Targeted delivery of edaravone by liposomes for the treatment of ischemic stroke.

Aim: To construct an edaravone-encapsulated liposomes (EDV-LIPs) formulation against acute ischemic stroke. Methods: EDV-LIPs were prepared by the film dispersion method. The biosafety was evaluated both in vitro and in vivo by flow cytometry and the histological staining method. Biodistribution and therapeutic effect of EDV-LIPs against acute ischemic stroke was investigated by fluorescent imaging, the behavior test, laser speckle imaging and triphenyltetrazolium chloride staining. Results: The nanoliposomes had a long circulation time and could accumulate in the brain lesion region in ischemic stroke rats. EDV-LIPs show good biosafety. EDV-LIPs could restore more cerebral blood flow, reduce infarct volume and decrease neuronal apoptosis. Conclusion: EDV-LIPs provide an effective alternative for drug-targeted delivery against acute ischemic stroke.

Folic acid modified Fe3O4 nanoclusters by a one-step ultrasonic technique for drug delivery and MR imaging.

Multifunctional nanoclusters based on Fe3O4 nanoparticles for magnetic resonance imaging (MRI) and drug delivery are reported here. At first, oleic acid (OA)-coated Fe3O4 nanoparticles were prepared. Then block copolymer Pluronic F127 or folic acid (FA) conjugated-Pluronic F127 was used to modify the hydrophobic nanoparticles to become hydrophilic Fe3O4@F127 nanoclusters via facile ultrasonic treatment. During this process, drug molecules can also be introduced into the nanoclusters and therefore the targeted drug delivery system was formed. Next, we verified the feasibility of the nanoclusters as drug delivery vehicles and magnetic contrast agents. The nanoclusters have an average size of 200 nm and remained stable in water for long periods. Folic acid-modified nanoclusters showed an enhanced intracellular uptake into HepG2 cells by using both cellular iron amount analysis and flow cytometry analysis. Besides, Fe3O4@F127@FA nanoclusters showed good compatibility in the tested concentration range and good sensitivity in T 2-weighted MRI. The magnetic nanoclusters combined with drug delivery properties have greatly increased the significance in the diagnosis and therapy of diseases, which are suitable for systematical administration of hydrophobic drugs and simultaneously MRI diagnosis.

Facile fabrication of water-dispersible nanocomposites based on hexa-peri-hexabenzocoronene and Fe3O4 for dual mode imaging (fluorescent/MR) and drug delivery.

The facile fabrication of multifunctional nanocomposites (Fe3O4/HBC@F127) consisting of superparamagnetic Fe3O4 nanoparticles and fluorescent organic hexa-peri-hexabenzocoronene (HBC) molecules incorporated in block copolymer diacylphospholipid-polyethyleneglycol F127 have been demonstrated for dual mode imaging (fluorescent/MR) and drug delivery. The obtained nanocomposites were water-dispersible, stable and biocompatible, as confirmed by dynamic light scattering (DLS) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Relativity measurements showed a T 2 relaxivity (r 2) of 214.61 mM-1 s-1, which may be used as T 2-weighted MR imaging agents. In vitro imaging studies indicated that the nanocomposites had good MR and fluorescence imaging effects with low cytotoxicity. Besides, the developed nanocomposites could also be applied as drug delivery vehicles. Doxorubicin (DOX) loaded Fe3O4/HBC@F127 nanocomposites significantly inhibited the growth of human hepatoma cells (HepG2). These findings suggested that the facile synthesized multifunctional nanocomposites may be used as a platform for dual mode imaging (both MR and fluorescence) and drug delivery.

Descending projections from the nucleus accumbens shell excite activity of taste-responsive neurons in the nucleus of the solitary tract in the hamster.

The nucleus of the solitary tract (NST) and the parabrachial nuclei (PbN) are the first and second relays in the rodent central taste pathway. A series of electrophysiological experiments revealed that spontaneous and taste-evoked activities of brain stem gustatory neurons are altered by descending input from multiple forebrain nuclei in the central taste pathway. The nucleus accumbens shell (NAcSh) is a key neural substrate of reward circuitry, but it has not been verified as a classical gustatory nucleus. A recent in vivo electrophysiological study demonstrated that the NAcSh modulates the spontaneous and gustatory activities of hamster pontine taste neurons. In the present study, we investigated whether activation of the NAcSh modulates gustatory responses of the NST neurons. Extracellular single-unit activity was recorded from medullary neurons in urethane-anesthetized hamsters. After taste response was confirmed by delivery of sucrose, NaCl, citric acid, and quinine hydrochloride to the anterior tongue, the NAcSh was stimulated bilaterally with concentric bipolar stimulating electrodes. Stimulation of the ipsilateral and contralateral NAcSh induced firings from 54 and 37 of 90 medullary taste neurons, respectively. Thirty cells were affected bilaterally. No inhibitory responses or antidromic invasion was observed after NAcSh activation. In the subset of taste cells tested, high-frequency electrical stimulation of the NAcSh during taste delivery enhanced taste-evoked neuronal firing. These results demonstrate that two-thirds of the medullary gustatory neurons are under excitatory descending influence from the NAcSh, which is a strong indication of communication between the gustatory pathway and the mesolimbic reward pathway.

Serum soluble ST2 is associated with ER-positive breast cancer.

BACKGROUND: ST2, a member of the interleukin (IL)-1receptor family, regulates Th1/Th2 immune responses in autoimmune and inflammatory conditions. However, the role of ST2 signaling in tumor growth and metastasis of breast cancers has not been investigated. This study investigated the possible role of soluble ST2 (sST2) in breast cancer. METHODS: The serum levels of IL-33, sST2, and vascular endothelial growth factor (VEGF) in 150 breast cancer patients and 90 healthy women were measured by enzyme-linked immunosorbent assay. Estrogen receptor(ER), progesterone receptor, human epithelial receptor (HER)-2, and cell cycle regulated protein Ki-67 were measured. Clinical stage, tumor size, lymph node metastasis, and histological type were also recorded. RESULTS: The serum levels of sST2, IL-33, and VEGF were significantly higher in breast cancer patients than in the control group (P < 0.05, each). Serum sST2 levels in ER-positive breast cancer patients were significantly associated with age, histological type, clinical stage, tumor size, and Ki-67 status (P < 0.05, each). Moreover, the serum levels of IL-33 and sST2 in breast cancers significantly correlated with VEGF levels (IL-33: r = 0.375, P < 0.0001; sST2: r = 0.164, P = 0.045). Serum levels of sST2, IL-33, and VEGF decreased after modified radical mastectomy in ER-positive breast cancers. Serum levels of IL-33, sST2, and VEGF and clinicopathological factors were not significantly correlated with disease-free survival and overall survival of ER-positive breast cancer women during follow-up. CONCLUSION: Serum sST2 levels in ER-positive breast cancer patients are significantly associated with factors that indicate poor prognosis.

Descending projections from the nucleus accumbens shell suppress activity of taste-responsive neurons in the hamster parabrachial nuclei.

The parabrachial nuclei (PbN), the second central relay for the gustatory pathway, transfers taste information to various forebrain gustatory nuclei and to the gustatory cortex. The nucleus accumbens is one of the critical neural substrates of the reward system, and the nucleus accumbens shell region (NAcSh) is associated with feeding behavior. Taste-evoked neuronal responses of PbN neurons are modulated by descending projections from the gustatory nuclei in the forebrain. In the present study, we investigated whether taste-responsive neurons in the PbN project to the NAcSh and whether pontine gustatory neurons are subject to modulatory influence from the NAcSh in urethane-anesthetized hamsters. Extracellular single-unit activity was recorded in the PbN, and taste responses were confirmed by the delivery of 32 mM sucrose, NaCl, quinine hydrochloride, and 3.2 mM citric acid to the anterior tongue. The NAcSh was then stimulated (0.5 ms, ≤100 µA) bilaterally using concentric bipolar stimulating electrodes. A total of 98 taste neurons were recorded from the PbN. Eighteen neurons were antidromically invaded from the NAcSh, mostly the ipsilateral NAcSh (n = 16). Stimulation of the ipsilateral and contralateral NAcSh suppressed the neuronal activity of 88 and 55 neurons, respectively; 52 cells were affected bilaterally. In a subset of pontine neurons tested, electrical stimulation of the NAcSh during taste stimulation also suppressed taste-evoked neuronal firing. These results demonstrated that taste-responsive neurons in the PbN not only project to the NAcSh but also are under substantial descending inhibitory influence from the bilateral NAcSh.

[Correlation between the expression of cytokeratin-18-mRNA and invasion and metastasis of oral squamous cell carcinoma].

OBJECTIVE: To investigate the relationship between the expression of cytokeratin (CK)-18 and biological behavior of oral squamous cell carcinoma (OSCC). METHODS: Twenty-three patients with OSCC were investigated for the expression of CK-18-mRNA by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Correlations between clinical stages, pathological differentiation, lymphatic metastasis and the expression of CK-18-mRNA were evaluated. CK-18-mRNA expression of peripheral blood from the 23 patients and 23 healthy people were also examined. During follow-up after operation, the peripheral blood was collected again for the expression of CK-19-mRNA. RESULTS: Expression of CK-18-mRNA was found in 16 patients. The expression of CK-18-mRNA was significantly associated with clinical stages, tumor differentiation and lymphatic metastasis. CK-18-mRNA was positive in 4 of 23 blood specimens before operation, but during follow-up only 1 of 23 patients was still positive in peripheral blood. CONCLUSIONS: CK-18 may provide additional information in forecasting the metastasis of OSCC and serve as a reference in monitoring recurrence.

[Immunofluorescence examination of CK-13 expression in cell line KB differentiated by all-trans retinoic acid or As2 O3].

OBJECTIVE: To examine the expression of cytokeratin-13 (CK-13) in oral squamous cell carcinoma (OSCC) and to discuss the effects of all-trans retinoic acid (ATRA) or arsenic trioxide (As2 O3) on the differentiation of human oral undifferentiated squamous cell carcinoma cell line KB cells. METHODS: The cultured KB cells were divided into three groups, ATRA group, As2 O3 group, and control. The expression of CK-13 in KB cells was detected using the immunofluorescence before and after KB cells were induced by ATRA or As2 O3. RESULTS: The expression rates of CK-13 in KB cells in the ATRA group and As2 O3 group were significantly higher than that in the control (P < 0.05), but there was no significant difference in the expression between ATRA and As2 O3 group(P > 0.05). CONCLUSIONS: ATRA and As2 O3 both have the ability to differentiate the KB cells, and the expression is associated with the degree of tumor differentiation. CK-13 may serve as a molecular marker to evaluate the effect of the differentiation treatment on OSCC.

[Cytokeratin 18 and their gene expression in jaw odontogenic keratocyst epithelial lining].

OBJECTIVE: To examine cytokeratin 18(CK18) and it's gene in jaw odontogenic keratocyst (OKC) epithelial lining. METHODS: The epithelial linings of 32 cases were subject to monoclonal antibody immunohistochemical staining for CK18, CK8 and CK19. RT-PCR and in situ hybridization for CK18 mRNA were conducted in 12 of 32 cases in keratocyst epithelial cell linings. RESULTS: In 17 cases, CK18 were observed in keratinized surface layers, though weakly positive. In 27 cases, CK18 were positive in the granular cell layers. CK18 were also positive in the spinous cell layers in 14 cases. In all cases, CK18 was negative in basal cell layers. By RT-PCR, 4 cases expressed CK18 strongly, 8 cases weakly. By in situ hybridization, 8 cases expressed CK18 mRNA positively in both spinous and granular cell layers, and 4 cases positively in basal and keratinized cell layers. CK8 were expressed in basal cell layers of keratocyst epithelial linings. In 23 cases, CK19 were expressed in surface cell layers of keratocyst epithelial linings. CONCLUSION: The expression of CK18 in keratocyst epithelial linings transfers from basal cell layer to spinous layer. The expression of CK18 immunohistochemical staining and CK18 mRNA in situ hybridization are different, which shows CK18 might be related to proliferation of OKC epithelial linings. That suggests the existence of regulation of CK18 and CK18 mRNA expression.

[Glandular odontogenic cyst: report of two cases with cytokeratin 18 expression].

OBJECTIVE: To report two cases of glandular odontogenic cyst and examine its cytokeratin 18,19 expression. METHODS: Two cases of glandular odontogenic cyst were reported and studied. The cytokeratin 18, 19 expression in these two cases were also investigated using immuno-histochemical staining as well as in the situ hybridization of the cyst epithelium. RESULTS: Histo-pathological examination revealed that ciliated columnar cells, squamous cells and low-columnar cells were found in the superficial layer of the lining epithelium. Several minor salivary glands, mainly composed of seromucous cells were observed near the satellite cyst. CK18 were expressed in all layers of the lining epithelium of varying intensity. CK18 was negative in lining epithelium of the daughter cyst, but CK19 was positive. CK18-mRNA was expressed in all the layers of the lining epithelium, the salivary glands and daughter cysts. CONCLUSIONS: Histological features and CK18 expression may be indicative of the possibility of salivary glandular and odontogenic differentiation.

[Cytokeratin18, 13 and their gene expression in post-operative maxillary cyst linings with metaplastic epithelium].

OBJECTIVE: To study the cytokeratin 18 and 13 and their gene (CK) expression in post-operation maxillary cyst linings with metaplastic epithelium. METHODS: CK expressions were examined with immunohistochemistry in 46 post-operative maxillary cyst (POMC) which were lined with pseudostriated columnar cells only (13 cases), both kinds of columnar and squamous cells (30 cases) and squamous cells only (3 cases). RESULTS: The expressions of CK8, CK13 and CK18 were observed in 39, 9 and all of the 43 columnar epithelial linings, respectively. Metaplastic squamous epithelia expressed more CK13 and less CK18 and CK8. Of the 33 metaplastic linings, 24 expressed CK8, 23 CK13 and 26 linings expressed CK18. The expression of CK13- and CK18-mRNA was generally correlated with the protein expression level. By in situ hybridization, CK18-mRNA expression was observed not only in 26 metaplastic linings which were positive for CK18 protein but also in five of the seven metaplastic linings which did not express CK18 protein. In addition, RT-PCR revealed an expression of CK18-mRNA in all metaplastic squamous linings although the expression level was weaker than that in the columnar epithelial linings. The CK13-mRNA was expressed in a fashion inverse to the CK18-mRNA. CONCLUSIONS: These results indicate that CK18-mRNA is preserved through metaplasia although the protein expression decreases and metaplastic squamous cells differentiate with a decrease of CK18 and an increase of CK13 expression.

Cytokeratin expression patterns in jaw cyst linings with metaplastic epithelium.

BACKGROUND: Cytokeratin (CK) expression patterns have been studied in numerous intact and diseased oral tissues. However, CK expression in metaplastic squamous cells has not been explored in depth and the origin of metaplastic epithelial linings of the jaw cysts has not been sufficiently investigated. METHODS: We examined CK expression in 46 postoperative maxillary cysts (POMCs) which were lined with pseudostratified columnar cells only, columnar and squamous cells, and squamous cells only, in 13, 30 and 3 cases, respectively. RESULTS: The expression of CK8, CK13 and CK18 were observed in 39, 9 and all 43 of the columnar epithelial linings, respectively. Metaplastic squamous epithelia expressed more CK13, and less CK18 and CK8. Of the 33 metaplastic linings, 24 expressed CK8, 23 CK13 and 26 linings expressed CK18. The patterns of expression of CK13 and CK18 observed were CK18(+)-CK13(-) in 10 metaplastic linings, CK18(+)-CK13(+) in 16, and CK18(-)-CK13(+) in 7. The expression of CK13- and CK18-mRNA was generally correlated with level of protein expressed. CK18-mRNA expression was observed by in situ hybridization, not only in the 26 metaplastic linings which were positive for CK18 protein, but also in five of the seven metaplastic linings which did not express CK18 protein. In addition, RT-PCR revealed an expression of CK18-mRNA in all metaplastic squamous linings, although the expression level was weaker than that in the columnar epithelial linings. The CK13-mRNA was expressed inversely to the CK18-mRNA. CONCLUSIONS: These results indicate that CK18-mRNA is preserved through metaplasia, although the protein expression decreased. Metaplastic squamous cells differentiate with a decrease of CK18 and an increase of CK13 expression.