Effects of Wuling mycelia on pentylenetetrazole-induced epilepsy in rats / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects and mechanisms of Wuling mycelia on seizure development and learning ability induced by pentylenetetrazole-kindling epilepsy in rats.</p><p><b>METHODS</b>SD rats were randomly divided into four groups: pentylenetetrazole-kindling model group (model group), low dose Wuling mycelia (0.3 g*kg(-1)) group (LD-WM group), high dose Wuling mycelia (0.6 g*kg(-1)) group (HD-WM group) and control group. The rats were intraperitoneal injected with a subconvulsive dose (35 mg*kg(-1)) of pentylenetetrazole (saline in control group) every 48 h for 12 times. Wuling mycelia was intragastrically applied 30 min before pentylenetetrazole injection. An 8-arm radial maze ( 4 arms baited) was used to measure the learning ability. Histamine was measured by chemical fluorometric enzyme immunoassay.</p><p><b>RESULTS</b>Compared with the model group, the kindling stage of LD-WM group degraded significantly after 7th injection, the latency to the onset of myoclonic jerks (LTMJ) and the latency to the onset of generalized seizures (LTGS) prolonged after the 6th and 7th injection, respectively (P<0.05). The kindling stage of HD-WM group also degraded markedly after the 6th to 8th injection, and the LTMJ and the LTGS extended after the 8th to 9th and 6th injection, respectively (P<0.05). Compared with the control group, the frequency of working memory error (WME) and reference memory error (RME) of the model group in the 8-arm radial maze increased through 3-d training (P<0.05). The memory tests showed that the impairment induced by pentylenetetrazole was partially reversed by Wuling mycelia. Compared with the control group, brain histamine contents (hippocampus, cortex, thalamus and hypothalamus) were significantly lower in model group (P<0.05). But compared with the model group, hippocampal histamine contents in LD-WM group and hippocampal, thalamic and hypothalamic histamine contents in HD-WM group were elevated (P<0.05).</p><p><b>CONCLUSION</b>Wuling mycelia can delay the kindling and ameliorate the ability of learning in rats with pentylenetetrazole-induced epilepsy and the enhancement of neuronal histamine activity may be one of possible mechanisms.</p>

Epidemiological and clinical analysis of Mycoplasma pneumoniae infection in children with acute respiratory tract infection / 南方医科大学学报

<p><b>OBJECTIVE</b>To summarize the epidemiology and clinical characteristics of Mycoplasma pneumoniae (MP) infection in children with acute respiratory tract infection (ARI) in Guangzhou.</p><p><b>METHODS</b>MP was detected using an indirect immunofluorescent method in 2084 children with ARI. The relations between MP infection rate and the gender, age, season, site of infection and wheezing diseases were analyzed.</p><p><b>RESULTS</b>A total of 433 children (20.8%) were positive for MP, including 222 boys (19.8%) and 211 girls (21.9%) without significant difference in the infection rate between the genders (P>0.05). In 0- to 3-year-old group, 106 children were positive for MP (15.0%), while in 3- to 5-year-old group and 5- to 14-year-old group, 163 (25.2%) and 164 (22.5%) were positive, respectively, showing a significant difference in the infection rate between the 3 groups (P<0.05). The MP infection rate was 18.0% in January to March, 25.1% in April to June, 17.7% in July to September, and 20.5% in October to December, showing significant differences between the periods (P<0.05). No significant difference was found in the infection rate between children with acute upper respiratory tract infection (URI) and those with lower respiratory tract infection (LRI) (P>0.05). Among the children with LRI, those having wheezing disease had significantly higher MP positivity rate than those without wheezing.</p><p><b>CONCLUSION</b>MP is a common causative agent for ARI in children. MP infection is not related to gender and infection site, but to age and season. Children over 3 years old are vulnerable to MP infection. MP infection can be associated with wheezing in LRI.</p>

Depressant Effect of siRNA on the Expression and Replication of Hepatitis B Virus in HepG2.2.15 Cell / 实用儿科临床杂志

Objective To explore the siRNA as a new antiviral therapy,evaluate the inhibition effect of siRNA based on vector on the HBV of HepG2.2.15 cell,and observe the side effect and toxicity of siRNA vector on cells and the off-target effect of siRNA.Methods Three pairs of siRNA duplexes targeting HBV C gene were designed as double strands,and the duplex were annealed and ligated into the p-Silencer-Cmv 4.1-hygro vector.The ligation products were used to transform JM109 cells.The clones with shRNA were obtained,and the vectors were purified.After the initial identification of the vector with agarose gel and the size of the inserted sequence got examined by native polyacrylamide gel electrophoresis,furthermore the sequencing was further carried out.The recombinant plasmids were purified with ultrapure Midipreps DNA Purification System.Then HepG2.2.15 cells were transfected with the plasmid mixed with siPort XP-1.The expression of HBsAg and HBeAg were detected by immunofluorescence and Western blot,and the HBV RNA was investigated by RT-PCR.Furthermore the real-time quantitive PCR was carried out to detect the changes of HBV DNA.In order to evaluate the toxicity of the shRNA,MTT was used to examine the growth rate and curve of cells.The ELISA was performed to detect the changes of interferon-? (IFN-?).Results The Western blot showed that the HBsAg and HBeAg protein were suppressed with (81.15?0.69)%,(88.12?0.92)% respectively by vector p-C2 on the third day of post-transfection.It had the similar result indicated by immunofluorescence.And the RT-PCR showed that the specific siRNA targeting HBV C gene could markedly suppress the expression of HBV mRNA and the HBV C gene mRNA was inhibited with 96.9%.The real-time quantitive PCR showed that the specific functional siRNA could markedly suppress HBV DNA copy with two orders of magnitude,while the siRNA vector had no effect on the growth of cell showed by MTT detection.Compared with the non-transfected group and p-NC group,the IFN-? level was almost the same with siRNA p-C1,p-C2,p-C3 groups.Conclusions The siRNA based on the expression vector can suppress the expression and replication of HBV in HepG2.2.15 cell.The inhibition effect was specific and had a certain dependency on siRNA concentration.No toxicity effect was found in the study.And the drug resistance wouldn′t happen because the silence was based on the split of gene.

Relationship of Neural Protection and Expression of P-CREB and C-Jun in Hippocam pal of Neonatal Rats after Hypoxia-Ischemia Brain Damage and Reperfusion / 实用儿科临床杂志

Objective To explore the survival mechanism of hippocampal ne urons after damage of hypoxia-ischemia and reperfusion of brain.Methods Seven days old SD rats(n=56) were randomly divided into hypoxia-ischemia br a in iniury(HIBD) group and sham group.The HIBD and reperfusion model was establis hed.The flowing of blood was de tected by multicolor Doppler.The p-CREB(phosphorylated c-AMP response element bi nding protein)and c-Jun were immunohistochemically evaluated in hippocampus.Thi onin staining was used to observe the apoptosis.Results The expression of p-CREB reache d the peak at 3,24 h postreperfusion in the right hippocampus of HIBD group,and then decreased to the normal level on the 7th day.In contral group the same reg ions showed basic immn-noreactivity.While c-Jun reached the peak at 6 h postreperfusion,then with a slightly decrease at 24 h;and at 48 h the other peak appeared,then with a gradual decline .On the 7th day the mumber of positive cells were still significanthy more than control group(P0.05).The sham animal showed very few apoptosis cells in the regio ns of hippocampus.Conclusions The persistent activation of CREB in the hippocampus regulates,the expression of c-Jun through the signal transductions and is involved in the course of neuron s′ survival and repair during the period of post hypoxia-ischemia reperfusion.I t is very important for the protection of the pyramidal hippocampal neurons on t he damaged side,especially for the sensitive region CA1. J Appl Clin Pediatr,2005,20(2):133-135