Objective To study the neuroprotective effect of erythropoietin(EPO) on neonatal rats model with hypoxia-ischemia encephalopathy(HIE).Methods HIE was induced in rats on 7th day of postnatal age by ligation of right common carotid artery,followed by 2 h of hypoxia(80 mL/L O2).The subjects were divided into sham-operated group,control group and EPO group.EPO 4 000 U/(kg?day) was injected daily from day 2 pre-surgery for 9 to 16 days and PBS was injected in the control group.The neuroprotective effect of EPO on HIE model was detected by brain weight,the difference in weights between the ipsilateral(right) and contralateral(left) brain and the function test.In vitro study,the neural progenitor cell line C17.2 under gone apoptosis following an ischemia-like metabolic inhibition.The effect of EPO on the cell line ischemia modle 17.2 was evaluated by detecting Annexin V with flow cytometry.Results The signi-ficant and sustained brain injury in the hypoxia-ischemia and vehicle-treated group was observed and measured by reduction in relative weights of ipsilateral to contralateral and compromised sensorimotor functions in response to postural reflex test,compared with those of sham-operated animals(Pa

This study was designed to explore the RNA interference technique in inhibition of the expression of the mouse fibrinogen like protein 2 (mfgl2), which has been reported to be involved in the development a variety of diseases including fulminant viral hepatitis. A plasmid named p-mfgl2shRNA,complementary to the sequence of mfgl2 was constructed, while another short hairpin RNA (shRNA)which was a mutated form of the mfgl2shRNA sequences was used as a control. A plasmid named pEGFP-mfgl2 expressing the mfgl2-EGFP fusion protein was also constructed for the screening of the effect of p-mfgl2shRNA on mfgl2 expression. By cotransfection of p-mfgl2shRNA and pEGFP-mfgl2 or pcDNA3.1-mfgl2 expression construct into CHO cells or HeLa cells, the inhibition of mfgl2 expression by mfgl2shRNA was analyzed by direct observation through fluorescent microscopy, FACS, RT-PCR and immunohistochemistry staining. The experiments showed the significant inhibitory effect of p-mfgl2shRNA on mfgl2 expression at 48h post-transfection in both CHO and Hela cell lines with the inhibitory efficiency as high as 80.1%. The study demonstrated that the construct of p-mfgl2shRNA successfully interfered with the mfgl2 expression in vitro.

BACKGROUNDReactive oxygen species (ROS) may play both physiological and pathophysiological roles. Transcription factor NF-E2-related factor 2 (Nrf2) regulates antioxidant response element (ARE)-mediated genes expression and coordinates induction of chemoprotective proteins in response to physical and chemical stresses. The exact role of Nrf2 in cellular responses to different levels of oxidative stresses remains unknown.

METHODSRat pulmonary microvascular endothelial cells were cultured and treated with 0 mmol/L, 0.125 mmol/L, 0.25 mmol/L, 0.5 mmol/L, 1.0 mmol/L and 2.0 mmol/L hydrogen peroxide solution for 2 hours. Nrf2 gene expression was assayed by reverse transcription-PCR, Nrf2-ARE binding activity was assayed with electrophoretic mobility shift assay (EMSA), and localization of Nrf2 was detected with immunohistochemistry.

RESULTSLow and moderate (0.125 mmol/L, 0.25 mmol/L and 0.5 mmol/L) doses hydrogen peroxide exposure of rat pulmonary microvascular endothelial cells led to the nuclear accumulation of Nrf2, increased activity of transcription regulation and up-regulation of ARE-medicated gene expression. In contrast, high doses of hydrogen peroxide (1 mmol/L, 2 mmol/L) exposure of the cells led to the nuclear exclusion of Nrf2, decreased activity transcription regulation and down-regulation of ARE-mediated gene expression.

CONCLUSIONLow and moderate doses of hydrogen peroxide play protective roles by increasing transcription activity of Nrf2, whereas high- dose hydrogen peroxide plays a deleterious role by decreasing transcription activity of Nrf2.

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Cells, Cultured ; Electrophoretic Mobility Shift Assay ; Gene Expression ; drug effects ; Hydrogen Peroxide ; pharmacology ; Immunohistochemistry ; NF-E2-Related Factor 2 ; genetics ; metabolism ; Rats ; Reverse Transcriptase Polymerase Chain Reaction

Purpose::Terminal performance of a bullet in human body is critical for the treatment of gunshot injury and optimization of bullet design. The effects of the impact velocity ( v0) and the impact attack angle ( δ0) of the bullet on its terminal performance was investigated, using a new evaluation method (called expansion method) based on the expansion of cracks and the permanent cavity wall in ballistic gelatin. Methods::Ballistic gelatin was used to simulate human body. The 7.62 mm × 39 mm rifle bullets with different v0 (600-760 m/s) and δ0 (0°-6°) were fired into the gelatin blocks. The gelatin block was cut into slices of about 20 mm thickness. The cracks and the permanent cavity on each slice were obtained manually. The damaged gelatin was determined using two methods: expanding the permanent cavity but ignoring the cracks, and expanding both the permanent cavity and the cracks. The relations between the damaged gelatin and v0 and δ0 were obtained using linear fitting method. Results::According to the distribution of the damaged gelatin along the penetration depth, the damaged gelatin block could be divided into two parts: the less damaged part and the severely damaged part. The length of the less damaged part depends mostly on δ0; while the average damaged area of this part depends on both δ0 as well as v0. The cracks contributed significantly to the total volume of damaged gelatin, particularly when the expansion was larger than 1.9 mm. The total damaged gelatin increases with v0, δ0 and the expansion extent. The average length of equivalent cracks grew with v0 and δ0 when considering the cracks, and decreased with v0 when ignoring the cracks. Conclusion::The expansion method is suitable to investigate the influence of different factors of bullets on their terminal performance. The characteristics of the damaged gelatin have a linear relationship with the initial attack angle ( δ0) and the initial velocity ( v0) of the bullet.

OBJECTIVETo investigate the effect of toluene diisocyanate (TDI) on the production of reactive oxygen species (ROS) and the permeability of human bronchial epithelial (HBE) cells.

METHODSTDI-human serum albumin (TDI-HSA) conjugate was prepared using a modified Son's method. MTT assay was used to assess HBE cell viability after exposure to different concentrations of TDI-HSA. The level of intracellular ROS of HBE cells was detected by flow cytometry with an oxidation-sensitive fluorescent probe 2',7'-dichlorofluorescein diacetate (DCFH-DA) uploading, and the permeability of cell monolayer was assessed by detecting the transepithelial electrical resistance (TEER).

RESULTSThe exposure to 120 µg/ml TDI-HSA did not obviously affect the cell viability. Compared with the control group, the intracellular fluorescent intensity increased significantly in the cells exposed to 20, 60, and 100 µg/ml TDI-HSA (P<0.05). The intracellular ROS production increased significantly after 100 µg/ml TDI-HSA treatment (P<0.05), but the increment in ROS production was significantly suppressed by pretreatment of the cells with N-acetylcysteine (NAC) (P<0.05), which also enhanced the TEER decreased by TDI-HSA treatment (P<0.05).

CONCLUSIONSTDI enhances the permeability of HBE cell monolayer partially through a ROS-mediated pathway, suggesting the importance of oxidative stress in TDI-induced pulmonary diseases.

Bronchi ; cytology ; Cell Line ; Cell Membrane Permeability ; drug effects ; Epithelial Cells ; cytology ; metabolism ; Humans ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Serum Albumin ; pharmacology ; Toluene 2,4-Diisocyanate ; pharmacology

OBJECTIVE: To investigate the difference of Pax2 and P53 expressions in children with primary nephritic syndrome (PNS) and the effect of Pax2 on glucocorsteroid (GC)-resistance. METHODS: Renal Pax2 and P53 expressions in children with PNS (40 patients) were detected by immunohistochemistry. A semiquantitative score was used to evaluate the injury degree of the glomeruli and the tubulointerstitium, and correlation analysis was done among Pax2, P53 and pathologic score. RESULTS: Pax2 and P53 expressions were not found in the control group. Pax2 expression of renal tubule epithelia exsisted in children with PNS and there was weak or no expression of Pax2 in the podocytes. Pax2 expressions in the proximal tubule and the distal tubule in the GC-resistant group were more intense than those in the GC-intensive group (P <0.01). The more the Pax2 expression in the tubule, the more abnormal structure such as dilation and atrophy. Pax2 expression in the tubule epithelia was positively correlated with pathologic score of tubulointerstitium (P < 0.01). There was no P53 expression in the GC-intensive group, but there exsisted P53 expression in parts of the patients from the GC-resistant group, mainly distributing in the renal tubular epithelia. P53 expression was positively correlated with P53 expression and the pathologic score of tubulointerstitium (P < 0.01). CONCLUSION Over-expression of Pax2 in the renal tubule epithelia may improve P53 expression to a certain degree, which may aggravate the lesion of the renal tubule. It may be one of the mechanisms resulting in GC-resistant in children with PNS.
Adolescent ; Child ; Child, Preschool ; Drug Resistance ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Male ; Nephrotic Syndrome ; drug therapy ; metabolism ; PAX2 Transcription Factor ; biosynthesis ; genetics ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

OBJECTIVE: To investigate the eliminating ability of catechin to eliminate O2-* and *OH. METHODS: The ability of catechin to clear away O2-* and *OH was respectively measured by faintness chemiluminescence and spin trapping assay. RESULTS: IC50 that catechin eliminated O2-* and *OH was 6.16, 0.59 g/mL respectively, and the eliminating ability of catechin was much stronger than that of the extract from liquorice, rosemary, grape pip, giant knotweed and ginkgo leaf. CONCLUSION Compared with several important natural plants of antioxidants, the eliminating ability of cathechin is the best.
Antioxidants ; pharmacology ; Catechin ; pharmacology ; Free Radical Scavengers ; pharmacology ; Hydroxyl Radical ; metabolism ; Luminescent Measurements ; Superoxides ; metabolism

OBJECTIVE: To evaluate the clinical and pathological features of 94 children suffering from IgA nephropathy (IgAN) while estimating the prevalent situation in Hunan province. METHODS: To summarize the annual number of hospitalized children, those with kidney diseases, those accepted biopsy, and those confirmed as IgAN in both Xiangya Hospital and Second Xiangya Hospital undertaking kidney biopsy in Hunan province during 1995 and 2004. RESULTS: In the past 10 years, as the hospitalized population in both hospitals accrued to 9.98% each year. The rate of 7.5% was seen in those with kidney diseases. Among whom 56.3% accepted kidney biopsy and 94 of them were confirmed as IgAN. Hematuria was the main clinical presentation, seen in 71 cases, accounting to 76%, and even to 98% after excluding those with nephrotic syndrome and isolating proteinuria type of IgAN. Inflammation infiltration (91%), renal tubule degeneration (81%), and renal interstitial fibrosis (31%) were the major pathological features of 94 children, especially in nephrotic syndrome IgAN. CONCLUSION The number of children with IgAN synchronously accrues as hospitalized population, those with kidney diseases, and those by kidney biopsy. Hematuria is the major symptom. To routinely perform urine analysis and kidney biopsy in asymptomatic hematuria may improve the diagnosis. Inflammation infiltration, renal tubule degeneration, and renal interstitial fibrosis are the major pathological features in IgAN children, especially in nephrotic syndrome IgAN, probably relating to continuous proteinuria. Early control of proteinuria may delay or decrease renal tubule fibrosis.
Adolescent ; Biopsy, Needle ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Glomerulonephritis, IGA ; complications ; epidemiology ; pathology ; Hematuria ; diagnosis ; etiology ; Hospitalization ; statistics & numerical data ; Humans ; Kidney ; pathology ; Male

OBJECTIVE: To investigate the efficacy and adverse effect of mycophenolate mofetil (MMF) in the treatment of frequently relapsing nephrotic syndrome in children. METHODS: The study population consisted of 37 children (24 simple nephrotic syndrome and 13 nephritis-type syndrome) suffering from frequently relapsing nephrotic syndrome. Patients received 20-30 mg/(kg d) of MMF in conjunction with 1 mg/(kg d) prednisone for 3-6 months. RESULTS: Out of 24 patients suffered from simple nephrotic syndrome, 17 patients (70.8%) with complete relief, 4 patients (16.7%) with partial relief and 3 patients (12.5%) with non-relief, whereas out of 13 patients suffered from nephritis-type syndrome 6 patients (46.2%) with complete relief, 3 patients (23.1%) with partial relief and 4 patients (30.7%) with non-relief. Eight patients with Minimal Change Disease (MCD) achieved complete relief. Of 23 patients with Mesangial Proliferative Glomerulonephritis (MsPGN) or Membranoproliferative Glomerulonephritis (MPGN), complete relief was observed in 17 patients (73.9%), partial relief in 4 patients (17.4%) and non-relief in 2 patients. CONCLUSION These Results suggest that MMF has better efficacy against simple renal disease than against nephritis-type syndrome, and MMF may be more suitable for the treatment of frequently relapsing nephrotic syndrome characterized by proliferative lesions.
Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Immunosuppressive Agents ; adverse effects ; therapeutic use ; Male ; Mycophenolic Acid ; adverse effects ; analogs & derivatives ; therapeutic use ; Nephrotic Syndrome ; drug therapy ; Recurrence ; Treatment Outcome

OBJECTIVETo investigate the expression of high mobility group box-1 (HMGB1) in the lung tissue and bronchoalveolar lavage fluid (BALF) of asthmatic mouse models and the influence of dexamethasone (DM).

METHODSEighteen female Balb/C mice were randomly divided PBS control group, OVA group and OVA/DM group, and asthmatic mouse models were established in the latter two groups. The airway responsiveness of the mice was assessed by whole-body plethysmography, and the cells in the BALF were counted and classified, with the supernatants of the BALF collected for detection of the level of HMGB1 by ELISA. The left lung of the mice was collected for HE staining, and the expression of HMGB1 in the right lung tissue was detected by Western blotting.

RESULTSAsthmatic mouse models were successfully established. The level of HMGB1 in the BALF was significantly higher in OVA group than in the control group (6.31 ± 4.05 ng/ml vs 2.59 ± 0.73 ng/ml, P = 0.017), but no significant difference was found between OVA/DM group (3.39 ± 0.50 ng/ml) and OVA group (PP = 0.052). The expression of HMGB1 relative to tubulin was significantly higher in OVA group than in the control group (2.08 ± 0.87 vs 0.85 ± 0.30, P = 0.032), but similar between OVA/DM group (1.15 ± 0.48) and OVA group (PP = 0.133).

CONCLUSIONThe expression of HMGB1 is obviously increased in the lung and BALF of asthmatic mice and DM produces no significant effect on HMGB1 expression, suggesting that HMGB1 may serve as a new therapeutic target for asthma treatment.

Animals ; Asthma ; drug therapy ; metabolism ; Bronchoalveolar Lavage Fluid ; chemistry ; Dexamethasone ; therapeutic use ; Female ; HMGB1 Protein ; genetics ; metabolism ; Lung ; metabolism ; Mice ; Mice, Inbred BALB C