Objective: To evaluate the protective effect of granulocyte-colony stimulating factor(G-CSF) on neonatal rats after hypoxic-ischemic brain damage(HIBD)and its effect on endoplasmic reticulum (ER) stress. Methods: According to the random number table, a total of 54 Sprague-Dawley (SD) rats aged 7 days were divided into 3 groups(18 rats in each group): Sham group, HIBD group and G-CSF group, and the improved Rice method was used to establish a neonatal rat model of HIBD.A dose of 50 μg/kg of G-CSF was administered intraperitoneally 1 hour after HIBD (G-CSF group), while the rats in HIBD group and Sham group received saline only.At 24 hours of HIBD, pups were euthanized to quantify brain infarct volume by using 2, 3, 5-Triphenyltetrazolium chloride.Hematoxylin-Eosin (HE) staining was used to observe the changes of brain structure.Neuronal cell death was determined by using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). Then the expressions of glucose-regulated protein 78 (GRP78), cysteinyl aspartate specific proteinase 12 (Caspase-12), CCAAT/enhancer binding-protein homologous protein (CHOP) were assessed by Western blot and immunofluorescence staining. Results: Twenty-four hours after operation, HE staining showed that no significant neuronal damage was observed in Sham group.The brain tissue structure of rats in the HIBD group was significantly damaged, while some improvement was observed in the G-CSF group.The infarction volume in HIBD group[(25.40±5.15)%] increased compared with that in the Sham group[(0.31±0.15)%] and the G-CSF group[(16.36±4.97)%], and the differences were statistically significant(all P<0.05). There was increased positive expression of GRP78 protein in HIBD group, compared with that in the Sham group and the G-CSF group[(49.38±10.06)% vs.(9.12±4.50)%, (30.61±6.35)%], and the differences were statistically significant (all P<0.05). The percentage of apoptosis in the hippocampal CA1 region and conex in HIBD group [(44.84±11.54)%, (48.23±14.07)%] were higher than those in the G-CSF group [(17.87±7.20)%, (26.18±9.96)%], and the differences were statistically significant (all P<0.05). The expression of GRP78, CHOP and Caspase-12 in the HIBD group (0.63±0.24, 0.72±0.21, 0.68±0.25) were higher than those in the Sham group (0.20±0.08, 0.28±0.08, 0.23±0.07), and the G-CSF group (0.39±0.13, 0.51±0.18, 0.48±0.16), and the differences were statistically significant (all P<0.05). Conclusions G-CSF exerts neuroprotective effect on the neonatal rats after HIBD.G-CSF may be an effective treatment of HIBD by reducing ER stress-induced neuronal apoptosis.

Objective: To investigate the role of granulocyte-colony stimulating factor (G-CSF) on the regulation of inflammatory cytokines in neonatal hypoxic-ischemic brain damage(HIBD) rat model, and to explore the possible mechanism involved in G-CSF neuroprotective effect via the mammalian target of Rapamycin/p70 ribosomal S6 protein kinase (mTOR/p70S6K) signaling pathway. Methods: A group of postnatal day 7 (P7) Sprague-Dawley rat pups (90 cases) were randomly divided into sham-operated group, hypoxia-ischemia(HI) group, G-CSF group, Rapamycin (RAP) group and control group, and the improved Rice method was used to establish a neonatal rat model of HIBD.One hour before HI induction, Rapamycin was administered intraperitoneally with a dose of 250 μg/kg, and the control group was given equal volume of ethanol injected intraperitoneally.One hour after HI, a dose of 50 μg/kg of G-CSF was injected intraperitoneally into the G-CSF group, Rapamycin group and control group.The same volume of normal saline was injected intraperitoneally into HI group and sham-operated group.Forty-eight hours after HI, Western blot was used to detect the protein levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-10, and the mTOR/p70S6K signaling pathway in brain tissue.Neuron injury of the hippocampal CA1 region and the cortex was assessed by Nissl staining, and infarct volume detected by 2, 3, 5-triphenyltetrazolium chloride staining. Results: The G-CSF group and control group were associated with significantly reduced infarction volume compared to the HI group [(12.87±1.54)%, (11.90±1.31)% vs.(24.21±3.28)%], and the differences were statistically significant(P<0.05). There was an increased positive neuron cell number in the ipsilateral hemispheres of the hippocampal CA1 region in the G-CSF group and the control group [(61.00±4.90) cell/field and (61.67±6.40) cell/field] and cortex [(92.67±6.68) cell/field and (90.17±4.45) cell/field] compared with those in HI group [(42.62±4.46) cell/field and (70.83±6.97) cell/field], and the differences were all statistically significant (all P<0.05). The expression levels of TNF-α and IL-1β were significantly decreased in the G-CSF group and the control group, compared with those in HI group(TNF-α: 0.67±0.07, 0.55±0.05 vs.0.86±0.05; IL-1β: 0.65±0.06, 0.52±0.10 vs.0.86±0.06), and the differences were all statistically significant (all P<0.05). There was increased expression levels of IL-10, p-mTOR/mTOR and p-p70S6K/p70S6K in the G-CSF group and the control group, compared with those in HI group (IL-10: 0.68±0.04, 0.62±0.05 vs.0.34±0.02; p-mTOR/mTOR: 0.53±0.02, 0.51±0.01 vs.0.26±0.01; p-p70S6K/p70S6K: 0.89±0.03, 0.90±0.03 vs.0.55±0.02), and the differences were all statistically significant(all P<0.05). There was an increased infarct volume in Rapamycin group [(25.70±1.50)%], compared with the G-CSF group and the control group, and there were decreased number of positive neuron cell count in the hippocampal CA1 region [(40.67±3.50) cell/field] and cortex [(68.33±8.17) cell/field], increased expression levels of TNF-α and IL-1β (0.97±0.06 and 0.98±0.10, respectively), decreased expression levels of IL-10, p-mTOR/mTOR and p-p70S6K/p70S6K (0.21±0.02, 0.30±0.01 and 0.55±0.01, respectively) in the Rapamycin group, and the differences were all statistically significant (all P<0.05). Conclusions G-CSF may inhibit inflammatory responses after HIBD by up-regulating the mTOR/p70S6K signaling pathway in neonatal HI encephalopathy.

OBJECTIVE:To systematically evaluate the efficacy and safety of metronidazole combined with hydrogen peroxide for uterine cavity lavage in the treatment of puerperal infection. METHODS:Retrieved from PubMed,Cochrane library,Wanfang database,VIP,CBM and CNKI,Meta-analysis was performed by using Cochrane collaboration Rev Man 5.3 software after data extraction and quality evaluation with modified Jadad scale. RESULTS:A total of 16 RCTs were included,involving 1 280 patients. Meta-analysis showed that total response rate of observation group was significantly higher than that of control group [OR＝5.76,95%CI(3.21,10.35),P＜0.001],but febrile duration [MD＝－2.08,95%CI(－2.32,－1.84),P＜0.001],duration of clinical symptom [MD＝－2.71,95%CI(－2.89,－2.53),P＜0.001] and average hospital stay [MD＝－3.05,95%CI(－3.32,－2.79),P＜0.001] are significantly shorter than control group; the incidence of ADR [OR＝0.27,95%CI(0.08,0.89),P＝0.03] was significantly lower than control group,with statistical significance. CONCLUSIONS:Metronidazole combined with hydrogen peroxide for uterine cavity lavage show good therapeutic efficacy and safety in the treatment of puerperal infection.

Objective To investigate the effects of granulocyte-colony stimulating factor (G-CSF) on neuronal apoptosis after hypoxia-ischemia brain damage (HIBD) and the possible role of the mTOR/p70S6K signaling pathway in neonatal rats.Methods Ninety seven-day-old Sprague-Dawley rats were assigned into 5 equal groups (n=18):sham group,HIBD group,G-CSF group,rapamycin group and ethanol group by random number table method.Pups were subjected to unilateral carotid artery ligation followed by 2hrs hypoxia or sham surgery.HIBD animals received normal saline,G-CSF (50 μg/kg),G-CSF combined with rapamycin (250 μg/kg) or ethanol (vehicle for rapamycin).Pups were euthanized 48hrs post-HIBD to quantify the percentage of brain infraction area.The pathomorphologic changes in the hippocampal CA 1 area and cortex were observed by Nissl staining.Neuronal cell death was determined using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL).mTOR,activated mTOR (p-mTOR),p70S6K,activated p70S6K (p-p70S6K),Cleaved Caspase-3 (CC3),Bax,and Bcl-2 were quantified using Western blot analysis.Results G-CSF treatment resulted in significantly reduced percentage of brain infraction area (P＜0.05) and neuronal apoptosis in the hippocampal CA1 area and cortex (P＜0.05) after HIBD in neonatal rats.However,rapamycin administration reversed the neuroprotective effect of G-CSF.G-CSF administration ameliorated the pathomorphologic damage in the ipsilateral hemisphere.Compared with the HIBD group,the Nissl stained neurons significantly increased in the G-CSF group (P＜0.05).Furthermore,G-CSF increased the expression ofp-mTOR,p-p70S6K and Bcl-2 but decreased the expression levels of CC3 and Bax in the ipsilateral hemisphere,which were all significantly reversed by rapamycin (P＜0.05).Conclusion G-CSF may attenuate caspase activation and reduce neuronal apoptosis by up-regulating the activity of mTOR/p70S6K signaling pathway after experimental HIBD in rat pups.

Granulocyte colony -stimulating factor, as a growth factor that stimulates bone marrow hematopoiesis, is clinically used to treat neutropenia caused by various causes. However, more and more in vitro and in vivo research experiments and clinical trials have shown that granulocyte colony-stimulating factor exerts neuroprotective effects in nervous system injury diseases by activating multiple signal transduction pathways, including mobilizing peripheral stem cells to migrate to the nervous system, reducing nerves apoptosis, balancing inflammatory responses, and promoting neural stem cell regeneration and angiogenesis, but there is also some controversy. Currently, the use of granulocyte colony-stimulating factor therapy in stroke, amyotrophic lateral sclerosis and spinal cord injury has entered clinical trials, and its efficacy has also been confirmed in a neonatal animal model of hypoxic-ischemic brain damage. The neuro-protective effects of granulocyte colony-stimulating factor and the safety and efficacy of clinical trials are reviewed.

Objective To explore the effect of docosahexaenoic acid (DHA) on long-term learning and memory disorders and potential mechanism in rats after hypoxic ischemic brain damage. Methods Sixty neonatal 7-day-old SD rats were ramdonly divided into three groups: group S (sham operation+vehicle treatment), group C (hypoxic-ischemic brain damage [HIBD]+vehicle treatment) and group D (HIBD+DHA treatment). After left common carotid artery was isolated and ligated for 2.5 h, rats of group C and group D were put into a condition which oxygen concentration was about 8%for 2 h;rats in the group S were only isolated the left carotid artery, without ligation or hypoxia treatment;rats in the group D were intraperitoneally injected DHA of 15 mg/kg after modeling, and rats in the group S and group C were intraperitoneally injected equivalent volume of vehcle, once a day for 10 consecutive days. The pathomorphology changes of the hypocampal CA1 area, and marginal division of striatum were observed by Nissl staining 48 h after modling; the apoptosis cells were measured by TUNEL;immunohistochemical method was used to detect the expressions of Bax and Caspase-3 positive cells in the two brain areas. Morris water maza test was used to evaluate the long-term lerning and momory functions of 2-month-old rats, and the expressions of N-methyl-D-aspartate receptor 1 (NMDAR1) positive cells were detected by immunohistochemical method. Results The pathomorphology damage was significantly improved, the expressions of Bax and Caspase-3 positive cells and the neuron apoptosis in hypocampal CA1 areas and marginal division of striatum in group D were all signficantly decreased as compared with those in the group C (P<0.05). Rats in group D had significantly decreased escape latency as compared with those in group C in Morris water maze test (P<0.05), and the expression of NMDAR1 positive cells in the two brain areas of group D was significantly increased as compared with that in the group C (P<0.05). Conclusion DHA has the ameliorative effect on long-term learning and memory disorders after hypoxic ischemic brain damage in rats, which may be associated with inhibitory action of cell apoptosis at early phase and up-regulation of expression of NMDA1 at the late phase.

Objective To investigate the effect of exogenous taurine (TAU) on amino acid content in the brains of rats suffered hypoxic-ischemic brain injury (HIBD).Methods The 7-day-old SD rats were randomly divided into sham-operated group, control group and TAU treatment group.HIBD models in the later two groups were established following Rice-Vannucci method.On the basis of breastfeeding, TAU at 300 mg/(kg ·d) via tube feeding to the TAU treatment group and an equal volume saline solution to the sham-operated group and control group were given.The aspartic acid (ASP), glutamic acid (GLY), glycine (GLY), TAU and γ-aminobutyric acid (GABA) contents in the rat brains were measured by high-performance liquid chromatography at 48 h and 14 d after HIBD.Results At 48 h after HIBD, the TAU, ASP and GLU contents in the TAU treatment group were significantly increased as compared with those in the control group (P＜0.05);the GLU/TAU ratio in the TAU treatment group was significantly decreased as compared with that in the control group (P＜0.05).Fourteen d after HIBD, the TAU, GABA and GLY contents were significantly increased, the GLU content was significantly decreased, and the ASP/GLY, GLU/GLY and GLU/TAU ratios were significantly deceased in the TAU treatment group as compared with those in the control group (P＜0.05).Conclusion Exogenous TAU addition during lactation of rats suffered HIBD may improve contents of inhibitory amino acids as TAU, GABA and GLY, and modulate the ratio of main excitatory/inhibitory amino acids neurotransmitters.

As an inhibitory amino acid similar to gama-aminobutyric acid,taurine can activate the corticostriatal pathway as an endogenous ligand for glycine receptors,establishing equilibrium between the excitatory and inhibitory processes in the brain.In mammalian brains,taurine concentrations increase during the developmental period of the brain until weaning,and subsequently decline reaching stable concentrations in adulthood.With abilities of anti-oxidative stress,anti-inflammatory and anti-apoptosis,taurine can improve the hypoxic-ischemic brain injury,promote the proliferation and differentiation of neurons and affect brain development,It needs more investigations to prove when and how taurine supplementation during gestation,baby,children or adult can assist the development of the brain and prevent the damage of the brain from hypoxic and ischemic damage.

Objective Marginal division (MrD) of striatum is a universal structure in the mammalian brain,and it plays an critical role in learning and memory.In the present study,we try to investigate the synaptic ultrastructure of Methionine enkephalin (MET-ENK) immunoreacted fibers connected with neurons in the marginal division of the striatum in the monkey brains to explore the ultrastructural basis of the mechanism of learning and memory function in MrD.Methods Six male monkeys (macaque) were perfused with paraformaldehyde through heart to fix the brain and the brains were sectioned by a cryostat.Sections of the brains were performed immunohistochemical staining to detect the MET-ENK expression in the stfiatum; the areas with positive immumohistochemical staining was performed ultrastructural observation for morphological characteristics of the MET-ENK synapses in the MrD.Results Immunohistochemistry staining showed a dense arrangement of MET-ENK immunoreactive cells between the putamen and globus pallidus.Five major types of MET-ENK synapses were identified in the MrD:the axo-dendritic synapses,the axo-spinous synapses,the axo-somatic synapses,the axo-axonic synapses and the compound synapses.Conclusion The MET-ENK synapses in the MrD are diverse and complex,and can be distinguished from the rest of the striatum.

Objective To investigate the neural representations of long-term digital memory in human brain by functional magnetic resonance imaging (fMRI) technique. Methods 22 right-handed normal volunteers were recruited to participate in a test of long-term digital memory while the fMRI data were recorded. Control tasks were performed for the block-design. SPM 99 was used to analyze the data and to obtain the activated brain regions.Numbers of activated voxels were used to calculate lateralization index (LI). Results When the threshold was set as P＜0. 0001 ,using a one-sample t -test,the middle gyrus of the left frontal lobe(t=9.68) and the right cerebellum ( t = 9.85 ) were activated remarkably during the memory task. The subcortical structures including the thalamus (t=6.72) and the caudate (t=6.58) were also obviously activated during the memory task. LI of the numbers of activated voxels was 0.51. Conclusions The subcortical structures and the cerebellum as well as the cerebral cortex are collaborative to contribute to long-term digital memory function in human brain. The results also reveal that the functional areas of long-term digital memory in human brain are localized with the functional lateralization in the left hemisphere.