Objective: Sialic acid (SA)-modified chlorogenic acid (CA) liposomes (CA-SAL) was prepared by response surface design to investigate its in vitro cytotoxicity and uptake. Methods: CA-SAL was prepared by a modified reverse-phase ethanol injection method. Sephadex G50 column was used to separate the CA-loaded liposomes and the free CA. The drug concentration was determined by HPLC method and the encapsulation efficiency was calculated. With encapsulation efficiency and drug loading as indicators, Box-Behnken response surface design experiments were used to optimize the prescription process of CA-SAL. The MTT method was used to evaluate the cytotoxicity of CA-SAL on human lung cancer cells A549. Inverted fluorescence microscope was used to investigate the uptake of CA-SAL by A549 cells. Results: The optimized preparation conditions: hydrogenated soybean lecithin-CA ratio at 15:1, hydration temperature 60 ℃, ultrasonic power 400 W. The average particle size of CA-SAL was (90.13 ± 0.51) nm, the polydispersity index (PDI) was 0.16 ± 0.01, the zeta potential was (-25.3 ± 0.5) mV, the encapsulation efficiency was 57.8%, RSD was 0.1%. MTT results showed that the inhibitory effect of CA-SAL on A549 cells was significantly greater than CA-CL. Greater cellular uptake of CA-SAL was observed compared with CA-CL. Conclusion: CA-SAL prepared by response surface optimization has a uniform particle size and good stability. SA-modified CA-loaded liposomes could enhance cellular uptake and cytotoxicity of human lung cancer cell A549 in vitro.

Objective To investigate the time course of cerebral infarction with diffusion kurtosis imaging-related parameters.Methods According to the time interval from symptom onset to MRI examination, 114 cases of cerebral infarction were divided into five groups:8 cases of hyperacute phase(less than 6 hours), 14 cases of acute(>6-24 h), 60 cases of early subacute(>24 h-7 d), 20 cases of late subacute(>7-14 d), and 12 cases of chronic phase ( >14 d-2 months).They underwent routine diffusion weighted imaging (DWI) and diffusion kurtosis imaging (DKI) scanning, and apparent diffusion coefficient ( ADC) and DKI-derived parameters were obtained from them.The derived diffusion parameters were compared among different phases in the patients , and the percent of changes in the infarcted regions were calculated.Paired t-test was used to compare the difference of each parameter between the infarcted region and contralateral normal region , and their correlation with time interval was tested using Pearson correlation analysis.Results Except for chronic phase , mean kurtosis ( MK) , axial kurtosis ( K∥) , radial kurtosis (K⊥)map showed uneven high signal in the infarcted regions , while mean diffusion(MD), axial diffusion(D∥), radial diffusion(D⊥) showed uniform low signal.MK values in the infarcted regions of hyperacute, acute, early subacute and late subacute phase (1.331 ±0.357,1.578 ±0.453,1.519 ±0.455, 1.403 ±0.275 ) increased significantly , compared with the contralateral normal mirror regions ( 0.850 ± 0.236,0.827 ±0.194,0.865 ±0.144,0.939 ±0.212) (t values were 5.242,6.907,12.416,5.629, respectively.P values were all less than 0.01 ).MK, K∥, K⊥ achieved a peak in the acute and early subacute phase , and showed more amplitude than the decrease of MD , D∥, D⊥.Then they gradually reduced, and tended to normalize.MK, MD, ADC had a significant correlation with the time onset of cerebral infarction ( r was 0.354, 0.747, 0.723, respectively, P values were all less than 0.05 ).Conclusion Diffusion kurtosis imaging can provide more diffusion information than conventional DWI , which can better reflect the microstructure changes in tissue.

This study was aimed to investigate the biological behavior of annonaceous acetogenin mimic AA005 in various kinds of leukemia cells and further elaborated its possible mechanisms in acute promyelocytic leukemia (APL) cell line NB4. The proliferative inhibition of leukemia cells was measured by CCK-8 method. Cell death was determined by trypan blue. Cell morphological features of NB4 treated with AA005 were examined by microscopy after Wright's staining. The form of cell death was measured by flow cytometry. Proteins PARP-1 and caspase-3 were detected by Western blot. Flow cytometry was used to detect the cell cycle arrest induced by AA005 of low concentration. The results showed that AA005 (> 200 nmol/L) significantly inhibited proliferation of all tested leukemia cell lines in a concentration-dependent manner. The vast majority of cells went to die after leukemia cell lines of NB4, U937 and K562 were treated with different concentration of AA005 for 48 h. Typical morphologic changes significantly appeared in NB4 cells after AA005 treatment. AA005 almost simultaneously induced early apoptosis and late apoptosis. The little cleavage of PARP-1 and activation of caspase-3 happened in AA005-induced cell death, and caspase-3 inhibitor Z-VAD-fmk could not block the cell death. The non-toxic concentrations of AA005 (< 200 nmol/L) caused NB4 cells G(2)/M-phase arrest. It is concluded that annonaceous acetogenin mimic AA005 induces significant proliferative inhibition of various leukemia cell lines in a concentration-dependent manner, which may be associated with cell death and G(2)/M-phase arrest induced by AA005.
Acetogenins ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Cycle Checkpoints ; Cell Proliferation ; drug effects ; Fatty Alcohols ; pharmacology ; HL-60 Cells ; Humans ; K562 Cells ; Lactones ; pharmacology ; Poly (ADP-Ribose) Polymerase-1 ; Poly(ADP-ribose) Polymerases ; metabolism

To investigate the relationship between the structures of methylhesperetin-7-alkyl ether analogues and their anti-inflammatory activities, nine new compounds, methyl-hesperetin (2), methylhesperetin-7-ethyl ether (3), 7-n-butyl ether (4), 7-n-hexyl ether (5), 7-n-octyl ether (6), 7-n-decyl ether (7), 7-n-dodecyl ether (8), 7-n-tetradecyl ether (9) and 7-n-hexadecyl ether (10), were synthesized with the lead compound of methylhesperidin (1). Their structures were confirmed by UV, 1H NMR, MS and HR-MS spectral data. The in vivo antiinflammatory activities of these compounds were tested on mouse paw edema induced by Freund's complete adjuvant (FCA) and mouse capillary permeability induced by acetic acid with po dose of 300 mg x kg(-1) x d(-1). The result indicated that the anti-inflammatory activities of the synthetic compounds increased firstly and then decreased with the elongating of the length of alkyl chain. After 25-day oral administration of compounds 6, 7 and 8, the inhibitory rates on mouse paw edema of adjuvant arthritis (AA) were 31.9%, 38.5%, 39.1%, respectively. They showed the concentrations of COX-2 in serum of AA mice respectively were 79.3, 75.4, 73.9 ng x L(-1) and the concentrations of PGE2 were in correspondence with 275.4, 258.9, 242.6 ng x L(-1). The inhibitory rates of compounds 6 and 7 on mouse capillary permeability induced by acetic acid were, respectively, 42.4% and 41.5% after 5-day oral administration. Compared with the lead compound of methylhesperidin, the anti-inflammatory activities of compounds 6, 7 and 8 were increased and showed an effective inhibition on the symptom of adjuvant arthritis and capillary permeability in mice.
Acetic Acid ; Animals ; Anti-Inflammatory Agents ; chemical synthesis ; chemistry ; pharmacology ; Arthritis, Experimental ; blood ; chemically induced ; drug therapy ; metabolism ; Capillary Permeability ; drug effects ; Cyclooxygenase 2 ; blood ; Dinoprostone ; metabolism ; Edema ; chemically induced ; drug therapy ; Female ; Freund's Adjuvant ; Hesperidin ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Male ; Mice ; Molecular Structure ; Random Allocation

Objective To investigate the effect on radioactivity in drinking water around Qinshan nuclear power station (QNPS)in normal operational condition.Methods The field monitoring and laboratory analysis methods were adopted to detect the total radioactivity level in drinking water in 2015,according to different distances from the nuclear island and different types of water.Results The total alpha and total beta radioactivity level in drinking water around QNPS were 0.027(0.098)Bq /L and 0.263(0.071)Bq /L respectively,which were obviously lower than the national health standard limits(total alpha and total beta are 0.5,1.0 Bq /L respectively).Total radioactivity level had no relation with the distance from the nuclear island (P >0.05).The total alpha radioactivity in deep well water was the highest among the investigated three types of drinking water,and the highest value was 0.224 Bq /L.The beta radioactivity level in river water was the highest,and the highest value was 0.408 Bq /L.The total alpha radioactivity level was 0.017 (0.013)Bq /L in 2015, higher than the average level during 2010—2014.The beta radioactivity average level was 0.319 (0.102)and 0.289 (0.055)Bq /L,also higher than the average level during 2010—2014.Conclusion The total radioactivity in drinking water among nuclear power plant is in normal background level,so at present there is no effect of the radioactive contamination on drinking water around QNPS in nuclear power plant's normal operational condition.

OBJECTIVETo explore the effect of Jianpi Tongluo Jiedu Recipe (JTJR) on protein expression levels of COX-2, NF-kappaBp65, Bcl-2, and Bax, mRNA expression levels of COX-2 and Bcl-2, and the apoptotic index (Al) in gastric mucosa of patients with precancerous lesions of gastric cancer (PL-GC).

METHODSTotally 65 PLGC patients were recruited and treated by JTJR (modified by syndrome typing), one dose per day for six successive months. Protein expression levels of COX-2, NF-KBp65, Bcl-2, and Bax were detected in 65 patients using immunohistochemical (IHC) assay before and after treatment. mRNA expression levels of COX-2 and Bcl-2 were detected in 54 patients using reverse transcription-polymerase chain reaction (RT-PCR). Meanwhile, changes of Al was detected in 65 patients using TdT-mediated dUTP-biotin nick end labeling (TUNEL) fluorescence method.

RESULTSAfter treatment with JTJR, positive protein expression levels of COX-2, NF-KBp65, and Bcl-2 were obviously decreased in the gastric mucosa of PLGC patients (P <0.01), but Bax positive protein expression was found to be higher (P < 0.05). At the same time mRNA expression levels of COX-2 and Bcl-2 were significantly lower after treatment than before treatment (P < 0.05, P < 0.01); Al also increased after treatment (P < 0.05).

CONCLUSIONJTJR could promote apoptosis possibly via NF-kappaBp65/COX-2, COX-2/Bcl-2, and NF-kappaBp65/Bcl-2 signaling pathways, thereby affecting PLGC patients.

Apoptosis ; Cyclooxygenase 2 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Gastric Mucosa ; metabolism ; Humans ; NF-kappa B ; metabolism ; Precancerous Conditions ; drug therapy ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Signal Transduction ; Stomach Neoplasms ; drug therapy ; metabolism ; bcl-2-Associated X Protein ; metabolism

Objective To observe the value of three-dimensional arterial spin labeling (3D-ASL) PWI in evaluating postoperative cerebral perfusion changes in patients with Moyamoya disease.Methods Totally 19 patients of Moyamoya disease confirmed with DSA were enrolled.All the patients received revascularization.Before and after operation,3D-ASL PWI and dynamic susceptibility contrast perfusion weighted imaging (DSC-PWI) were performed.ROI was located in the region with obvious perfusion changes supplied by middle cerebral artery on the operating side.Then the cerebral blood flow (CBF) was measured on 3D-ASL images,and time to peak (TTP) was measured on DSC-PWI images before and after operation.The differences of CBF and TTP before and after operation were compared,as well as the improvement rate of CBF,TTP and clinical symptoms.Results Before and after operation,CBF was (41.40±11.36) ml/(100 g · min) and (54.10±16.69) ml/(100 g · min),respectively,and the difference was statistically significant (t=-4.273,P＜0.01).TTP was (28.66 ± 3.21) s and (26.44 ± 3.93) s,respectively,and the difference was also statistically significant (t =-2.936,P＜0.01).The improvement rate of clinical symptoms was 84.21％ (16/19),of CBF was 78.95％ (15/19) and of TTP was 68.42％ (13/19),the differences of improvement rate had no statistically significant (P=0.625).Conclusion 3D-ASL PWI is noninvasive,no contrast agent need to be used,and can be used to evaluate perfusion changes after operation of revascularization in patients with Moyamoya disease.

BACKGROUNDPrevious studies have indicated that the cognitive deficits in patients with Alzheimer's disease (AD) may be due to topological deteriorations of the brain network. However, whether the selection of a specific frequency band could impact the topological properties is still not clear. Our hypothesis is that the topological properties of AD patients are also frequency-specific.

METHODSResting state functional magnetic resonance imaging data from 10 right-handed moderate AD patients (mean age: 64.3 years; mean mini mental state examination [MMSE]: 18.0) and 10 age and gender-matched healthy controls (mean age: 63.6 years; mean MMSE: 28.2) were enrolled in this study. The global efficiency, the clustering coefficient (CC), the characteristic path length (CpL), and "small-world" property were calculated in a wide range of thresholds and averaged within each group, at three different frequency bands (0.01-0.06 Hz, 0.06-0.11 Hz, and 0.11-0.25 Hz).

RESULTSAt lower-frequency bands (0.01-0.06 Hz, 0.06-0.11 Hz), the global efficiency, the CC and the "small-world" properties of AD patients decreased compared to controls. While at higher-frequency bands (0.11-0.25 Hz), the CpL was much longer, and the "small-world" property was disrupted in AD, particularly at a higher threshold. The topological properties changed with different frequency bands, suggesting the existence of disrupted global and local functional organization associated with AD.

CONCLUSIONSThis study demonstrates that the topological alterations of large-scale functional brain networks in AD patients are frequency dependent, thus providing fundamental support for optimal frequency selection in future related research.

Aged ; Aged, 80 and over ; Alzheimer Disease ; diagnosis ; Brain ; pathology ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged

Objective:To observe the effects of miR-146a on human retinal endothelial cell (HREC) under high glucose condition.Methods:Total of 57 cases diagnosed as diabetic mellitus and 40 cases with diabetic retinopathy (DR) in Wuxi People's Hospital Affiliated to Nanjing Medical University from October to December 2013.Forty-one healthy volunteers were enrolled and served as control group.The clinical data and venous blood samples of subjects were collected.HRECs were cultured in normal glucose (5.5 mmol/L) or high glucose medium (30 mmol/L). Real-time PCR was used to detect the expression of miR-146a.The cultured HRECs were transfected with miR-146a mimic, mimic negative control, inhibitor and inhibitor negative control by lipofectamine2000, respectively.The expression of miR-146a and intercellular cell adhesion molecule-1 (ICAM-1) mRNA was examined by real-time PCR and the expression of nuclear factor-кB (NF-кB) p65 and NF-кB p65 Ser536 was detected by Western blot assay. Results:The relative expression of miR-146a mRNA in the diabetic mellitus group and DR group was 0.36±0.08 and 0.27±0.08, respectively, which were significantly lower than 1.00±0.16 in the control group (both at P<0.01). The expression of miR-146a mRNA was 0.37±0.11 in the high glucose group, which was lower than 1.00±0.18 in the normal control group ( t=5.57, P<0.01). The relative expression of miR-146a mRNA in the miR-146a mimic group was 2 540.00±105.00, which was significantly higher than 61.00±17.90 in the miR-146a mimic control group; The relative expression of miR-146a mRNA in the miR-146a inhibitor group was 0.04±0.01, which was significantly lower than 0.88±0.04 in the miR-146a inhibitor control group ( t=23.23, 17.12; both at P<0.01). The relative expression of ICAM-1 mRNA in the miR-146a mimic group was 0.35±0.12, which was significantly lower than 1.00±0.13 in the miR-146a mimic control group; The relative expression of ICAM-1 mRNA in the miR-146a inhibitor group was 2.74±0.48, which was significantly higher than 1.00±0.16 in the miR-146a inhibitor control group ( t=3.58, 3.37; both at P<0.05). The relative expression of NF-кB p65 Ser536 in the miR-146a mimic group was 0.43±0.03, which was significantly lower than 1.07±0.09 in the miR-146a mimic control group ( t=6.74, P<0.01). The relative expression of NF-кB p65 Ser536 in the miR-146a inhibitor group was 2.08±0.12, which was significantly higher than 1.00±0.01 in the miR-146a inhibitor control group ( t=8.76; P<0.01). Conclusions:miR-146a can reduce inflammation of HREC in high glucose condition through inhibiting ICAM-1 expression and NF-кB phosphorylation.

AIM To investigate the effects of different compatibility ratios and processing method on the content of rutin,isoquercetin,ferulic acid,quercetin,isotoosendanin,kaempferol,toosendanin,α-pinene,trans-anethole in the combination use of Toosendan Fructus and Foeniculi Fructus,and to explore the optimal compatibility ratio for its use.METHODS The analysis of HPLC-DAD was performed on a 30℃thermostatic ZORBAX SB C18 column(4.6 mm×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1%phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the use of DAD detector.SPSS 24.0 software was used to analyze the data differences.RESULTS Nine constituents showed good linear relationships within their own ranges(r≥0.999 1),whose average recoveries were 96.19%-103.13%with the RSDs of 1.86%-2.67%.Generally higher total content of nine constituents were detected in the combination use groups when Toosendan Fructus-Foeniculi Fructus were at ratios of 1 ∶ 1,1 ∶ 2,and 2 ∶ 1 than those single uses(P<0.05),and among which the 1 ∶ 1 ratio contributed the highest total content.After salt processing,decreased content of toosendanin and isotoosendanin,α-pinene and trans-anethole(P<0.05,P<0.01)),increased isoquercetin content(P<0.01),and no significant content changes of other ingredients were detected.CONCLUSION Through this method of high accuracy and good reproducibility,we learn that the combination use of Toosendan Fructus and Foeniculi Fructus promotes the dissolution of the nine constituents,and the maximum content is achieved at ratio of 1 ∶ 1.