Objective To investigate the changes of antibrain-antibody (AB-Ab) in blood serum and the effect of AB-Ab on blood-brain barrier (BBB) and cerebral edema after the traumatic brain injury (TBI) in rabbits.Methods Forty-five New Zealand rabbits were randomly divided into control group (n =5),sham operation group (fenestration only without injury,n =15) and TBI group (severe lateral fluid percussion brain injury,n =15) according to the random number table.Sham and TBI animals were subdivided at postoperative 1,3,7 and 14 days.Level of AB-Ab,BBB permeability and brain water content were determined after operation.Results At each time point,serum AB-Ab level,BBB permeability and brain water content were higher in sham operation group and TBI group than in control group (P ＜0.05).At postoperative 3,7 and 14 days,serum AB-Ab level,BBB permeability and brain water content were higher in TBI group than in sham operation group (P ＜ 0.05).Conclusion Serum level of AB-Ab is significantly increased after TBI along with synchronous changes in BBB permeability and brain water content,indicating that AB-Ab may be associated with the increase of BBB permeability and brain edema.

Objective To observe the dynamic change of brain natriuretic peptide (BNP) in plasma of elderly patients with hy-pertensive intracerebral hemorrhage ,and to explore the relationship between BNP levels and cerebral edema .Methods 56 elderly patients with hypertensive intracerebral hemorrhage were recruited and divided into the operation group (n=26) and the non-opera-tion group(n=30) ,25 hypertensive patients were collected as the control group .ELISA was applied to detect the levels of plasma BNP at 6 ,24 ,72 ,168 h after the onset of hypertensive intracerebral hemorrhage .The non-invasion brain edema monitor was utilized to dynamically monitor the cerebral edema degrees .Results The plasma BNP levels in the operation group and the non-operation group at 6 ,24 ,72 ,168 h were higher than those in the control group ,the differences showed statistical difference (P<0 .05) ,but which at 168 h in the operation group was lower than that in the non-operation group ,cerebral edema appeared at 24 h in the two groups ,and reached the peak value at 72 h in the operation group and at68 h in the non-operation group;the changes of plasma BNP levels had positive correlation with cerebral edema degree (r=0 .685 ,P<0 .05) .Conclusion Dynamic monitoring of plasma BNP levels is an effective indicator for early diagnosis and monitoring of acute brain edema in elderly patients with hypertensive in-tracerebral hemorrhage .

Objective To investigate relationship of immunoglobulin concentrations (IgG,IgA,and IgM)and decrease of serum Mg2+ concentration in patients with severe traumatic brain injury (sTBI).Methods Sixty patients with sTBI were randomly divided into control group and study group.Intravenous injection of MgSO4 was given to study group.Levels of Mg2+ in serum and immunoglobulin (IgG,IgA,and IgM) in both groups were measured at 1,3,7,and 14 days after injury so as to analyze correlations of serum Mg2+ concentration with each humoral immune indices.Results IgG concentration at days 3 and 7 in study group were (10.79 ± 2.65) g/L and (10.2 1 ± 2.54) g/L respectively,higher than that in control group (P ＜ 0.05).IgM at days 3 and 7 in study group were (1.27 ± 0.32) g/Land (1.31 ± 0.25) g/L respectively,higher than that in control group (P ＜ 0.05).Difference of IgA level between the two groups was not significant.Level of Mg2 + was positively related to levels of IgM and IgG,but not to level of IgA.Pathological amelioration at day 14 between the two groups showed insignificant difference.Conclusion Intravenous replenishment of MgSO4 for patients with sTBI at acute phase can improve humoral immunosuppressive state,but can not ameliorate prognosis.

Objective To investigate the effect of neutrophil elastase (NE) inhibitor on the blood-brain barrier(BBB) permeabili-ty and hydrocephalus in rats with traumatic brain injury (TBI) .Methods 99 SD rats were randomly divided into the control group , the TBI group and the intervention group(dividing into 5 sub-groups:6 ,24 ,48 ,72 ,168 h) .The hydraulic impact model of rats was duplicated .Sivelestat sodium was given in the intervention group .The NE concentration in the brain tissue ,BBB permeability and brain water content were detected in each group and performed the comparative analysis .Results The NE concentration in the brain tissue ,BBB permeability and brain water content at each timepoint in the TBI group and the intervention groups were higher than those in the control group .The NE concentration at 24 ,48 ,72 ,168 h in the intervention group was lower than that in the TBI group .The BBB permeability and the brain water content at 48 ,72 ,168 h in the intervention group were lower than those in the TBI group(P<0 .05) .Conclusion Sivelestat sodium can inhibit the NE release in TBI rat brain tissue ,reduce the BBB permeability and the occurrence of hydrocephalus ,which indicating that sivelestat sodium has the protective effect on TBI secondary lesion in rat .

AIM: To observe the change of subunit of NADPH oxidation enzyme complex nox - 1 protein in cardiocyte hypoxia - reoxygenation injury and the role of cardiotrophin -1.METHODS: Cardiomyocytes from the hearts of 1 -3 d old neonatal rats were prepared by a modified method. Five groups were included in the study: control; hypoxia/ reoxygenation; hypoxia/reoxygenation + CT - 1; CT - 1 + hypoxia/reoxygenation + LY294002 (PIK3/Akt inhibitor) ; CT -1 + hypoxia/reoxygenation + PD98059 (ERK inhibitor) ; CT - 1 + hypoxia/reoxygenation + DMSO. The concentration of CT -1 was 10 μg/L. The survival rate of myocytes was evaluated by MTS method. Apoptosis, mitochondrial permeability transition pore ( △ψm) and reactive oxygen species ( ROS) were detected by flow cytometry. Nox - 1 protein was determined by Western blotting. RESULTS: Apoptosis of cardiomyocytes and the level of ROS (19.7% ±1.4% vs 2.1% ± 0.5% , 14.07% ± 1.25% vs 3.54% ± 0.86% , P < 0.05 ) increased markedly after hypoxia/reoxygenation, but cardio-myocyte survival rate and the level of△ψm (40.55% ±4.25% vs 86.28% ±7.15% , P <0.01) decreased significantly. The expression of nox - 1 protein was upregulated markedly. With CT - 1 intervention, cardiomyocyte survival rate increased markedly, apoptosis, both ROS and expression of nox - 1 protein reduced significantly. The level of△ψm increased obviously. The effect of CT - 1 was inhibited by LY294002.No significant effect was observed on cells survival in DMSO group, which confirmed that LY294002 was specifically involved in blocking the protective effect of CT - 1.CONCLUSION : The expression of subunit of NADPH oxidation enzyme complex nox - 1 protein is upregulated markedly in cardiocyte hypoxia - reoxygenation injury.CT - 1 protects cardiac cells against hypoxia - reoxygenation injury by downregulating the expression of nox -1 protein to decrease the level of ROS.

Objective To study the effect of Cardiotrophin-1 (CT-1) on cardiocyte hypoxia-reoxygenation injury,and to investigate the signaling pathways involved in the protective effect. Method This study was carried out in Key Lab of Molecular Medicine in Jiangxi Province. Cardiomyocytes from the hearts of 2-day-old Sprague-Dawley neonatal rats were prepared by a modified method. Five groups were included in the study. Group (ⅰ): control, Group (ⅱ): hypoxia/reoxygeuation, Group (ⅲ): hypoxia / reoxygenation + CT-1, Group (iv) : CT- 1 + hypoxia/ reoxygenation + LY294002 (PIK3/Akt inhibitor), Group (ⅴ): CT-1 + hypoxia / reoxygenation +DMSO. The concentration of CT-1 was 10 ng/mL. Myocytes survival rote was evaluated by MTS method, apopto-sis, mitochondrial permeability transition pore (△ψm) and reactive oxygen species(ROS) were detected by flow cy-tometer, phosphorylased GSK-3β and PI3K protein by western blotting. Analysis of variance and q test as statistical methods was used to analyze the data. Results Cardiomyocyte apoptosis and ROS increased markedly after hy-poxia/reoxygenation,but cardiomyocyte survival rate and the level of △ψm [(40.55±4.25) vs. (86.28±7.15), P < 0.01]decreased significantly. With CT-1 intervention, cardiomyocyte survival rate increased markedly (87%),apoptosis and ROS reduced significantly. The level of △ψm increased, the level of phosphorylased GSK-3β and phosphorylased PI3K protein obviously increased. The effect of CT-1 was inhibited by LY294002, but no significant effect was observed on ceils survival in DMSO group, which confirmed that LY294002 specifically in-volved blocking the protective effect of CT-1. Conclusions CT-1 can protect cardiac cells against hypoxia- reoxy-genation injury, these effects are dependent upon its ability to activate the PI3K/GSK-3β pathway.

Objective To purify native and recombinant heparin-binding hemagglutinin(HBHA)protein,and investigate the activity of HBHA polyclonal antibody against aggregation of Bacillus CalmetteGuerin(BCG)induced by HBHA.Methods After growing BCG to the stationary phase in the 7H9 liquid medium,the native HBHA protein(nHBHA)was obtained by CL-6B column chromatography.At the same time,the HBHA gene fragment was cloned and expressed by transforming Escherichia coli BL-21.Then the polyclonal antibody against rHBHA was prepared by immunizing rabbit.Different comcentration of the HBHA protein was added to the BCG liquid medium,and the aggregation of the BCG was observed.Then,add the HBHA protein that incubated with anti-HBHA antibodies to the BCG culture medium and observe the aggregation of BCG.Results The purity of native HBHA was 99% and the concentration was 1.016 mg/ml.The expressed product contained 36% of total somtic protein.After purified,the purity of the recombinant HBHA protein was 97.1% and the concentration was 10.98 mg/ml.Both the rHBHA and nHBHA could induce the aggregation of BCG.When then concentration of nHBHA is 0.2μg/ml,BCG could be induced to aggregate,while the rHBHA concentration is 2μg/ml could induce the aggregation.Both aggregations could be suppressed by the polyclonal antibody against rHBHA.Conclusions The native and recombinant HBHA are successfully obtained.It is proved that the rHBHA could induce the aggregation of BCG similar as nHBHA,and polyclonal antibody against rHBHA could also suppress the activity of nHBHA.It suggested that rHBHA could be further used in clinical diagnosis and vaccination.

OBJECTIVETo study the expression and significance of nestin (a type of cytoskeletal protein) in normal and diseased human kidneys.

METHODSDiseased kidney tissues were obtained from needle biopsies in 32 patients with glomerulonephritis (including 8 cases of membranous glomerulopathy, 3 cases of focal segmental glomerulosclerosis, 17 cases of IgA nephropathy with proteinuria and 4 cases of IgA nephropathy without proteinuria). Control kidney tissues were obtained from nephrectomy specimens for renal tumors. The expression of nestin in the control kidney tissues was studied using immunoelectronic microscopy and immunohistochemistry. The expression of nestin in the diseased kidney tissues was detected by immunohistochemistry and real-time reverse transcription-polymerase chain reaction.

RESULTSIn normal kidney tissues, nestin was detected at the periphery of glomerular capillary loops. Semi-quantitative morphometric analysis showed that the glomerular nestin expression level in cases of IgA nephropathy without proteinuria did not differ from that in the normal controls. However, the glomerular nestin expression levels in cases of IgA nephropathy with proteinuria, membranous glomerulopathy and focal segmental glomerulosclerosis were significantly lower than those in the normal kidneys and IgA nephropathy without proteinuria. The glomerular nestin expression levels inversely correlated with the 24-hour urine protein results.

CONCLUSIONNestin may play an important role in maintaining the normal function of podocytes in human kidney.

Adult ; Gene Expression Regulation ; Glomerulonephritis, IGA ; immunology ; Humans ; Immunohistochemistry ; Intermediate Filament Proteins ; genetics ; metabolism ; Kidney ; Kidney Diseases ; metabolism ; pathology ; Kidney Glomerulus ; metabolism ; pathology ; Middle Aged ; Nephrectomy ; adverse effects ; Nerve Tissue Proteins ; genetics ; metabolism ; Nestin ; Proteinuria ; metabolism

This study was purposed to investigate the effects of 2-deoxy-D-glucose (2-DG) on sensitizing HL-60 cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis and its possible mechanism. The proliferative inhibition of HL-60 cells treated with different concentrations of 2-DG and TRAIL was measured by MTT assay. The cells were treated with 2-DG, TRAIL, and 2-DG combined with TRAIL at the concentration < IC50 value, i.e. 10 mmol/L for 2-DG and 100 ng/ml for TRAIL. Apoptosis was analyzed by flow cytometry with PI staining; the expression of RIP1, GRP78, and PARP was analyzed by Western blot; the activity of caspase-3 was detected by special detection kit. The results showed that the combined treatment of HL-60 cells for 48 h induced an apoptotic rate of (45.1 ± 4.3)%, which was significantly higher than that of treated with 2-DG or TRAIL alone; at the same time, the combined treatment potentiated the expression of GRP78 and caspase-3 activity, and down-regulated the expression of RIP1. It is concluded that 2-DG can sensitize HL-60 cells to TRAIL-induced apoptosis, which may be correlated with excessive endoplasmic reticulum stress response, down-regulation of RIP1, and increase of caspase-3 activity.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Deoxyglucose ; pharmacology ; HL-60 Cells ; Heat-Shock Proteins ; metabolism ; Humans ; Nuclear Pore Complex Proteins ; metabolism ; RNA-Binding Proteins ; metabolism ; TNF-Related Apoptosis-Inducing Ligand ; metabolism ; pharmacology

Objective To make an etiology study on children with unexplained mental retardation (MR) by using combined multiplex ligation-dependent probe amplification (MLPA),and to explore associations between subtelomeric aberrations and phenotypes in local children.Methods Sixty-seven children with unexplained MR were enrolled in study group from Jul.2009 to Dec.2011 in Guangdong Women and Children's Hospital.Peripheral blood of patients and their parents were collected as samples of subtelomeric test by MLPA.Two kinds of probes of MLPA were combined to verify the aberration results.After confirmed test the parent of positive children were tested by the same way,then to analyze associations between test data and the clinical feature.Results Among sixty-seven children enrolled in the study aged 6 months to 15 years,where were 42 male and 25 female ;the intelligence of 47 children belonged to mild degree(IQ≥50 scores),that of 20 children belonged to severe degree(IQ ＜50 scores) ;7 patients had convulsion history,24 patients had malformation,18 cases had idiopathic organ aberrations.Four patients had aberration copies in subtelomeric region by MLPA test,the detection rate was 5.97％,4 patients were novel cases.There was no significant differences in genders,age and convulsion history between positive and negative children (all P ＞ 0.05).The aberration rate in moderate to severe degree group was higher than those mild degree group.The rates of features including physical developmental retardation,malformation and organ aberrations in positive children were higher than those of the negative cases.There were significant differences between the 2 severity groups (all P ＜ 0.05).Conclusions Aberrations in subtelomeric region can be one of the important causes of unexplained MR.The MR patients were supposed to have subtelomeric region tested so as to provide the evidence for diagnosis and genetic counseling.