Objective To observe the change of neuron-specific enolase (NSE) an d glial fibrillary acidic protein (GFAP) in neonatal rats' cerebral cortex with hypoxic-ischemic brain damage (HIBD). Methods The 7 day- old SD rats were subjected to the ligation of right carotid artery, then were pu t into a hypoxic box to establish a HIBD model. The immunohistochemical method w as used to detect the expression of NSE and GFAP in rats' cerebral cortex. Results ① The NSE decreased in damaged cerebral cortex in HIBD 24 h group, after 7 days it gradually increased but was still lower than that of the controls. ② The expression of GFAP was limited and scarce in control and it did not change in HIBD 24 h group, while in HIBD 7 d group GFAP expression was increased and spread widely in the damaged cerebral cortex. Conclu sion ① The NSE decreases in damaged cerebral cortex in early stage of neonatal rat HIBD, suggesting that NSE is a specific marker for neuron damage . ② The GFAP increases in damaged cerebral cortex in the recovery stage of neon atal rat HIBD, suggesting that GFAP participates the repair of lesion region.

Objective To investigate relationship of serum insulin like growth factors(IGFs) and catch up growth. Methods The model of rats intrauterine growth retardation (IUGR) was built by clamping bilateral uterine arteries and veins of pregnant rats. The growth parameters (body weight and length), organs weights (brain, lungs and liver) and serum concentrations of IGF Ⅰ, Ⅱ of all neonatal rats was measured at birth and postnatal day 3 and day 5, respectively. Results (1)At birth, the serum levels of IGF Ⅰ, Ⅱ and the average growth parameters, organs weights in IUGR rats were significantly lower than those in control group( P 0.05). The serum of IGF Ⅱ in high catch up group was significantly higher than that at birth( P 0.05). Conclusion Both IGF Ⅰ, Ⅱmay play an important role in the regulation of catch up growth and the decrease of IGF Ⅰ, Ⅱlevels may be the internal factors contributing to the absence of postnatal catch up growth in IUGR rats.

Objective To investigate the expression of two long non-coding RNAs ( lncRNAs ) during HIV-1 infection, which were nuclear-enriched autosomal transcript 1 (NEAT1) and metastasis asso-ciated lung adenocarcinoma transcript 1 ( MALAT1 ) , and their relationships with disease progression . Methods Fifty-nine patients with HIV-1 infection and 21 healthy subjects were recruited in this study , of which 31 patients were highly active antiretroviral therapy ( HAART)-na?ve and 28 patients received HAART for more than one year with undetectable viral loads .Total RNAs were extracted from PBMC and plasma samples, respectively.The levels of NEAT1 and MALAT1 were detected by quantitative real time polymer-ase chain reaction .Results The levels of NAET1 and MALAT1 in PBMC from HAART na?ve patients were 3 to 5 times higher than those in healthy subjects (P<0.01).The levels of NAET1 and MALAT1 in PBMC from HAART treated patients were similar to those in healthy subjects .The levels of NEAT1 in plasma sam-ples from patients with HIV-1 infection were lower than those in healthy subjects , and were positively corre-lated with CD4+T cell counts (P<0.01).No significance differences in the levels of MALAT1 in plasma samples were found between those with and without HIV-1 infection (P>0.05).Conclusion This study suggested that NEAT 1 and MALAT1 might be involved in the disease progression in patients with HIV-1 in-fection.The level of NEAT1 in plasma could be used as a potential biomarker of HIV-1 infection.

BACKGROUND:The discovery and concept of pulp tissue-derived stem cells is beneficial to the understanding of tooth development and regeneration and repair mechanisms from the cellular level. OBJECTIVE:To understand the induced differentiation capacity and induced conditions in vitro of human dental pulp stem cells into neuron-like cells. METHODS:Pulp tissue was separated from human healthy third molars. Single cellsuspensions were prepared and seeded into 6-wel plates containing alpha-modified minimum essential medium supplemented with 15%fetal bovine serum. Subconfluent cultures (first passage) of colony forming cells were induced with butylhydroxy anisole, forskolin,β-mercaptoethanol, basic fibroblast growth factor. RESULTS AND CONCLUSION:Immunofluorescence and reverse transcription-PCR assay showed that human dental pulp stem cells positively expressed stro-1, Col-I, dentin sialoprotein after 2 weeks of induction. Nestin and neuron-specific enolase were strongly expressed, but the gingival fibroblasts were negatively expressed. It indicates that adult stem cells in human dental pulp have a high neuron-like celldifferentiation potential under a certain inductive condition.

BACKGROUND:Hyperbaric oxygen(HBO)treatment promotes the proliferation and differentiation of endogenous neural stem cells in neonatal rats following hypoxic/ischemic brain damage(HIBD).The Wnt signaling pathway is associated with neurogenesis.However,there are few data recording the role of HBO in the differentiation of neural stem cells in vitro.OBJECTIVE:To observe the effect of HBO on differentiation and Wnt3 expression of bone marrow mesenchymal stem cells(BMSCs).METHODS:BMSCs were isoiated and cultured.The rat BMSCs of passages 3-5 were cultured in DMEM/F12(1:1)medium with basic fibroblast growth factor,epidermal growth factor and B27 for 24 hours.The induced BMSCs were randomly divided into two groups:control group(no treatment)and HBO group(HBO,0.10 MPa,60 minutes stabilizing pressure with at least 90% oxygen).The neuron specific encloase(NSE),glial fibrillary acidic protein(GFAP)and 04 marked oligodendrocyte immunocytochemistry were detected by immunofluorescent staining,and Wnt3 protein expression was detected by Western-blot.RESULTS AND CONCLUSION:BMSCs cultured in classic medium of neural stem cells could significantly induce the expression of nestin.The expression of NSE and 04 of HBO group was greater than control group(P＜0.01),but GFAP expression displayed no significant difference between the groups(P＞0.05).Western blot showed HBO could enhance the Writ3 expression (P＜0.05).Results show that HBO can induce BMSCs to differentiate into neural cells and oligodendrocyte,which is correlated with the activation of the Wnt3 protein.

BACKGROUND: The researches on the onset mechanism and intervention of periventricular leukomalacia (PVL) are affected due to few cases of generally-acknowledged animal model, so it is necessary to establish a reliable animal model for the study of PVL.0BJECTIVE: To establish PVL animal model of the 2-day-old SD rats.DESIGN: Randomized control animal trial.SETTING: Department of Pediatrics, West China Second University Hospital, Sichuan University.MATERIALS: A total of 36 2-day-old SD rats of cleaning grade and either gender, weighing 6-8 g, were provided by the West China Experimental Animal Center of Sichuan University; Mice anti-O4 was purchased by Chemicon Company,rabbit anti-glial fibrillary acidic protein (GFAP), rabbit anti-β-amyloid precursor protein (β-APP), rabbit anti-myeline basic protein (MBP), SABC immunohistochemical kit and DAB color reagent were all offered by Wuhan Boster Biological Technology Co., Ltd. Rabbit anti-mice IgG-FITC was obtained from Zhongshan Golden Bridge BioTechnology Co., Ltd.METHODS: The experiment was accomplished in the Laboratory of Women and Children, West China Second Hospital of Sichuan University between May and December in 2005. Totally 36 rats Were randomly divided into experimental PVL group and control group, 18 in each. The experimental PVL group was subjected to unilateral carotid ligation (UCL), and then they were put into a box filled with 6% oxygen and 94% nitrogen for 4 hours. Six rats were executed at ischemic 72hours, 14 days and 28 days respectively. Meanwhile sham surgeries were performed on the control group without ligation or exposure to hypoxia. And the time segment was identical with that of experimental group. ①Histopathological examination: Rat hearts were fixed by perfusion and stained with hematoxylin-eosin (HE). Light and electronic microscopy were used to observe the brain pathological and ultrastructure changes, ②Immunohistochemistry method was used to detect the distribution and expression of GFAP, β-APP, MBP and O4 in the white matter of both experimental and the control groups 72 hours post-operation. ③Neuroethology examination: Hanging test (rats were forced to hold the horizontal glass rod with forelegs, and the time of dropping was recorded in the distance of 45 cm.Scoring: 1 point: ＜ 10 s; 2 points: 10-30 s; 3 points: 30 s-2 minutes; 4 points: 2-5 minutes; 5 points: ＞ 5 minutes),inclined plane test (rats were laid on the inclined plate at the angle of 45° while rat heads turning upwards at the angle of more than 135°), open field test (square box without summit was divided into 9 equal grills at the bottom, rats were placed in the central grill to observe the activity within 30 seconds. Scoring: 1 point as rats entered the neighbor cage above half the body; 1 point as standing by hind limbs; total scores were the addition of the two), and cylinder test (rats were put in the cylinder of 20 cm×30 cm×5 cm to record the time of initial forepaw of each weight-bearing contact with the wall during a full rear, right (ipsilateral) or left (contralateral) percentage of total forepaw contacts at initiation was calculated.) were tested on the SD rats at 28 days post-operation. Then statistical management was conducted.MATN OUTCOME MEASURES: ①HE stain and electronic microscope were used to detect the histopathology changes after ischemia and hypoxia.②lmmunohistochemistry method was used to detect the expressions of GFAP, β-APP, MBP and O4 in the corresponding cell and tissue after ischemia and hypoxia. ③Neuroethology examination was used to evaluate the rats after ischemia and hypoxia by scores of each test.RESULTS: All 36 rats were involved in the result analysis. ①White matter damage was observed in the periventricular white matter in the PVL group by light and electronic microscopy at the early stage of post-operation; Ventricular dilatation and the loss of medullary sheath were detected in the white matter at the latter stage. ②The integrated optical density (IOD) of GFAP in PVL group was stronger than that of the control group (6 566.93±455.56, 1 069.32±791.71,P ＜ 0.05), and the mean diameter of GFAP-immunoreactive cells was increased in the brain tissue of PVL group compared with those of the control group [(11.69±0.97), (8.24±0.22), P ＜ 0.05]; β-APP immunohistochemistry demonstrated the IOD of PVL group was stronger than that of the control group [(59 304.07±6 864.03), (15 132.29±2 455.52),P＜ 0.05]; MBP IOD of the PVL group was decreased compared with the control group [(21 764.29±1 981.63), (69 174.72±3 199.90), P ＜ 0.05]; The density of O4-immunoreactive pyknotic cells was dramatically increased in the PVL group compared with the control group [(54.08±11.99), (1.25±0.51), P＜ 0.05].③In PVL group, the hanging time was shorter in the hanging test than that of the control group [(1.27±0.14), (4.24±0.59) minutes, P ＜ 0.05]; The turning-around time was longer in the inclined plane test than that of the control group [(7.17±2.32), (3.27±0.82) s, P ＜ 0.05]; The score in the open field test was decreased than that of the control group [(3.68±0.82), (12.67±1.00) s, P ＜ 0.05]; In the cylinder test the activity of the left limb was less than that of the right limb [(19.25±2.77), (64.55±0.36)%, P ＜ 0.05].CONCLUSION: PVL animal model can be successfully established by the method of UCL-hypoxia using the 2-day-old SD rat, and appears the obvious white atter damage, abnormal neurobehavior, reasonable pathological and behavior change.

Humans ; Infant, Newborn ; Microbiological Techniques ; Sepsis ; blood ; microbiology

Objective To separate NK cells of mice from NK cell separation medium and study inhibitory effect of proliferated NK cell induced by low dose radiation on the leukemia model of K562 cells.Methods Flow cytometry and 3H-TdR methods were respectively used to measure proliferation index and activity of NK cells treated with low-dose radiation( which means exposure dose in 20 cGy low LET beam or 5 cGy high LET beam).CD13 + cells were measured by flow cytometry and TNF-α content in blood-serum was detected by ELISA.In vivo,peripheral blood leucocyte count,index of liver,indexes of spleen and kidney were observed in control group and experimental group.Results The purity of NK cell separation was (82.54 ± 0.18)%.The proliferation index of NK cells at 24 hours after 80 mGy irradiated was 36.31 ± 1.32% ,(t =24.69,P ＜0.05).Killing activity of NK cell induced by low dose radiation to K562 cell was (12.59±0.63)%(t=6.63,P＜0.05)and the inhibition ratio was 29.52%.Conclusion The injection of proliferated NK cell induced by low dose radiation demonstrated significant inhibitory effect on the growth of leukemia nude mouse.

Female ; Humans ; Infant, Newborn ; Liver Diseases ; surgery ; Liver Transplantation ; Pregnancy ; Pregnancy Complications ; surgery ; Pregnancy Outcome

Objective To investigate the effect of sepsis on vecuronium-induced inhibition of acetylcholine release in neuromuscular junction in rats.Methods Thirty-six adult male SPF SpragueDawley rats,aged 2-3 months,weighing 200-220 g,were randomly divided into 3 groups (n=12 each) using a random number table:control group (group C),sham operation group (group S) and sepsis group (group Sep).Sepsis was induced by cecum ligation and puncture (CLP) in rats anesthetized with intraperitoneal chloral hydrate 350 mg/kg.At 12 h after CLP,the sciatic nerve-pretibial muscle was prepared.Vecuronium was added to the culture medium with the final concentration of 0.08 μg/ml,and the sciatic nerve-pretibial muscle was incubated for 15 min.Before and after administration,evoked endplate potentials (EPPs) and miniature endplate potentiais (MEPPs) were recorded by using intracellular microelectrode.EPP/MEPP ratio was calculated.Results Compared to C and S groups,EPPs,MEPPs and EPP/MEPP ratio were significantly increased before and after administration in group Sep.EPPs,MEPPs and EPP/MEPP ratio were significantly lower after administration than before administration in the three groups.Conclusion Sepsis can promote acetylcholine release in neuromuscular junction,thus weakening vecuronium-induced inhibition of acetylcholine release in neuromuscular junction in rats.