuncta and LC3II turnover observed after exhaustive exercise were significantly mitigated by EP in the late phase. These results suggest that EP alleviates myocardial injury induced by exhaustive exercise through the downregulation of cardiac KATP channels and autophagy.

Cationic nanomicelles derived from Pluronic F127 as delivery vehicles of Chinese herbal medicine active components of ursolic acid for colorectal cancer treatment.

Ursolic acid (UA) has shown great potential in cancer therapy but their efficacy is seriously compromised by poor water-solubility and limited cellular uptake. In this paper, cationic nanomicelles self-assembled from Pluronic F127 with the cationic polymer of C18-polyethylenimine (C18-PEI) as a functional component are fabricated as delivery vehicles of Chinese herbal medicine active components of ursolic acid (UA) for colorectal cancer treatment. The inhibition effects of this drug loaded cationic nanomicelles (named as FUP) on cell viability and cell colony formation were more significant than the free UA, due to their cationic surface which can increase UA uptake by colorectal cancer cells. Cell cycle analysis showed that this inhibition effect was mediated by a cell cycle arrest at the G1 checkpoint, and the cell death induced by these nanomicelles occurred via apoptosis, which was detected by annexin V antibody and propidium iodide staining. Further western blot analysis demonstrated the apoptosis mechanism was associated with the regulation of Fas/FasL and activation of caspase-8 and caspase-3. Therefore, our cationic nanomicelles can potentially be used to enhance the therapeutic effect of UA for colorectal cancer treatment.

Magnetite nanocluster@poly(dopamine)-PEG@ indocyanine green nanobead with magnetic field-targeting enhanced MR imaging and photothermal therapy in vivo.

Multifunctional nanomaterials with the magnetic resonance imaging (MRI) guided tumor photothermal ablation ability have been extensively applied in biomedical research as one of the most exciting and challenging strategies for cancer treatment. Nevertheless, most of these nanomaterials still suffer from low accumulation in tumor tissues and insufficient photothermal ablation of tumors so far. Here, we report a novel approach to overcome these limitations using a core-shell magnetite nanocluster@poly(dopamine)-PEG@ICG nanobead compositing of magnetite nanocluster core with coating of poly(dopamine), then further conjugating with polyethylene glycol (PEG) and adsorbing indocyanine green (ICG) on the surface. The adsorbed ICG in the nanobead displays a higher photostability and photothermal conversion ability than free ICG, as well as additional photothermal effect rather than magnetite nanocluster and poly(dopamine), which endow the nanobead with enhanced photothermal killing efficiency against cancer cells under near-infrared (NIR) laser irritation. Furthermore, it is proved that these nanobeads have excellent biocompatibility, T2-weighted MR imaging and magnetic field targeting ability. By applying an external magnetic field (MF) focused on the targeted tumor, a magnetic targeting mediated enhanced accumulation is observed at tumor site as proved by a darker T2-weighted MR image. Utilizing the magnetic targeting strategy, enhanced photothermal tumor ablation was achieved under laser irradiation in vivo, which is reflected by the degree of tumor tissue damage and tumor growth delay. Therefore, this nanobead integrates the abilities of magnetic field-targeting, MR imaging and photothermal cancer therapy, and might be a promising theranostic platform for tumor treatment.

Chlorin e6 Conjugated Poly(dopamine) Nanospheres as PDT/PTT Dual-Modal Therapeutic Agents for Enhanced Cancer Therapy.

Photodynamic therapy (PDT), using a combination of chemical photosensitizers (PS) and light, has been successfully applied as a noninvasive therapeutic procedure to treat tumors by inducing apoptosis or necrosis of cancer cells. However, most current clinically used PS have suffered from the instability in physiological conditions which lead to low photodynamic therapy efficacy. Herein, a highly biocompatible poly(dopamine) (PDA) nanoparticle conjugated with Chlorin e6 (referenced as the PDA-Ce6 nanosphere) was designed as a nanotherapeutic agent to achieve simultaneous photodynamic/photothermal therapy (PDT/PTT). Compared to the free Ce6, the PDA-Ce6 nanosphere exhibited significantly higher PDT efficacy against tumor cells, because of the enhanced cellular uptake and subsequently greater reactive oxygen species (ROS) production upon laser irradiation at 670 nm. Meanwhile, the PDA-Ce6 nanosphere could be also used as a photoabsorbing agent for PTT, because of the excellent photothermal conversion ability of PDA nanoparticle under laser irradiation at 808 nm. Moreover, our prepared nanosphere had extremely low dark toxicity, while excellent phototoxicity under the combination laser irradiation of 670 and 808 nm, both in vitro and in vivo, compared to any single laser irradiation alone. Therefore, our prepared PDA-Ce6 nanosphere could be applied as a very promising dual-modal phototherapeutic agent for enhanced cancer therapy in future clinical applications.

Detection and mechanism of action of ESM-1 in rat kidney transplantation under various immune states.

OBJECTIVE: To investigate whether ESM-1 expression change reflects the impairment of endothelial cells and rejection after kidney transplantation, ESM-1 expression was detected under various immune states in this study. METHODS: Kidney transplantations were performed from BN to LEW rats. Syngenic LEW-LEW grafts were used as controls. The LEW recipient rats were divided into acute rejection (AR) group, ciclosporin A (CsA) group and control group. In each group, 10 rats were sacrificed at 1, 5, and 7d after operation, respectively, and blood and kidney samples were collected. In the rat model of kidney transplantation, ESM-1 mRNA and ESM-1 protein expression were detected in various immune states to verify if ESM-1 can reflect endothelial cell impairment sensitively. RESULTS: ESM-1 mRNA (1d vs. 3d, P<0.01;3d vs. 7d, P=0.018) and ESM-1 protein expression was upregulated significantly in the AR group (P<0.01, 5 and 7d), when compared to CsA group and control group. In CsA group, the cell apoptosis rate decreased when compared to AR group (P<0.01). Pathological impairment was more serious in AR group than in CsA group (P<0.01). CONCLUSIONS: Peripheral blood ESM-1 mRNA and ESM-1 protein expression in kidney grafts can reflect the severity of endothelial cell impairment. Thus, ESM-1 may be used as a new indicator for AR prediction and diagnosis. Nevertheless, further investigation is required to test if it meets the criteria for clinical utility.

Preparation of novel alumina nanowire solid-phase microextraction fiber coating for ultra-selective determination of volatile esters and alcohols from complicated food samples.

A novel alumina nanowire (ANW) solid-phase microextraction (SPME) fiber coating was prepared by a simple and rapid anodization-chemical etching method for ultra-selective determination of volatile esters and alcohols from complicated food samples. Preparation conditions for ANW SPME fiber coating including corrosion solution concentration and corrosion time were optimized in detail for better surface morphology and higher surface area based on scanning electron microscope (SEM). Under the optimum conditions, homogeneous alumina nanowire structure of ANW SPME fiber coating was achieved with the average thickness of 20 µm around. Compared with most of commercial SPME fiber coatings, ANW SPME fiber coatings achieved the higher extraction capacity and special selectivity for volatile esters and alcohols. Finally, an efficient gas sampling technique based on ANW SPME fiber coating as the core was established and successfully applied for the ultra-selective determination of trace volatile esters and alcohols from complicated banana and fermented glutinous rice samples coupled with gas chromatography/mass spectrometry (GC/MS) detection. It was interesting that 25 esters and 2 alcohols among 30 banana volatile organic compounds (VOCs) identified and 4 esters and 7 alcohols among 13 identified VOCs of fermented glutinous rice were selectively sampled by ANW SPME fiber coatings. Furthermore, new analytical methods for the determination of some typical volatile esters and alcohols from banana and fermented glutinous rice samples at specific storage or brewing phases were developed and validated. Good recoveries for banana and fermented glutinous rice samples were achieved in range of 108-115% with relative standard deviations (RSDs) of 2.6-6.7% and 80.0-91.8% with RSDs of 0.3-1.3% (n=3), respectively. This work proposed a novel and efficient gas sampling technique of ANW SPME which was quite suitable for ultra-selectively sampling trace volatile esters and alcohols from complicated food samples.

microRNA response elements-regulated TRAIL expression shows specific survival-suppressing activity on bladder cancer.

BACKGROUND: Bladder transitional cell carcinoma greatly threatens human health all over the world. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) shows a strong apoptosis-inducing effect on a variety of cancer cells including bladder cancer. However, adenovirus-mediated TRAIL expression still showed cytotoxicity to normal cells mainly due to lack of tumor specificity. METHODS: To solve the problem, we applied miRNA response elements (MREs) of miR-1, miR-133 and miR-218 to confer TRAIL expression with specificity to bladder cancer cells. RESULTS: Expression of miR-1, miR-133 and miR-218 was greatly decreased in bladder cancer than normal bladder tissue. Luciferase assay showed that application of the 3 MREs was able to restrain exogenous gene expression to within bladder cancer cells. Subsequently, we constructed a recombinant adenovirus with TRAIL expression regulated by MREs of miR-1, miR-133 and miR-218, namely Ad-TRAIL-MRE-1-133-218. qPCR, immunoblotting and ELISA assays demonstrated that Ad-TRAIL-MRE-1-133-218 expressed in bladder cancer cells, rather than normal bladder cells. The differential TRAIL expression also led to selective apoptosis-inducing and growth-inhibiting effect of Ad-TRAIL-MRE-1-133-218 on bladder cancers. Finally, bladder cancer xenograft in mouse models further confirmed that Ad-TRAIL-MRE-1-133-218 effectively suppressed the growth of bladder cancers. CONCLUSIONS: Collectively, we demonstrated that MREs-based TRAIL delivery into bladder cancer cells was feasible and efficient for cancer gene therapy.

Detection on dynamic changes of endothelial cell specific molecule-1 in acute rejection after renal transplantation.

OBJECTIVE: To investigate the value of dynamic monitoring of endothelial cell specific molecule-1 (ESM-1) in diagnosing acute rejection after renal transplantation. METHODS: ESM-1 expression was observed in peripheral blood circulating endothelial cells and renal allografts after renal transplantation, and was compared to the flow cytometry (FCM) results of urine HLA-DR(+) lymphocytes. RESULTS: In patients with acute rejection, ESM-1 mRNA and protein expression increased significantly (P < .01). Real-time polymerase chain reaction detection of ESM-1 mRNA in peripheral blood cells was more sensitive and specific than FCM detection of urine HLA-DR(+) lymphocytes. Moreover, real-time PCR detection is characterized by convenient sampling and good reproducibility. CONCLUSION: ESM-1 is a new marker for endothelial cell activation. However, its significance during acute rejection in renal transplantation is still unclear. Our study demonstrates that ESM-1 may reflect the degree of endothelial cell injury in renal allografts, and that it may serve as a highly sensitive and specific marker for acute rejection after renal transplantation.

Fabrication of novel nanoporous array anodic alumina solid-phase microextraction fiber coating and its potential application for headspace sampling of biological volatile organic compounds.

In the study, nanoporous array anodic alumina (NAAA) prepared by a simple, rapid and stable two-step anodic oxidization method was introduced as a novel solid-phase microextraction (SPME) fiber coating. The regular nanoporous array structure and chemical composition of NAAA SPME fiber coating was characterized and validated by scanning electron microscopy and energy dispersive spectroscopy, respectively. Compared with the commercial polydimethylsiloxane (PDMS) SPME fiber coating, NAAA SPME fiber coating achieved the higher enrichment capability (1.7-4.7 folds) for the mixed standards of volatile organic compounds (VOCs). The selectivity for volatile alcohols by NAAA SPME fiber coating demonstrated an increasing trend with the increasing polarity of alcohols caused by the gradually shortening carbon chains from 1-undecanol to 1-heptanol or the isomerization of carbon chains of some typical volatile alcohols including 2-ethyl hexanol, 1-octanol, 2-phenylethanol, 1-phenylethanol, 5-undecanol, 2-undecanol and 1-undecanol. Finally, NAAA SPME fiber coating was originally applied for the analysis of biological VOCs of Bailan flower, stinkbug and orange peel samples coupled with gas chromatography-mass spectrometry (GC-MS) detection. Thirty, twenty-seven and forty-four VOCs of Bailan flower, stinkbug and orange peel samples were sampled and identified, respectively. Moreover, the contents of trace 1-octanol and nonanal of real orange peel samples were quantified for the further method validation with satisfactory recoveries of 106.5 and 120.5%, respectively. This work proposed a sensitive, rapid, reliable and convenient analytical method for the potential study of trace and small molecular biological VOCs by the novel NAAA SPME fiber coating.

[Efficacy of compound Xuanju capsule in the treatment of chronic prostatitis with erectile dysfunction].

OBJECTIVE: To evaluate the efficacy and safety of Compound Xuanju Capsule (CXC) in the treatment of chronic prostatitis with erectile dysfunction (ED). METHODS: We obtained NIH-CPSI and IIEF-5 scores from 132 chronic prostatitis patients with ED and divided the patients into a control (n = 70) and a treatment group (n = 62), the former treated with oral levofloxacin 0.2 g bid for 4-6 weeks and oral Terazosin at 2 mg qd for 2 months, and the latter with oral CXC once 2 capsules tid for 2 months in addition to the above. RESULTS: None of the patients had serious medication-related adverse reactions. After treatment, the control group showed significantly decreased NIH-CPSI scores and slightly increased IIEF-5 scores as compared with the baseline (16.5 +/- 5.9 vs 25.1 +/- 5.5, P < 0.05 and 13.1 +/- 5.2 vs 11.3 +/- 4.5, P > 0.05), while the treatment group exhibited significant improvement in both NIH-CPSI (13.4 +/- 5.7 vs 25.5 +/- 5.3, P < 0.05) and IIEF-5 scores (17.5 +/- 6.5 vs 10.8 +/- 3.8, P < 0.05). The total effectiveness rate for ED was significantly higher in the treatment than in the control group (74.2% vs 20%, P < 0.05). CONCLUSION: Compound Xuanju Capsule can significantly alleviate both the symptoms of chronic prostatitis and ED in the treatment of chronic prostatitis patients with ED.

ZnO nanorod array polydimethylsiloxane composite solid phase micro-extraction fiber coating: fabrication and extraction capability.

ZnO nanorod array coating is a novel kind of solid-phase microextraction (SPME) fiber coating which shows good extraction capability due to the nanostructure. To prepare the composite coating is a good way to improve the extraction capability. In this paper, the ZnO nanorod array polydimethylsiloxane (PDMS) composite SPME fiber coating has been prepared and its extraction capability for volatile organic compounds (VOCs) has been studied by headspace sampling the typical volatile mixed standard solution of benzene, toluene, ethylbenzene and xylene (BTEX). Improved detection limit and good linear ranges have been achieved for this composite SPME fiber coating. Also, it is found that the composite SPME fiber coating shows good extraction selectivity to the VOCs with alkane radicals.

Treatment of urinary lithiasis following kidney transplantation with extracorporeal shock-wave lithotripsy.

BACKGROUND: The incidence of urinary lithiasis following kidney transplantation is very low, and decision-supporting data are not available. The aim of this study was to review the diagnosis and treatment of urinary lithiasis following kidney transplantation, which is of realistic significance to reduce urinary lithiasis following kidney transplantation, prolong the survival of renal allografts. METHODS: The incidence, diagnosis and treatment of urinary lithiasis in ten patients following kidney transplantation were analyzed retrospectively. Seven out of these patients had stones sized approximately 0.4 - 1.1 cm, and they were treated with low-voltage, low-frequency extracorporeal shock-wave lithotripsy (ESWL). Two patients had stones sized < 0.3 cm and they underwent cystoscopy and ureteroscopy. The ureteral catheter endoscopes were inserted in a retrograde manner to mobilize stones repeatedly. After elimination of obstruction, a ureteral double J stent was indwelt. One patient had a pelvic stone (1.2 cm), which was removed surgically. RESULTS: The major clinical manifestations were hematuria, oliguria or anuria. Some patients were asymptomatic and they were diagnosed through laboratory tests and imaging examinations, e.g., ultrasonography. After elimination of obstruction, subjective symptoms disappeared in all patients, and the function of renal allografts recovered. A six-month follow-up indicated no remnant stones or lithiasis relapse. CONCLUSIONS: The diagnosis and treatment of renal allograft lithiasis are challenging. After prompt and appropriate treatment, the prognosis was satisfactory, and permanent renal functional impairment did not occur in most patients.

Increased antitumor capability of fiber-modified adenoviral vector armed with TRAIL against bladder cancers.

Adenoviral vectors are widely used for cancer therapy and show a tumor-suppressing effect. However, bladder cancers are found to be resistant against infection of Ad5-derived adenoviral vector, limiting the application of the existing strategy of gene therapy. Therefore, efforts to develop novel types of adenoviral vector aimed for improving the viral infection and enhancing expression level of tumor-inhibiting transgene is urgently required. We constructed a 5/35 fiber-modified E1A-deleted adenoviral vector armed with TRAIL gene. Its ability to express this gene for inhibition of bladder cancer cell growth was investigated in our work. The results showed that this modification in fiber region facilitates adenoviral infection to bladder cancer, perhaps due to high expression of CD46 on target cell surface. Subsequently, we found an enhanced expression level of TRAIL mediated by 5/35 fiber-modified adenoviral vectors in bladder cancer cells, leading to an increased tumor-inhibiting capability of 5/35 adenoviral vector against bladder cancer cells. Consistently, growth of xenograft tumors in mice was also effectively inhibited by 5/35 fiber-modified vector-mediated gene therapy strategy. The 5/35 fiber-modified adenoviral vector-based gene transfer shows an improved efficacy against bladder cancers. The application of this novel gene therapy vector may benefit the patients in clinical bladder cancer treatment.

[Expression of hypoxia-inducible factor 1 alpha and vascular endothelial growth factor in prostate cancer and its significance].

OBJECTIVE: To study the expression of hypoxia-inducible factor 1 alpha (HIF-1 alpha) and vascular endothelial growth factor (VEGF) in prostate cancer (PCa) and its biological significance. METHODS: Specimens from 75 patients with PCa (32 cases), benign prostatic hyperplasia (BPH, 16 cases), high-grade prostate intraepithelial neoplasia (PIN, 15 cases) and normal prostate (NP, 12 cases) were comprised in the study. The PCa was divided into high Gleason score group (Gleason score > or = 7) and low Gleason score group (Gleason score < 7). Immunohistochemistry was performed with antibodies against HIF-1 alpha, VEGF and CD34. Microvessel density (MVD) was analyzed by staining with antibodies to CD34. RESULTS: The positive rates of HIF-1alpha were significantly higher in PCa (62.5%) and PIN (60.0%) than those in BPH (6.3%) and NP(0) (P < 0.05), respectively. The positive rates of VEGF were significantly higher in PCa (78.1%) and PIN (73.3%) than those in BPH (18.7%) and NP (8.3%) (P < 0.05), respectively. A higher concentration of MVD was observed in PCa (66.9 +/- 18.0) compared with those in BPH (28.3 +/- 6.9) and NP (15.3 +/- 2.9) (P < 0.05). MVD and the positive rates of HIF-1alpha and VEGF were significantly higher in high Gleason score group than those in low Gleason score group. CONCLUSION: The results suggest that the overexpression of HIF-1alpha and VEGF are closely related to PCa, and the son score group. up-regulation of HIF-1alpha and VEGF are early events in PCa.