Death of nerve cells after cerebral ischemia have a variety of forms,including cell necrosis occurs immediately in ischemic core area and the subsequent apoptosis and autophagy induced by oxidative stress and inflammatory response in the course of reperfusion.After cerebral ischemia,a variety of different molecular mechanisms eventually lead to cell death,and the process involves several signaling pathways.Intervention of different forms and mechanisms of cell death may alleviate cell death after cerebral ischemia.

As an intracellular energy sensor, AMP-activated protein kinase (AMPK) plays an important role in maintaining the energy balance of the cells and organisms. Initially, the effects of AMPK on the processes of pathophysiology in diabetes, obesity and other metabolic diseases were well studied. In recent years, the roles of AMPK in the pathophysioiogical processes, including distribution and acidosis, oxidative stress injury and apoptosis in brain tissue have received increasing attention. At the same time, it also found that artificially regulates the AMPK activity after stroke may change the outcome of neurons. Therefore, AMPK is expected to become a new target in the treatment of ischemic cerebrovascular disease.

As an important component of the “neurovascular unit”,astrocytes provide protective effect for nervous through intaking excessive excitatory amino acids,providing energy substances,maintaining extracellular K + and water balance,scavenging oxygen free radicals and secreting neurotrophic factor during ischemic stroke.This article elaborates the mechanisms of astrocytes participating in ischemic stroke in recent years.

Bone Diseases, Metabolic ; physiopathology ; Bone and Bones ; metabolism ; physiopathology ; Child ; Child Nutrition Disorders ; physiopathology ; Child Nutritional Physiological Phenomena ; physiology ; Humans ; Infection ; physiopathology ; Kidney Failure, Chronic ; physiopathology

Objective To investigate the effects of adenosine monophosphate-activated protein kinase (AMPK) on high-mobility group box 1 (HMGB1) release from PC12 cells after oxygen-glucose deprivation and reoxygenation (OGD/R) and its mediated inflammatory response in BV2 cells.Methods PC12 and BV2 cells were cultured,respectively.The PC12 cells were used to induce a model of oxygen glucose deprivation for 12 h and reoxygenation for 24 h.After giving 5-aminoimidazole-4-carboxamide (AICAR) 5,50 and 100 μmol/L as well as Compound C 0.1,1 and 10 μmol/L activation or inhibition of AMPK phosphorylation,respectively,methyl thiazolyl tetrazolium (MTT) was used to detect the PC12 cell activity.Enzyme-linked immunosorbent assay was used to detect the HMGB1 release level in the PC12 cell culture media.After OGD/R in each group,the PC12 culture media were acted on normal cultured BV2 cells for 24 h respectively.Westem blotting and Enzyme-linked immunosorbent assay were used to detect the NFκB inhibitory protein (inhibitor of NFκB,IκB) phosphorylation level and TNF-α release level in BV2 cells,respectively.Results After OGD/R,the PC12 cell activity was decreased significantly (68.84％±6.60％ vs.100.04％ ± 8.82％;P ＜ 0.01);the AMPK phosphorylation level was increased significantly (1.95 ±0.39 vs.1.00 ±0.20;P＜0.05),and the extracellular HMGB1 release was increased significantly (287.66 ± 26.42 pg/μl vs.53.05 ± 9.11 pg/μl;P ＜ 0.01).Compared with the OGD/R group,AICAR 100 μmol/L significantly increased the survival rate of PC12 cell after OGD/R (78.6％ ± 3.75％ vs.68.84％ ± 6.60％;P ＜ 0.05),promoted AMPK phosphorylation (3.32 ± 0.66 vs.1.95 ± 0.39;P ＜ 0.01),and reduce the release of extracellular HMGB1 (164.06 ± 12.77 pg/μl vs.287.66 ± 26.42 pg/μl;P ＜0.01).In contrast,Compound C 10 μmol/L significantly reduced the cell survival rate of PC12 (40.44％ ±3.79％ vs.68.84％ ±6.60％;P ＜0.01),inhibited AMPK phosphorylation (1.07 ± 0.21 vs.1.95 ± 0.39;P＜0.05),and increased the release of HMGB1 (337.97 ± 18.9 pg/μlvs.287.66 ± 26.42 pg/μl;P＜0.01).The conditioned medium from the AICAR 100 μmol/L group significantly inhibited IκB phosphorylation (1.68 ±0.51 vs.3.09 ± 0.10;P ＜ 0.05) and reduced the release of TNF-α (669.53 ±38.58 pg/μlvs.841.76 ± 45.82 pg/μl;P＜ 0.05) in BV2 cells.The conditioned medium from the compound C 10 μmol/L group significantly promoted IκB phosphorylation (4.98 ± 1.24 vs.3.09 ± 0.10;P ＜0.01) and increased the release of TNF-α (1 035.32 ± 128.06 pg/μl vs.841.76 ± 45.82 pg/μl;P ＜0.05) in BV2 cells.Conclusions Promoting AMPK phosphorylation activation may reduce the release of HMGB1 from PC12 cells after OGD/R,and inhibit its mediated NF-κB inflammatory pathway and reduce the release of TNF-αin BV2 cells,and thus reducing neuroinflammatory injury.On the contrary,inhibiting AMPK phosphorylation may promote the release of HMGB1 from PC12 cells after OGD/R and aggravate its mediated inflammatory reaction in BV2 cells.

Objective To investigate the changes in the concentration of myocardin in children with lupus nephritis (LN) under different degree of vessel damage.Methods Forty-nine children diagnosed with LN by routine tissue immunolfuorescence, light microscope, and electron microscope were included, and 30 healthy children were included as control group. The pathological classiifcations were performed according to the ISN/RPS 2003 LN pathological classiifcation criterion. According to the Katafuchi evaluation method, the semi quantitative assessment of glomerular and kidney tubule damage was carried out, and the degree of vascular damage was evaluated at the same time. Double antibody sandwich method was used to detect the concentration of serum myocardin.Results The glomerular and kidney tubules damage in children with LN were signiifcantly aggravated with higher pathological classification (P<0.05). Glomerular damage was positively correlated with renal interstitial damage (r=0.96, P<0.01). The degree of vascular damage was related to the degree of glomerular injury and renal interstitial injury, while it was no related with the results of clinical tests. There were different concentrations of myocardin among mild-, moderate-, severe-vessel damage and control groups (F=378.61,P<0.001), and the concentration of myocardin in moderate- and severe-vessel damage groups were obviously lower than those in control group and mild-vessel damage group (P<0.01) while there was no difference between control group and mild-vessel damage group (P>0.05). According to pathological type, there were signiifcant differences in the concentration of myocardial between control group and different pathological types (F=626.793,P<0.01). FromⅡ,Ⅲ,Ⅲ+Ⅴ,Ⅳ toⅣ+Ⅴ, the concentrations of myocardial were decreased systematically, and there were statistic differences between groups (P all<0.05).Conclusion The concentration of myocardin in children with LN can relfect the renal vascular damage to a certain extent. Elevation of myocardin concentration may be helpful for the repair of vascular damage.

Objective To establish a cell line with overexpression of rat serotonin1A receptor (5-HT1AR).Methods Human neuroblastoma cells-SH-SY5Y were donated by cancer institute attached to the 4 th Affiliated Hospital, Hebei Medical University. Total RNA was extracted from brain tissues of male SD rats and rat 5-HT1A R was obtained by RT-PCR. Plasmid pc-DNA3. 1/hisC containing the rat 5-HT1AR (pc-DNA3.1/hisC-Rat-5-HT1AR)was constructed and transfected into SH-SY5Y cells. The transfected cells were isolated by G418 selection and SH-SY5Y-Rat-5-HT1A R cells were obtained. Expression of 5-HT1A R was detected by Western blot analysis. Cell viability was evaluated by MTT assay. SH-SY5Y-Rat-5-HT1AR cells were further observed for 5-HT1AR by immuno-fluorescence staining. Results Plasmid pc-DNA3. 1/hisC-Rat-5-HT1AR was successfully constructed by linking Rat-5-HT1A R with pc-DNA3.1/hisC and transfected into SH-SY5Y. The SH-SY5Y-Rat-5-HT1A R cells were more slender than SH-SY5Y cells with less and longer processes. MTT showed that the viability of SH-SY5Y-Rat-5-HT1A R cells was much lower than SH-SY5Y. Rat 5-HT1A R was expressed efficiently on the membrane of SH-SY5Y-Rat-5-HT1A R cells. Conclusion A cell line with overexpress of rat 5-HT1A R is successfully established.

Objective To research Henoch-Schonlein purpura purpura (HSP) and renal pathology in children.Methods 31 hospitalized HSP children that with normal urine routine and accepted renal biopsy in our hospital.Results There were different levels of kidney pathological damage in this group of 31 cases,the results of light microscope were from grade Ⅱ to grade Ⅵ The proportion was grade Ⅱ(35.48％,11 of 31),grade Ⅲ (54.83％,17 of 31),and grade Ⅳ,Ⅴ and Ⅵ (each 1 case of 31,3.23％ ).lmmunofluorescence pathology results were showed as following:merely IgA depositional (48.38％,15 of 31 ),IgA + IgG depositional ( 19.36％,6 of 31 ),IgA + IgM depositional ( 19.36％,6of 31 ),IgA + igG + IgM depositional ( 12.90％,4 of 31 ).Microalbuminuria had been founded in 14 cases,and the microalbuminuria level of 10 cases were higher than normal value( 10 of 14,71.43％ ).Conclusions HSP children had renal pathologic dysfunction,even the urine routine were normal,and the detection of urine microalbumin was a significant marker in the early stage.

ObjectiveStudy the relationship among CaSR expression, tubulointerstitial damage,metabolic disturbance of calcium and phosphorus and microvascular density around the tubulointerstitium in children with steroid-resistant nephrotic syndrome.Methods36 cases of children with primary nephrotic syndrome were divided into hormone-sensitive group and steroid-resistant group.Semi-quantitative scores for tubulointerstitial pathological evaluation of the extent of damage, automatic biochemical analyzer for the determination of serum calcium (Ca), phosphorus (P) concentration of renal tubular epithelial CaSR expression and microvessel microvascular density around the tubulointerstitium were determined by immunohistochemical assay.ResultsMore severe the tubulointerstitial damage, lower level of serum Ca and higher level of serum P were observed [(2.26 ± 0.15) mmol/L]in children of the steroid-resistant group and the steroid-sensitive group [(1.90 + 0.12) mmol/L, P ＜ 0.05].CaSR expression (4.63 + 0.78) of renal tubular epithelial cells in the steroid- sensitive group was significantly lower than that in the steroid-resistant group (6.56 + 1.22, P ＜ 0.05), but microvascular density was significantly higher in the steroid- sensitive group(2.98 +0.35 vs 2.02 +0.24, P ＜0.05).When the tubulointerstitial damage was mild, CaSR expression (4.15 +0.58) in renal tubular epithelial cells in the steroid- sensitive group (4.26 ±0.61) was lower than the steroid-resistant group(3.12 ± 0.33; 3.01 ± 0.21), and microvascular density was higher,but the difference was not significant(P ＞0.05).In the moderate tubulointerstitial damage, CaSR expression in renal tubular epithelial cells in the steroid- sensitive group (5.35 ± 0.64) was significantly lower than the resistant group (7.37 +0.81, P ＜0.01), and microvascular density was significantly higher than the resistant group (2.81 ±0.16, 2.02 ±0.14, P ＜0.05).Compared by mild and moderate tubulointerstitial damage in children with the steroid-resistant, CaSR expression (11.46 ± 1.38) in children with severe tubulointerstitial damage was significantly increased, and microvascular density (1.15 ± 0.11) was significantly decreased (all P ＜ 0.01).ConclusionsCaSR expression was increased and microvascular density around the tubulointerstitium was decreased in children with steroid-resistant nephrotic syndrome.Dut to steroid resistance, the cytotoxic of steroid damaged the renal tubular epithelial cells, the metabolic disturbance of calcium and phosphorus and the damage of blood vessel endothelium finally resulted in severe tubulointerstitial damage.

Objective To explore the relationship between vascular endothelial growth factor (VEGF) concentration in urine and renal vascular damage in children with Henoch Schonlein purpura nephritis (HSPN).Methods The kidney pathological lesion of 78 biopsy-proven HSPN children was assessed with renal vascular damage, glomerular pathological damage, and tubulointerstitial pathological damage semi-quantitative points. The children were divided into 3 groups (light, medium, and heavy group) according to the renal vascular, glomerular, tubulointerstitial, glomerular and tubulointerstitial total pathological points. Blood and urine vascular endothelial growth factor concentration was detected by enzyme linked immunosorbent assay;the localized renal VEGF expression and microvessel density were detected by immunohistochemistry assay in the kidneys. Results The semi-quantitative points of glomerular, tubulointerstitial, renal vascular, and glomerular and tubulointerstitial total points in different groups had significant difference (all P＜0.01);the minor renal vascular damage, the higher light microvessel density, blood and kidney concentration of VEGF, and the VEGF excretion in the urine were also lower in different groups, and there were significant differences (all P＜0.01). Glomerular points were positively related with tubular points, vascular points, kidney total score (r=0.596,0.612, and 0.728;P＜0.05, 0.05, and 0.01 respectively). Microvessel density was highly positively related with blood VEGF and renal VEGF, and negatively rela-ted with urine VEGF (r=0.601, 0.696, and -0.639,all P＜0.01). Conclusion The urinary excretion of VEGF leads to the decrease of local kidney VEGF concentration resulting in the renal vascular injury, which may be the important reason for renal vascular damage and pathology chronic progress in HSPN children.