Objective To observe the change of brain iron content in deep gray nucleus using susceptibility weighted imaging (SWI) in patients with Parkinson's diseases (PD). Methods The SWI examination was performed in 40 PD patients (10 patients with Hoehn-Yahr stage Ⅰ , 9 patients with Hoehn-Yahr stage Ⅱ , 9 patients with Hoehn Yahr stage Ⅲ , 6 patients with Hoehn-Yahr stage Ⅳ, 6 ptients with Hoehn Yahr stage Ⅴ ) and 33 gender- and age- matched controls, after conventional brain magnetic resonance imaging examination on a 3.0T magnetic resonance imaging.The signal values of substantia nigra zona compacta (SNc), substantia nigra zona reticulate (SNr),red nucleus (RN), putamen (Pu), globus pallidus (GP) and caudate nucleus (CN) were assessed.Results Compared with the controls, the PD patients had statistically significance of signal value differences of SNc (P=0.002), SNr (P=0.043). RN (P= 0.003), Pu (P=0.023). GP (P=0.001) andCN (P=0.033). The more significant differences of SNc(P=0.001), SNr (P=0.010),RN (P＜0. 001 ), Pu (P=0. 008), GP (P＜0. 001) and CN (P=0. 011) were observed between more severe PD lesion and control. The signal values of SNc and GP showed obviously negative correlations with Hoehn-Yahr grading (SNcr=-0.943. P＜0.001; GPr=-0.923, P＜0.001). But there was weakly correlation of the signal values of SNr, RN, Pu, CN with Hoehn Yahr grading (SNr r=0. 496. P=0.001; RN r=-0. 480. P=0.002; Pu r=-0. 494, P=0.001; CN r=-0.471, P=0.002) Conclusions Measurement of the brain iron content of SNc and GP using SWI on MRI is a reliable means of diagnosing PD, and it has significant correlation with Hoehn-Yahr grading, It could evaluate the severity of PD.

In order to explore the underlying mechanism of high effects and low toxicity of trichostatin A (TSA), the effect of TSA on growth inhibition, histone acetylation level and apoptosis in HL-60 cells and normal human peripheral blood mononuclear cells (NPBMNC) were examined using MTT method, immunocytochemistry technology, and Annexin-V-FITC/PI double staining flow cytometry. The results showed that TSA inhibited growth of HL-60 in time- and dose-dependent manners, and the IC(50) of 36 hours was 100 ng/ml. The apoptosis induction effect of TSA in HL-60 cells was also time- and dose-dependent. Besides, the dose of TSA showing significant apoptotic cytotoxicity in HL-60 cells did not demonstrate apparent apoptosis induction in NPBMNC within definite dose and time range. The histone acetylation level in HL-60 cells and NPBMNC both showed remarkable increase (P < 0.05) after incubated with 100 ng/ml TSA for 4 hours without statistical difference between them is detected (P > 0.05). It is concluded that TSA shows effects of definite and significant growth inhibition and apoptosis induction on HL-60 cells in time- and dose-dependent manners. TSA is able to selectively induce apoptosis in HL-60 cells with low toxicity in NPBMNC at the same time. The mechanism of this selectivity can not be ascribed to the differential regulation of histone acetylation level between HL-60 cells and NPBMNC.
Acetylation ; Apoptosis ; drug effects ; Cell Division ; drug effects ; DNA-Binding Proteins ; HL-60 Cells ; drug effects ; Histone Deacetylases ; physiology ; Histones ; metabolism ; Humans ; Hydroxamic Acids ; pharmacology ; RNA, Messenger ; analysis ; Telomerase ; genetics

Objective To explore the feasibility and efficacy of constructing tissue engineered cartilage in vitro(using) a perfusion-hydrodynamic pressure bioreactor. Methods Chondrocytes isolated from swine's auricular cartilage were seeded onto polyglycolic acid(PGA) to be cultured in a three dimensional environment for 1 week.Then the chondrocyte-polymer constructs were divided into two groups: the experimental group and control group(8 constructs in each group).The experimental group was put into the perfusion-hydrodynamic pressure bioreactor to be cultured for another 3 weeks.The parameters of bioreactor were set as follows: flow rate of 100 mL/min,clockwise and anticlockwise 30 min respectively,on/off 8 h/16h,hydrodynamic pressure of 100 kpa with 0.5 Hz for 4 h/d.The control group was cultured with the routine method.Specimens were harvested and analyzed by gross observation,histology,typeⅡcollagen immunohistochemistry and biochemistry after 4 weeks. Results After 4 weeks,gross observation showed cartilage-like tissue was formed in both groups,and tissue wet weight of experimental group and control group were(191.03?18.55) mg and(130.78?10.33) mg,respectively(P

Objective To study the pharmacokinetics of sodium aesci-nate after intravenous infusion in Chinese healthy volunteers. Methods Sodium aescinate for injection were given to 9 healthy volunteers of a single dose of 10 mg, the concentrations of aescinate in human plasma and urine were determined by HPLC-MS/MS, the main pharmacokinetic parameters were calculated with WinNonlin 5. 0. Results The main pharmacokinetic parameters of A ingredient of aescinate and B ingredient of aescinate were as follows: C_(max) were ( 283. 00 ± 70. 53 ), (206. 33 ± 57.20) ng · mL~(-1); AUC_(0-t) were (1008.05 ±396.49), (638.96 ± 259. 48) ng · h · mL~(-1);t_(1/2) were(3. 72 ±0. 44), (3.57 ±0.48) h;V_z were (19. 39 ±7. 05), (23.82 ±11.43) L;CL were (3. 66 ± 1. 36), (4. 55 ± 1. 86) L · h~(-1) ;36 hour accumulative urine excretion rates were (4. 91 ± 1. 38) % , (2. 80 ± 0. 71 ) % , respectively. Conclusion The disposing process of A ingredient of aescinate and B ingredient of aescinate in healthy subjects were resemble, the elimination half life was about 3. 6 h, the accumulative urine excretion rate was low.

OBJECTIVE Regulating P2x7R- NLRP3 inflammasome activation might be a potentialtherapeutic strategy to treat alcoholic hepatosteatosis. We investigated whether this process would be modulated by gentiopicroside (GPS), which is attributed to the bitterness of gentian root extract. METHODSAn in vivo model was established by intragastrically treating mice with ethanol, and an in vitro modelwas created by treating HepG2 cells with ethanol or treating RAW 264.7 macrophages and murinebone marrow- derived macrophages (BMDMs) with lipopolysaccharides (LPS) plus adenosine triphos-phate (ATP). RESULTS In alcoholic hepatosteatotic mice model, GPS decreased serum aminotrans-ferase and triglyceride accumulation. GPS regulated sterol regulatory element-binding protein-1 (Srebp1),peroxisome proliferators- actived receptors α (PPARα) and acetyl CoA carboxylase (ACC) expressionvia elevating liver kinase B1 (LKB1)/AMP-activated Kinase (AMPK). Suppression of nucleotide-bindingoligomerization domain-like receptor protein 3 (NLRP3), caspase-1 and expression by GPS resulted inthe inhibition of interleukin-1β (IL-1β) production. In ethanol-exposed HepG2 cells, GPS reduced lipo-genesis and promoted lipid oxidation via P2x7R- NLRP3 inflammasome activation. P2x7R silencingenhanced AMPK activity, and reduced Srebp1 expression in ethanol-treated hepatocytes. GPS down-regulated P2x7R-mediated inflammatory response to extracellular ATP in LPS-primed RAW 264.7 macro-phages and BMDMs. Additionally, P2x7R deficiency attenuated IL- 1β cleavage in RAW 264.7 macro-phages, and GPS further suppressed IL-1β cleavage. CONCLUSION Activation of LKB1/AMPK signalingby GPS might be mediated by P2x7R-NLRP3 inflammasome, suggesting a therapeutic utility of P2x7Rblockade in alcoholic hepatosteatosis treatment.

OBJECTIVE The current study was designed to investigate the anti-steatosis effect of Pleurotus citrinopileatus extract (PC) and the underlying mechanism in vivo and in vitro. METHODS Acute and chronic alcoholic hepatosteatosis murine models and ethanol-treated HepG2 cells were applied. RESULTS In vitro,the anti-steatosis effect of PC was further confirmed via Nile red staining in HepG2 cells treated with ethanol.Both of acute and chronic alcohol-induced mice hepatosteatosis model,PC decreased serum aminotransferase and triglyceride accumulation. Upregulated sterol-regulatory element binding protein1(Srebp1),purinergic ligand-gated ion channel 7receptor(P2X7R)and downregulated sirtuin1 (SIRT1), adenosine 5′-monophosphate (AMP)-activated protein kinase α (AMPKα) caused by acute and chronic alcohol intake were modulated by PC.In ethanol-exposed HepG2 cells,PC reduced lipid accumulation in a concentration-dependent manner and exhibited superior ability in controlling lipid accumulation compared with metformin. CONCLUSION PC could abolish hepatic lipid accumulation through regulating SIRT1-AMPKα signaling in acute and chronic alcohol-induced hepatic steatosis.

OBJECTIVE Dihydroquercetin(TAX)is the most abundant dihydroflavone found in on-ions,milk thistle and Douglas fir bark.We investigated whether TAX could inhibit the lipid accumulation in alcoholic liver steatosis in vivo and in vitro.METHODS An in vivo model was established by intragas-trically treating mice with ethanol,and an in vitro model was created by treating HepG2 cells with etha-nol.RESULTS TAX regulated Sterol Regulatory Element-binding Protein-1(SREBP1)and Acetyl CoA Carboxylase (ACC) expression via elevating Liver Kinase B1 (LKB1)/ AMP-activated Kinase (AMPK) phosphorylation. Also, TAX upregulated SIRT1 expression, which suppressed by ethanal intake. Suppression of Purinergic 2X7 receptor (P2x7R), nucleotide-binding oligomerization domain-like re-ceptor protein 3(NLRP3)and Cysteine protease-1(caspase-1)cleavage by TAX resulted in the inhibi-tion of Interleukin-1β(IL-1β) production and release. Additionally, TAX reduced lipogenesis and pro-moted lipid oxidation via the regulation of AMPK and ACC in ethanol-treated steatotic HepG2 cell.TAX downregulated IL-1β cleavage response to Lipopolysaccharides (LPS) plus adenosine triphosphate(ATP) stimulation in HepG2 cells. P2x7R deficiency attenuated lipid accumulation with increasing AMPK activity and decreasing SREBP1 expression in ethanol-treated HepG2 cells.CONCLUSION Our data showed that TAX exhibited the inhibitory properties on lipogenesis and hepatoprotective ca-pacity,indicating that TAX has therapeutic potential for preventing alcoholic liver steatosis.

Objective To assess the value of cardiac magnetic resonance (CMR) imaging in left ventricular structure and function in patients with end stage renal disease (ESRD).Methods Twenty-five patients with ESRD and 10 healthy subjects underwent CMR.Left ventricular end diastolic volume(EDV),end-diastolic diameter(EDD),end-systolic volume(ESV),end-systolic diameter(ESD),stroke volume(SV),ejection fraction(EF),LVM and interventricular septum (IVS) thickness were measured and compared.The parameters from CMR and 2DTTE were compared.Results The EF in patients with ESRD was significantly lower than that in controls (P＜0.001),while ESV,ESD,IVS and LVM were respectively higher than these in controls (P＜0.05).There was no significant difference (P＞0.05) in ESV between CMR and 2DTTE,but EF of CMR was significantly higher than this of 2DTTE (P＜0.05).There was no significant difference (P =0.296) in left ventricular systolic functional category.Bland-Altman plots showed a good agreement between the two methods.Conclusion CMR is a helpful tool to assess left ventricular structure and function in patients with ESRD.

BACKGROUNDActivation of c-Jun NH(2)-terminal kinase (JNK) has been implicated in neuron apoptosis as well as autophagy in response to various stressors after traumatic brain injury (TBI). However, the underlying molecular pathway remains unclear. Our study assessed whether JNK-mediated p53 phosphorylation might be an important mechanism for enhancing neuron autophagy in response to TBI.

METHODSA total of 186 male Sprague-Dawley (SD) rats (300 - 350 g) were used in this study. By randomized block method rats were randomly divided into four groups: sham-operated (n = 46), TBI (n = 60), TBI + dimethyl sulfoxide (DMSO) (n = 40), and TBI + SP600125 (n = 40). JNK was treated with SP600125, a specific JNK inhibitor. JNK, p-P53, Beclin-1, damage-regulated autophagy modulator (DRAM) and p-bcl-2 were evaluated by Western blotting analysis. The cellular localization and expression of Beclin-1 and DRAM was observed by immunofluorescence and immunohistochemistry, and the expression of Beclin-1-Bcl-2/Bcl-xL complexes was evaluated by immunoprecipitation. Multiple-group comparisons were conducted using analysis of variance (ANOVA). P values of less than 0.05 were considered statistically significant.

RESULTSIt was observed that the expression of JNK, p-P53, Beclin-1, DRAM and p-bcl-2 was increasing after TBI, and the expression of Beclin-1 and DRAM was mainly located in the cytoplasm of neurons. But these were significantly inhibited in SP600125 group compared with sham group and TBI + SP600125 group (P < 0.05). The expression of Beclin-1-Bcl-2/Bcl-xL complexes was reduced after TBI.

CONCLUSIONJNK-mediated p53 phosphorylation might be an important mechanism for enhancing neuron autophagy in response to TBI.

Animals ; Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; Beclin-1 ; Blotting, Western ; Brain Injuries ; metabolism ; Fluorescent Antibody Technique ; Hippocampus ; cytology ; metabolism ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Male ; Microscopy, Fluorescence ; Neurons ; cytology ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tumor Suppressor Protein p53 ; metabolism ; bcl-X Protein ; metabolism

OBJECTIVETo study the tumor cell killing function of T lymphocytes stimulated by dendritic cells (DC) and to analyze the differences of protein contents of exosomes in each type of cell.

METHODSThe exosomes of hepatic cell lines with high (P group) or low (F group) metastatic potentials were isolated by a process of four-step centrifugation and the collected exosomes were observed under an electron microscope (EM). The tumor cell killing experiment was performed by adding T lymphocytes activated by DC loaded with exosomes from corresponding P and F group cells and was studied using 3H-TdR experiments. The proteomic analysis was performed by surface-enhanced laser desorption/ ionization time of flight mass spectrometry (SELDI-TOF-MS ) on the exosomes of P and F group cells.

RESULTSThe density distribution and content of exosomes in the P group were not equal to those in the F group observed by EM. The CD80, CD86, MHC-I and MHC-II in the P group were 64.27+5.00, 44.89+10.11, 84.35+19.89 and 59.03+19.37, and those in the F group were 71.53+4.85, 50.01+9.50, 80.68+29.87 and 58.86+21.11, respectively (P>0.05, compared with the control group). The counts per minute value in the P group was 528.40+179.06 and 78.80+24.44 in the F group after being loaded with exosomes (P<0.01, compared with the control group). There were significant differences between the proteins in the exosomes of hepatic cancer cell lines with high or low metastatic potentials.

CONCLUSIONExosomes have potential values of application in immunotherapy and in biotherapy for recurrences and metastases of hepatic carcinomas.

Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Dendritic Cells ; immunology ; metabolism ; Exosomes ; Liver Neoplasms ; metabolism ; pathology ; Lymphocyte Activation ; Male ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes ; immunology ; metabolism