A simple and efficient method for fabricating a novel surface-enhanced Raman scattering ( SERS) substrate with good reproducibility and high SERS activity was reported. Cu2 O was prepared by mixing CuCl2 ·2H2 O with ascorbic acid, which was then used as the templates for depositing of gold nanoparticles ( AuNPs) on their surfaces, forming Cu2 O@ Au with heterostructures. Transmission electron microscopy, scanning electron microscopy and X-ray diffraction observation revealed that Cu2 O had polyhedral structure and smooth surface, and AuNPs were closely deposited on the surface of Cu2 O. It was used as SERS substrate for detection of Rhodamine B with linear detection range of 1 × 10-2-5 × 10-6 mol/L, and detection limit of 3 ×10-7 mol/L. Cu2 O@Au showed good chemical stability, remained stable in acid, PBS and river samples, and could be used in the SERS detection of target in water sample.

Objective To explore the application of diffusion weighted imaging(DWI) in diagnosis of viral encephalitis in children.Methods Conventional MR imaging(T2WI and T1WI) and DWI were performed on 20 patients with viral encephalitis diagnosed clinically.Location and number of lesions demonstrated on these imagings and percents in their abnormality were compared.Results Percen-(tage of abnormality) demonstrated on DWI was significantly higher than that on T1WI(?~2=4.44 P

OBJECTIVETo compare the features of brain injury in neonatal rats with different severities of hypoxia-ischemia (HI), and explore the role of microglial activation and cytokines.

METHODSOne hundred and twenty 7-day-old rats were randomized to three groups: sham control, mild HI, and severe HI. The rats in the HI groups were subjected to right carotid artery occlusion and 8% oxygen hypoxia exposure (40 minutes, 34.5 Celsius degree in the mild HI group; 65 minutes, 35.5 Celsius degree in the severe HI group). MRI, microtubule associated protein (MAP2) and TUNEL staining were used to confirm the severity of brain injury. Changes in expression of activated microglia (ED1) and signs of cytokine involvement or oxidative stress (TNF-alpha, nitrotyrosine) were assessed immunohistochemically.

RESULTSIn the mild HI group, MRI scans demonstrated increased T2 values in the ipsilateral subcortical white matter and a slight loss of T2 values in the cortex, corresponding to a medium loss of MAP2 in the ipsilateral cortex. There was an increase in the number of TUNEL positive cells compared to the control group within the subcortical white matter. In the severe HI group, the T2 value increased in the majority of the hemisphere, corresponding to a severe loss of staining for MAP2 in the ispilateral hemisphere. The number of TUNEL positive cells significantly increased in the ipsilateral cortex and white matter. In the mild HI group, ED1, TNF-alpha and nitrotyrosine expression increased only in the acute stage and was only observed in subcortical white matter. In contrast, after severe HI, the increase in ED1, TNF-alpha and nitrotyrosine expression was observed in the whole ipsilateral hemisphere and prolonged for weeks.

CONCLUSIONSFollowing a mild HI a relatively selective white matter injury compares to the pannecrosis in the cortex and white matter following a severe HI. Microglial activation and over-expression of cytokines might contribute to the development of hypoxic-ischemic brain damage.

Animals ; Animals, Newborn ; Apoptosis ; Hypoxia-Ischemia, Brain ; etiology ; metabolism ; pathology ; Magnetic Resonance Imaging ; Microglia ; pathology ; Microtubule-Associated Proteins ; analysis ; Rats ; Tumor Necrosis Factor-alpha ; analysis ; Tyrosine ; analogs & derivatives ; analysis

BACKGROUNDLipid abnormalities are often complicated by renal dysfunction. 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) are the first-line choice for lowering cholesterol levels. The present study was designed to investigate whether statins could prevent and invert the development of renal injury in cholesterol-fed rabbits and to find the possible mechanism of their effects by detecting gene and protein expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) in the renal artery.

METHODSTwenty-four male New Zealand white rabbits were divided into three groups: (1) control group, regular granules chow; (2) HC-diet group, granules chow with 1% cholesterol and 5% lard oil; and (3) fluvastatin group, 1% cholesterol and 5% lard oil diet plus fluvastatin [10 mg.kg(-1).d(-1)]. After 16 weeks, serum total cholesterol (TC), low-density lipoprotein (LDL) and creatinine (Cr) levels were measured. Renal hemodynamics and function, mainly including glomerular filtration rate (GFR) in vivo were quantified using (99m)Tc-DTPA single photon emission computed tomograph ((99m)Tc-DTPA SPECT). The thickness of the renal artery intima was quantitated in HE-stained segments by histomorphometry. Gene expression of LOX-1 in the renal artery was examined by semi-quantitative RT-PCR and its protein expression was evaluated by immunohistochemistry.

RESULTSHigh cholesterol diet induced hypercholesterolemia (HC) complicated by renal dysfunction with increased levels of serum lipid and Cr, decreased GFR and delayed excretion and extensively thickened renal arterial intima in the HC-diet group. Rabbits in the control group showed a minimal LOX-1 expression (mRNA and protein) in the endothelium and neointima of the renal artery. Intimal proliferation of the renal artery in the HC-diet group was associated with a marked increase of LOX-1 expression (protein and mRNA). Treatment with fluvastatin improved renal function, attenuated intimal proliferation of the renal artery and markedly decreased the enhanced LOX-1 expression in the endothelium and neointima of the renal artery in rabbits.

CONCLUSIONSFluvastatin treatment could prevent the development of renal injury in patients with HC and early atherosclerosis (AS). This beneficial effect might be mediated by its pleiotropic effects including a decrease in total cholesterol exposure level and prevention of LOX-1 expression in atherosclerotic arteries.

Animals ; Cholesterol ; blood ; Creatinine ; blood ; Fatty Acids, Monounsaturated ; pharmacology ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; pharmacology ; Hypercholesterolemia ; drug therapy ; metabolism ; pathology ; Immunohistochemistry ; Indoles ; pharmacology ; Kidney ; drug effects ; pathology ; Male ; RNA, Messenger ; analysis ; Rabbits ; Receptors, LDL ; analysis ; genetics ; Receptors, Oxidized LDL ; Scavenger Receptors, Class E ; Tomography, Emission-Computed, Single-Photon

This paper is to report the study of the metabolism of lidamycin in vitro including in plasma and microsomes to guide clinical therapy. Lidamycin was quantified by detecting its active ingredient using HPLC-MS/MS. The metabolic stability of lidamycin in rat, Beagle dog, monkey and human plasma and liver microsomes, and its inhibition to cytochrome P450 isoforms in human liver microsomes were studied. Results showed that lidamycin was metabolized in the four species of plasma, and the sequence of metabolic rates in plasma were in rat > in dog > in human > in monkey. But among the four species of liver microsomes, lidamycin was metabolized only in monkey liver microsomes. There was almost no inhibition to cytochrome P450 isoforms at the concentrations of between 0.0005 and 10 ng x mL(-1). Therefore, the property of lidamycin metabolism in human is similar with that in dog, and metabolism of other drugs would not be decreased by cytochrome P450 as used along with lidamycin in clinic.
Aminoglycosides ; blood ; metabolism ; Animals ; Antibiotics, Antineoplastic ; blood ; metabolism ; Chromatography, High Pressure Liquid ; Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Dogs ; Enediynes ; blood ; metabolism ; Enzyme Activation ; Humans ; Macaca ; Microsomes, Liver ; metabolism ; Rats ; Tandem Mass Spectrometry

OBJECTIVETo investigate the effect of angiotensin II and angiotensin1-7 on alpha-smooth muscle actin (alpha-SMA)-induced Ca(2+)-independent pathways mediated by Rho kinase2 in hepatic stellate cells (HSCs).

METHODSHSC-T6 cells were treated with 10 micromol/L of AngII, Ang1-7, AngII +Ang1-7, and Ang1-7+A779. RT-PCR was used to detect the expression of Rho kinase2 (Rock2) in Ca(2+)-independent pathways, and alpha-SMA protein expression was detected by Western blotting.

RESULTSThe mRNA expression of Rock2 increased significantly in the cells after AngII treatment (P<0.01), but decreased following Ang1-7 treatment. Ang1-7 treatment significantly reduced alpha-SMA level in AngII-induced cells (P<0.01).

CONCLUSIONAng1-7 can inhibit AngII-induced activation of Rock2 and reduce alpha-SMA expression in HSCs.

Actins ; metabolism ; Angiotensin I ; pharmacology ; Angiotensin II ; pharmacology ; Animals ; Calcium ; metabolism ; Cell Line ; Gene Expression Regulation ; drug effects ; Hepatic Stellate Cells ; cytology ; drug effects ; metabolism ; Peptide Fragments ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Signal Transduction ; drug effects ; rho-Associated Kinases ; genetics

OBJECTIVETo investigate the effect of angiotensin II (AngII) type 1 (AT-1) receptor and angiotensin-converting enzyme (ACE) gene silencing on nuclear factor-kappaB (NF-kappaB) activity in hepatic stellate cells (HSCs).

METHODSpSilencer/AT-1 alpha receptor siRNA and pSilencer/ACE siRNA plasmids were transfected into cultured HSC-T6 cells, which were subsequently stimulated by 10(-6) mol/L AngII or ACE inhibitor (ACEI). The DNA binding activity of NF-kappaB in the transfected cells was analyzed using electrophoretic gel mobility shift assay (EMSA).

RESULTSs Gel shift studies showed that stimulation of the HSCs by AngII markedly increased the DNA-binding activity of NF-kappaB, which was inhibited by the transfection with pSilencer/ AT-1 alpha receptor siRNA plasmid or pSilencer/ACE siRNA plasmid.

CONCLUSIONAT-1 alpha receptor and ACE gene silencing result in inhibition of NF-kappaB activity in HSCs in vitro.

Cell Line ; Hepatic Stellate Cells ; cytology ; metabolism ; Humans ; NF-kappa B ; genetics ; metabolism ; Peptidyl-Dipeptidase A ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Receptor, Angiotensin, Type 1 ; genetics ; Transfection

OBJECTIVETo observe the effect of diet-control only, diet-control with swimming training or with polysaccharide sulfate (PSS), a kind of blood lipid-lowering drug on the serum lipid level and vascular endothelial function in obese rats fed by fat-rich-diet.

METHODSForty Wistar rats were randomly divided into the following 5 groups: group F (n = 8), group N (n = 8), group A (n = 8), group B (n = 8) and group C (n = 8), where the rats were given fat-rich-diet, basic-diet, 12 weeks of diet-control after 8 weeks of fat-rich-diet, 12 weeks of diet-control with swimming training after 8 weeks of fat-rich-diet and 12 weeks of diet-control with PSS after 8 weeks of fat-rich-diet, respectively. All rats were sacrificed after 12 weeks of intervention. Then the levels of Lee index, serum total cholesterol (TC), triglyceride (TG), plasma endothelin (ET), nitric oxide (NO) and von Willebrand Factor (vWF) were measured. The protein expression of intercellular adhesion molecule-1 (ICAM-1) in artery endothelium was evaluated by immunohistochemistry (IHC) and the gene expression of ICAM-1 was examined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSAfter the interventions for 12 weeks, the levels of serum TC, TG and ET decreased in group A (P < 0.05). The levels of Lee index, TC, TG, ET, vWF, ICAM-1 protein and ICAM-1 mRNA decreased in group B and C (P < 0.05). Three interventions increased serum NO production (P < 0.05) in group B.

CONCLUSIONSDiet-control could a meliorate the hyperlipidemia and vascular function. Diet-control with swimming training and diet-control with PSS could result in weight loss of rats and meliorate the hyperlipidemia, vascular endothelial function, coagulatory activities and adhesive dysfunction. The effects of diet-control with swimming on vascular endothelial function were prominent.

Animals ; Cholesterol ; blood ; Combined Modality Therapy ; methods ; Diet ; Dietary Fats ; adverse effects ; Disease Models, Animal ; Endothelins ; blood ; Endothelium, Vascular ; cytology ; drug effects ; metabolism ; Immunohistochemistry ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Nitric Oxide ; blood ; Obesity ; diet therapy ; metabolism ; therapy ; Polysaccharides ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Swimming ; Treatment Outcome ; Triglycerides ; blood ; von Willebrand Factor ; analysis

There are sex differences in some brain areas in mammalians. Parkinson's disease (PD), caused by the mesencephalic substantia nigra (SN) dopaminergic neuronal loss, displays sexual difference, i.e., the incidence is higher and the symptoms are more intense in males than that in females. However, it has not been known whether sexual dimorphisms exist in the SN. Sixty adult Sprague-Dawley rats were randomly divided into 5 groups: (1) Female intact group (F-INT group); (2) Male intact group (M-INT group); (3) Ovariectomized group (OVX group); (4) Castration group (CAST group); (5) Ovariectomized + estrogen-replaced group (OVX-E(2) group): The rats received sequentially physiological dose of estrogen for 3 d from the 7th day after ovariectomization. P50 auditory evoked potential (P50) was recorded for 14 d from electrodes inserted in the rat right SN in quiet and awake state. After recording, the brain tissues were dissected and the tyrosine hydroxylase (TH)-expressing neurons in the compact zone of the SN were counted using immunohistochemical method. The results showed that the number of TH-positive (TH(+)) cells in the SN of normal male animals was less than that in normal female rats (P<0.05), and the T/C ratio of P50 in normal males was significantly less than that in normal females (P<0.01), indicating that there exists sexual difference in function and structure in the SN. No differences in the T/C ratio of P50 or the number of TH(+) cells were found between M-INT and CAST groups. The T/C ratio of P50 and the number of TH(+) cells in the SN in OVX group were reduced significantly compared with those in F-INT group (P<0.01). There was no significant difference in the T/C ratio of P50 and the number of TH(+) cells in the SN between OVX- E(2) and F-INT groups 15-20 d after estrogen replacement, suggesting that estrogen can promote the survival and functional recovery of dopaminergic neurons in the SN. The results suggest that there exist sex-specific differences in the dopaminergic neurons in the SN structurally and functionally. The difference of estrogen level in cerebra between male and female animals may account for the sexual differences. Endogenous estrogen plays an important role in maintaining the integrity and modulating the functional activity of dopamine system in the SN.
Animals ; Dopaminergic Neurons ; cytology ; Estrogens ; pharmacology ; Evoked Potentials, Auditory ; Female ; Male ; Orchiectomy ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Sex Characteristics ; Substantia Nigra ; cytology

OBJECTIVETo investigate the signal transduction mechanism underlying the effects of angiotensin II (AngII) on extracellular signal-regulated kinase 1/2 (ERK1/2), early growth response-1 (EGR-1) and platelet-derived growth factor-B (PDGF-B) in hepatic stellate cells (HSCs).

METHODSHSC-T6 cells treated with AngII for 10 or 30 min were examined for phospho-P42/44 protein expression using Western blotting. In another experiment, the cells were preincubated for 1 h in the presence of U0126 (an inhibitor of the MAPK/ERK kinase), irbesartan (an AT-1 receptor blocker), or antioxidant-N-acetylcysteine (NAC) prior to AngII exposure, and the protein expression of phospho-P42/44 and PDGF-B were measured with Western blotting. The DNA binding activity of EGR-1 was analyzed using electrophoretic gel mobility shift assay (EMSA), and the expression of PDGF-B was detected immunohistochemically.

RESULTSAngII induced phospho-P42/44 expression in HSC-T6, which was abrogated by U0126 or irbesartan. NAC did not inhibit phospho-P42/44 expression. EMSA showed that AngII exposure of the HSC cells markedly increased EGR-1 DNA binding activity, reaching the maximum after 60 min of exposure followed by progressive declination; irbesartan and U0126 significantly suppressed AngII-induced EGR-1 activity enhancement. ACEI at 1 micromol/L and 10 nmol/L inhibited EGR-1 activity, but ACEI at the concentration of 0.1 nmol/L resulted in enhanced EGR-1 activity. NAC showed no obvious effect in suppressing EGR-1 activity. AngII increased PDGF-B protein level in the HSCs, the effect of which was inhibited by irbesartan. U0126, NAC and ACEI did not attenuate PDGF-BB protein level in the HSCs.

CONCLUSIONStimulation of the HSCs with AngII results in EGR-1 activation via the ERK1/2 pathway, leading to up-regulation of PDGF-B expression.

Angiotensin II ; pharmacology ; Blotting, Western ; Cells, Cultured ; Early Growth Response Protein 1 ; metabolism ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Hepatic Stellate Cells ; cytology ; drug effects ; metabolism ; Humans ; Immunohistochemistry ; Platelet-Derived Growth Factor ; biosynthesis ; Proto-Oncogene Proteins c-sis ; Signal Transduction ; drug effects