Objective To establish a better method of isolating andculturing ofneural stem cells(NSCs) in neonatal rat brain. Methods Tissue of brain was isolated from neonatal rats. Different medium and culture concentration were used toculture NSCs of neonatal rat. The culture concentration used were 1×10 4, 1×105, 1×106and 1×107/ml respectively. Ingredient of medium was classified into group 1 to 8 respectively according to whether to add 2% B27, epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) as well as the difference in culture concentration. The cells were induced to differentiate asto be confirmed as NSCs, and then were checked by phase contrast microscopy and identified by immunocytochemistry. Results The cells isolated and cultured gathered into neurospheres. The cells were capable of proliferating and maintaining longterm survival in vitro. The cells could be differentiated into neurons and glia.It was to the benefit of the survival of NSCs to add 5% fetal bovine serum(FBS)into the medium at the beginning of the culturing. When 10% FBS was added intothe medium, the neurospheres differentiated quickly. When concentration 1×106/ ml was used, the growth rate of the cells was the highest of all the concentrations. Reasonably higher cell concentration promoted the proliferation of NSCs. It was necessary to add 2% B27, EGF, and bFGF into the medium. The cells had the best growth when 2% B27, 20 ng/ml bFGF and 20 ng/ml EGF were added into the culture medium. EGF and bFGF had cooperative effect. Conclusion A better method of isolating and culturing of NSCs in neonatal rat brain is established and the foundation for future research is laid.
Objective To observe the expression of related proteins of retina after subretinal implantation with inactive chips.Methods A total of 27 healthy adult New Zealand white rabbits were randomly divided into three groups: operation group (12 rabbits) in which the rabbits were implanted with inactive chips into the interspace beneath retina;shamoperation group (12 rabbits) in which the rabbits were implanted with inactive chips into the interspace beneath retina which was taken out immediately;the control group (3 rabbits). Animals were sacrified for immunohistological study 7,15,30 and 60 days after surgery.The rabbits in control group group were sacrified for immunohistological study after bred for 30 days.The expressions of glial fibrillary acidic protein (GFAP) and brain derived neurotrophic facor (BDNF) were observed.Results In operation group, the outer nulear layer of retina thinned, and the cells in the inner nulear layer was disorganized 7,15,and 30 days after the surgery;glial cells proliferated 60 days after surgery; the positive expression of BDNF and GFAP was more than that in the shamoperation and control group.In shamoperation group, the positive expression of BDNF and GFAP was more than that in the control group.No obvious difference of expression of BDNF and GFAP between each time point groups was found.Conclusions The expression of neroprotective related proteins increased after subretinal implantation with inactive chips suggests that limited neuroprotective effects might be led by the implantation.
Objective To investigate the perioperative changes of serum brain natriuretic peptide (BNP) concentrations in patients undergoing cardiac valve replacement. Methods There were 20 patients admitted to the study, the serum BNP concentrations were measured before cardiac surgery, 24 hours, 7days, 14 days, and 30 days after operation. The preoperative NYHA cardiac function and the left ventricular ejection fraction(LVEF) were measured by echocardiogram. Results The preoperative BNP level was the baseline, it elevated markedly and acutely to a peak value 24 hours after operation ( P =0.003), then the BNP decreased 7 days later, but was still higher than the concentration before operation ( P =0.015), 14 days later it reached to the concentration before operation, 30 days later it was mild lower than preoperative BNP level, but there was no significant difference. There was a positive correlation between NYHA and BNP ( r =0.69, P lt;0.05), but no correlation between LVEF and BNP( r =0.29, P gt;0 05). Conclusion The preoperative serum BNP concentration can reflect the preoperative cardiac function in patients undergoing cardiac valve replacement, the high BNP level indicates the poor cardiac function. The BNP sharply elevated in the early time after operation, then gradually decreased in the late phase postoperation.
Objective To explore the pulmonary arterial pressure level in patients with predialysis chronic kidney disease ( CKD) and its relationship to cardiac structure and function. Methods 397 patients with predialysis CKD and 50 healthy subjects were enrolled. Cardiac structure was evaluated by Doppler echocardiography. Glomerular filtration rate ( GFR ) were assessed by radiant 99mTc-DTPA.Differences of PAP, BNP, LA, IVST, LVDd, LVDs, LVEF, LVMI and the correlation of PAP with cardiac structure and function were examined. Results The PAP level in the predialysis CKD patients was much higher than that in the healthy subjects [ ( 33. 13 ±9. 00) mm Hg vs. ( 29. 43 ±3. 71) mmHg, P lt;0. 01] .18. 9% of the CKD patients were complicated with pulmonary hypertension. PAP was higher in the CKD patients in stages 4-5 than those CKD patients in stages 1-3 [ ( 35. 90 ±9. 34) mmHg vs. ( 32. 08 ±8. 62)mmHg, P lt;0. 01) ] , so as to the prevalene of pulmonary hypertension ( 21. 60% vs. 13. 47% , P lt;0. 01) .Compared with the healthy, the level of lnBNP [ ( 3. 59 ±1. 63) pg/mL vs. ( 2. 88 ±1. 51) pg/mL, P lt;0. 01] , LA [ ( 40. 42 ±6. 77) mmvs. ( 36. 75 ±4. 94) mm, P lt; 0. 01) ] , LVPW [ ( 9. 55 ±1. 96) mm vs.( 8. 54 ±0. 88) mm, P lt; 0. 01) ] , IVST [ ( 9. 76 ±1. 75) mm vs. ( 8. 71 ±0. 90) mm, P lt; 0. 01) ] , LVMI[ ( 105. 61 ±36. 47) g/m2 vs. ( 87. 41 ±17. 08) g/m2 , P lt; 0. 01) ] were all much higher. There was a negative correlation between PAP and GFR( r = - 0. 461, P lt;0. 01) , and positive correlations between PAP and LA ( r=0. 491, P lt; 0. 01) , LVPW ( r =0. 298, P lt;0. 01) , IVST ( r = 0. 613, P lt;0. 01) , lnBNP ( r =0. 536, P lt;0. 01) , LVMI ( r = 0. 382, P lt;0. 01) . LVMI and lnBNP were both independent risk factors of PAP. The regression equation: y = 16. 447 + 0. 105x1 + 1. 724x2 ( F = 23. 482, P = 0. 000) , y: PAP( mm Hg) , x1 : LVMI( g/m2 ) , x2 : lnBNP( pg/mL) . Conclusions Pulmonary hypertension is a common morbidity of predialysis CKD patients, and deteriorates with degression of renal function. PAP is related to indexes of cardiac structure ( LVMI, LA, LVPW, IVST) and index of cardiac function ( lnBNP) . LnBNP and LVMI are independent risk factors of PAP.
Aiming at the problem of low recognition accuracy of motor imagery electroencephalogram signal due to individual differences of subjects, an individual adaptive feature representation method of motor imagery electroencephalogram signal is proposed in this paper. Firstly, based on the individual differences and signal characteristics in different frequency bands, an adaptive channel selection method based on expansive relevant features with label F (ReliefF) was proposed. By extracting five time-frequency domain observation features of each frequency band signal, ReliefF algorithm was employed to evaluate the effectiveness of the frequency band signal in each channel, and then the corresponding signal channel was selected for each frequency band. Secondly, a feature representation method of common space pattern (CSP) based on fast correlation-based filter (FCBF) was proposed (CSP-FCBF). The features of electroencephalogram signal were extracted by CSP, and the best feature sets were obtained by using FCBF to optimize the features, so as to realize the effective state representation of motor imagery electroencephalogram signal. Finally, support vector machine (SVM) was adopted as a classifier to realize identification. Experimental results show that the proposed method in this research can effectively represent the states of motor imagery electroencephalogram signal, with an average identification accuracy of (83.0±5.5)% for four types of states, which is 6.6% higher than the traditional CSP feature representation method. The research results obtained in the feature representation of motor imagery electroencephalogram signal lay the foundation for the realization of adaptive electroencephalogram signal decoding and its application.
Fifteen rabbits were given a second degree scald on the back, then a large amount of fluid were given rapidly. In this way an acute diluted hyponatremia and a brain edema model were produced. In one group, the magnetotherapy was instituted immediately following the acut injury, while in the other group, it was started after the brain edema being developed. After giving a large quantity of fluid, the Na~+, CI~-, and K~+ concentrations in the blood were decreased and the quantity of water in the brain tissue was increased evidentely. The phenomena of neuron edema and widening of the perivascular space were also found. It was proved that the model of experimental brain edema could be produced on animals and the treatment and the prevention for brain edema could be obtained to advantage to certaiin degree. Also,it was better to use the magnetotherapy in brain edema as early as possidle.
ObjectiveTo investigate the effect of intravitreal injection of neural stem cells (NSC) derived from human umbilical cord mesenchymal stem cells (hUCMSC) on the expression of brain-derived neurotrophic factor (BDNF) and the number of retinal ganglion cells (RGC). MethodsFifty-two adult male Sprague-Dawley rats were randomly divided into normal group (group A) and diabetes mellitus group which received intraperitoneal injection of streptozocin to make diabetic rat models. One month after the diabetic rat models were confirmed successfully, diabetic rats were randomly divided into diabetic group (group B), hUCMSC group (group C) and hUCMSC-induced NSC group (group D). And thirteen diabetic rats were included in each group. Immuno-cytochemistry was applied to observe BDNF and thymosin-1(Thy-1) staining in the retina. Then mean integrated absorbance of the staining region on the retina slices were analyzed by Image-Pro Plus 6.0. The number of Thy-1 labeled RGC was record. ResultsBDNF and Thy-1 were positive on the retina slices from group A. The staining intensity from group B became weak and the expression of BDNF and Thy-1 gradually decrease with time (P < 0.05), and those from group C and group D were positively (P < 0.05), especially in group D (P < 0.05). The BDNF expression and Thy-1 labeled RGC were the same between group B and C (P > 0.05) at 2 weeks after injection, but were significant different for other time points (P < 0.05).Significant positive correlation between the expression of BDNF and the number of RGC were found by the Pearson correlation analysis (r=0.964, P < 0.05). ConclusionIntravitreal injection of hUCMSC-derived NSC to diabetic rat may protect the retina by promoting the expression of BDNF and increasing the number of RGC.
Objective To study the relationship between the expression ratio of induced nitric oxide synthase (iNOS) over glial fibrillary acidic protein (GFAP) and the time of injury after brain concussion in rat, in order to acquire a new visual angle for determining injury time of cerebral concussion. Methods Eighty-five healthy Sprague-Dawley rats were divided into three groups randomly: model group (n=25), experimental group (n=55), and control group (n=5). The rats in the model group were used to confirm the attack hight to make the model of brain concussion; according to the time of execution, rats in the experimental group were then subdivided into 11 groups with 5 rats in each subgroup, and their execution time was respectively hour 0.5, 1, 3, 6, 12, 24, 48, 96, 168, 240, and 336; the rats in the control group were executed after fed for 24 hours. After the model of cerebral concussion was established through freefalling dart method, hematoxylin-eosin staining and immunohistochemistry staining of iNOS and GFAP were conducted for the brain of the rats. All related experimental results were studied by using microscope with image analytical system and homologous statistics. Results The ratio of positive expression of iNOS over that of GFAP increased gradually during hour 0.5- 3 after injury in brain (from 5.03 to 10.47). At the same time, the positive expression of iNOS increased significantly (from 14.61% to 37.45%). However, the increase of the positive expression of GFAP was not obvious. Between hour 3 and 12, the ratio began to decline to 4.98, which was still at a high level, and during the same time period, the positive expressions of iNOS and GFAP also experienced the same change pattern. Later, the ratio began to decline between hour 12 and 336 after injury (from 4.98 to 0.95). All ratios at this time were lower than those between hour 0.5 and 12. The positive expression of iNOS and GFAP both increased to a climax before declining. Conclusions The ratio of positive expression of iNOS over GFAP and the respective change pattern of iNOS and GFAP can be used as the evidence of estimating the injury time of cerebral concussion. We can use the ratio of two or more markers to provide a new visual angle for concluding the concussion injury time.
Objective To investigate present status of health care in peri-brain-death and analyze its effectiveness and health economic characteristics. Method Retrospective analysis of case series was conducted and a total of 940 patients from surgical intensive care unit (SICU) were reviewed on treatment and part of direct medical expenditure. The patients admitted from Jun. 1999 to Dec. 2000 and Nov. 2001 to Jun. 2002 were included in this study. Data were processed by SPSS 10.0. Results Patients were included if they had two of the three symptoms for at least one hour: deep coma, pupillar light reflex disappear, and no autonomic respiratory. Ultimately 115 patients were included, with a total cost of ￥2 515.9 per day for each case, whereas mortality was 99.10%. Mortality increased with the state of peri-brain-death prolonged. Eighty percent of patients included were dead within 72 hours after admission. Conclusions Attempts to resuscitate patients of peri-brain-death have been the most widely applied in China, however, it resulted in great unnecessary consumption of health resources. It is of great importance to promote legislation of brain death in China.
【摘要】 目的 探讨颅脑损伤(BI)死亡的法医病理学特点,以及继发性脑干损伤、并发症的发生与死亡之间的因果关系。方法 从性别、年龄、致伤方式、损伤类型、生存时间、死亡原因等方面,对四川大学华西法医学鉴定中心1998年1月-2008年12月127例BI死亡尸检案例进行回顾性统计研究分析。结果 127例法医病理学检案中,原发性BI死亡51例(402%),继发性脑干损伤死亡61例(480%),并发症死亡15例(118%),其中伤后12 h内死亡者直接死因均为严重原发性脑损伤,存活12 h～1周者直接死因以继发性脑干损伤居多,生存时间超过1周者约半数死于并发症。结论 在BI案例的死亡原因确定时,应在全面系统的病理学检验基础上,结合案情及临床资料进行综合分析。【Abstract】 Objective To explore the characteristics of forensic pathology in traumatic brain injury and the relationships between secondary brainstem damage, complications and the causes of death. Methods 127 cases were reviewed from gender, age, manner of injury, survival time and the direct causes of death from January 1998 to December 2008. Results Of the 127 cases, the key direct cause of death was secondary brainstem damage, followed by severe primarily brain injury and complications. For those who died within 12 hours after injury, the direct cause was severe primarily brain injury; for those who survived between 12 hours to one week, secondary brainstem damage was in the majority of the causes and for those who survive more than one week time, complication was an important cause. Conclusion In the cases of traumatic brain injury, we should take comprehensive and systematic examination of forensic pathology, and refer to clinical data at the same time to determine the direct cause of death.