Objective To construct the multi-epitope antigen(mea) gene of G2 glycoprotein of Hantavirus SEO type L99 strain.Methods The B cell epitopes was chosen and connected by three-peptide GPG after the amino acid sequence of G2 protein was analyzed and predicted by bioinformatical soft wares.The corresponding gene mea was constructed by overlap PCR,and cloned into the prokaryotic expression plasmid pET32a(+).Results The mea gene was successfully constructed by the five epitopes being chosen.The recombinant plasmid pET32a-mea was acquired by directional cloning.Conclusion The mea and its expression system E.coli BL21/pET32a-mea were constructed for the first time.The foundation was laid for the expression of the mea and its immunological applications.

Objective To observe the results of crown lengthening surgery on the teeth with subgingival defect.Methods Crown lengthening surgery was performed on 54 terior teeth,which were divided into two groups based on probing depth of pre-operation:group A (<3 mm) and group B (3-4 mm).All teeth were restored 8 weeks after operation and followed-up to 1 year.The parameters of plaque index (PLI),suleus bleeding index (SBI),maximal defect probing depth (PD) and mobility degree (MD) at different times were measured,respectively.Results PLI,BI and PD were significantly improved during the follow-up period (P<0.01).The success rates of both groups were 96.8% and 69.6%, respectively (P<0.01).No significant difference about MD in the group A one year after restoration (P>0.05),but a significant improvement in the group B (P<0.01).Conclusion Crown lengthening surgery is a good method on the teeth with subgingival defect,but one must select the right indication.

Objective To explore neurobehavioral changes in rats with recurrent seizures and the prevention effect of melatonin.Methods 6-day-old (P6) SD rats were randomly divided into four groups of 24 (n =6):the control group (CONT),melatonin per se group (MEL),recurrent neonatal seizure group (RS) and melatonin administration prior to RS group (RS + MEL).Rats in RS group were subjected to 5 seizures with flurothyl during the first 14 days of life.In RS + MEL group,melatonin was injected at 8:00 before seizures were induced.Neurobehavioral tests including Plane righting experiment,Cliff avoidance test,the grip-strength test and negative geotaxis test were implemented on P24,while open field test on P35.Results (1) Plane righting experiment:the time of plane righting in RS group ((0.33 ± 0.51)s) was significantly shorter than that in the CONT group ((1.17 ± 0.40) s) and RS + MEL group ((0.50 ± 0.54) s) (P ＜ 0.05).(2) Cliff avoidance test:the time of cliff avoidance in RS group ((16.00 ± 6.32) s) was significantly longer than that in CONT group ((4.00 ± 2.60) s)(P ＜ 0.01),while the time of cliff avoidance in RS + MEL group ((7.67 ± 3.26) s) was shorter than that in the RS group (P ＜ 0.05).(3) The grip-strength test:compared with CONT group ((49.50 ± 28.96) s),the time needed to hold on wire in RS group((11.67 ± 7.58)s)was significantly shorter (P ＜ 0.05) and longer in RS+ MEL group ((24.83 ± 6.61) s) (P ＜ 0.05).(4) Negative geotaxis test:the time for rats to turn 180° upward in RS group((7.67 ± 1.36) s) was longer than that in the CONT group ((4.50 ± 2.66) s) and RS + MEL group ((6.17 ± 0.75) s) (P ＜ 0.05).(5) Open field test:the time for rats to begin to run in the RS group ((8.17 ± 3.86) s) was longer than that in the CONT group ((3.00 ± 1.41) s) (P ＜ 0.05).Conclusion The neurobehaviors are damaged following flurothyl-induced recurrent neonatal seizures,and melatonin can reduce the neurobehavioral injury.

Objective To investigate dynamic expressions of cortex clusterin (CLU) and intervention effect of ketogenic diet (KD) on neonatal rats with recurrent seizures.Methods Thirty-six-8-day postnatal SD rats were randomly divided into 3 groups:normal control group (NS + ND group,n =12),and the recurrent-seizure and normal diet group (RS + ND group,n =12),and the recurrent-seizure and KD group (RS + KD group,n =12).From 9 d,rats in RS + ND group and RS + KD group were subjected to recurrent seizures induced by volatile flurothyl 30 min each day for consecutive 8 days.Rats in NS + ND group were placed into the container for an equal amount of time to their counterpart without exposure to flurothyl.Scores on neurological behaviors at 35 days postnatally were examined.CLU protein levels in cerebral cortex were determined by Western blot at 58 days postnatally.Results Neurodevelopmental indicators analysis:in the plane righting experiment,there were significant differences between NS + ND group [(1.03 ± 0.54) s],R S + KD group [(0.89 ± 0.16) s] and RS + ND group [(0.64 ± 0.30) s] about the time of plane righting (all P ＜ 0.05) ; in the negative geotaxis reaction experiment,the rats of NS + ND group [(1.92 ± 0.90) s],and RS + KD group [(5.17 ± 0.72) s] about the time of negative geotaxis reaction were significantly different compared with RS + ND gouup [(7.33 ± 0.65) s] (all P ＜ 0.01).In the cliff avoidance test,there were significant differences between NS + ND group,R S + KD group [(4.33 ± 2.54) s,(8.75 ± 2.26) s] and R S + ND group [(16.58 ± 4.25) s] about the time of cliff avoidance (all P ＜ 0.01).Western blot showed that the expression of CLU in cerebral cortex of the RS + ND group [(2.24 ± 0.53) s] was obviously increased compared with NS + ND group [(1.44 ± 0.11) s] (P ＜0.01),and there also had significant difference between RS + KD group [(1.56 ±0.24) s] and RS + ND group (P ＜ 0.05).Conclusions It shows that the up-regulated expression of CLU in cerebral cortex may be associated with recurrent neonatal seizure-induced brain damage,while KD may protect them from recurrent neonatal seizure-induced brain damage by down-regulating expression of CLU.

Objective To systematic evaluation the antiepileptic drug valproic acid and topiramate effects on body weight and plasma leptin levels.Methods By searching the Cochrane Library,PubMed,EMBASE,CNKI,VIP Chinese Scientific Journals Database and Articles Database (from building a database to August 1,2014),qualified RCT was chosen from those articles according to inclusion and exclusion criterias and evaluated their quality.Then the meta-analysis was performed by Rev Man 5.3.Results (1) A total of 70 documents retrieved for the evaluation,12 of the literatures could be incorporated into the Meta-analysis,including a total of 634 participants,9 literatures using valproate to treat epilepsy,4 literatures used topiramate.(2)Meta-analysis:compared with other drugs,valproic acid treatment increased body mass index and leptin level in patients with epilepsy,BMI combined MD=0.88(95％ CI:0.45-1.31),leptin level in the combined MD=0.58(95％ CI:0.07-1.09);compared with other drugs,topiramate affect body mass index and leptin level in patients with epilepsy had no statical difference,BMI combined MD=-0.02 (95％ CI:-0.62-0.58),leptin levels in the combined MD =-0.05(95％ CI:-0.31-0.20).Conclusion Antiepileptic drug valproic acid may increase patients' BMI and serum leptin levels,yet topiramate in do not have influence in patients BMI and serum leptin levels;more basic and clinical researches are needed to explore serum leptin levels and the exact therapy mechanisms of antiepileptic drugs for clinicians to select antiepileptic drugs.

Objective To establish RT-PCR-RFLP method for studying the genotype of wild mea-sles virus strains isolated from Tianjin area from 2002 to 2008. Methods Isolations of measles virus were carried out by tissue culture method from urine and throat swab specimens collected from suspected cases. RNA were extracted from the virus specimens. The 594 bp fragment of C terminal of the N (nucleoprotein) gene was amplified by one-step RT-PCR, then the PCR products were digested with Bcn I , separated on agarose gel electrophoresis and then analyzed by the method of RFLP (restriction fragment length polymor-phism). In addition, above results were compared with DNA sequencing. Phylogenetic tree was plotted based on the results for the genetic relationship and distance analysis. Results Sixty-nine measles virus strains were isolated from 189 specimens from 2002 to 2008, of which the C terminals of N gene were all de-tected positive. Among the 69 strains of measles virus isolates, 98.55% (68/69) belonged to Hla sub-geno-type which was the predominant sub-genotype, and only one strain (1.45%) belonged to H1b sub-genotype by RFLP analysis which was in accordance with the results by DNA sequencing method. Phylogenetic tree analysis indicated the H1a sub-genotype measles virus strains should be further divided into 2 clades, and the variation fluctuated between 0.2% and 3.8%. There were transmission chains caused by different virus strains co-cireulation. Conclusion A genotype, H1a and H1b sub-genotype can be identified by RT-PCR-RFLP assay specically based on the restriction enzyme Bcn I .The RT-PCR-RFLP assay can be a rapid, simple, accurate and efficient method for large-scale surveillance of measles virus strains in China.

Objective To investigate the intervention effect of ketogenic diet (KD) on neurobehavioral demages after flurothyl-induced recurrent neonatal seizures in rats and on the expression of ApoE.Methods Postnatal day 8 (P8) SD rats (quantity:48) were randomly divided into two groups:the non-seizure group (NS group,n =24) and the recurrent-seizure group (RS group,n =24).From P9,rats in RS group were subjected to recurrent seizures induced by volatile flurothyl 30 min each day for consecutive 8 days.While rats in NS group were placed into the container for an equal amount of time to their counterpart without exposuring to flurothyl.At P28,each group was divided into two groups again:non-seizure and normal diet group(NS + ND group,n =12),non-seizure and ketogenic diet group(NS + KD group,n =12),recurrent-seizure and normal diet group (RS + ND group,n =12),recurrent-seizure and ketogenic diet group(RS + KD group,n =12).At P42,neurodevelopmental indicators were monitored.ApoE protein levels in cerebral cortex were determined by western blot at P58.Results Neurodevelopmental indicators were analyzed at P42:in the plane righting experiment,the rats of group NS + ND (1.0 ±0.14) about the time of plane righting was significant different comparing with group RS + ND ((0.75 ±0.32) s) (P ＜ 0.05).There was no significant difference between group RS + KD and group RS + ND about the time of plane righting(P＞ 0.05).In the negative geotaxis reaction experiment,the rats of groups NS + KD and RS + ND((3.17 ± 0.58)s,(6.75 ± 0.75)s) about the time of negative geotaxis reaction were significant different comparing with group NS + ND ((1.58 ±0.52)s) (P＜0.05).Compared with group RS + ND,the group RS + KD in the time of negative geotaxis reaction was obviously shortened (P ＜ 0.05).In the cliff avoidance test,there were significant differences between group NS + ND、RS + KD ((5.75 ± 2.90) s,(9.50 ± 4.36) s) and group RS + ND ((14.00 ± 4.79) s) about the time of cliff avoidance (P ＜ 0.05).In western blot,the expression of ApoE in cerebral cortex in the RS + ND group (1.26 ± 0.30) was obviously increased compared with group NS + ND (0.78 ±0.12) (P＜0.05),and there had also significant difference between group RS + KD (0.89 ±0.10) and group RS + ND (P ＜ 0.05).Conclusion The neuroprotective effects of ketogenic diet on recurrent neonatal seizure-induced neurobehavioral demages may be associated with the reduction of ApoE in cerebral cortex.

Objective To compare the effect of routine ketogenic diet and every other day ketogenic diet on neurobehavioral damage induced by recurrent seizures in neonatal rats.Methods 48 postnatal day 8 (P8) SD rats were randomly divided into four groups with 12 in each group:the control group (CONT),the recurrent seizure group(RS+ND),recurrent seizure + routine ketogenic diet group(RS+KD) and recurrent seizure+ every other day ketogenic diet group(RS+KOD).The recurrent seizures model was induced by flurothyl at p9 and last for 8 days.After a day of fasting the postnatal 28 day rats were placed on either ordinary or ketogenic diet according to packet design.Plane righting experiment,cliff avoidance test and negative geotaxis test were used to assess the neurobehavioral performance at p35.Results (1) Plane righting experiment:the plane fighting time of RS+ ND group ((0.17±0.39) s) was significantly shorter than that of NS+ND group ((0.67 ±0.49) s) (P＜0.05) ; and the plane righting time of RS+KD group((0.58±0.52) s) was significantly longer than that of RS+ND group ((0.17±0.39) s) (P＜0.05).There was no statistical significance between RS+KOD group((0.17±0.39) s) and RS+ND group ((0.17±0.39) s) (P＞0.05).(2) Cliff avoidance test:the cliff avoidance time of RS+ND group ((12.58±4.83) s)was significantly longer than that of NS+ND group ((1.92±0.90) s),RS+KD group((3.33± 1.50)s) and RS+ KOD group (P＜0.05) ;and the cliff avoidance time of RS+KOD group((5.58± 1.93)s) was significant longer than that of RS+KD group ((3.33± 1.50) s) (P＜0.05).(3) Negative geotaxis test:the negative geotaxis time of RS+NDgroup((3.17±1.70)s) was significantly longer than that of NS+ND group((1.42±0.67) s) and RS+KD group ((1.42±0.52)s) (P＜0.05).There was no statistical significance between RS+KOD group and RS+ND group(P＞0.05).Conclusion The ketogenic diet can improve neurobehavioral damage caused by flurothyl-induced recurrent seizures in neonatal rats.The every other day KD group shows a weak intervention effect comparing with the routine KD group.

Objective To explore the intervention effect of ketogenic diet (KD) on neurobehavioral damages after flurothyl-induced neonatal recurrent seizures in rats and on the expression of ZIP7.Methods Postnatal day 8 SD rats (n=24) were divided into four groups randomly:normal group (NS+ND group),non-seizure and ketogenic diet group (NS+KD group),seizure and normal diet group (RS+ND group),seizure and ketogenic diet group (RS+KD group),n=6 in eacb group.At postnatal day 31,the grip-strength test and open field test were monitored.At postnatal day 32,rats were sacrificed and the expression of ZIP7 protein level in cerebral cortex was detected with Western blot.Results (1) The grip-strength test:compared with NS+ND group ((32.67±2.42) s),the time needed to hold on glass bar in RS+ND group ((19.17±2.48) s) was shorter significantly (P＜0.05).Compared with RS+ND group,the time needed to hold on glass bar in RS+KD group ((26.25±2.87) s) was significantly longer (P＜0.05).(2) Open field test:compared with NS+ ND group ((2.00± 0.63) times),the times of grooming in RS+ND group ((4.00±0.63) times) were more (P＜0.05).Compared with RS+ND group,the times of grooming in the RS+KD group ((2.17±0.75) times) were fewer (P＜0.05).(3)Western blot:compared with NS +ND group,the level of ZIP7 of the RS+ND group in cerebral cortex were lower (P＜0.05).Compared with RS+ND group,the level of ZIP7 of the RS+KD group in cerebral cortex were higher (P＜0.05).Conclusion Neonatal recurrent seizures may damage neurobehavior,and the neuroprotective effects of ketogenic diet may be associated with the increasing of ZIP7 in cerebral cortex.

Objective:To investigate the drug resistance and pathogenic mechanism of a uropathogenic Escherichia coli (UPEC) strain UPEC132 at the genome-wide level. Methods:The susceptibility of UPEC132 strain to 16 antimicrobial agents was determined by minimum inhibitory concentration (MIC) assay. The UPEC132 strain was genotyped by multilocus sequence typing (MLST). The three-generation sequencing platform was used to sequence and assemble the whole genome of the UPEC132 strain. Drug resistance and virulence gene function annotations were predicted by Prodigal software and screened by using genome database. Genome sequences of the UPEC132 strain and 23 other UPEC strains collected from GenBank were phylogenetically analyzed, and a phylogenetic tree was constructed by RAxML software.Results:The UPEC132 strain was resistant to ampicillin, ampicillin/sulbactam, tetracycline, erythromycin, chloramphenicol and cefazolin. Its genotype was ST10522 by MLST. The whole genome of the UPEC132 strain included one complete genome (chromosome) and two plasmid sequences. The sequence sizes of the chromosome and plasmids 1 and 2 were 5 234 468 bp, 117 139 bp and 101 356 bp, and the guanine-cytosine (GC) content was 50.48%, 49.05%, and 54.04%, respectively. There were 4 856, 140 and 116 genes annotated in the chromosome and plasmids 1 and 2, respectively. Drug resistance genes were mainly distributed in the chromosomal genome, mainly including the multidrug resistance efflux pump gene clusters. Only blaTEM-1 and tetG genes were carried in the plasmid 2. Virulence genes were also mainly distributed in the chromosome genome, including nine pilus adhesins, five iron uptake systems and three secretory toxins. Gene clusters encoding Afa and type Ⅳ fimbriae were located on plasmids 1 and 2, respectively. The phylogenetic tree showed that the UPEC132 strain was not in the same branch with either of the 23 UPEC strains. Conclusions:The UPEC132 strain belonged to ST10522, which was a newly named ST type of Escherichia coli and first reported at home and abroad. The genome-wide genetic information of the UPEC132 strain was fully revealed. The multidrug resistance genes and virulence genes carried by the UPEC132 strain were associated with its drug resistance and pathogenicity.