Agrin expression is correlated with tumor development and poor prognosis in cholangiocarcinoma.

OBJECTIVE: This study examined the role of agrin in the development of cholangiocarcinoma (CCA). METHODS: Western blotting was performed to detect the expression of target genes. The correlation between agrin expression and prognosis was analyzed using the Kaplan-Meier method. Proliferation, migration, invasion, and tumorigenesis were examined in CCA cells and tissues using the Cell Counting Kit-8 assay, cell cycle analysis, transwell migration assay, and nude mouse tumorigenicity assay in vivo, respectively. RESULTS: Agrin expression was significantly upregulated in CCA tissues compared with that in adjacent non-tumor tissues, and agrin expression was correlated with poorer tumor characteristics such as portal vein tumor thrombus, intrahepatic metastasis, and worse survival. Forced agrin expression in CCA cells apparently promoted proliferation, colony formation, migration, invasion, and cell cycle progression, but agrin depletion had the opposite effects. Furthermore, agrin-depleted CCA cells developed fewer and smaller tumors than control cells in vivo. Mechanistic analyses indicated that agrin activated the Hippo signaling pathway and induced the translocation of YAP to the nucleus. CONCLUSIONS: Agrin promoted CCA progression by activating the Hippo signaling pathway, suggesting its promise as a target for CCA therapy.

An optically active isochroman-2H-chromene conjugate potently suppresses neuronal oxidative injuries associated with the PI3K/Akt and MAPK signaling pathways.

Increasing evidence suggests that the use of potent neuroprotective agents featured with novel pharmacological mechanism would offer a promising strategy to delay or prevent the progression of neurodegeneration. Here, we provide the first demonstration that the chiral nonracemic isochroman-2H-chromene conjugate JE-133, a novel synthetic 1,3-disubstituted isochroman derivative, possesses superior neuroprotective effect against oxidative injuries. Pretreatment with JE-133 (1-10 µM) concentration-dependently prevented H2O2-induced cell death in SH-SY5Y neuroblastoma cells and rat primary cortical neurons. Pretreatment with JE-133 significantly alleviated H2O2-induced apoptotic changes. These protective effects could not be simply attributed to the direct free radical scavenging as JE-133 had moderate activity in reducing DPPH free radical. Further study revealed that pretreatment with JE-133 (10 µM) significantly decreased the phosphorylation of MAPK pathway proteins, especially ERK and P38, in the neuronal cells. In addition, blocking PI3K/Akt pathway using LY294002 partially counteracted the cell viability-enhancing effect of JE-133. We conclude that JE-133 exerts neuroprotection associated with dual regulative mechanisms and consequently activating cell survival and inhibiting apoptotic changes, which may provide important clues for the development of effective neuroprotective drug lead/candidate.

[Effect of injecting mouse nerve growth factor in different ways on motor development, cerebral hemodynamics and biochemical metabolism in children with cerebral palsy].

OBJECTIVE: To compare the effect of acupoint injection and intramuscular injection with mouse nerve growth factor (mNGF) on gross motor function development of children with cerebral palsy (CP), and explore the treatment mechanism. METHODS: A total of 63 children with CP were randomly divided into an observation group (32 cases, 4 cases dropped off ) and a control group (31 cases, 3 cases dropped off). Based on the routine rehabilitation therapy, the control group received intramuscular injection of mNGF(18 µg/2 mL), and the observation group received acupoint injection of mNGF at Xinshu (BL 15), Ganshu (BL 18), Pishu (BL 20), Shenshu (BL 23), Sanjiaoshu (BL 22), Shenting (GV 24), Baihui (GV 20), Fengfu (GV 16), Dazhui (GV 14), etc. Of them, 5-6 acupoints alternately were selected each time, and each acupoint was given 0.3-0.5 mL, totally 18 µg/2 mL. Both treatment were carried out once every other day for six months. Before and after treatment, the children's development of brain function was assessed using gross motor function classification system (GMFCS). Before treatment (T0), after 2 (T2), 4 (T4) and 6 (T6) months of treatment, the motor function was evaluated by gross motor function measure (GMFM-88). The systolic peak velocity (Vs), mean velocity (Vm) and vascular resistance index (RI) of anterior cerebral artery (ACA) and middle cerebral artery (MCA) were measured, and the level of N-acetyl aspartate acid (NAA), choline (Cho), lactate (Lac) and creatine (Cr) from the basal ganglia, thalamus and periventricular white mater were detected by magnetic resonance spectroscopy (MRS) technology with MAGNETOM Skyra3.0T magnetic resonance imaging system before and after treatment. RESULTS: Compared with before treatment, the GMFCS classification of the observation group after treatment was significantly improved (P<0.05); after treatment, the difference of GMFCS classification between the two groups was not significant (P>0.05), however, the observation group had a 3.142 times of feasibility for good gross motor function development by more than level 1 compared to the control group (P<0.05). After 2, 4, and 6 months of treatment, the GMFM-88 scores of the two groups showed an upward trend (P<0.01), and the increase of the observation group was greater than that of the control group (P<0.05). Compared with before treatment, in the ACA and MCA, the Vs and Vm increased, RI decreased in both groups after treatment (P<0.01), and in the brain, NAA/Cr increased, Cho/Cr and Lac/Cr decreased (P<0.01), and after treatment, the Vs, Vm of ACA and MCA and NAA/Cr of brain in the observation group were higher than those in the control group (P<0.05), and the RI of ACA and MCA and Cho/Cr and Lac/Cr of brain in the observation group were lower than those in the control group (P<0.05). CONCLUSION: The mNGF acupoint injection has a better effect on the gross motor function in the children with cerebral palsy compared with the intramuscular injection, and the mechanism may be associated with exhibiting the double effects of acupoint effect and the targeting therapy of drug, which can effectively improve the cerebral hemodynamics and the metabolism of cerebral nervous substances.

LncRNA SNHG14 promotes the progression of cervical cancer by regulating miR-206/YWHAZ.

Accumulating evidence suggests that lncRNAs play key roles in many cancers. It has been reported that long non-coding RNA SNHG14 promotes cell proliferation and metastasis in multiple cancers. However, the role and underlying molecular mechanism of SNHG14 in cervical cancer (CC) remain largely unclear. In this study, we discovered that the relative expression of SNHG14 was significantly upregulated in CC tissues and cells, and associated with the overall survival of CC patients. Moreover, knockdown of SNHG14 significantly inhibited cell proliferation, migration and invasion, and promoted cell apoptosis in CC. Molecular mechanism explorations revealed that SNHG14 acted as a sponge of miR-206 and that YWHAZ was a downstream target gene of miR-206 in CC. Spearman's correlation analysis uncovered a significantly negative correlation between SNHG14 (or YWHAZ) and miR-206 expression, while a significantly positive correlation between SNHG14 and YWHAZ expression in CC tissues. We also found that the effect of SNHG14 knockdown on the CC progression could be partly rescued by overexpression of YWHAZ at the same time. Our findings revealed that SNHG14 acted as a sponge of miR-206 to regulate the expression of YWHAZ in CC, hinting the promising therapeutic target role of SNHG4 for CC patients.

2-(2-benzofuranyl)-2-imidazoline (2-BFI) improved the impairments in AD rat models by inhibiting oxidative stress, inflammation and apoptosis.

Alzheimer's Disease (AD) is one of the commonest neural degeneration in aging population, and has become a global health challenge. 2-(2-benzofuranyl)-2-imidazoline (2-BFI) was reported to effectively improved the damage of patients with neuropathological disorders. In the present study, we investigated the effect of 2-BFI on the improvement of antioxidative, inflammation, and apoptosis in AD rats. Sprague-Dawley rats (2 months old, n=40) were used in this study and after injection of Aß1-42 into hippocampal CA1 (Cornu Ammonis) region, the rats were given high, moderate and low dose of 2-BFI though intraperitoneal (i.p.) injection. Then spatial memory and navigation ability were analyzed by Morrize Water Maze. For the molecular testing, chemical colorimetry, ELISA and immunoblotting were performed to measure the activities of antioxidative enzymes, the abundance of immune cytokines and expression of apoptotic proteins, respectively. Hematoxylin and Eosin staining was used to analyze the pathological changes. We observed that 2-BFI significantly ameliorated the learning and memory abilities in rat models with AD by dosage treatment, as demonstrated by the shorten learning latency and greater times of travel across the platform quadrant. Additionally, reactive oxygen species (ROS) and malondialdehyde (MDA), were decreased after treatment of 2-BFI with dosage dependency, while the activities of superoxidase dismutase (SOD) and (GPX) Glutathione peroxidase were in turn enhanced, suggesting that 2-BFI could protect the antioxidative enzymes and reduce the oxidative stress in the hippocampus. Moreover, the expression of inflammatory factors including TNF-a and IL-1ß were decreased after 2-BFI treatment. Additionally, the neuronal apoptosis was also attenuated, as shown by Western blot results. Taken together, the cognitive impairment in AD rats could be significantly improved by 2-BFI in a dose-dependent manner through suppressing oxidants accumulation, inhibiting of inflammatory response, as well as enhancing the neural viability.

Design, synthesis and biological evaluation of novel chiral oxazino-indoles as potential and selective neuroprotective agents against Aß25-35-induced neuronal damage.

A series of chiral oxazino-indoles have been synthesized via a key intermolecular oxa-Pictet-Spengler reaction. These compounds exhibited significant and selective neuroprotective effects against Aß25-35-induced neuronal damage. This is the first report of evaluating the influence of chiral diversity of oxazino-indoles on their neuroprotective activities, with the structure-activity relationship been analyzed. The highly active compounds 3f, 3g, 4g, 4h, and 6b all performed over 90% cell protection, providing a new direction for the development of neuroprotective agents against Alzheimer's disease.

[Degradation of PCDD/Fs by the Mixture of V2O5-WO3/TiO2 Catalyst and Activated Carbon].

The mixture of V2O5-WO3/TiO2 catalyst and two kinds of Activated Carbons (AC) (AC-1: based on lignite; AC-2: based on coconut shell) was used to destroy gas phase PCDD/Fs with high concentration (9. 80 ng.m-3, evaluated by international toxic equivalence quantity (I-TEQ) under low thermal temperature (160°C) based on a dioxin generating system. After mixing with AC, removal efficiency (RE) and destruction efficiency (DE) of PCDD/Fs increased by 20% compared with only catalyst condition. In comparison with mixture of AC based on coconut shell, mixture of AC based on lignite had lower RE-values and higher DE-values. The adjustments of the ratio of catalyst and AC could cause the different degradation effects, and RE-values increased and DE-values decreased with increasing proportions of catalyst. When the volume fraction of oxygen was 0% in experimental atmosphere, catalyst could lose its activity and most PCDD/Fs were not oxidized but adsorbed by the mixture. RE and DE-values increased with increasing content of oxygen. The addition of ozone (concentration of 200 mg.m-3) could improve catalytic oxidation effects to a certain degree. However, ozone might react with AC, which could influence the lifetime of the mixture. Under 200°C, the mixture with proportion of AC: catalyst = 1:1 and in the present of 200 mg.m-3 ozone conditions, the highest RE and DE-value were obtained with 98. 0% and 94. 8% respectively, and the concentration of PCDD/Fs residual in off-gas was only 0. 51 ng.m-3 evaluated by I-TEQ.

Treating PCDD/Fs by combined catalysis and activated carbon adsorption.

V2O5-WO3/TiO2 catalysts are used to destroy dioxins present in the gas phase, yet both their removal efficiency (RE) and destruction efficiency (DE) decrease with rising initial concentration (IC). Therefore, activated carbons (AC-1: based on lignite; AC-2: based on coconut shell) were mixed with the catalyst to tackle these high IC gases. A gas phase dioxin-generating system was used to supply three different stable IC-values. When the highest IC is used (20.5 ng I-TEQ Nm(-3)) without AC, at 200°C, the RE and DE-value of PCDD/Fs reaches only 76% and 64%, respectively. At the same conditions, using a mix of catalyst and AC-2, these RE and DE-values rise to 90.1% and 82.0%, respectively. The mix catalyst/AC also shows better performance at low temperature (160 and 180°C). The AC characteristics influence upon the adsorption and degradation abilities of the mixtures.

[Optimize extraction technology of Fufang Bajitian Shenggu Particles by central composite design and response surface method].

OBJECTIVE: To optimize extraction process of Fufang Bajitian Shenggu Particles by central composite design and response surface method. METHODS: The preparation of Fufang Bajitian Shenggu Particles was designed according to the test. With the overall desirability (OD) of Icariin content and the dry extract yield in medicinal materials extract concentrate as examining indexes, immersion time, material liquid ratio, extraction time and stress concentration temperature were investigated. Based on the single factor tests, material liquid ratio, extraction time and stress concentration temperature which impacted response values significantly were investigated by three factors and five levels of central composite design. RESULTS: Optimum extraction technology of Fufang Bajitian Shenggu Particles were as follows: material liquid ratio was 1:11.33, extraction time was 39 min, and stress concentration temperature was 66 °C. Bias of the lcariin content and the dry extract yield between observed and predicted values were 1.60% and 1.55% ,respectively. CONCLUSIONS: Using central composite design and response surface method to optimize extraction of Fufang Bajitian Shenggu particles has a good prediction.

[COX-2 inhibitor celecoxib can suppress the proliferation of FLT3-ITD positive acute myeloid leukemia cells with prominent down regulation of MEK/MCL-1 expression in vitro].

The purpose of this study was to investigate the effects of Celecoxib on the proliferation of the FLT3-ITD positive and negative acute myeloid leukemia cells and its mechanism. The proliferation inhibition effect of Celecoxib with different doses on the FLT3-ITD positive cells MV4-11 and the FLT3-ITD negative K562 cells was detected by CCK-8 method, the cell apoptosis was determined by flow cytometry, and the MEK, Mcl-1, pAKT expression was tested by Western blot. The results showed that Celecoxib inhibited the proliferation of both MV4-11 and K562 cells, but the IC50 for MV4-11 was (29.14 ± 2.4) µmol/L, which was significantly lower than that of K562 cells (39.84 ± 1.0) µmol/L (P < 0.05); The induced apoptosis rate of Celecoxib at 20-80 µmol/L on MV4-11 was not observed, but there was apparent influence on K562 at the same concentration. Western blot showed that Celecoxib down-regulated the expression of MEK and Mcl-1 but did not change the expression of pAKT obviously on MV4-11 cells, while the expression of Mcl-1 was reduced a little, but no obvious change were found in the expression of MEK and pAKT on K562 cells. It is concluded that the Celecoxib can inhibit the proliferation of FLT3-ITD positive AML cells distinctly, and the potential mechanism may be related to the inhibition of the MEK/Mcl-1 signaling pathway.

Ozone-enhanced oxidation of PCDD/Fs over V(2)O(5)-TiO(2)-based catalyst.

The catalytic oxidation of PCDD/Fs (polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans) vapors was studied in a temperature range of 180-220°C on a honeycomb V2O5-TiO2-based catalyst, in the presence and the absence of ozone. A stable dioxin-generating system was established to support the experimental program and this system could adjust the concentration of PCDD/Fs by injecting appropriate mother liquors. At 220°C the removal efficiency(1) (RE) of PCDD/Fs reaches up to 97% and the degradation efficiency (DE) up to 90%. Both values diminish at lower operating temperatures. In the presence of ozone, however, these values rise to 99% and 98% at 220°C. Especially at low temperatures the effect of ozone is obvious. Catalytic oxidation with ozone thus offers a low-temperature solution to achieve higher rates and low activation energies. The morphology and microstructure of the catalysts changes after ozone treatment and some of their characteristics seem closely related with DE-values.

COX-2 inhibitor celecoxib can suppress the proliferation of FLT3-ITD positive acute myeloid leukemia cells with prominent down regulation of MEK/MCL-1 expression in vitro / 中国实验血液学杂志

The purpose of this study was to investigate the effects of Celecoxib on the proliferation of the FLT3-ITD positive and negative acute myeloid leukemia cells and its mechanism. The proliferation inhibition effect of Celecoxib with different doses on the FLT3-ITD positive cells MV4-11 and the FLT3-ITD negative K562 cells was detected by CCK-8 method, the cell apoptosis was determined by flow cytometry, and the MEK, Mcl-1, pAKT expression was tested by Western blot. The results showed that Celecoxib inhibited the proliferation of both MV4-11 and K562 cells, but the IC50 for MV4-11 was (29.14 ± 2.4) µmol/L, which was significantly lower than that of K562 cells (39.84 ± 1.0) µmol/L (P < 0.05); The induced apoptosis rate of Celecoxib at 20-80 µmol/L on MV4-11 was not observed, but there was apparent influence on K562 at the same concentration. Western blot showed that Celecoxib down-regulated the expression of MEK and Mcl-1 but did not change the expression of pAKT obviously on MV4-11 cells, while the expression of Mcl-1 was reduced a little, but no obvious change were found in the expression of MEK and pAKT on K562 cells. It is concluded that the Celecoxib can inhibit the proliferation of FLT3-ITD positive AML cells distinctly, and the potential mechanism may be related to the inhibition of the MEK/Mcl-1 signaling pathway.

[Low-temperature thermal treatment of dioxin in medical waste fly ash under unert atmosphere].

Two kinds of fly ash (AG and BG) from hazardous waste rotary kiln were investigated as the targets of thermal treatment. AG was sampled after fabric filter with activated carbon spray and BG was sampled in pipe before fabric filter. The effects of temperature and time on PCDD/Fs degradation rate in fly ash were investigated in quartz tubular. Under nitrogen atmosphere, low-temperature thermal treatment for dioxin degradation had obvious effect. For sample BG, the degradation rate of dioxin in solid phase was 82.8%-99.9%, and the degradation rate of I-TEQ was 77.3%-99.8%. For sample AG, the degradation rate was 66.8% - 99.8%, and the degradation rate of I-TEQ was 43.5% - 99.6%. Although dioxin in solid phase was reduced, it was generated in gas phase, and among all the different temperatures tested, dioxin was generated at highest amounts at 300 degrees C - 350 degrees C, and among all the toxic congeners, OCDD was detected as the most abundant in this experiment. The best conditions of thermal treatment were: heating time 60 min, temperature 400 degrees C, under nitrogen atmosphere. The results indicate that low-temperature thermal treatment under inert atmosphere has good effect on treating dioxin present in medical fly ash, it can be applied in practical projects in large scales.

2-(-2-benzofuranyl)-2-imidazoline induces Bcl-2 expression and provides neuroprotection against transient cerebral ischemia in rats.

Stroke is the third leading cause of death and disability in North America and is becoming the most frequent cause of death in the rapid developing China. Protecting neurons in order to minimize brain damage represents an effective approach towards stroke therapeutics. Our recent study demonstrated that 2-(-2-benzofuranyl)-2-imidazoline (2-BFI), a ligand for imidazoline I(2) receptors, is potently neuroprotective against stroke, possibly through transiently antagonizing NMDA receptor activities. In this study, we further investigated the characteristics and mechanisms of 2-BFI-mediated neuroprotection using a rat stroke model of transient occlusion of the middle cerebral artery. Here, we show that 2-BFI was most effective at the dose of 3mg/kg in vivo, with significantly reduced brain infarct size and improved neurological deficits. Lower doses of 2-BFI at 1.5mg/kg, or higher dose of 2-BFI at 6 mg/kg, were either not effective, or toxic to the brain, respectively. Treating stroke rats with 3mg/kg 2-BFI significantly reduced the number of TUNEL positive cells and preserved the integrity of subcellular structures such as nuclear membranes and mitochondria as shown under the electron microscope, confirming neuroprotection. Most interestingly, 2-BFI-treated brains exhibited significant expression of Bcl-2, a gene with a known function in neuroprotection. Taken together, these studies not only demonstrated that 2-BFI at 3mg/kg was effective in neuroprotection, but also, for the first time, showed that 2-BFI provided neuroprotection through up-regulating the neuroprotective gene Bcl-2. 2-BFI can be further developed as a therapeutic drug for stroke treatment.