Biomass-based controllable morphology of carbon microspheres with multi-layer hollow structure for superior performance in supercapacitors.

The electrochemical properties of corn starch (CS)-based hydrothermal carbon microsphere (CMS) electrode materials for supercapacitor are closely related to their structures. Herein, cetyltrimethyl ammonium bromide (CTAB) was used as a soft template to form the corn starch (CS)-based carbon microspheres with radial hollow structure in the inner and middle layers by hydrothermal and sol-gel method. Due to the introduction of multi-layer hollow structure of carbon microsphere, more micropores were produced during CO2 activation, which increased the specific surface area and improved the capacitance performance. Compared to commercial activated carbon, the four different morphologies of corn starch CMS had better electrochemical performances. Consequently, the proposed CO2-(CTAB)-CS-CS exhibits a high discharge specific capacitance of 242.5F/g at 1 A/g in three-electrode system with 6 M KOH electrolyte, better than commercial activated carbon with 208.5F/g. Moreover, excellent stability is achieved for CO2-(CTAB)-CS-CS with approximately 97.14 % retention of the initial specific capacitance value after 10,000 cycles at a current density of 2 A/g, while the commercial activated carbon has 86.96 % retention. This implies that the corn starch-based multilayer hollow CMS could be a promising electrode material for high-performance supercapacitors.

Changes of serum metabolites levels during neoadjuvant chemoradiation and prediction of the pathological response in locally advanced rectal cancer.

INTRODUCTION: Previous studies have explored prediction value of serum metabolites in neoadjuvant chemoradiation therapy (NCRT) response for rectal cancer. To date, limited literature is available for serum metabolome changes dynamically through NCRT. OBJECTIVES: This study aimed to explore temporal change pattern of serum metabolites during NCRT, and potential metabolic biomarkers to predict the pathological response to NCRT in locally advanced rectal cancer (LARC) patients. METHODS: Based on dynamic UHPLC-QTOF-MS untargeted metabolomics design, this study included 106 LARC patients treated with NCRT. Biological samples of the enrolled patients were collected in five consecutive time-points. Untargeted metabolomics was used to profile serum metabolic signatures from LARC patients. Then, we used fuzzy C-means clustering (FCM) to explore temporal change patterns in metabolites cluster and identify monotonously changing metabolites during NCRT. Repeated measure analysis of variance (RM-ANOVA) and multilevel partial least-squares discriminant analysis (ML-PLS-DA) were performed to select metabolic biomarkers. Finally, a panel of dynamic differential metabolites was used to build logistic regression prediction models. RESULTS: Metabolite profiles showed a clearly tendency of separation between different follow-up panels. We identified two clusters of 155 serum metabolites with monotonously changing patterns during NCRT (74 decreased metabolites and 81 increased metabolites). Using RM-ANOVA and ML-PLS-DA, 8 metabolites (L-Norleucine, Betaine, Hypoxanthine, Acetylcholine, 1-Hexadecanoyl-sn-glycero-3-phosphocholine, Glycerophosphocholine, Alpha-ketoisovaleric acid, N-Acetyl-L-alanine) were further identified as dynamic differential biomarkers for predicting NCRT sensitivity. The area under the ROC curve (AUC) of prediction model combined with the baseline measurement was 0.54 (95%CI = 0.43 ~ 0.65). By incorporating the variability indexes of 8 dynamic differential metabolites, the prediction model showed better discrimination performance than baseline measurement, with AUC = 0.67 (95%CI 0.57 ~ 0.77), 0.64 (0.53 ~ 0.75), 0.60 (0.50 ~ 0.71), and 0.56 (0.45 ~ 0.67) for the variability index of difference, linear slope, ratio, and standard deviation, respectively. CONCLUSION: This study identified eight metabolites as dynamic differential biomarkers to discriminate NCRT-sensitive and resistant patients. The changes of metabolite level during NCRT show better performance in predicting NCRT sensitivity. These findings highlight the clinical significance of metabolites variabilities in metabolomics analysis.

High-Voltage Redox Mediator of an Organic Electrolyte for Supercapacitors by Lewis Base Electrocatalysis.

Redox electrolytes for supercapacitors (SCs) have recently sparked widespread interest. Due to the redox reactions within electrolytes, they can achieve high capacitance and long cycle stability. However, the energy density of SCs with redox electrolytes is limited by the narrow applied electrochemical window due to the irreversible side reaction of redox mediators at high potential. To overcome this issue, a redox mediator with a high redox potential, tetrachloridehydroquinone (TCHQ), is added to organic electrolytes to obtain a broad electrochemical window. TCHQ is designed to undergo a dehydrogenation reaction catalyzed by N-doped activated carbon to provide capacitance. The pyrrole N atoms have the highest electrocatalytic activity based on the theoretical calculation of reaction overpotential with predicted reaction pathways due to their Lewis basicity. Benefitting from that, TCHQ shows promising reversibility with a larger electrochemical window (up to 2.7 V). As a result, a higher energy density is obtained when compared to commercial SCs. This study proposes a strategy for designing redox mediators and interfaces of SCs with high energy density and a calculation method of dehydrogenation reaction electrocatalysis.

Bruton's tyrosine kinase knockout in macrophages attenuates diabetic kidney disease in the streptozotocin-induced mice / 中华肾脏病杂志

Objective@#To investigate whether Bruton's tyrosine kinase knockout (Btk-/-) in macrophages attenuates diabetic kidney disease in the streptozotocin (STZ)-induced mice.@*Methods@#Macrophages-specific Btk-/- mice and control mice (C57BL/6N) were randomly divided into WT group, diabetic group, Btk-/- group and Btk-/- diabetic group. The diabetic models were induced by STZ (50 mg/kg). After 12 weeks, relevant biochemical parameters and the histological changes of kidneys were detected. The expression of macrophages marker CD68 were detected by immunofluorescence, and the immunohistochemistry was employed to detect the expression of WT1 and Nephrin on renal podocytes. In addition, the expression of fibronectin (FN), collagen type IV (IV-Col), transforming growth factor-β1 (TGF-β1), iNOS, phospho (p)-Btk, interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), MAPK and NF-κB signaling pathway were detected by Western blotting. RT-PCR was used to detect the mRNA of IL-1β, TNF-α and monocyte chemotactic protein-1 (MCP-1).@*Results@#Compared with diabetic group, the mice in Btk-/- diabetic group had reduced albuminuria and attenuated kidney histopathology significantly, significantly increased WT1 and Nephrin, significantly decreased expression of CD68, FN, IV-Col and TGF-β1, and these changes were correlated with decreased of renal inflammatory cytokines such as IL-1β, TNF-α, MCP-1 and down-regulating MAPK and NF-κB signaling pathway (all P<0.05).@*Conclusion@#Macrophages-specific Btk-/- may protect the kidney of diabetic mice by reducing the expression of renal inflammatory cytokines in MAPK and NF-κB signaling pathway.

Bruton's tyrosine kinase knockout in macrophages attenuates diabetic kidney disease in the streptozotocin-induced mice / 中华肾脏病杂志

Objective:To investigate whether Bruton's tyrosine kinase knockout (Btk -/-) in macrophages attenuates diabetic kidney disease in the streptozotocin (STZ)-induced mice. Methods:Macrophages-specific Btk -/- mice and control mice (C57BL/6N) were randomly divided into WT group, diabetic group, Btk -/- group and Btk -/- diabetic group. The diabetic models were induced by STZ (50 mg/kg). After 12 weeks, relevant biochemical parameters and the histological changes of kidneys were detected. The expression of macrophages marker CD68 were detected by immunofluorescence, and the immunohistochemistry was employed to detect the expression of WT1 and Nephrin on renal podocytes. In addition, the expression of fibronectin (FN), collagen type IV (IV-Col), transforming growth factor-β1 (TGF-β1), iNOS, phospho (p)-Btk, interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), MAPK and NF-κB signaling pathway were detected by Western blotting. RT-PCR was used to detect the mRNA of IL-1β, TNF-α and monocyte chemotactic protein-1 (MCP-1). Results:Compared with diabetic group, the mice in Btk -/- diabetic group had reduced albuminuria and attenuated kidney histopathology significantly, significantly increased WT1 and Nephrin, significantly decreased expression of CD68, FN, IV-Col and TGF-β1, and these changes were correlated with decreased of renal inflammatory cytokines such as IL-1β, TNF-α, MCP-1 and down-regulating MAPK and NF-κB signaling pathway (all P<0.05). Conclusion:Macrophages-specific Btk -/- may protect the kidney of diabetic mice by reducing the expression of renal inflammatory cytokines in MAPK and NF-κB signaling pathway.

Melatonin inhibits high glucose-induced cell proliferation and expressions of inflammatory factor via Toll-like receptor 4 signaling pathway in mouse mesangial cells / 中华肾脏病杂志

Objective To investigate effects of melatonin (MT) on high glucose-induced cell proliferation,Toll-like receptor 4 (TLR4) signaling pathway and expressions of inflammatory factor in mouse mesangial cells (SV40).Methods SV40 cells were divided into mannitol control group (30 mmol/L mannitol),normal control group (5 mmol/L glucose),control (5 mmol/L glucose)+ 1000 μmol/LMT group,high glucose group (25 mmogL glucose),high glucose +10,100,1000 μmol/L MT group and high glucose + TLR4 inhibitor (TAK242) group.(1) The cell viability was measured by CCK-8 cytotoxicity kits,and cell proliferation was measured by EdU kits.The expression of TLR4 and the nuclear translocation of nuclear factor-κB (NF-κB p65) were observed by immunofluorescence.(2) Realtime quantitative PCR was used to detect TLR4 mRNA expression.Real-time quantitative PCR and ELISA were used to determine the mRNA and protein secretion levels of the downstream inflammatory factors,such as monocyte chemoattractant-1 (MCP-1),interleukin-1β (IL-1β) and tumor necrosis factor of α (TNF-α);Western blotting was used to detect TLR4 pathway proteins,such as TLR4,myeloid differentiation factor 88 (MyD88),β interferon TIR domain adaptor (Trif),phosphorylated interferon regulatory factor 3 (p-IRF3) and phosphorylated NF-κB inhibitory protein (p-IκB).Results High glucose stimulated mesangial cell proliferation,promoted TLR4 expression and NF-κB p65 transcription activity.Both MT and TAK242 inhibited the above reactions,and the effects of MT was concentration-dependent.Compared with the normal control group,high glucose group had up-regulated expressions of TLR4,MCP-1,IL-1β and TNF-α mRNA (all P ＜ 0.05),but also significantly increased the protein expressions of MyD88,Trif,p-IRF3 and p-IκB (all P ＜ 0.05).Compared with those in the high glucose group,the expression of TLR4 was down-regulated in the high glucose+ 10,100,1000 μmol/L MT group and the high glucose+TAK242 group (all P ＜ 0.05),while the expressions of MyD88,Trif,p-IRF3,p-IκB,MCP-1,IL-1β and TNF-α decreased (all P ＜ 0.05).The effects of MT was concentration-dependent.Conclusions High glucose stimulates the proliferation of SV40,and MT can inhibit the proliferation of mesangial cells and the expressions of inflammatory factors through TLR4 signaling pathway.

Contrast dose reduction with shortened injection durations in coronary CT angiography on 16-cm Wide-detector CT scanner.

OBJECTIVES:: To investigate the use of shortened contrast injection with late triggering in coronary CT angiography (CCTA) for decreasing contrast dose and maintaining image quality. METHODS:: 106 patients for CCTA on a 16-cm wide-detector CT were prospectively enrolled into groups A (n = 50) and B (n = 56) randomly. Patient weight-dependent contrast medium (Iopamiro, 370 mgI ml-1) at dose rate of 25 mgI/kg/s was used with 8 s and the standard 10 s injection time in groups A and B, respectively. CT values of the aortic sinus (AS), right coronary artery, left anterior descending and left circumflex at the proximal, middle and distal segments were measured and compared. Subjective image quality was evaluated and analyzed with Fisher exact test. Contrast dose, injection rate and enhancement duration (between the start of enhancement in AS and scan finish) were also compared. RESULTS:: There was no difference in the injection rate and enhancement duration between the two groups (p > 0.05), while the total contrast dose in group A (36.2 ± 5.7 ml) was significantly lower than in group B (46.4 ± 6.3 ml) (p < 0.001). There was no difference for CT values in all major coronary vessels between the two groups and no difference in subjective image quality scores (all p > 0.05). CONCLUSION:: It is feasible to shorten contrast injection to 8 s in CCTA on wide-detector CT systems to significantly reduce contrast dosage, maintain adequate enhancement and reduce contrast-related artifacts. ADVANCES IN KNOWLEDGE:: (1) Coronary CT angiography (CCTA) scans with shortened contrast medium injection duration and late triggering are feasible with a 16-cm wide-detector CT system (2) Compared with the conventional CCTA with 10 s contrast injection duration, the new contrast injection protocol of using shortened injection duration (to 8 s) and late triggering reduces contrast dose to 36.2 ml, while maintaining adequate enhancement in vessels and reducing contrast-related artifacts.

LncRNA Neat1 is upregulated in P19 cell line processed with all-trans-retinoic acid / 基础医学与临床

Objective To investigate the expression of Neat1 in the differentiation of P19 cells induced by all-trans-retinoic acid(atRA) and to explore the effect of histone modification on its expression. Methods The differentiation of P19 cells was induced with the α-MEM culture media containing 0.5 μmol/L all-trans-retinoic( atRA) acid and the expression Mash1 which is the neural differentiation maker gene was measured. The expression of Neat1 was measured with RT-qPCR. The cells were treated with TSA, NAM or AdOx respectively to investigate the effect of the histone modifier inhibitor on the expression of Neat1. Results The model of differentiation of P19 cells induced by atRA was successfully constructed. Mash1 was significantly upregulated in the process of P19 cells differentiation.Neat1 was significantly upregulated with the induction of P19 cells treated with atRA(P＜0.01). The TSA but not the NAM or AdOx could induce the expression of the Neat1.Conclusions The expression of Neat1 is significantly upregulated in the process of P19 differentiation induced by atRA and the the high level of histone acetylation maintained by TSA potentially induce the expression of Neat1.

The role and mechanism of macrophage activation induced by exosomes from high glucose-treated renal tubular epithelial cells / 中华肾脏病杂志

Objective To investigate the role and mechanism of macrophage activation induced by exosomes from high glucose-treated renal tubular epithelial cells.Methods (1) The supernatant of renal tubular epithelial cells which were cultured in normal glucose control group (5.5 mmol/L D-glucose) or high glucose group (30.0 mmol/L D-glucose) for 48 h were collected and ultracentrifuged to harvest exosomes.Exosomes were identified by transmission electron microscope and Western blotting.(2) Exosomes were labeled with the green lipophilic fluorescent dye PKH67 and cultured with THP-1 macrophage to investigate whether HK2-derived exosomes could be internalized by THP-1 macrophage.Observing the morphology microscopically and detecting the chemotaxis function of THP-1 macrophages in Transwell chamber after co-cultured with exosomes.The expression of inducible nitric oxide synthase (iNOS),tumor necrosis factor-α (TNF-α),Interleukin-1β (IL-1β),monocyte chemoattractant protein-1 (MCP-1) in cells and supernatants were separately detected by quantitative Real-time PCR (qRT-PCR) and enzyme linked immunosorbent assay (ELISA),and the expression of p-c-Jun NH2-terminal kinase (p-JNK),mitogen-activated protein kinase p-p38 (p-p38MAPK) and nuclear factor κB p65 (NF-κB p65) in THP-1 macrophages were detected by Western blot.Results (1) Vesicles that harvested by ultracentrifugation ranged in size from 30 nm to 100 nm and expressed exosomal marker CD63,TSG101 but absence of calnexin which is a marker of endoplasmic reticulum,suggesting that the exosomes were not contaminated with cells.(2) Results from laser scanning confocal microscope showed that each group of exosomes can be internalized by THP-1 macrophages.Compared with normal glucose exosomes group,high glucose exosomes had increased the expression of iNOS,TNF-α,IL-1β and MCP-1 in THP-1 macrophages (all P ＜ 0.01),moreover,p-JNK,p-p38 MAPK and NF-κB p65 proteins level also increased significantly (all P ＜ 0.01).Conclusions Exosomes from high glucose-treated HK2 cells can induce THP-1 macrophage activation and functional changes through MAPK/NF-κB pathway.

Grey Turner's and Cullen's signs induced by spontaneous hemorrhage of the abdominal wall after coughing

Grey Turner's and Cullen's signs are rare clinical signs, which most appear in patients with severe acute pancreatitis. The present patient complained of abdominal pain after coughing. However, contrast-enhanced CT revealed a hemorrhage of the abdominal wall. Therefore, spontaneous hemorrhage of the abdominal wall was diagnosed. The patient recovered through immobilization and hemostasis therapy. This case report and literature review aims to remind clinicians of manifestations and treatment of spontaneous hemorrhage.

A canonical correlation analysis based EMG classification algorithm for eliminating electrode shift effect.

Motion classification system based on surface Electromyography (sEMG) pattern recognition has achieved good results in experimental condition. But it is still a challenge for clinical implement and practical application. Many factors contribute to the difficulty of clinical use of the EMG based dexterous control. The most obvious and important is the noise in the EMG signal caused by electrode shift, muscle fatigue, motion artifact, inherent instability of signal and biological signals such as Electrocardiogram. In this paper, a novel method based on Canonical Correlation Analysis (CCA) was developed to eliminate the reduction of classification accuracy caused by electrode shift. The average classification accuracy of our method were above 95% for the healthy subjects. In the process, we validated the influence of electrode shift on motion classification accuracy and discovered the strong correlation with correlation coefficient of >0.9 between shift position data and normal position data.

Influence of Baihe Zhimu Decoction in levels of key factors of CaM signaling pathway in hippocampus tissue of rats with depression and its antidepressant mechanism / 吉林大学学报(医学版)

Objective:To study the antidepressant effect of Baihe Zhimu decoction (BZD)and its influence in the key factors (CaM,CaMKⅡ,CREB)of CaM signaling pathway in hippocampus of the rats with depression,and to explore the antidepressant effect of BZD. Methods:Fifty rats were divided into control group,model group, fluoxetine group,low and high doses of BDZ groups (n = 10).Expect for control group,all the rats in other groups were made depression models by means of chronic unpredictable mild stress along with isolated raising,for 21 d.Then the rats were fed with NS, fluoxetine (1.8 mg · kg-1 ), and BZD (1.5 and 3.0 g · kg-1 ), respectively;for 28 d.The learning and memory ability,autonomous activities and the fixed time in 5 min of the rats were tested by Morris water amaze,Open-field Test and Forced Swimming Test respectively. The damage and repair status of hippocampal neurons were observed by Nissl staining method;the expression levels of CaM,CaMKⅡ protein,CREB mRNA in hippocampus of the rats were detected by Western blotting and RT-PCR method. Results:Compared with model group,the total time of rats in the platform quadrant of Morris water maze in BZD groups and fluoxetine group,the total distance and the number of crossing platform were increased (P <0.05 or P <0.01),and the time of first crossing platform were shortened (P <0.01);the total scores in open field test were increased (P <0.01),the fixed time with 5 min in the forced swimming test was shortened (P <0.05 or P <0.01).Compared with fluoxetine group,the fixed time within 5 min of the rats in swimming test was shortened (P <0.05).The result of Nissl staining showed that the hippocampal neuron injury in BZD groups and fluoxetine group was improved compared with model group.The molecular test results showed that the CaM and CaMKⅡprotein expression levels in hippocampus of the rats in BZD groups and fluoxetine group were increased compared with model group (P < 0.05 or P < 0.01).Compared with model group,the CREB mRNA expression levels in fluoxetime group and BZD groups were increased (P < 0.05 or P < 0.01).Conclusion:BZD has antidepressant effect and can improve the hippocampal neuron injury of the rats with depression and its mechanism is related to increasing the expression levels of CaM,CaMKⅡ and CREB in hippocampus CAM signaling pathway of the rats.

Regulation of melatonin on Toll-like receptor 4 signaling in diabetic db/db mice kidneys / 中华肾脏病杂志

Objective To investigate the regulation of melatonin (MT) on Toll-like receptor 4 (TLR4) signaling in diabetic db/db mice kidneys.Methods The 48 10-week-old male db/db mice were randomly divided into db/db group,db/db+MT 50 μg/kg group,db/db+MT 100 μg/kg group and db/db+MT 200 μg/kg group,each consisting of 12 mice.These mice received i.p.injections of MT These mice received i.p.injections of MT [dissoved in phosphate buffer solution (PBS)/ dimethylsulfoxide (DMSO) solution,given every day].Alternatively,12 db/m mice served as the control group.db/m and db/db group were injected i.p.with the same volume of PBS/DMSO solution.The animals were sacrificed after 12 weeks of dosage administration.Blood glucose (BG),body weight (BW),kidney weight (KW) and 24 h urinary albumin excretion rate (UAER) were determined;Kidney pathological lesions were evaluated by renal pathological staining.Immunohistochemistry of renal TLR4,NF-κB p65,and ED-1 was performed to determine the immunoreactivity.Western blotting was used to detect the expression of renal TLR4,myeloid differentiation factor 88 (MyD88),TIR-domaincontaining adaptor inducing interferon-β (TRIF),interferon regulatory factor 3 (IRF-3) and NF-κB p65,while the mRNA expressions of renal tumor necrosis factor-α (TNF-α) and monocyte chemotactic protein-1 (MCP-1) were evaluated by real-time PCR.Results Compared with control group,the levels of BG,BW,KW and UAER were much higher in db/db mice group (P ＜ 0.01),while KW in db/db+MT (100,200 μg/kg) groups and UAER level in db/db+MT (50,100,200 μg/kg) groups were distinctly decreased compared with those in db/db group (P ＜ 0.01).In week 12 db/db mice,the glomerular mesangial expansion index and tubulointerstitial injury index were increased compared with those in db/m mice (P ＜ 0.01).The above kidney histopathologic lesions were distinctly ameliorated by 50,100,200 μg/kg MT (P ＜ 0.05).Immunohistochemistry intensity of renal TLR4,NF-κB p65 and ED-1 displayed obvious differences between db/m mice and db/db mice (P ＜ 0.01),and that were remarkably decreased in db/db+MT (50,100,200 μg/kg) mice compared with db/db mice (P ＜ 0.05).Western blotting showed that the protein expression of renal TLR4,MyD88,TRIF,IRF-3 and NF-κB p65 were stronger in db/db group compared with those in db/m group (P ＜ 0.05) and weaker in db/db+ MT (50,100,200 μg/kg) groups compared with those in db/db group (P ＜ 0.05).Futhermore,the mRNA expressions of renal MCP-1 and TNF-α were higher in db/db group compared with those in db/m group (P ＜ 0.01) and lower in db/db+MT (50,100,200 μg/kg) groups compared with those in db/db group (P ＜ 0.01).Conclusion Melatonin may partly down-regulate TLR4 signaling pathway to inhibit Inflammatory reaction and alleviate kidney injury in diabetic db/db mice.

Variation of extralaryngeal furcation of the recurrent laryngeal nerve in total thyroidectomy / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore the extralaryngeal furcation variation of the recurrent laryngeal nerve (RLN) in total thyroidectomy.@*METHOD@#The clinical data of 216 RLNs from 108 patients undergone total thyroidectomy were retrospectively analyzed.@*RESULT@#RLN was found during every operation and exposed in whole course until access into larynx. Twenty (9.26%) pieces of RLNs showed bifurcated or trifurcated RLNs before access into larynx. Ratio of furcation is lower than that reported before internationally. Bifurcations of RLNs on the left were more than that on the right.@*CONCLUSION@#The protection of RLN is important for thyroid operation, especially in total thyroidetomy. Variation of extralaryngeal furcation of RLN usually leads to injury of RLN. Understanding of variation of RLN could decrease nerve function related complication.

Multifunctional envelope-type mesoporous silica nanoparticles for tumor-triggered targeting drug delivery.

A novel type of cellular-uptake-shielding multifunctional envelope-type mesoporous silica nanoparticle (MEMSN) was designed for tumor-triggered targeting drug delivery to cancerous cells. ß-Cyclodextrin (ß-CD) was anchored on the surface of mesoporous silica nanoparticles via disulfide linking for glutathione-induced intracellular drug release. Then a peptide sequence containing Arg-Gly-Asp (RGD) motif and matrix metalloproteinase (MMP) substrate peptide Pro-Leu-Gly-Val-Arg (PLGVR) was introduced onto the surface of the nanoparticles via host-guest interaction. To protect the targeting ligand and prevent the nanoparticles from being uptaken by normal cells, the nanoparticles were further decorated with poly(aspartic acid) (PASP) to obtain MEMSN. In vitro study demonstrated that MEMSN was shielded against normal cells. After reaching the tumor cells, the targeting property could be switched on by removing the PASP protection layer via hydrolyzation of PLGVR at the MMP-rich tumor cells, which enabled the easy uptake of drug-loaded nanoparticles by tumor cells and subsequent glutathione-induced drug release intracellularly.

Effects of fluoride on fibronectin expression of rats osteoblasts / 中国地方病学杂志

Objective To observe the expression of fibronectin in bone of fluorosis rats and in vitro cultured osteoblast,and to study the role of fibronectin in pathogenesis of chronic fluorosis.Methods Male and female Wistar rats 144 were randomly divided into four groups,which were designated as the control group(normal diets,n =36),fluoride group(normal diets + 100 mg/L fluoride,n =36),lower calcium monophagia group (synthetic diets,n =36) and lower calcium monOphagia with fluoride group(synthetic diets + 100 mg/L fluoride,n =36).Rats were sacrificed 4 and 8 months after beginning of the experiment,respectively,and femur tissue was fixated and paraffin-embedded.The osteoblast isolated from calvaria of neonatal rats was treated with different dose of fluoride(0,1,2,4 mg/L fluoride,respectively) for 48 and 72 h,cell culture supernatant and cells were collected,respectively.The cranial osteoblasts were cultured in vitro and divided into four groups according to different concentration of fluoride added,which were 0(control group),0.01,1.00,and 10.00 mg/L groups.These cells were treated with mineralized induced medium at day 2 and cultured for 3 more weeks whereafter,and then the slides were fixed in alcohol.The expression of fibronectin in rat femur tissue was detected by immunohistochemistry (IHC),and fibronectin mRNA expression was determined by in situ hybridization; the fibronectin levels in supernatant of cultured osteoblast was examined by enzyme-linked immunosorbent assay(ELISA),and the expression of fibronectin mRNA in osteoblasts was detected with RT-PCR; skull mineralized nodule formation of osteoblasts was observed under a light microscopy after stained with 0.1％ red alizarin liquid.Results Little expression of fibronectin (brown granules under light microscope) could be seen in femur tissue of fluorosis rats of control group and lower calcium monophagia group; but abundantly expressed in fluoride group and lower calcium monophagia with fluoride group; the fibronectin was also expressed in osteoblasts,bone cells and bone marrow cells with less red particles in the control group and lower calcium monophagia group,but more in the fluoride group and lower calcium monophagia with fluoride group.The expression of fibronectin protein in supernatant of cultured osteoblasts was significantly increased in the group of 4 mg/L fluoride at 48 h(0.108 ± 0.042,t =0.764,P＜ 0.05) compared with control group(0.081 ± 0.010); the value was also significantly increased in 1,2,4 mg/L groups at 72 h(0.089 ± 0.010,0.087 ± 0.012,0.098 ± 0.023; t =0.765,0.704,0.996; all P ＜ 0.05) compared with control group (0.070 ± 0.014) ; the expression of fibronectin mRNA was much higher in 1,2,4 mg/L groups at 48 h (0.61 ±0.06,0.77 ± 0.07,0.77 ± 0.07) and 72 h(1.61 ± 0.14,2.54 ± 0.20,2.75 ± 0.22) compared with control group [0.48 ± 0.04(t =0.111,0.182,0.182,all P ＜ 0.05),0.97 ± 0.08(t =0.093,0.109,0.108,all P＜ 0.05) ].A lot of mineralized nodules could be seen under light microscope in 1.00 and 10.00 mg/L groups.Conclusions The expression of fibronectin in bone of fluorosis rats and in vitro cultured osteoblasts are increased,and fluoride also promotes the mineralization nodules formation of osteoblasts.These results suggest that fibronectin may regulate the process of bone mineralization,and possibly play a role in the development of skeletal fluorosis.

Self-assembled micellar aggregates based monomethoxyl poly(ethylene glycol)-b-poly(ε-caprolactone)-b-poly(aminoethyl methacrylate) triblock copolymers as efficient gene delivery vectors.

Amphiphilic triblock copolymers monomethoxyl poly(ethylene glycol) (mPEG)-b-poly(ε-caprolactone) (PCL)-b-poly(aminoethyl methacrylate)s (PAMAs) (mPECAs) were synthesized as gene delivery vectors. They exhibited lower cytotoxicity and higher transfection efficiency in COS-7 cells in presence of serum compared to 25 kDa bPEI. The influence of mPEG and PCL segments in mPECAs was evaluated by comparing with corresponding diblock copolymers. The studies showed the incorporation of the hydrophobic PCL segment in triblock copolymers affected the binding capability to pDNA and surface charges of complexes due to the formation of micelles increasing the local charges. The presence of mPEG segment in gene vector decreased the surface charges of the complexes and increased the stability of the complexes in serum because of the steric hindrance effect. It was also found that the combination of PEG and PCL segments into one macromolecule might lead to synergistic effect for better transfection efficiency in serum.

Influence of hydroxyl groups on the biological properties of cationic polymethacrylates as gene vectors.

In this study poly(aminoethyl methacrylate) (PAEMA), poly(3-amino-2-hydroxypropyl methacrylate) (PAHPMA), poly(2-(2-aminoethylamino)ethyl methacrylate) (PAEAEMA) and poly(3-(2-aminoethylamino) 2-hydroxypropyl methacrylate) (PAEAHPMA) were synthesized using atom transfer radical polymerization to evaluate the effect of hydroxyl groups on the relative properties of cationic polymeric gene vectors. The results of heparin displacement assays showed that PAHPMA possessed a stronger binding capacity than PAEMA. PAHPMA/DNA complexes and PAEAHPMA/DNA complexes had lower zeta potentials than those of PAEMA and PAEAEMA. MTT assay results indicated that PAHPMA and PAEAHPMA exhibited obviously lower cytotoxicities than PAEMA and PAEAEMA. Subsequently, in vitro gene transfection studies in 293T cells without serum showed that PAHPMA exhibited a lower transfection efficiency than PAEMA and PAEAHPMA/DNA complexes possessed a similar transfection efficiency to PAEAEMA/DNA complexes. Moreover, PAHPMA and PAEAHPMA retained similar transfection efficiencies in DMEM with 10% serum, but PAEMA and PAEAEMA showed slightly lower transfection efficiencies than in the absence of serum. The reason for these phenomena might be attributed to the introduction of hydroxyl groups into PAHPMA and PAEAHPMA, i.e. the existence of hydroxyl groups might increase the binding capacity to DNA and at the same time decrease the surface charge of the polymer/DNA complexes due to the formation of hydrogen bonds between the polymers and DNA. Therefore, a lower zeta potential and stronger binding ability may result in a lower gene transfection efficiency. This effect of hydroxyl groups decreased with increasing amino group density on the polymer.

Influence of fluoride on osteoprotegerin mRNA and protein expression in rat osteoblasts: an in vitro study / 中国地方病学杂志

Objective To study the influence of fluoride on the expression of osteoprotegerin(OPG) mRNA and protein in suckling rat osteoblasts. Methods Osteoblasts obtained from calvarial of suckling Wistar rats were cultured in vitro in the media supplemented with NaF at a series of doses[O(control), 1,2 and 4 mg/L groups], and OPG mRNA expression and protein were evaluated by RT-PCR and ELISA methods, respectively. Results OPG mRNA expression in suckling rat osteoblasts cultured in vitro significantly increased after exposure to NaF for 48 h and 72 h(F=333.48,808.34,P<0.05). OPG mRNA expression in suckling rat osteoblasts cultured in vitro after exposure to NaF for 48 h at different doses(0.810±0.003, 0.819±0.031 and 0.870±0.044 for 1,2 and 4 mg/L groups, respectively) compared with that of control (0.800±0.040, all P<0.05). OPG mRNA expression further increased for 72 h exposure to NaF(0.933±0.047,1.031±0.051,1.240±0.062 for 1,2 and 4 mg/L, respectively), significantly higher than that of the control (0.805±0.020,all P<0.05) and corresponding groups at 48 h. NaF doses and time exposure exhibited a significant synergistic effect on OPG mRNA expression(F=2004.16, P<0.05). NaF also enhanced OPG protein expression in suckling rat osteoblasts cultured in vitro. Significant differences were observed only in 4 mg/L group(0.228±0.014,0.277±0.048) and control(0.205±0.012,0.229±0.010) at 48 h and 72 h (P<0.05). In addition, OPG protein expression at 72 h post-exposure was higher than that at 48 h,but there was no synergistic effect between concentration and time(F=1.21,P>0.05). Conclusions The results suggested that NaF could increase OPG mRNA and protein expression in suckling rat osteoblasts with a synergistic effect between the doses and exposure time.

Influence of fluoride on Runx2 mRNA and protein expression in rat osteoblasts / 中国地方病学杂志

Objective To study the influence of fluoride on the expression of Runx2 in suckling rat osteoblasts. Methods Osteoblasts obtained from calvarium of suckling Wistar rats were cultured in the media supplemented with NaF at different doses(0, 1,2 and 4 rag/L), and Runx2 Mrna expression and protein expression were evaluated by RT-PCR and ELISA, respectively. Results Runx2 Mrna expression in suckling rat osteoblasts cultured in vitro significantly increased after exposure to NaF for 48 h at different doses (0.613±0.055, 0.773±0.070 and 0.775±0.070 for 1,2 and 4 mg/L,respecfively) compared to the control (0.482±0.043 ,P< 0.05). Runx2 Mrna expression further increased after 72 h exposure to NaF(0.969±0.048,1.229±0.061,1.255± 0.063 for 1,2 and 4 mg/L, respectively) ,which is significantly higher than the control(0.724±0.036,P<0.05) and corresponding groups at 48 h. NaF doses and exposure time exhibited a significant synergistic effect on Runx2 Mrna expression (P<0.05). Similarly, NaF also enhanced Bunx2 protein expression in suckling rat osteoblasts cultured in vitro. Significant differences were observed between groups exposed to NaF (1,2 and 4 rag/L) and control at48 h post-exposure (0.141±0.007, 0.143±0.008, 0.143±0.011 vs 0.129±0.012, P<0.05) as well as 72 h post-expesure(0.156±0.014, 0.168±0.018, 0.162±0.0100 vs 0.137±0.016, P<0.05). In addition, Runx2 protein expression at 72 h post-exposure was significantly higher than that at 48 h. Conclusions The results suggested that NaF could increase Runx2 expression in suckling rat osteoblasts with a synergistic effect between the doses and exposure time.