Objective • To detect the effects of propofol sedation on cognitive function in rats and its mechanism. Methods • Forty-eight SD rats were randomly divided into three groups, i.e. control group, 100 mg/kg group and 300 mg/kg group. Rats were administrated intraperitoneally with propofol (10 mg/mL, 100 mg/kg or 300 mg/kg). The mRNA levels of brain derived neurotropic factor (BDNF)-TrkB/p75 signal molecules in rat hippocampus were evaluated by realtime PCR 45 min after propofol treatment. Learning and memory ability was examined by inhibitory avoidance (IA) test after propofol treatment. Results • The mRNA levels of BDNF in the hippocampal tissue were (1.20±0.13) fold (P=0.002) and (88±12)% (P=0.044) of that in control group, respectively, in 100 mg/kg group and 300 mg/kg group after injection of propofol. The mRNA levels of TrkB were (1.01±0.11) fold (P=0.982) and (86±11)% (P=0.018) of that in control group, respectively, in 100 mg/kg group and 300 mg/kg group. The mRNA levels of p75 were (1.02±0.10) fold (P=0.778) and (1.59±0.18) fold (P=0.000) of that in control group, respectively, in 100 mg/kg group and 300 mg/kg group. There was no significant difference of the 24 h IA memory retention latency between 100 mg/kg group and control group. The 24 h IA memory retention latency in 300 mg/kg group was significantly decreased compared with control group (P=0.028) and 100 mg/kg group (P=0.020). Conclusion • Propofol dose-dependently regulates the expression of BDNF-TrkB/p75 signal molecules, and high dose propofol may reduce cognitive function via BDNF-TrkB/p75 signal.

Objective: To detect the effects of propofol on rat hippocampal astrocytes and clarify its mechanism. Methods: According to the time after propofol injection, twenty-four SD rats were randomly divided into three groups, i. e. 0 min, 45 min and 90 min group. Rats were administrated intraperitoneally with propofol (10 mg/mL, 100 mg/kg body weight). The levels of glial fibrillary acidic protein (GFAP) and S100β mRNA in rat hippocampus were evaluated by realtime PCR. And cell viabilities and levels of GFAP mRNA were examined in primary cultured hippocampal astrocytes induced by 10 μmol/L propofol with or without 10 μmol/L extracellular signal-regulated kinase (ERK) inhibitor PD98059 pretreatment. Results: The mRNA levels of GFAP in the hippocampal tissue were (1.32±0.12) times (P=0.000) and (1.12±0.09) times (P=0.012) that in 0 min group, respectively, 45 min and 90 min after injection of propofol. The mRNA levels of S100β in the hippocampal tissue were (1.14±0.11) times (P=0.005) and (1.05±0.10) times (P=0.284) that in 0 min group, respectively, 45 min and 90 min after injection of propofol. The mRNA levels of GFAP and S100β were timedependently altered, first increasing, and then decreasing. In vitro, the cell viabilities (P=0.041) and levels of GFAP mRNA (P=0.026) in primary cultured hippocampal astrocytes were significantly elevated after propofol treatment, and these effects of propofol were reversed by ERK inhibitor PD98059. Conclusion: Propofol time-dependently upregulated the expression of GFAP and S100β via ERK signaling pathway in rat hippocampal astrocytes, so as to activate astrocytes.

Objective To study the status of preoperation requirements and its fulfillment of parents of congenital heart disease children.Methods A total of 105 parents of congenital heart disease children were recruited and were investigated with the Critical Care Family Needs Inventory (CCFNI) and the Needs Met Inventory ( NMI ) on one day before children received the operation.Results Totals of 100 questionnaires were collected and the total average score of the patients' preoperation requirements was( 3.24 ±0.46)and the mean score of fulfillment was( 2.26 ± 0.41 ).The high requirements dimensions were the requirement of enhancing the confidence and reducing anxiety ( 3.70 ± 0.29 ),the requirement of getting information ( 3.58 ± 0.45 ),the requirement of accessing to children ( 3.48 ± 0.67 ),while the fulfillment score of the requirement of getting information and accessing to children was (2.24 ±0.47) and (2.28 ±0.46).Multiple stepwise regression analysis showed that the patients' preoperation requirements was related with their age and education background (r=0.196,0.248;P＜0.05).Conclusions The requirements of parents of congenital heart disease children before operation is high and its fulfillment level is low.Nurses should know the needs of parents and support parents in time by effective communication and corresponding nursing to improve the nursing quality.

Objective To construct the recombinant virus-like particles containing HCV envelope glycoprotein E1E2 based on sindbis virus vector.Methods The gene encoding HCV envelope glycoprotein E1F2 was cloned into sindbis virus vector to construct recombinant plasmids pBR-XJE1E2 and pVAXJE1E2,and transfect them into BHK-21 cells to obtain recombinant virus-like particles.The expression of E1 and E2 protein were verified by Western Blot and indirect immunofluorescenct assay (IFA).Results The results of restriction enzyme digestion,PCR and sequencing analysis showed that the recombinant plasmids were constructed successfully.And the results of RT-PCR,Western blotting and IFA detection showed that the transfect cells could package HCV-like particles of expressing structural proteins E1E2.Conclusion The recombinant expression plasmids pBR-XJE1E2 and pVA-XJE1E2 based on sindbis virus vector could package HCV-like particles in eukaryotic cell,which provides a foundation for further study of its in vivo animal immune response.

OBJECTIVETo report the results of clinical characteristics, enzyme activity determination and mutation analysis of GLB1 gene in a Chinese patient with mucopolysaccharidosis (MPS) type IVB (Morquio B disease).

METHODA 14-year-old Chinese boy with MPS type IVB was firstly diagnosed by blood leucocytes galactosamine-6-sulfate sulfatase (GALNS) and β-galactosidase (GLB1) determination, who was characterized by short stature, multiplex skeletal abnormalities, difficulty in walking. PCR-sequencing analysis was applied to detect the mutations in GLB1 of the patient.

RESULTThe patient was characterized by dwarfism, pectus carinatum, kyphosis, normal intelligence, and no neurologic damage of spasms, linguistic capacity and so on. The patient had normal GALNS enzyme activity and very low GLB1 enzyme activity [5.03 nmol/(h·mg) vs. normal value 118 - 413 nmol/(h·mg) ] in leukocytes. A compound heterozygous missense mutations c.442C > T(p.R148C)/c.1454A > G(p.Y485C) in GLB1 gene were detected in this patient. The mutation p.Y485C is a novel variant. With the method of gene analysis of new variant, the mutation p.Y485C was considered to be a pathogenic mutation.

CONCLUSIONThe MPS IVB patient showed severe multiple skeletal deformities, normal intelligence, no neurologic damage and very low GLB1 enzyme activity, who carries compound heterozygous mutations p.R148C/p.Y485C. The mutation p.Y485C in GLB1 gene may be a novel pathologic mutation of MPS type IVB.

Adolescent ; Amino Acid Sequence ; Asian Continental Ancestry Group ; genetics ; Chondroitinsulfatases ; genetics ; metabolism ; DNA Mutational Analysis ; Humans ; Joints ; pathology ; Male ; Molecular Sequence Data ; Mucopolysaccharidosis IV ; enzymology ; genetics ; pathology ; Mutation, Missense ; Pedigree ; Polymerase Chain Reaction ; Radiography ; Spine ; diagnostic imaging ; pathology ; beta-Galactosidase ; genetics ; metabolism

OBJECTIVETo evaluate the effects of glial cell line-derived neurotrophic factor (GDNF) and memantine on the long-term prognosis in neonatal rats with ischemia-induced periventricular leukomalacia (PVL).

METHODSThirty-two 5-day-old neonatal rats were randomly divided into 4 groups: sham-operated, PVL, GDNF-treated and memantine-treated. PVL was induced by right carotid artery ligation and hypoxia in the PVL, GDNF-treated and memantine-treated groups. GDNF (100 μg/kg) or memantine (20 mg/kg) was injected in the two treatment groups immediately after PVL inducement. The weight of the rats was measured immediately before and after hypoxia ischemia (HI). Both of Morris water maze test and Rivlin inclined plane test were performed at 26 days old (21 days after HI). The values of the escape latency (EL) and swimming distance, and the maximum inclined plane degree which the rats could stand at least 5 seconds were compared among the four groups.

RESULTSThe lower weight, the prolonged mean values of EL and swimming distance and the reduced maximum inclined plane degree were observed in the PVL group compared to those in the sham-operated, GDNF-treated and memantine-treated groups. There were no significant differences in the weight, the values of EI and swimming distance and the maximum inclined plane degree between the two treatment groups and the sham-operated group.

CONCLUSIONSThe administration of either GDNF or memantine can markedly increase the abilities of spatial discrimination,learning and memory, and motor coordination, promote weight gain, and improve long-term prognosis in rats with PVL.

Animals ; Animals, Newborn ; Body Weight ; Excitatory Amino Acid Antagonists ; therapeutic use ; Glial Cell Line-Derived Neurotrophic Factor ; therapeutic use ; Humans ; Infant, Newborn ; Leukomalacia, Periventricular ; drug therapy ; psychology ; Maze Learning ; drug effects ; Memantine ; therapeutic use ; Motor Activity ; drug effects ; Rats

OBJECTIVETo compare brain lead accumulation and neurotoxicity induced by lead under drinking purified water and tap water on rat.

METHODSAll 104 male weaning SD rats were randomly divided into eight groups, matched-four pairs according to drinking water: tap water, purified water, tap water with lead 50 mg/L(lead acetate water-solution), purified water with lead 50 mg/L, tap water with lead 200 mg/L, purified water with lead 200 mg/L, tap water with lead 800 mg/L. All were fed with normal food and environmental cognitions kept consistent Morris water maze(including Place Navigation, Spatial Probe Test, Visible Platform Trial) was measured to test rat spatial learning at the 12 and 24 week. At the end of the experiment (28 week), rats were killed and the lead of brain and blood was measured by Graphite furnace atomic absorption spectrometric method; the NR1, NR2A, NR2B of NMDAR (N-methyl-D-aspartame receptor) in hippocampus were analyzed by RT-PCR.

RESULTSUnder the same lead exposure, no significant differences were observed in blood lead, however, brain lead level showed higher in drinking purified water group than that in tap water group. Expression of NR1, NR2A and NR2B in hippocampus of the rats drinking purified water was lower than those drinking tap water, especially at low lead exposure (50 mg/L) (P < 0.05). In the 24 week Morris water maze, place navigation test's escape latency showed significantly prolonged at the rats drinking purified water as compared with those drinking tap water on the pairs of 50 mg/L and 200 mg/L pb2+ groups (P < 0.05), and the differences occurred in early 1-2 days.

CONCLUSIONCompared with drinking tap water, drinking purified water might increase the accumulation of brain lead, lower NR1, NR2A, NR2B expression and delay the spatial learning and memory ability under chronic lead exposure in water.

Animals ; Drinking ; Intelligence ; drug effects ; Lead ; toxicity ; Male ; Maze Learning ; drug effects ; Memory ; drug effects ; N-Methylaspartate ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; drug effects

OBJECTIVETo study in vivo the endogenous self-repair mechanism in immature white matter induced by ischemia in neonatal rats with periventricular leukomalacia (PVL).

METHODSFive-day-old neonatal Sprague-Dawley (SD) rats were randomly divided into sham and PVL groups. Rat model of PVL was prepared by ligation of the right common carotid artery following 2 hours of exposure to 8% oxygen. Pathological changes and myelination in the white matter were assessed under light and electron microscopy at 7 and 21 days after PVL. O4-positive OL precursor cells in the white matter were determined with immunofluorescence staining. Activation, proliferation, migration and differentiation of glial progenitor cells in SVZ were observed using immunofluorescent double labeling of either NG2 (marker of progenitor cells) and 5-bromodeoxyuridine (BrdU), or O4 (marker of OL precursor cells) and BrdU.

RESULTSAll rats in the PVL group manifested either mild or severe white matter injury under light microscopy, and had higher pathological scores of white matter compared with the sham group at 7 and 21 days after PVL (P<0.05). Electron microscopy showed that the number and thickness of myelin sheath in the PVL group were significantly reduced compared with the sham group (P<0.01). O4-positive OL precursor cells in the white matter observed under fluorescence microscopy were significantly reduced in the PVL group compared with the sham group (P<0.05). BrdU/NG2-positive cells in the SVZ increased significantly in the PVL group 48 hours after PVL and migrated into the periventricular area, reaching a peak on day 7 after PVL. BrdU/O4-positive newborn cells began to appear in the periventricular area 72 hours after PVL, and the number of BrdU/O4-positive cells in the PVL group was statistically more than in the sham group on day 21 after PVL (P<0.05).

CONCLUSIONSIschemia may induce brain self-repair in neonatal rats, resulting in activation and proliferation of NG2 glial progenitor cells in the SVZ migration and differentiation into OL precursor cells in periventricular white matter.

Animals ; Animals, Newborn ; Brain ; pathology ; Brain Ischemia ; pathology ; Bromodeoxyuridine ; metabolism ; Cell Differentiation ; Disease Models, Animal ; Humans ; Infant, Newborn ; Leukomalacia, Periventricular ; pathology ; Myelin Sheath ; physiology ; Neuroglia ; pathology ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; pathology

In order to solve the problem of selection and in vivo delivery problem in siRNA treatment, hepatitis B virus (HBV) HBx gene which could be targeted by siRNA was studied. The siRNA expression plasmid which specific inhibits HBx expression was obtained by in vitro selection via a dual-luciferase plasmid including HBx-Fluc fusion protein expression domain. The selected siRNA expression plasmid was then encapsulated in PEG-modified cationic liposome, which was devoted into pharmacodynamic studies at both cellular and animal level. The results illustrated that the cationic liposome which encapsulated siRNA expression plasmid could effectively inhibit HBx gene expression both in vitro and in vivo.
Cations ; Gene Expression Regulation, Viral ; drug effects ; Hepatitis B virus ; genetics ; Liposomes ; chemistry ; Plasmids ; RNA, Small Interfering ; chemistry ; Trans-Activators ; genetics ; metabolism