The objectives of this study are to prepare resveratrol loaded mixed micelles composed of poloxamer 403 and poloxamer 407, and optimize the formulation in order to achieve higher drug solubility and sustained drug release. Firstly, a thin-film hydration method was utilized to prepare the micelles. By using drug-loading, encapsulation yield and particle size of the micelles as criteria, influence of three variables, namely poloxamer 407 mass fraction, amount of water and feeding of resveratrol, on the quality of the micelles was optimized with a central composite design method. Steady fluorescence measurement was carried out to evaluate the critical micelle concentration of the carriers. Micelle stability upon dilution with simulated gastric fluid and simulated intestinal fluid was investigated. The in vitro release of resveratrol from the mixed micelles was monitored by dialysis method. It was observed that the particle size of the optimized micelle formulation was 24 nm, with drug-loading 11.78%, and encapsulation yield 82.51%. The mixed micelles increased the solubility of resveratrol for about 197 times. Moreover, the mixed micelles had a low critical micelle concentration of 0.05 mg · mL(-1) in water and no apparent changes in particle size and drug content were observed upon micelles dilution, indicating improved kinetic stability. Resveratrol was released from the micelles in a controlled manner for over 20 h, and the release process can be well described by Higuchi equation. Therefore, resveratrol-loaded poloxamer 403/407 mixed micelles could improve the solubility of resveratrol significantly and sustained drug release behavior can be achieved.
Drug Carriers ; chemistry ; Fluorescence ; Kinetics ; Micelles ; Particle Size ; Poloxamer ; chemistry ; Solubility ; Stilbenes ; chemistry ; Water

Aged ; Humans ; Laryngectomy ; instrumentation ; Larynx, Artificial ; Male ; Middle Aged

Objective To analyze if there are membrane estrogen receptors(ER)in endometrial carcinoma and if there is some relationship between membrane ER and nuclear ER.Methods The cell membrane and total cell ER?and ER?expressions of high and moderate differentiation endometrial carcinoma cells(Ishikawa and HEC-1A cells)were analyzed.Intermittent immunofluorescence dyeing and fluorescent microscopy were carried out with the ceils treated with polyformaldehyde and Triton X-100. Intermittent immunofluorescence dyeing and flow cytometry were carried out with the live cells and the cells treated with Triton X-100 respectively.Results There were fluorescences on the membrane of the Ishikawa and HEC-1A cells which were treated with polyformaldehyde.When the cells were treated with Triton X- 100,the fluorescences were also seen inside the cells.The fluorescence intensity of ER?and ER?in Ishikawa cell membrane(1.09?0.21,1.27?0.33)was stronger than the control,but there were no significant differences(P＞0.05).When treated with Triton X-100,the total cell fluorescence intensity of ER?and ER?in Ishikawa cell(4.21?0.34,4.69?1.96)was stronger than the membrane(P＜0.05). The ER?and ER?fluorescence intensity of HEC-1A cell membrane(1.58?0.13,1.49?0.04)were stronger than the control(P＜0.05).The fluorescence intensity of ER?and ER?of the HEC-1A cell (2.34?0.33,2.52?0.15)was stronger than the membrane also(P＜0.05).The membrane ER?fluorescence intensity of Ishikawa was lower than HEC-1A(P=0.028).But the total cell ER?fluorescence intensity of Ishikawa was higher than HEC-1A(P=0.002).Conclusions There are membrane ER on endometrial carcinoma cells Ishikawa and HEC-1A.The membrane ER must have some similarity to the nuclear receptor.There is no direct correlation between the quantity of the membrane ER and nuclear ER.

AlM: To describe the expression of hypoxia-inducible factor-1α ( HlF-1α) and erythropoietin ( EPO ) in rats' corneal and evaluate its potential effect on corneal neovascularization ( CNV) growth.
METHODS:The young SD rats (3mo) was chosen and randomly divided into 2 groups, which were experimental group and normal control group. CNV model was established by alkali burn, and the length and area of CNV was observed everyday after operation by slit lamp. After that, the expression of HlF-1α and EPO was measured by SABC and RT-PCR methods at 1, 3, 5, 7, and 14d after alkali burn. The data was analyzed by SPSS 20. 0.
RESULTS:The area of CNV was increasing at 1, 3, 5, 7, and 14d after alkali burn, and the peak point appear at 7d. The growth speed was decreased after 14d. SABC method told us that no HlF-1αand very tiny amount EPO was detected at normal rats' corneal. The expression of the two factors increased at 1d after alkali burn in corneal epithelium and endoderm. The results of RT - PCR showed that a few amounts of HlF-1α and EPO mRNA were detected at normal group. The expression of the two factors was increased at 3d after alkali burn, and the peak value was found at 7d, however, it was decreased at 14d. Statistical difference was found at different time (P<0. 05).
CONCLUSlON: HlF- 1α and EPO is closely related to CNV.

To analyze the relationship among serum heart type fatty acid binding protein (HFABP) , ho‐mocysteine (Hcy) levels and ventricular remodeling , prognosis in patients with chronic heart fail (CHF).Methods :A total of 128 patients with CHF admitted to our hospital from May 2013 to Nov 2015 were enrolled .According to the New York Heart Association (NYHA) cardiac function grading , patients were divided into three groups : grade II (n＝48) , grade III (n＝42) and grade IV (n＝38) , at the sane time 35 patients with heart function grade I as a control group .Cardiac examination was performed by echocardiography to obtain parameters related to left ventric‐ular remodeling .The levels fo serum H‐FABP and Hcy were measured on the next day of admission , and correlation analysis was performed .All patients were treated with targeted cardiac therapy and followed up for 24 months.The cardiac events were used as the end point of the study , indicating poor prognosis .The relationship among serum H‐FABP and Hcy levels and prognosis was analyzed .Results : Left ventricular end diastolic diameter (LVEDd ) [ (40.40 ± 4.41) mm vs .(42. 64 ± 3.22) mm vs.(45.20 ± 3. 41 ) mm vs.(46. 94 ± 4.22) mm] , left ventricular posterior wall thickness (LVPW) [ (8.45 ± 1.15) mm vs.(9. 04 ± 1. 21) mm vs.(10.05 ± 1.35) mm vs.(11. 94 ± 1.27) mm] , and interventricular septum thickness (IVS) [ (7. 01 ± 0.92) mm vs.(8.93 ± 1. 12) mm vs.(10. 09 ± 1.29) mm vs.(11. 93 ± 1. 32) mm] were significantly increased in patients with grade I , II , III , and IV , and left ventricular ejection fraction ( LVEF) [ (52.16 ± 4.02)% vs.(50. 32 ± 3.29)% vs .(48.16 ± 3. 32)% vs.(45.32 ± 2.29)%] decreased significantly ( P＜0. 05 or P＜0. 01) , and serum H‐FABP [ (3. 36 ± 0.71 ) ng／L vs.(4. 13 ± 1.13) ng／L vs .(5. 65 ± 1.42) ng／L vs.(6.49 ± 1. 69) ng／L] and Hcy [ (9. 46 ± 1.55) μmol／L vs.(14. 49 ± 2. 60) μmol／L vs.(17.71 ± 3. 61) μmol／L vs.(20. 26 ± 3. 37 ) μmol／L ] levels increased significantly ( P＜ 0.05 or P＜0.01).Pearson correlation analysis showed that serum H‐FABP and Hcy levels were positively correlated with LVEDd , IVS and LVPW ( r＝0.312～0. 392 , P＜0.05) , and negatively correlated with LVEF ( r＝ －0.453 ,－0.484 , P＜0.05).During 24 months of follow‐up , 56 patients (43. 75%) with CHF had a poor prognosis , com‐pared with the prognosis group , serum H‐FABPP [ (4. 74 ± 1.43) ng／L vs.(6.27 ± 1.28 ) ng／L ] and Hcy level [ (11.35 ± 2.03) μmol／L vs.(18.33 ± 3.46) μmol／L] in the poor prognosis group were significantly increased (P＝0.001 ).Conclusion : There is certain correlation between serum H‐FABP and Hcy levels and ventricular remodeling in patients with CHF , which plays an important role in the assessment of disease and prognosis .

Objective To investigate the effects of cognitive interventions (CIs) in the context of communi- ty based rehabilitation (CBR) on cognitive deficits (CDs) in stroke patients.Methods Ninety-two stoke patients with CDs were randomly divided into a CI group and a control group.All patients were treated with conventional CBR.In addition,the patients in the CI group were also treated with special intervention therapy.The patients in both groups were assessed with the neurological and cognitive status examination (NCSE) for cognitive functioning, the FCA for motor function and the BI for their ability in the activities of daily living.Results The NCSE,FCA and BI scores in the cognitive intervention group after treatment were significantly higher than those before treatment and also significantly higher than those in the control group after treatment.Conclusion CIs can not only improve CDs,but also enhance recovery of motor function and ADL.

OBJECTIVETo analyze the clinical and pathological features of 40 patients with secondary syphilis.

METHODSA total of 40 cases of secondary syphilis confirmed by serology were collected during 1994-2004 and were first diagnosed on presentation with oral lesions.

RESULTSThe white patch in oral mucosa was found in 32 cases with painless or slight pain in most cases. The most common site of the lesion was the tongue. The histological examination on eight cases was initially misdiagnosed as oral candidosis or lichen planus, but confirmed as syphilis after serology revealed nonspecific inflammation with intraepithelial microabscess and dense perivascular infiltration of lymphocytes and plasma cells in connective tissue. The symptoms showed dramatic improvement in 16 cases after benzathine penicillin treatment.

CONCLUSIONSThe oral manifestations of syphilis have specific clinical and pathological feature and attention should be paid to the suspicious oral lesions when patients are first presented in a dental office.

Adult ; Female ; Humans ; Male ; Middle Aged ; Mouth Mucosa ; pathology ; Oral Ulcer ; etiology ; Syphilis ; complications ; diagnosis ; pathology

OBJECTIVETo construct the sodium channel gene SCN5A-delQKP1507-1509 mutation associated with congenital long QT syndrome, and its eukaryotic expression vector, and to examine the expression of mutation protein in human embryonic kidney (HEK) 293 cells.

METHODSEukaryotic expression vector PEGFP-delQKP-hH1 for SCN5A-delQKP1507-1509 mutation was constructed by rapid site-directed mutagenesis. HEK293 cells were transfected with the wild or mutant vector using lipofectamine, and then subjected to confocal microscopy. The transfected cells were immunostained to visualize intracellular expression of the mutant molecules.

RESULTSDirect sequence and electrophoresis analysis revealed 9 basic group absences at position 1507-1509. The delQKP1507-1509 mutation eukaryotic expression vector was expressed in HEK293 cells. Immunostaining of transfected cells showed the expression of both wild type and mutant molecules on the plasma membrane and there was no difference in the amount of protein, which suggested that the mutant delQKP1507-1509 did not impair normal protein expression in HEK293 cells.

CONCLUSIONSSuccessful construction of mutant SCN5AdelQKP1507-1509 eukaryotic expression vector and expression of SCN5A protein in HEK293 cells provides a basis for further study on the functional effects of congenital long QT syndrome as a cause of SCN5A mutation.

Blotting, Western ; HEK293 Cells ; Humans ; Long QT Syndrome ; congenital ; genetics ; Mutagenesis, Site-Directed ; NAV1.5 Voltage-Gated Sodium Channel ; analysis ; genetics ; physiology

OBJECTIVETo evaluate the effect of adriamycin (ADM) in enhancing the sonodynamic effect of chlorin e6 against the proliferation of human breast cancer MDA-MB-231 cells in vitro.

METHODSMDA-MB-231 cells were treated with ultrasound/Chlorin e6 alone or in combination with ADM, and the changes in the cell proliferation was determined by MTT assay.

RESULTSUltrasound (1.0 MHz) at the power intensity of 0.5-2.0 W/cm2 inhibited the proliferation of MDA-MB-231 cells in an intensity-dependent manner, and chlorin-e6 (0.05-1.6 mg/ml) and ADM (0.1-0.4 g/ml) alone both inhibited the proliferation of MDA-MB-231 cells dose-dependently. Compared with ultrasound (0.5 W/cm2, 1.0 MHz, 60 s) or chlorin-e6 (0.05-0.2 mg/ml) alone, a combined treatment with ultrasound and chlorin e6 significantly enhanced the inhibitory effect on the proliferation of MDA-MB-231 cells (P<0.05). ADM significantly enhanced the sonodynamic effect of chlorin e6 (0.1 mg/ml) against the cell proliferation of MDA-MB-231 cells (P<0.05), and the effect was schedule-dependent, which was greater when ADM was added after the sonodynamic treatment (P<0.05).

CONCLUSIONADM can enhance the sonodynamic effect of chlorin e6 against the proliferation of MDA-MB-231 cells in vitro.

Breast Neoplasms ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Doxorubicin ; pharmacology ; Female ; Humans ; Porphyrins ; therapeutic use ; Ultrasonic Therapy

OBJECTIVE Angiogenesis therapy has attracted interest as a potential treatment for hepatocellular carcinoma (HCC). In this study, we investigated the anti-proliferative activities and anti-angiogenesis effects of saikosaponins (SS)-b on hepatocellular carcinoma (HCC) and its regulation on VEGF/ERK/HIF-1α signal pathway. METHODS H22 hepatoma-bearing mice model and HepG-2 cells were used to study the anti-tumor and anti-angiogenesis effects of SS-b in vivo and in vitro. Pathological change of tumor tissue was observed by HE staining, the microvascular changes were detected by immunohistochemical method. The effects of SS-b on angiogenesis were examined by using the chick embryo chorioallantoic membrane (CAM) model. The effects of SS- b on proliferation, migration and invasion were investigated by MTT assay, scratch wound healing assay and transwell assay inhuman umbilical vein endothelial cell (HUVEC) and HepG2 cells in vitro. Vascular endothelial growth factor (VEGF), matrix metalloproteinase-2/9(MMP-2/9), hypoxia-inducible factor-1α (HIF-1α) expression and the phosphorylation of extracellular regulated kinase(ERK) were analyzed using RT-PCR and Western-blot. RESULTS SS-b effectively inhibited the tumor growth of H22 mice in vivo. The inhibitory rate of tumor was 49.1%, 50.7%, 66.1% in SS-b 5, 10 and 20 mg·kg-1 group respectively. HE staining results showed that SS-b induced tumor necrosis and nuclear dissolution in H22 mice. Moreover, SS-b also reduced the number of microvessels of tumor tissue in H22 mice significantly and suppressed the angiogenesis of CAM induced by b-FGF. SS-b had an obvious inhibitory effect on cell proliferation, migration and invasion of HUVEC cells and HepG-2 cells. These effects were associated with down-regulation of the expression of MMP2/9 and suppression of VEGF/ERK/HIF-1α signaling in H22 mice and Hep-G2 cells. CONCLUSION Our findings showed that SS-b exerts anti-tumor effects by inhibit?ing tumor angiogenesis via regulating VEGF/ERK/HIF-1α signal pathway in vivo and in vitro.