Search

The present study used comparative proteomic analysis of cerebrospinal fluid (CSF) in amyotrophic lateral sclerosis (ALS) patients in order to identify proteins that may act as diagnostic biomarkers and indicators of the pathogenesis of ALS. This analysis was performed using isobaric tags for relative and absolute quantitation (iTRAQ) technology, coupled with 2-dimensional liquid chromatography/mass spectrometry. Database for Annotation, Visualization and Integrated Discovery software was utilized for bioinformatic analysis of the data. Following this, western blotting was performed in order to examine the expression of 3 candidate proteins in ALS patients compared with healthy individuals [as a normal control (NC) group] or patients with other neurological disease (OND); these proteins were insulin-like growth factor II (IGF-2), glutamate receptor 4 (GRIA4) and leucine-rich α-2-glycoprotein 1 (LRG1). Clinical data, including gender, age, disease duration and ALS functional rating scale (ALSFRS-R) score, were also collected in the ALS patients. Multiple linear regression analysis was performed between the clinical data and the results of western blot analysis. A total of 248 distinct proteins were identified in the ALS and NC groups, amongst which a significant difference could be identified in 35 proteins; of these, 21 proteins were downregulated and 14 were upregulated. These differentially-expressed proteins were thus revealed to be associated with ALS. The western blot analysis confirmed a proportion of the data attained in the iTRAQ analysis, revealing the differential protein expression of IGF-2 and GRIA4 between the ALS and NC groups. IGF-2 was significantly downregulated in ALS patients (P=0.017) and GRIA4 was significantly upregulated (P=0.016). These results were subsequently validated in the 35-patient ALS and OND groups (P=0.002), but no significant difference was identified in LRG1 expression between these groups. GRIA4 protein expression was higher in male than female patients and was positively correlated with the ALSFRS-R score, meaning that GRIA4 expression was negatively correlated with the severity of ALS, while IGF-2 and LRG1 expression did not correlate with any clinical data. The present study thus demonstrated that GRIA4 expression levels, as a marker of severity, may be used as a reference for the timing of treatment, and that IGF-2 may serve as an effective biomarker of ALS progression.

Increasing atmospheric CO2 concentrations ([CO2]) in agricultural and natural ecosystems is known to reduce plant stomatal opening, but it is unclear whether these CO2-induced stomatal alterations are associated with foliar pathogen infections. In this study, tomato plants were grown under ambient and elevated [CO2] and inoculated with Pseudomonas syringae pv. tomato strain DC3000, a strain that is virulent on tomato plants. We found that elevated [CO2] enhanced tomato defence against P. syringae. Scanning electron microscopy analysis revealed that stomatal aperture of elevated [CO2] plants was considerably smaller than their ambient counterparts, which affected the behaviour of P. syringae bacteria on the upper surface of epidermal peels. Pharmacological experiments revealed that nitric oxide (NO) played a role in elevated [CO2]-induced stomatal closure. Silencing key genes involved in NO generation and stomatal closing, nitrate reductase (NR) and guard cell slow-type anion channel 1 (SLAC1), blocked elevated [CO2]-induced stomatal closure and resulted in significant increases in P. syringae infection. However, the SLAC1-silenced plants, but not the NR-silenced plants, still had significantly higher defence under elevated [CO 2] compared with plants treated with ambient [CO2]. Similar results were obtained when the stomata-limiting factor for P. syringae entry was excluded by syringe infiltration inoculation. These results indicate that elevated [CO 2] induces defence against P. syringae in tomato plants, not only by reducing the stomata-mediated entry of P. syringae but also by invoking a stomata-independent pathway to counteract P. syringae. This information is valuable for designing proper strategies against bacterial pathogens under changing agricultural and natural ecosystems.

Accumulation of α-synuclein (α-Syn) is a common pathology for both familiar and sporadic Parkinson's disease (PD), enhancing its clearance might be a promising strategy for treating PD. To assess the potential of trehalose in this regard, we investigated its effect on the PC12 cells overexpressing wild type (WT) or A53T mutant α-Syn and the implicated pathway it might mediated. We observed that trehalose promoted the clearance of A53T α-Syn but not WT α-Syn in PC12 cells, and confirmed the increased LC3 and Lysotracker RED positive autolysosomes by using lysotracker and LC3 staining, the enhanced expression of LC3-II in Western blot, and more autophagosomes under Transmission Electron Microscope in a dose dependent manner after the trehalose treatment. The activation of autophagy can be alleviated by applying macroautophagy inhibitor 3-methyladenine (3-MA). In addition, degradation of A53T and WT α-Syn was blocked after Ubiquitin Proteasome System (UPS) inhibitor (MG132) was applied in those PC12 cells overexpressing A53T or WT α-Syn, suggesting that A53T α-Syn could be degraded by both UPS and macroautophagy. But the effect of trehalose on A53T α-Syn is mainly mediated through the macroautophagy pathway, which is not a dominant way for WT α-Syn clearance. Further in vivo research will be needed to verify the effectiveness of trehalose in treating PD.

Few studies have been performed on the individual-specific trajectory of left ventricular aging as assessed by echocardiography in an asymptomatic elderly cohort. In the present study, a representative cohort of elderly men, who were long-term asymptomatic for cardiovascular issues, were selected from an ongoing observational aging study. Annual echocardiographic data were used to establish an age-dependent hierarchical model. Based on two-level linear regression results, four echocardiographic indexes [left ventricular mass (LVmass; −1.872 g/yr), posterior ventricular wall thickness (−0.048 mm/yr), fraction shortening (0.097/yr), and transmitral peak A velocity (−0.006 m·s−1·yr−1)] changed significantly with increasing age and were age- and subject-dependent. The most characterized results of the study were the significant, age-related, within-individual variances in echocardiographic results, which were observed using the likelihood ratio test at an occasion-dependent level. Of these, fluctuated amplitudes of two systolic variables [i.e., LVmass (con/age = −0.012 ± 0.004; P = 0.0007) and fraction shortening (con/age = −0.001 ± 0.004; P = 0.05)] were significantly attenuated with increasing age within individuals. On the other hand, the age-related variability of four diastolic Doppler variables [i.e., peak A velocity (con/age = 0.003 ± 0.002; P = 0.0009), peak E velocity (con/age = 0.004 ± 0.003; P = 0.01), E/A ratio (con/age = 0.007 ± 0.003; P = 0.0002), and deceleration time of E wave (con/age = 0.025 ± 0.007; P < 0.0001)] significantly increased with increasing age within individuals. The age-related individual variability of left ventricular indexes observed in this continuous asymptomatic cohort may reflect the mechanism of preclinical, individualized heart aging. In conclusion, successfully fitted multilevel models were applied as a valuable tool to determine the mechanism of individual cardiac aging in the elderly.

To study the PCO of high-salinity organic wastewater, nano-sized TiO2 was used for PCO of MB in solutions with different pH values and high concentrations of Na2SO4 or NaCl. The rates of PCO were monitored by total organic carbon (TOC) measurement. The results showed that the degradation rates increased with increasing pH and decreased as the concentration of Na2SO4 or NaCl increased. The difference of degradation rates in Na2SO4 or NaCl aqueous solutions increased as the ion concentration increased. In the orthogonal experiments, the rates decreased remarkably under certain ratios of the ions concentration. The mechanisms were explored in some detail.

CO2 enrichment and brassinosteroids (BR) both have positive impacts on photosynthesis and plant growth. To examine the interactive effect of CO2 enrichment and BR on photosynthesis and plant growth, CO2 assimilation, chlorophyll fluorescence quenching, carbohydrate metabolism, photosynthetic gene transcript and enzyme activity were analyzed in leaves of young plants of cucumber (Cucumis sativus L.) in response to a doubling of growth CO2 level, foliar BR application alone or in combination. Both CO2 elevation and application of BR increased shoot biomass, leaf area, CO2 assimilation, total soluble sugar and starch contents, transcript for photosynthetic gene and activity for enzymes involved in Benson–Calvin cycle but a combination of the two treatments resulted in a more significant effect. Although an elevation of CO2 level had little effects on quantum efficiency of PSII (ФPSII), it significantly increased the electron flux for photosynthetic carbon reduction [Je(PCR)] but decreased electron flux for photorespiratory carbon oxidation [Je(PCO)]. In contrast, BR treatment increased ФPSII and this increase in ФPSII was associated with increased Je(PCR) and Je(PCO). Furthermore, a combined treatment of CO2 elevation and BR resulted in an additive effect on PSII electron flux. However, alternative electron flux was almost unaltered after CO2 enrichment and BR treatment. Thus, short term CO2 elevation did not induce a down-regulation of photosynthesis and there was an additive effect between BR and CO2 on the enhancement of CO2 assimilation in leaves of young cucumber plants.

Chlorothalonil (CHT) and carbendazim (CAR) are two widely used fungicides in agriculture. Despite their agronomic importance in pest control, little is known about their detoxification in the plant. In this study, we investigated the effects of these fungicides on glutathione (GSH) content, GSH-dependent enzyme activities and gene expression in tomato leaves. Results showed that exposure to CHT resulted in increases in GSH content, activities of glutathione S-transferases (GSTs) and glutathione reductase (GR), as well as the transcriptional levels of glutathione S-transferase genes (GST1, GST2 and GST3), glutathione synthetase gene (GSH), glutathione reductase gene (GR) and glutathione peroxidase gene (GPX) in tomato leaves, but such increases were not observed in leaves exposed to CAR. In addition, GSTs, GR, peroxidase (POD) activities and most of GSH-dependent gene expression were induced by CHT in a concentration- and time-dependent manner. These results suggest that GSH-dependent pathway plays an important role in the CHT detoxification but not in the CAR detoxification in tomato leaves.

To determine whether light quality affects the incidence of disease, we exposed cucumber (Cucumis sativus L. cv. Jinyan No. 4) plants at the 4-leaf stage to white and other monochromatic lights and tested the effects on plant response to Sphaerotheca fuliginea, defence-related gene expression and metabolic changes. Exposure to red light resulted in higher levels of H2O2 and salicylic acid (SA), and stronger expression of defence genes such as PR-1 than exposure to white or other monochromatic lights. In comparison, plants grown under purple and blue light had higher activities of phenylalanine ammonia-lyase (PAL) and polyphenoloxidase (PPO) and higher level of flavonoids than plants grown under other lights. Furthermore, plants grown under red light were more resistant whilst plants grown under other monochromatic lights were less resistant to Sphaerotheca fuliginea than plants grown under white light. These results suggest a role of red light in light-enhanced resistance, which correlates with enhanced SA-dependent signaling pathway.

The potential value of glial cell line-derived neurotrophic factor (GDNF) in treating Parkinson's disease (PD) remains controversial. In order to evaluate the therapeutic effect of GDNF-engineered bone marrow stromal cells (BMSCs) in parkinsonian rat model, GDNF-BMSCs and LacZ-BMSCs were transplanted into striatum and followed by Lactacystin lesioning at median forebrain bundles 1 week later. We observed that the intrastriatal transplantation of GDNF-BMSCs could significantly rescue the dopaminergic neurons from lactacystin-induced neurotoxicity with regard to behavioral recovery, tyrosine hydroxylase level in nigra and striatum, and striatal dopamine level. We interpret the outcomes that intrastriatal transplantation of GDNF-BMSCs might be beneficial in the treatment of PD.

Dopamine transporter (DAT) mediates the regulation of dopaminergic function. Two agents of TRODAT-1 and FP-CIT were observed in evaluating DAT change of Parkinson's disease (PD). The relationship between them was also evaluated. The results suggested that 99m Tc-TRODAT-1 SPECT and 131 I-FP-CIT SPECT may serve as sensitive and objective in vivo markers to reflect the severity of PD. The 99c Tc-TRODAT-1 image is more accurate and clearer compared with 131 I-FP-CIT.

The inclusion behavior between Costa-type complexes and cyclodextrins (CDs) was studied by 1H NMR in aqueous solution. The results indicated that 1:1 inclusion complex was formed, in which the alkyl group of the guest was included in the cavity of CDs. The stability constants of the inclusion complexes were determined by the quantitative 1H NMR method. The effects on stability constants were discussed when various host and guest compounds were used.

Spatial variability and management zone of soil major nutrients in tobacco fields in Qian-nan mountainous region were analyzed using geostatistics and fuzzy c-mean algorithm. Results indicated that the level of soil organic matter (OM) was moderate, and alkalytic nitrogen (AN), available phosphorus (AP) and available potassium (AK) were rich according to tobacco soil nutrient classification standards. Coefficients of variation (CV) of OM, AN, AP and AK were moderate. Contents of OM, AN, AP and AK fitted log-normal distributions. Correlation analysis showed moderate correlations between OM and AN, AP and AK. OM and AN were best described by Gaussian semivariogram models, while AP and AK were described by exponential models. The four nutrients displayed moderate spatial autocorrelation. There were significant differences among lag distances of four soil nutrients. OM, AN, AP and AK in the majority of studied regions varied at moderate to very rich levels, and deficiencies of OM, AN, AP and AK only accounted for 0.93%, 0.53%, 0.24% and 7.91% of the total studied region, respectively. Based on the results, the studied region was divided into two management zones (MZ), namely MZ1 and MZ2, accounting for 69. 8% and 30. 2% of the studied region respectively. The soil levels of OM, AN, AP and AK in MZ1 were significantly lower than those in MZ2 (P0.01).

To investigate the effects of neopterin on ABCA1 expression and cholesterol efflux in human THP-1 macrophage-derived foam cells, and to explore the role of the liver X receptor alpha (LXRα) involved. In the present study, THP-1 cells were pre-incubated with ox-LDL to become foam cells. The protein and mRNA expression were examined by Western blot assays and real-time quantitative PCR, respectively. Liquid scintillation counting and high performance liquid chromatography assays were used to test cellular cholesterol efflux and cholesterol content. Neopterin decreased ABCA1 expression and cholesterol efflux in a time- and concentration-dependent manner in THP-1 macrophage-derived foam cells, and the LXRα siRNA can reverse the inhibitory effects induced by neopterin. Neoterin has a negative regulation on ABCA1 expression via the LXRα signaling pathway, which suggests the aggravated effects of neopterin on atherosclerosis.

Brassinosteroids (BRs) are potent regulators of photosynthesis and crop yield in agricultural crops; however, the mechanism by which BRs increase photosynthesis is not fully understood. Here, we show that foliar application of 24-epibrassinolide (EBR) resulted in increases in CO(2) assimilation, hydrogen peroxide (H(2)O(2)) accumulation, and leaf area in cucumber. H(2)O(2) treatment induced increases in CO(2) assimilation whilst inhibition of the H(2)O(2) accumulation by its generation inhibitor or scavenger completely abolished EBR-induced CO(2) assimilation. Increases of light harvesting due to larger leaf areas in EBR- and H(2)O(2)-treated plants were accompanied by increases in the photochemical efficiency of photosystem II (Φ(PSII)) and photochemical quenching coefficient (q(P)). EBR and H(2)O(2) both activated carboxylation efficiency of ribulose-1,5-bisphosphate oxygenase/carboxylase (Rubisco) from analysis of CO(2) response curve and in vitro measurement of Rubisco activities. Moreover, EBR and H(2)O(2) increased contents of total soluble sugar, sucrose, hexose, and starch, followed by enhanced activities of sugar metabolism such as sucrose phosphate synthase, sucrose synthase, and invertase. Interestingly, expression of transcripts of enzymes involved in starch and sugar utilization were inhibited by EBR and H(2)O(2). However, the effects of EBR on carbohydrate metabolisms were reversed by the H(2)O(2) generation inhibitor diphenyleneodonium (DPI) or scavenger dimethylthiourea (DMTU) pretreatment. All of these results indicate that H(2)O(2) functions as a secondary messenger for EBR-induced CO(2) assimilation and carbohydrate metabolism in cucumber plants. Our study confirms that H(2)O(2) mediates the regulation of photosynthesis by BRs and suggests that EBR and H(2)O(2) regulate Calvin cycle and sugar metabolism via redox signaling and thus increase the photosynthetic potential and yield of crops.

Brassinosteroids (BRs) play important roles in plant growth, development, photosynthesis and stress tolerance; however, the mechanism underlying BR-enhanced photosynthesis is currently unclear. Here, we provide evidence that an increase in the BR level increased the quantum yield of PSII, activities of Rubisco activase (RCA) and fructose-1,6-bisphosphatase (FBPase), and CO 2 assimilation. BRs upregulated the transcript levels of genes and activity of enzymes involved in the ascorbate–glutathione cycle in the chloroplasts, leading to an increased ratio of reduced (GSH) to oxidized (GSSG) glutathione in the chloroplasts. An increased GSH/GSSG ratio protected RCA from proteolytic digestion and increased the stability of redox-sensitive enzymes in the chloroplasts. These results strongly suggest that BRs are capable of regulating the glutathione redox state in the chloroplasts through the activation of the ascorbate–glutathione cycle. The resulting increase in the chloroplast thiol reduction state promotes CO 2 assimilation, at least in part, by enhancing the stability and activity of redox-sensitive photosynthetic enzymes through post-translational modifications.

The aim of this study was to examine the role of brassinosteroids (BRs) in protecting the photosynthetic apparatus from cold-induced damage in cucumber (Cucumis sativus) plants. Recovery at both high light (HL) and low light (LL) after a cooling at 10/7°C induced irreversible inhibition of CO2 assimilation, photoinhibition at photosystem I (PSI) and inhibition of enzyme activities of Calvin cycle and ascorbate (AsA)-reduced glutathione (GSH) cycle, followed by accumulation of H2 O2 and malondialdehyde. However, cold-induced photoinhibition at PSII was fully recovered at LL but not at HL. Meanwhile, recovery at HL increased electron flux to O2 -dependent alternative pathway [Ja(O2 -dependent)]. Foliar application of 24-epibrassinolide (EBR) accelerated recovery from photoinhibition of PSII but not of PSI. EBR also significantly increased CO2 assimilation, activity of Calvin cycle enzymes and electron flux to carbon reduction [Je(PCR)], with a concomitant decrease in Ja(O2 -dependent); meanwhile EBR increased the activity of enzymes in AsA-GSH cycle and cellular redox states. However, the positive effect of EBR on plant recovery was observed only at HL, but not LL. These results indicate that BR accelerates the recovery of photosynthetic apparatus at HL by activation of enzymes in Calvin cycle and increasing the antioxidant capacity, which in turn mitigate the photooxidative stress and the inhibition of plant growth during the recovery.

Brassinosteroids (BRs) play a vital role in plant growth, stress tolerance and productivity. Here, the involvement of BRs in the regulation of CO(2) assimilation and cellular redox homeostasis was studied. The effects of BRs on CO(2) assimilation were studied in cucumber (Cucumis sativus) through the analysis of the accumulation of H(2)O(2) and glutathione and photosynthesis-related enzyme activities using histochemical and cytochemical detection or a spectrophotometric assay, and Rubisco activase (RCA) using western blot analysis and immunogold labeling. Exogenous BR increased apoplastic H(2)O(2) accumulation, the ratio of reduced to oxidized glutathione (GSH:GSSG) and CO(2) assimilation, whereas a BR biosynthetic inhibitor had the opposite effects. BR-induced CO(2) assimilation was decreased by a H(2)O(2) scavenger or inhibition of H(2)O(2) generation, GSH biosynthesis and the NADPH-generating pentose phosphate pathway. BR-, H(2)O(2) - or GSH-induced CO(2) assimilation was associated with increased activity of enzymes in the Benson-Calvin cycle. Immunogold labeling and western blotting showed that BR increased the content of RCA and this effect was blocked by inhibitors of redox homeostasis. These results strongly suggest that BR-induced photosynthesis involves an H(2)O(2) -mediated increase in the GSH:GSSG ratio, which may positively regulate the synthesis and activation of redox-sensitive enzymes in carbon fixation.

The role of mitochondrial alternative oxidase (AOX) and the relationship between systemic AOX induction, ROS formation, and systemic plant basal defense to Tobacco mosaic virus (TMV) were investigated in tomato plants. The results showed that TMV inoculation significantly increased the level of AOX gene transcripts, ubiquinone reduction levels, pyruvate content, and cyanide-resistant respiration (CN-resistant R) in upper, un-inoculated leaves. Pretreatment with potassium cyanide (KCN, a cytochrome pathway inhibitor) greatly increased CN-resistant R and reduced reactive oxygen species (ROS) formation, while application of salicylhydroxamic acid (SHAM, an AOX inhibitor) blocked the AOX activity and enhanced the production of ROS in the plants. Furthermore, TMV systemic infection was enhanced by SHAM and reduced by KCN pretreatment, as compared with the un-pretreated TMV counterpart. In addition, KCN application significantly diminished TMV-induced increase in antioxidant enzyme activities and dehydroascorbate/total ascorbate pool, while an opposite change was observed with SHAM-pretreated plants. These results suggest that the systemic induction of the mitochondrial AOX pathway plays a critical role in the reduction of ROS to enhance basal defenses. Additional antioxidant systems were also coordinately regulated in the maintenance of the cellular redox homeostasis.

We performed a six-week study of pramipexole vs. a placebo in Chinese restless legs syndrome patients.Overall, 305 enrolled patients were assigned randomly in a 2:1 ratio to the pramipexole group (N=202) and the placebo group (N=103).Of 287 patients in the full analysis set, the pramipexole group showed significant improvement compared with the placebo group in the change of their International Restless Legs Syndrome Study Group Rating Scale of Severity (IRLS) total score from baseline to week 6 after adjustment of centers and baseline characters (-15.87±0.66 vs. -11.35±0.92, p0.0001) and in the proportion of patients who were "much improved" and "very much improved" when measured by Clinical Global Impressions-Improvement (81.9% vs. 54.3%, p0.0001). At week 6, the IRLS responder rate was 73.8% (pramipexole) and 48.9% (placebo) (p0.0001) and the patient global impression responder rate was 68.6% (pramipexole) and 43.5% (placebo) (p0.0001). The proportion of adverse events was 62.9% in the pramipexole group and 43.7% in the placebo group, respectively. No deaths occurred.Pramipexole was effective and well-tolerated in Chinese patients with restless legs syndrome.

According to the law of circulation of meridians and the locations of acupoints, the opposite acupoints were proposed. It facilitates comprehension of the routes of meridians and the locations of acupoints. Application of opposite needling or penetrative needling, it is easy to practice and the effectiveness is significant. Promoting this concept into acupuncture training, can expand out from acupoint to meridian, from one meridian to other meridians, it will generate good rewards.

To investigate the therapeutic effect of microencapsulated porcine retinal pigmented epithelial cells (RPE-M) transplantation on rat model of Parkinson's disease (PD).Primary porcine RPE cells were harvested by enzyme digestion and expanded in culture medium. Determine the levels of dopamine (DA) and homovanillic acid (HVA) by high performance liquid chromatography electrochemical (HPLC) assay, and the levels of brain-derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF) were detected by ELISA. Alginate-polylysine-alginate (APA) microencapsulated cells were produced by using a high voltage electrostatic system. PD rat model was established by unilateral injection of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle (MFB). After that, the RPE-M was transplanted into the corpus striatum of PD rat, and then the rotation test scores were recorded and biochemical changes of the corpus striatum were tested.The levels of DA, HVA, BDNF and GDNF secreted by RPE were stable in the RPE culture supernatant and were not changed by the microencapsulation. Eighty-three percent rats developed PD by unilateral lesion of 6-OHDA in the MFB. The RPE-M transplantation had therapeutic effect on 33% PD rats.Porcine RPE cells grow actively in vitro and could secrete DA, HVA, BDNF, and GDNF constantly, which does not be affected by the passage culture and the APA miroencapsulation. RPE-M transplantation of may be a curative therapy for PD.

In a previous study, we have verified that CXCR4 expression is correlated with tumor aggressive progression and poor prognosis in patients with epithelial ovarian cancer. The aim of this study was to explore the effect of CXCL12-CXCR4 axis on the metastasis of human ovarian cancer.The expressions of CXCR4 and CXCL12 mRNA and protein in human ovarian cancer cell line CAOV-3 was detected by RT-PCR and immunocytochemistry. Methythiazolyltetrazolium (MTT) was used to analyze the effect of different concentrations of CXCL12 on the proliferation of CAOV-3 cells. Transwell invasion chamber and matrigel were used to evaluate the effect of various concentrations of CXCL12 and ascites on the migration and invasion of CAOV-3 cells. The expressions of integrin beta(1) and vascular endothelial growth factor-C (VEGF-C) mRNA were detected by RT-PCR. Data were analyzed using ANOVA by SAS 6.12.Under serum-free suboptimal culture conditions, CXCL12 (100 ng/ml) significantly enhanced the proliferation of CAOV-3 cells compared with the control and 10 ng/ml CXCL12 groups (0.428 +/- 0.051 vs. 0.325 +/- 0.045 and 0.328 +/- 0.039, P0.05). This enhancing effect of CXCL12 was significantly inhibited by 10 microg/ml neutralizing CXCR4 antibody or 1 microg/ml CXCR4 antagonist AMD3100. However, 10 microg/ml neutralizing CXCR4 antibody could not inhibit cell proliferation without CXCL12. The levels of migration and invasion of the CAOV-3 cells treated with 100 ng/ml CXCL12 were significantly higher than those in the control (migration: 523.3 +/- 25.2 vs 108.0 +/- 7.2; invasion: 39.3 +/- 4.0 vs. 4.0 +/- 1.0). The enhancing effect of CXCL12 on cell migration and invasion increased with the concentration of CXCL12 (100 ng/ml vs10 ng/ml: migration, 523.3 +/- 25.2 vs 211.7 +/- 24.7; invasion, 39.3 +/- 4.0 vs 15.7 +/- 3.1, P0.05), and was strongly inhibited by 10 microg/ml neutralizing CXCR4 antibody or 1 microg/ml AMD3100. The number of migrated and invading cells in the CAOV-3 added with ascites was significantly higher than those in the 100 ng/ml CXCL12 group (migration: 706.6 +/- 30.6 vs 523.3 +/- 25.2, invasion: 61.7 +/- 7.6 vs 39.3 +/- 4.0, P0.05). The level of integrin beta(1) mRNA was greatly increased at 3 hours after being treated with CXCL12 (0.53 +/- 0.10 vs. 1.53 +/- 0.16, P0.05), and VEGF-C mRNA displayed significant augment at 24 hours after being treated with CXCL12 (0.52 +/- 0.09 vs 1.11 +/- 0.15, P0.05).CXCL12 and its receptor CXCR4 can promote the proliferation, migration, invasion of ovarian cancer cell line CAOV-3 and enhance its secretion of integrin beta(1) and VEGF-C. These effects can be inhibited by neutralizing CXCR4 antibody or AMD3100. CXCL12-CXCR4 axis plays an important role in ovarian cancer growth and metastasis.

To explore the effect of chemokine CXCL12 and its receptor CXCR4 on proliferation, migration and invasion of epithelial ovarian cancer cells.CXCR4 and CXCL12 mRNA and protein expression of human ovarian cancer cell line CAOV3 was detected by RT-PCR and immunocytochemistry. Integrin beta1 and vascular endothelial growth factor-C (VEGF-C) mRNA expression were detected in CAOV3 cells stimulated by CXCL12. The CAOV3 cells were divided into 6 groups: control group (un-stimulated), experimental group 1 (stimulated by 100 ng/ml CXCL12), experimental group 2 (stimulated by 10 ng/ml CXCL12), experimental group 3 (100 ng/ml CXCL12 and 10 microg/ml neutralizing CXCR4 antibody), experimental group 4 (100 ng/ml CXCL12 and 1 microg/ml CXCR4 antagonist AMD3100), experimental group 5 (10 microg/ml neutralizing CXCR4 antibody or ascites). Methyl thiazolyl tetrazolium (MTT) was used to analyze the effects of different concentrations of CXCL12 on CAOV3 cell proliferation. Transwell invasion chamber and reconstructed basement membrane (Matrigel) were used to evaluate effect of various concentrations of CXCL12 and ascites on CAOV3 cell migration and invasion.CAOV3 cells expressed CXCR4 mRNA (0.70 +/- 0.10) and protein, but did not express CXCL12 mRNA or protein. Immunostaining of CXCR4 was mainly located in cytoplasm. CXCR4 mRNA was up-regulated after 100 ng/ml CXCL12 stimulation (1.24 +/- 0.14; t = -7.1088, P = 0.0021). Integrin beta1 mRNA was greatly increased at 3 hours by stimulation of 100 ng/ml CXCL12 (before and after stimulation 0.53 +/- 0.10, 1.53 +/- 0.16; P0.01), and VEGF-C mRNA showed significant increase at 24 hours by treatment with CXCL12 (before and after stimulation 0.52 +/- 0.09, 1.11 +/- 0.15; P0.05). Under serum-free sub-optimal culture conditions, experimental group 1 greatly enhanced cell proliferation in CAOV3 cells compared with control group and experimental group 2 (respectively 0.428 +/- 0.051, 0.325 +/- 0.045, 0.328 +/- 0.039; P0.05). Experimental group 1 was strongly inhibited compared with experimental groups 3 and 4 (the latter two groups respectively 0.356 +/- 0.031, 0.373 +/- 0.029; P0.05). There was no significant difference between experimental group 5 (0.349 +/- 0.038) and control group (P0.05). Experimental group 1 stimulated the migration and invasion of CAOV3 cells in chemotaxis assay compared with control group and experimental group 2 (number of cell migration respectively 523.3 +/- 25.2, 108.0 +/- 7.2, 211.7 +/- 24.7, number of cell invasion respectively 39.3 +/- 4.0, 4.0 +/- 1.0, 15.7 +/- 3.1; P0.01). This enhancing effect of experimental group 1 was strongly inhibited compared with experimental groups 4 and 5 (P0.05). The number of migrating and invading cells in experimental group 5 (migration: 706.6 +/- 30.6, invasion: 61.7 +/- 7.6) was significantly higher than that of experimental group 1 (P0.05).This in vitro study shows CXCL12 promote proliferation, migration, invasion of ovarian cancer cell line CAOV3, and up-regulate integrin beta1 and VEGF-C expression, and these effects are strongly inhibited by neutralizing CXCR4 antibody. It suggests CXCL12 and its receptor CXCR4 may play important roles in ovarian cancer growth and metastasis.

To construct recombinant lentiviral vectors for gene delivery of the glial cell line-derived neurotrophic factor (GDNF), and evaluate the neuroprotective effect of GDNF on lactacystin-damaged PC12 cells by transfecting it into bone marrow stromal cells (BMSCs).pLenti6/V5-GDNF plasmid was set up by double restriction enzyme digestion and ligation, and then the plasmid was transformed into Top10 cells. Purified pLenti6/V5-GDNF plasmids from the positive clones and the packaging mixture were cotransfected to the 293FT packaging cell line by Lipofectamine2000 to produce lentivirus, then the concentrated virus was transduced to BMSCs. Overexpression of GDNF in BMSCs was tested by RT-PCR, ELISA and immunocytochemistry, and its neuroprotection for lactacystin-damaged PC12 cells was evaluated by MTT assay.Virus stock of GDNF was harvested with the titer of 5.6 x 100,000 TU/mL. After transduction, GDNF-BMSCs successfully secreted GDNF to supernatant with higher concentration (800 pg/mL) than BMSCs did (less than 100 pg/mL). The supernatant of GDNF-BMSCs could significantly alleviate the damage of PC12 cells induced by lactacystin (10 micromol/L).Overexpression of lentivirus-mediated GDNF in the BMSCs cells can effectively protect PC12 cells from the injury by the proteasome inhibitor.

Previous studies suggest that chemokine CXCL12 and its receptor CXCR4 play important roles in embryonic development, stem cell trafficking, and inflammatory reactions. It is a central factor in directing cell movement of many physiologic and pathologic processes. The mechanisms of cancer cell metastasis and white blood cell trafficking are similar. CXCL12-CXCR4 axis exerts important effects in regulating growth, invasion, metastasis, and secretion of malignant cells. Data from animal experiments suggest that CXCR4 may be an important therapeutic target in inhibiting malignant growth and metastatic behavior of tumor cells. This review focused on the role of CXCL12-CXCR4 axis in regulating tumor metastasis and progression, and the molecular mechanisms that are essential to this process.

To investigate the usefulness of 18F-FP-CIT PET for assessing the severity of Parkinson's disease (PD) at various clinical stages, 41 patients with PD were divided into early (Hoehn&Yahr I-II, n = 23) and advanced (Hoehn & Yahr III-IV, n = 18) subgroups. 18F-FP-CIT PET was performed in these patients and 12 normal subjects. 18F-FP-CIT uptake in striatal subregions and its correlation with UPDRS were first evaluated by ROI analysis, and between-group differences were also analyzed by Statistical Parametric Mapping (SPM). Our results showed that striatal 18F-FP-CIT binding were significantly reduced to 70.9% (caudate), 46.8% (anterior putamen) and 24.0% (posterior putamen) in early PD compared with that of the control, and to 52.0%, 34.5% and 16.5% correspondingly in advanced PD, respectively. There was significant negative correlation between total motor UPDRS score of all parkinsonian patients and 18F-FP-CIT uptake in caudate nucleus (r = -0.53, p < 0.001), anterior putamen (r = -0.53, p < 0.001) and posterior putamen (r = -0.61, p < 0.001). SPM comparison of 18F-FP-CIT uptake between early or advanced PD and the control group showed significant decline in striatum, predominantly localized on the contralateral side and in the dorsal-posterior putamen. These results indicate that 18F-FP-CIT PET can serve as a suitable biomarker to represent the severity of PD in early and advanced stages.

Argyrophilic grain disease (AGD) is a progressive degenerative disease of the human brain, the prevalence of which increases with advancing age. The features of AGD in autopsied brains from 32 centenarians were studied using phosphorylated tau (AT8) immunostaining combined with Gallyas–Braak staining and 4R tau-specific antibody (RD4) immunostaining. Ten of 32 centenarians were diagnosed as AGD, yielding an overall frequency of 31.3%. In the demented group, nine (39.1%) of 23 cases were found with argyrophilic grains (AGs), while in the non-demented group, AGs were found in only one (11.1%) of nine cases, the difference between them being significant (P

Objective – The clinical differentiation between early Parkinson's disease (PD) and essential tremor (ET) could be difficult, therefore, the aim of this study was to investigate 99mTc-TRODAT-1 SPECT as an applicable tool in this field. Methods – 99mTc-TRODAT-1 single photon emission computed tomography (SPECT) was performed in 10 healthy volunteers, 27 patients with idiopathic PD (Hoehn and Yahr 1–1.5) and 12 patients with ET. The ratio of striatal 99mTc-TRODAT-1 binding was calculated as the index of (striatum − occipital cortex)/occipital cortex. Results – Compared with the striatal 99mTc-TRODAT-1 uptake in the ET group (0.49 ± 0.07) or healthy controls (0.54 ± 0.18), there was a significant decrease in the bilateral striatums of early PD, with a greater reduction in the contralateral striatum (0.27 ± 0.08) than ipsilateral one (0.36 ± 0.10, P

Objectives. Chemokine CXCL12 and its unique receptor CXCR4 have been recently implicated in cancer metastasis. Our goal was to explore expression of CXCL12 and CXCR4 protein in normal ovarian surface epithelium, primary tumors and paired metastases of epithelial ovarian cancer as well as its association with clinicopathological features. We also wanted to test if expression of CXCR4 has prognostic value in epithelial ovarian cancer patients. Method. Sections from 6 normal ovarian surface epithelium, 44 primary epithelial ovarian tumors and 30 paired metastatic tumors in omentum were evaluated for CXCL12 and CXCR4 expression using immunohistochemistry (IHC). Results. All samples of normal ovarian surface epithelium were negative for CXCL12 and CXCR4 protein. Ovarian cancer cells mainly showed cytoplasmic staining of CXCL12 and CXCR4. CXCL12 and CXCR4 staining were detected in 40/44 (91%) and 26/44 (59%) patients with primary epithelial ovarian tumors respectively. CXCR4 expression in primary tumors had no significant correlation with lymph nodes metastasis. However, if we combined CXCR4 expression in primary tumors with metastatic tumors, a significant correlation with lymph nodes metastasis was found ( P = 0.018). The intensity of CXCL12 staining correlated with ascites ( P = 0.014). The rate of CXCR4 expression in refractory and recurrent group (81% versus 28%, P = 0.0008) was significantly higher than that in no-recurrent group. After a median follow-up of 37 months, CXCR4 expression was found associated with an unfavorable prognosis with significantly reduced median disease progression-free survival and overall survival of 15 and 27 months ( P = 0.0004, P = 0.017) respectively. Median time-to-event was not reached in patients with negative CXCR4 staining. In multivariate analysis, CXCR4 expression and residual tumor size emerged as independent prognostic factors in epithelial ovarian cancer patients. Conclusions. This article provides the first evidence that CXCR4 expression could be an independent prognostic factor for epithelial ovarian cancer patients.

To investigate the characteristics of alpha-synucleinopathy in the brains of centenarians, the autopsied brains and spinal cords from 23 cases were studied. Coronal slices were prepared from a section of the cerebral hemisphere, following the guidelines of the Consortium to Establish a Registry for Alzheimer’s Disease (AD) (CERAD) and the consensus guidelines for the clinical and pathologic diagnosis of dementia with Lewy bodies (DLB). Spinal cord specimens were prepared at each segment from the third cervical to the third sacral segment. In all cases, we performed standard stainings of hematoxylin–eosin, Kluver–Barrera, and Gallyas–Braak combined with Luxol fast blue/cresyl violet, and alpha-synuclein (AS), phosphorylated tau (AT8) and beta-amyloid protein immunostainings. One-way ANOVA analysis, Chi-square or Fisher exact test were used for statistical analysis. Overall, AS-positive structures were found in 8 (34.8%) of our 23 centenarians, 6 (35.3%) of 17 demented patients, and four (40%) out of ten AD patients. The frequencies of AS lesions in the brains with senile plaque (SP) stage 0–A, B, and C were 27.7, 33, and 50%, respectively. No statistical differences were found among the frequencies of AS lesions in the subgroups of NFT stages I–II, III–IV, and V–VI (P=0.478). Most cases showed a widespread distribution of AS-positive structures except for one patient, in whose brain only the medulla was involved. The distribution pattern of AS-positive lesions was similar to that in Parkinson’s disease or DLB, but the pigmented neurons in substantia nigra were relatively well preserved. Our findings indicate that there is a high frequency of alpha-synucleinopathy in centenarians, SP-positive and AS-positive lesions may involve a synergistic interaction.