The Orally Active Noncompetitive AMPAR Antagonist Perampanel Attenuates Focal Cerebral Ischemia Injury in Rats.

Inhibition of ionotropic glutamate receptors (iGluRs) is a potential target of therapy for ischemic stroke. Perampanel is a potent noncompetitive α-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor (AMPAR) antagonist with good oral bioavailability and favorable pharmacokinetic properties. Here, we investigated the potential protective effects of perampanel against focal cerebral ischemia in a middle cerebral artery occlusion (MCAO) model in rats. Oral administration with perampanel significantly reduced MCAO-induced brain edema, brain infarct volume, and neuronal apoptosis. These protective effects were associated with improved functional outcomes, as measured by foot-fault test, adhesive removal test, and modified neurological severity score (mNSS) test. Importantly, perampanel was effective even when the administration was delayed to 1 h after reperfusion. The results of enzyme-linked immunosorbent assay (ELISA) showed that perampanel significantly decreased the expression of pro-inflammatory cytokines IL-1ß and TNF-α, whereas it increased the levels of anti-inflammatory cytokines IL-10 and TGF-ß1 after MCAO. In addition, perampanel treatment markedly decreased the expression of inducible nitric oxide synthase (iNOS) and neuronal nitric oxide synthase (nNOS), and also inhibited nitric oxide (NO) generation in MCAO-injured rats at 24 and 72 h after reperfusion. In conclusion, this study demonstrated that the orally active AMPAR antagonist perampanel protects against experimental ischemic stroke via regulating inflammatory cytokines and NOS pathways.

[A Preliminary Analysis of Bifidobacterial Species Dependent Influence on the Health of the Elderly].

OBJECTIVE: To analyze the species dependent effect of bifidobacteria to the heath of the middle and elderly. METHODS: Total 148 healthy volunteers living in Chengdu with 5074 years old were recruited in 2015. The blood samples were collected from them and analyzed physiologically and immunologically. The fecal bifidobacterial were also analyzed by realtime fluorescence quantitative PCR (qPCR) using 16S rRNA targeting designed gene and species specific primers. RESULTS: Bifidobacterium can be detected in feces of the elderly,the detection rate was 100%,108/g fecal. Especially,more species predominated in the infants were found the tested subjects. Bifidobacterium was positively associated with urea nitrogen ( r=0.214, P<0.05). B. adolescentis was negatively associated with body mass index (BMI) ( r=-0.311, P<0.05),while B.catenulatum was positive to BMI ( r=0.167, P<0.05). B.breve and high density lipoproteincholesterol (HDLC) were negatively correlated ( r=-0.247, P<0.05). Bifidobacterium infantis and HDLC were positively correlated ( r=0.350, P<0.05). Among tested immune parameters,only B.bifidum was found to be positive associated with IgA ( r=0.365, P<0.05). CONCLUSION: Bifidobacterium might affect the host physiologically and immunology in the species dependent manner. Keeping intestinal bifidobacteria in the ideal species composition might be one of effective ways to slow immune senescence,and promote the health of the elderly.

[A preliminary analysis of changes in composition of intestinal microbiota during infancy using polymerase chain reaction-denaturing gradient gel electrophoresis].

OBJECTIVE: To investigate the composition of bacteria in the stools of infants and the colonization of intestinal microbiota during infancy. METHODS: Fresh stools were collected from 15 healthy infants at 0, 2, 4, 7, 10, 14, and 28 days and 3, 6, and 12 months after birth. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) was used to analyze the composition of intestinal microbiota, perform sequencing of dominant bacteria, and to analyze the changes in the composition of intestinal microbiota during infancy. RESULTS: DGGE fingerprint showed that the composition of intestinal microbiota during infancy changed significantly over time after birth. The cloning and sequencing results indicated that Proteobacteria colonized the earliest, mainly the obligate aerobes Enterobacter and Pseudomonas, followed by the obligate anaerobes (Clostridium hathewayi and Veillonella parvula) and the facultative anaerobe Clostridium ramosum in Firmicutes, and Verrucomicrobia. Actinobacteria colonized the latest, mainly Bifidobacterium, and gradually became dominant bacteria. CONCLUSIONS: During infancy, obligate aerobes colonize the intestinal tract the earliest, followed by obligate anaerobes and facultative anaerobes. Proteobacteria colonizes the earliest, followed by Firmicutes and Verrucomicrobia, and Actinobacteria, mainly Bifidobacterium, colonizes the latest.

[Analyzing Colonization of Bifidobacteria in Infants with Real-time Fluorescent Quantitative PCR].

OBJECTIVES: In order to know how intestinal Bifidobacteria community could be built in the infants and whether the environmental factors could affect them, the present study was conducted to characterizethe species composition and trace the quantitative changes of intestinal Bifidobacteria of the infants in their early stages with non-culture dependent molecular method. The possible association of Bifidobacteria community of the infants with their health was also discussed. METHODS: Total 16 of full-term newborn infants born between March and April 2013 were recruited for the present study. Fecal samples were collected from them at 1 day, 2 days, 4 days, 7 days, 10 days, 14 days, 28 days, 3 months, 6 months and 1 year after birth. Real-time fluorescent quantitative PCR with genus and species specific premiers was used to detect Bifidobacteria and 8 predominate species in human intestine qualitatively and quantitatively present in these collected fecal samples. RESULTS: Total 136 fecal sample were collected and Bifidobacteria were detected from 93.4% (127/136) of them with the concentration of 1.0×10 5 to 1.0×10 11 CFU/g. Bifidobacteria were found in 83.3% of the fecal samples collected from the first day after birth with more than about 10 5 CFU/g. However, Bifidobacteria were detected relative low until 14 days and were taxonomically belonged only to one or two species. Bifidobacteria were found in almost 100% of the fecal samples collected after birth 28 days with more than 108 CFU/g, and the detected species of Bifidobacteria was increased to 3 species after 28 days to 6 months. All of the fecal samples collected from one year had more than 3 species of Bifidobacteria with high cell counts. Among the detected Bifidobacteria were B.breve 92.1%, B.infantis 66.1%, B. catenulatum 59.8%, B. bifidum 25.2%, B. longum 24.4%, B.dentium 13.4%, B.angulatum 5.5% and B.adolescentis 1.6%, respectively. CONCLUSIONS: The detected Bifidobacteria greatly varied qualitatively and quantitatively after birth to one year which could be considered as the important and sensitive period for Bifidobacteria to colonize and built its communityin the infants. Different from previous studies, the colonization of Bifidobacteria in the tested infants was found delayed and the composition and diversity of Bifidobacteria species was different from other studies. These might result from different deliveryway, feeding pattern and other environmental factors related to the tested infants.

[Colonization and development of intestinal bifidobacteria in early infancy].

OBJECTIVE: To study the characteristics of the colonization of 8 species of bifidobacteria by systematically profiling fecal bifidobacterial community in the early life of infants. METHODS: Fresh fecal samples including meconium samples were collected for culture and isolation of fecal bifidobacteria from 16 cases of full-term newborn infants born between March and April 2013 at their life of 2, 4, 7, 10, 14, 28, and 90 days. The isolated fecal bifidobacteria were taxonomically identified to genus and 8 species with PCR analysis. RESULTS: One hundred and fifty-two predominant bifidobacteria strains were detected in the fecal samples, the detection rate of B. breve (22.4%) were the highest. Bifidobacteria were found in the feces of 8% infants 4 days after birth. The colonization rates increased to 54% and 60% at 28 days and 3 months respectively, significantly exceeding the colonization rate at 4 days after birth (P<0.05). Adult-type bifidobacteria B. catenulatum were found in the infants 10 days after birth, and infant-type bifidobacteria B. infantis were found at 14 days after birth, but infant-type bifidobacteria B. infantis were detected at a high level until 3 months after birth. The most tested infants had 2 species or less of bifidobacteria. CONCLUSIONS: Intestinal bifidobacteria in infants might have less diversity in early infancy. Infant-type bifidobacteria appear late, while adult-type bifidobacteria colonize earlier.