This study investigated a nano drug delivery system built by one sort of modified trimethyl chitosan (TMC). The TMC was modified by cRGDyk, ligand of integrin receptor avβ3. Single factor screening was used to optimize the prescription in which the particle sizes of TMC nanoparticle (TMC NPs) and cRGDyk modified TMC nanoparticle (C-TMC NPs) were (240.3 ± 4.2) nm and (259.5 ± 3.3) nm. Electric potential of those two nanoparticles were (33.5 ± 0.8) mV and (25.7 ± 1.6) mV. Encapsulation efficiencies were (76.0 ± 2.2) % and (74.4 ± 2.0) %. Drug loading efficacies were (50.1 ± 2.1) % and (26.1 ± 1.0) %. Then the cellular uptake, uptake mechanism and transport efficacy of TMC NPs and C-TMC NPs were investigated using Caco-2 cell line. The uptake rate and accumulating drug transit dose of C-TMC NPs were 1.98 and 2.84 times higher than TMC NPs, separately. Mechanism investigations revealed that caveolae-mediated endocytosis, clathrin-mediated endocytosis and macropinocytosis were involved in the intercellular uptake of both TMC NPs and C-TMC NPs. What is more, free cRGDyk could remarkably inhibit the uptake of C-TMC NPs.
Biological Transport ; Caco-2 Cells ; Caveolae ; Chitosan ; chemistry ; Clathrin ; Endocytosis ; Humans ; Integrin alphaVbeta3 ; chemistry ; Nanoparticles ; Particle Size ; Pinocytosis

ObjectiveTo study the effect of sub-chronic exposure to deltamethrin(DM) on neural behavior and expression of NMDA receptor in hippocampus of mice.Methods 60 Female SPF Kunming mice were divided into 4 groups and given DM 60 days by gavage.Hot-plate,rotarod,grip strength,hing limb landing foot splay were used to examine the sensory and motor change of mice.Autonomic activity test was used for detecting the functional status of the central nervous system in mice.Passive avoidance test for detection of the behavior changes of learning and memory,and RQ-PCR was employed to measure the expression of NMDA receptor in hippocampus of mice.ResultsThe behavior of sensory and motor of mice sub-chronic exposure to deltamethrin did not have changes significantly(P ＞ 0.05 ).In the test of autonomic activity test,the average of autonomic activity times were (93 ± 18) times,(107 ± 13) times,(105 ±22) times.Compared with the control group,the average of autonomic activity times in middle-and high-dose groups were increased significantly (P ＜ 0.05 ).The latent periods in poisoning groups were (175.4 ±38.4) s,(146.4 ±51.2)s,(132.3 ±45.0) s,and the error times were (0.7 ±0.3)times,( 1.4 ± 0.5 ) times,( 1.8 ± 0.9) times.Compared with the control group,latent periods of high-dose group were decreased and the error times of middle-and high-dose groups were increased significantly (P ＜ 0.05 ).Compared with the control group,the relative expression levels of NR1 and NR2A mRNA in hippocampus of middle and high-dose groups were increased significantly (P＜ 0.05 ),and the relative expression level of NR2B mRNA in highdose group was decreased significantly(P ＜ 0.05).ConclusionSub-chronic exposure to DM could increase the excitability of mice,damage the function of learning and memory,and influence the expression of NMDA receptor in hippocampus of mice.

Objective To observe the effects and security of rapamycin drug-eluting stents implanting in eld-eay patients with coronary heart disease(CHD).Methods 107 patients with CHD treated with Firebird rapamycin drug-eluting stents,the immediate angiographic outcome,complication in hospitalization.six months'follow-up results were assessed.Results 175 Firebird stents implanted successfully,the successful stenting procedure achieved in 98.9% patients,1 patient(0.9%) died of thrombosis instent during hospitalization.there were no myocardial infarction occurred during six months'follow-up,angina recrudesced in 6 patients,and the ratio of instent restenosis was 2.6%,the MACE rate during 6 months follow-up was 2.8%.Conclusion Firebird rapamycin drug-eluting stents implanta-tion in CHD in elderly was safe and effective.

Objective To evaluate the accuracy of core needle biopsy (CNB) in diagnosing breast masses and its coherence with immunohistochemical (IHC) examination results of estrogen receptor (ER), progesterone receptor (PR) and Her2 protein between pre-and post-chemotherapy in invasive breast cancer. Methods The results of 516 CNB cases from June, 2005 to April, 2008 were analyzed retrospectively. The pathological examination was performed by two pathologists independently. Results 484 cases of malignant tumor, carcinoma in situ and phyllodes tumor were found in this group with the sensitivity of 96.7%. Sixteen cases of false negative (3. 3% ) were demonstrated by surgical biopsy. The accurate rate of CNB was not influenced by the maximum diameter of masses ( P = O. 423 ). The agreement rate of IHC results of ER, PgR and Her2 between pre- and post-chemotherapy were 90. 3%, 76. 8% and 82.5%, respectively. Conclusion CNB is a useful diagnostic method with a satisfactory accuracy in any size of breast masses. Given the histological heterogeneity of invasive breast cancer and the influence of ehemotherapy, the coherence of prechemotherapy IHC for ER, PgR and Her2 is not optimal with that of post-chemotherapy.

Objective To investigate the effect of high mobility group box-1 protein (HMGB1) on cytokine expression in splenic dendritic cell (DCs). Method DCs isolated from the spleens of male Wistar rats were seed-ed on 96-well (1×10s cells/well) cell culture plates, and the cells were stimulated with HMGB1 for various length of time or in different concentrations. (1) The time-dependent response between HMGB1 and tumor necrosis factor-α(TNF-α) as well as interleukin-12 (IL-12) gene/protein expressions: 24 wells of DCs were dividedly into six groups including 24 h-normal controls (n=4), 48 h-normal controls (n=4), 72 h-normal controls (n=4), and 24 h-HMGB1 treated group (n=4), 48 h-HMGB1 treated group (n=4) as well as 72 h-HMGB1 treated group (n=4), respectively. Among three HMGB1-treated groups, DCs were stiraulated by 1 μg/mL HMGB1. DCs were denatured in cell culture plates to determine gene expression of IL-12 as well as TNF-α, and supernatants were harvested to determine Il-12 as well as TNF-α protein levels. (2) The dose-dependent response between HMGB1 and TNF-α as well as IL-12 gene/protein expressions: 16 wells of DCs were dividedly into four groups in-cludingnormal controls (n=4), 0.1 μg/mL HMGB1 treated group (n=4), 1 μg/mL HMGB1 treated group (n =4), and 10 μg/mL HMGB1 treated group (n=4), respectively. After stimulated for 48 h, DCs were dena-tured in cell culture plates to determine gene expression of IL-12 as well as TNF-α, and supernatants were harvest-ed to determine IL-12 as well as TNF-α protein levels. Total RNA was extracted from cells using the single-step technique of acid guanidinium thiocyanate-chloroform extraction according to the manufacturer' s instruction (Promega, Madison, WI). mRNA for TNF-α and IL-12 were quantified by SYBR Green two-step, real-time re-verse transeription-polymerase chain reaction taking glyceraldehyde-3-phesphate dehydrogenase (GAPDH) as an internal standard. Levels of IL-12 and TNF-α in cell culture supernatants were determined with ELISA, strictly fol-lowing the protocols provided by the manufacturer. Data were analyzed with a one-way ANOVA. A P-values <0.05 were considered statistically significant. Results After stimulation with 1 μg/mL HMGB1, IL-12 and TNF-α protein and gene expressions in rat splenic DCs were markedly up-regulated at 24 h to 72 h (P<0.05 or P<0.01), and the expression levels of IL-12 and TNF-α peaked at 48 h (P<0.01). When DCs were cultured in the presence of 0.1 μg/mL, 1 μg/mL, and 10 μg/ml HMGBI for 48 h, expressions of IL-12 and TNF-α were also significantly up-regulated (P<0.01), and values of these cytokines were highest in 1 μg/mL HMGB1-treated group (P<0.01). Conclusions These data suggest that HMGB1 appears to be a potential immunostimulatory signal that induced DC maturation, and HMGB1 stimulation can result in marked up-regulation of IL-12 as well as TNF-α synthesis and release in splenic DCs.

Objective To examine the effectiveness of self-efficacy training on fluid intake in maintenance hemodialysis patients.Methods Fifty-four hemodialysis patients participated in the study.The intervention was based on Bandura's self-efficacy theory.The outcome measure was self-efficacy for managing chronic disease,the mean interdialysis weight gain,dialysis-related complications and patient satisfaction.Data were collected at baseline and 3 months after the intervention.Results The mean score of selfefficacy,the mean interdialysis weight gain,dialysis-related complications and patient satisfaction after the intervention were all statistically significant compared with those bcfore the intervention.Conclusions The study supports the effectiveness of the self-efficacy training in improving the fluid intake compliance in maintenance hemodialysis patients.So it is worth being popularized in clinic.

Objective To establish embryonic stem cells (ESC) that can express green fluorescent protein (CFP) and differentiate them into neurons. It would provide tagging neurons for clinical transplantation to cure neural system diseases. Methods ESC (R1) was transfeeted with a plasmid containing the GFP by electroporation. A transgeuic cell line was obtained after selection with G418. The ESCs were characterized by AKP staining. Monolayer differentiation method was used to induce neural differentiation derived from GFP-ESC and immunofluorescence method was used to identify Tuj1 positive cells. Results There was no significant difference(X2=3.14,P0.05) in transfect rates between liposome and electroporation (65% vs 79%). The AKP staining of GFP-ESC was positive. GFP-ESC could be differentiated into neural cells. Conclusions These results show that ESC expressing GFP has been estabhshed, which can be differemiated into neurons.

AIM To study the chemical constituents from Nepeta angustifolia C.Y.Wu.METHODS The ethyl acetate fraction of N.angustifolia 70％ ethanol extract was isolated and purified by silica,Sephadex LH-20,ODS and semi-preparative HPLC column,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Ten compounds were isolated and identified as oleanolic acid (1),betulinic acid (2),ursolic acid (3),3-sitosterol (4),angelicin (5),bakuchiol (6),blumenol C glucoside (7),psoralen (8),methyl rosmarinate (9),hesperidin (10).CONCLUSION Compounds 5-8are isolated from genus Nepeta for the first time,compounds 1,4-10 are first isolated from this plant.

Minimal access spinal surgery(MASS),a new minimally invasive technique,has been developed on the basis of classic minimally invasive spinal surgery(MISS).With some special equipments,this procedure can reduce approach-related morbidity and collateral damage to surrounding tissues,and can be performed under direct visualization of the spine.The MASS combines the merits of MISS and traditional open procedures,and comparable results can be obtained while decreasing approach-related complications and learning curve.Now,the application of MASS has become more frequent in clinical practice due to its favourable characteristics in curative effect,postoperative recovery and economic efficacy.

Extracellular single-unit discharges were obtained from 106 spontaneously active neurons in the region of the rostral ventrolateral medulla (RVLM) by glass microelectrode from 73 brain slices of Sprague-Dawley rats. Exogenous γ-aminobutyric acid (GABA, 0.1 ～ 3.0 mmol/L) inhibited the electrical activity of 84 out of 106 RVLM neurons dose-dependently. The inhibitory effect of GABA could be blocked by GABAA with BMI or PTX alone, the firing rates of most of the RVLM neurons were significantly increased. In 41 neurons responding to GABA, baclofen (0.1 ～ 3.0 μmol/L), a GABAB receptor selective agonist, inhibited the discharges of 33 of the neurons dose-dependently. GABAB receptor antagonist CGP35348 ( n = 13) blocked the inhibition due to baclofen ( n = 21). After perfused with CGP35348 alone, the firing rates of most of the RVLM neurons were significantly increased. Taken together, the inhibition of GABA on RVLM neurons is mediated through either GABAA or GABAB receptors and some intrinsic GABA neurons exert tonic inhibition activity.